Lighting Strategies for the Flowering Stage of Indoor Cannabis Production

by

Victoria A. Rodriguez Morrison

A Thesis
presented to
The University of Guelph

In partial fulfilment of requirements

for the degree of
Master of Science
in
Environmental Sciences

Guelph, Ontario, Canada

© Victoria A. Rodriguez Morrison, May 2021

 ABSTRACT

LIGHTING STRATEGIES FOR THE FLOWERING STAGE OF INDOOR CANNABIS

PRODUCTION

Victoria A. Rodriguez Morrison Advisor:

University of Guelph, 2021 Dr. Youbin Zheng

Given the paucity of scientific research regarding lighting in cannabis production, this thesis

investigated the effects of light intensity (LI) and ultraviolet (UV) radiation on indoor cannabis

production during the flowering stage. When plants grew under LI ranging from 120 to 1800

μmol·m–2·s–1 provided by light emitting diodes (LEDs), inflorescence yield increased linearly as

LI increased up to 1800 μmol·m–2·s–1. When plants were grown under 400 μmol·m–2·s–1

supplemented with UV (peak wavelength of 287 nm) levels from 0.01 to 0.8 μmol·m–2·s–1, for

3.5 h·d–1, there were no changes in total Δ9-tetrahydrocannabinol or total cannabidiol

concentrations. The severity of UV-induced cannabis morphology and physiology symptoms

worsened as UV exposure level increased. The light response models developed in this thesis can

be used to determine the optimum LI for a production environment, but caution should be used

when exposing cannabis to UV radiation.

 iii

ACKNOWLEDGEMENTS
I would like to thank my advisor, Dr. Youbin Zheng for the wisdom you have shared with me
throughout my graduate studies. I will carry the academic, professional, and personal life lessons
you have taught me through all my next chapters in life. I am so grateful to have had an advisor
that values a holistic education, allowing me to gain practical experience in the industry, expand
my knowledge of plant science, and learn from your experience.

I would sincerely like to thank David Llewellyn for the endless technical support, writing
feedback, and encouragement. Your passion for research, attention to specificity, and
compassionate guidance have made a lasting impression on me. I will always be deeply grateful
for the amount of time and effort you have put into teaching me to be a better researcher, and that
you somehow always knew when words of encouragement would be helpful. I would also like to
thank my advisory committee, Dr. Nigel Gale and Dr. Gale Bozzo, for providing guidance
throughout my studies.

I would like to thank the Natural Sciences and Engineering Research Council for funding this
research, and Green Relief Inc., for providing the research facility, cannabis plants, experimental
materials, and logistical support for the experiments. Thank you to the Green Relief Inc. staff:
Derek Bravo, Tim Moffat, Dane Cronin and Madeline Baker for the technical support during my
trials. I also thank Angus Footman and Erica Emery for supporting the experiment through a
change of management.

Thank you to the Dr. Zheng lab, for providing feedback for presentations, answering academic
questions, providing graduate advice and helping with long harvests. Thank you Qinglu Ying,
Brandon Yep, Melissa Moher, Devdutt Kamath, James Nesbitt, Yun Kong, Chase Jones-
Baumgardt and Lewys Bevan. Lab trivia nights will be missed!

And finally, thank you to my supportive friends and loved ones, who have all contributed to
getting me here and keeping me grounded along the way. Thank you to my mom, dad and
brother for providing me with a lifetime full of love and encouragement, which helped make all
of my accomplishments possible.
 iv

TABLE OF CONTENTS

ABSTRACT ................................................................................................................................................................. II
ACKNOWLEDGEMENTS.......................................................................................................................................III
TABLE OF CONTENTS........................................................................................................................................... IV
LIST OF TABLES ..................................................................................................................................................... VI
LIST OF FIGURES ................................................................................................................................................. VII
LIST OF SYMBOLS, ABBREVIATIONS OR NOMENCLATURE ................................................................... IX
CHAPTER ONE........................................................................................................................................................... 1
INTRODUCTION ........................................................................................................................................................ 1
1.1 INTRODUCTION TO CANNABIS ...................................................................................................................... 1
1.2 HISTORY OF CANNABIS ................................................................................................................................ 3
1.2.1 Medicinal uses through history .............................................................................................................. 3
1.2.2 Prohibition ............................................................................................................................................. 3
1.2.3 Legalization and usage .......................................................................................................................... 4
1.3 CANNABIS CULTIVATION ............................................................................................................................. 5
1.3.1 Current practices ................................................................................................................................... 5
1.4 LIGHT INTENSITY IN CANNABIS PRODUCTION .............................................................................................. 6
1.4.1 Effects of light intensity on cannabis photosynthesis, growth and yield ................................................ 8
1.5 ULTRAVIOLET RADIATION IN PLANT PRODUCTION .................................................................................... 10
1.5.1 Effects of ultraviolet radiation on cannabis photosynthesis, growth and secondary metabolites ....... 14
1.6 THESIS STRUCTURE AND OBJECTIVES ........................................................................................................ 17
CHAPTER TWO ....................................................................................................................................................... 18
CANNABIS YIELD, QUALITY, AND LEAF PHOTOSYNTHESIS RESPOND DIFFERENTLY TO
INCREASING LIGHT LEVELS IN AN INDOOR ENVIRONMENT ................................................................ 18
ABSTRACT ................................................................................................................................................................ 18
2.1 INTRODUCTION .......................................................................................................................................... 18
2.2 MATERIALS AND METHODS........................................................................................................................ 22
2.2.1 Experimental Design ............................................................................................................................ 23
2.2.2 PPFD Levels ........................................................................................................................................ 25
2.2.3 Plant Culture ........................................................................................................................................ 26
2.2.4 Leaf Photosynthesis.............................................................................................................................. 27
2.2.5 Leaf Morphology .................................................................................................................................. 29
2.2.6 Yield and Quality.................................................................................................................................. 29
2.2.7 Data Processing and Analysis ............................................................................................................. 30
2.3 RESULTS .................................................................................................................................................... 31
2.3.1 Leaf Photosynthesis.............................................................................................................................. 33
2.3.2 Chlorophyll Content Index and Plant Morphology ............................................................................. 36
2.3.3 Yield and Quality.................................................................................................................................. 41
2.4 DISCUSSION ............................................................................................................................................... 41
2.4.1 Cannabis Inflorescence Yield is Proportional to Light Intensity ......................................................... 41
2.4.2 Increasing Light Intensity Enhances Inflorescence Quality ................................................................ 43
2.4.3 Plasticity of Cannabis Leaf Morphology and Physiology Responses to LI and Over Time ................ 45
2.5 CONCLUSIONS ............................................................................................................................................ 49
 v

CHAPTER THREE ................................................................................................................................................... 50

CANNABIS INFLORESCENCE YIELD AND CANNABINOID CONCENTRATION IS NOT IMPROVED
WITH LONG-TERM EXPOSURE TO UV RADIATION.................................................................................... 50
ABSTRACT ................................................................................................................................................................ 50
3.1 INTRODUCTION .......................................................................................................................................... 51
3.2 MATERIALS AND METHODS........................................................................................................................ 53
3.2.1 Plant culture ......................................................................................................................................... 53
3.2.2 Experimental setup............................................................................................................................... 53
3.2.3 Growth measurements.......................................................................................................................... 56
3.2.4 Leaf chlorophyll and fluorescence measurements ............................................................................... 56
3.2.5 Leaf gas exchange measurements, leaf size and specific leaf weight .................................................. 57
3.2.6 Visual observations .............................................................................................................................. 58
3.2.7 Yield and quality .................................................................................................................................. 58
3.2.8 Statistical analysis................................................................................................................................ 59
3.3 RESULTS .................................................................................................................................................... 59
3.3.1 UV-induced cannabis morphology and physiology changes ............................................................... 60
3.3.2 Growth responses to UV ...................................................................................................................... 67
3.3.3 Responses of inflorescence yield, quality and cannabinoid and terpene concentrations to UV.......... 69
3.4 DISCUSSION ............................................................................................................................................... 71
3.4.1 UV radiation alters cannabis morphology and physiology ................................................................. 72
3.4.2 UV radiation suppresses cannabis growth and yield........................................................................... 74
3.4.3 UV radiation alters the secondary metabolite composition of cannabis inflorescences ..................... 75
3.4.4 Implications of UV in indoor cannabis production and future research directions ............................ 77
3.5 CONCLUSIONS ............................................................................................................................................ 80
CHAPTER FOUR ...................................................................................................................................................... 81
GENERAL DISCUSSION AND CONCLUSIONS................................................................................................. 81
REFERENCES ........................................................................................................................................................... 85
APPENDIX A ........................................................................................................................................................... 103
SUPPLEMENTARY INFORMATION: PLANT GROWTH RESPONSE TO INCREASING LI .................................................. 103
Method .............................................................................................................................................................. 103
Results............................................................................................................................................................... 103
Discussion......................................................................................................................................................... 105
APPENDIX B ........................................................................................................................................................... 106
SUPPLEMENTARY INFORMATION: PHOTOBLEACHING AT HIGH LI .......................................................................... 106
Methods ............................................................................................................................................................ 106
Results............................................................................................................................................................... 106
Discussion......................................................................................................................................................... 106
APPENDIX C ........................................................................................................................................................... 108
 vi

LIST OF TABLES
Table 2.1. Cannabinoid concentration in apical inflorescences of Cannabis sativa L. ‘Stillwater’.
....................................................................................................................................................... 39

Table 2.2. The relationships between average photosynthetic photon flux density (APPFD)
applied during the flowering stage (81 days) and terpene concentration in apical inflorescences
of myrcene, limonene and total terpenes, and the mean concentration for terpenes with no
APPFD treatment effects, of Cannabis sativa L. ‘Stillwater’. ..................................................... 40

Table 3.1. Range of canopy-level UV photon flux density, UV biologically-effective photon flux
density and daily UV biologically-effective dose from UV LEDs with a peak wavelength of 287
nm and a daily 3.5 h photoperiod.................................................................................................. 56

Table 3.2. Minimum UV-PFD (µmol·m–2·s–1) where symptoms were observed in Cannabis
sativa L. ‘Low Tide’ (LT) and ‘Breaking Wave’ (BW) cultivars (CV) in each week after the
initiation of UV treatments, regardless of whether or not all plants above the minimum UV-PFD
presented the observed symptom. ................................................................................................. 66

Table 3.3. The effects of UV-PFD (µmol·m–2·s–1) applied during the flowering stage on
physiological, morphological and yield parameters of Cannabis sativa L. ‘Low Tide’ and
‘Breaking Wave’. .......................................................................................................................... 67

Table 3.4. The effects of UV-PFD (µmol·m–2·s–1) applied during the flowering stage on
cannabinoid and terpene concentrations (mg·g–1) in the apical inflorescence of Cannabis sativa
L. ‘Low Tide’ and ‘Breaking Wave’. ........................................................................................... 71
 vii

LIST OF FIGURES
Figure 2.1. Relative spectral photon flux distribution of Pro-650 (Lumigrow) LED fixtures...... 22

Figure 2.2. Schematic of a single light rack (8 LED fixtures, in magenta) above one third of a
deep-water culture basins (CB). The entire growing area consists of 6 of these light racks. Within
each light rack, each of the 8 target PPFD levels (i.e., the “treatments”) were randomly assigned
to one fixture (i.e., plot). This resulted in a randomized complete block type of experimental
layout, comprised of 8 treatments × 6 replications. However, each treatment plant (in blue) was
assigned an average photosynthetic photon flux density (APPFD) as LI treatment levels,
reflecting the average canopy-level light intensity measured throughout the trial. The APPFD
levels were used as the independent variable in subsequent analyses of plant growth, physiology
and harvest metrics. Each plot was surrounded by non-treatment plants (diagonal lines) to ensure
uniform growing environment and normal planting density. ....................................................... 24

Figure 2.3. Typical light response curves [net CO2 exchange rate (NCER) response to light
intensity] of two youngest fully-expanded fan leaves of Cannabis sativa L. ‘Stillwater’ grown
under either low or high localized photosynthetic photon flux densities (LPPFD). The low and
high LPPFD were 91 and 1238 μmol·m–2·s–1, respectively. Measurements were made during
week 5 after the initiation of the 12-h photoperiod....................................................................... 32

Figure 2.4. The light-saturated net CO2 exchange rate (Asat) (A), the light saturation point (LSP)
(B), the localized net CO2 exchange rate (LNCER) (C), and the Fv/Fm (D) of the youngest fully-
expanded fan leaves of Cannabis sativa L. ‘Stillwater’ at the localized photosynthetic photon
flux densities (LPPFD) that the respective leaves were growing under when the measurements
were made, during weeks 1, 5, and 9 after initiation of the 12-h photoperiod. Each datum is a
single plant. Regression lines are presented when P ≤0.05. ......................................................... 33

Figure 2.5. The specific leaf weight (SLW; on a dry weight basis) of young, fully-expanded
Cannabis sativa L. ‘Stillwater’ leaves in response to the average photosynthetic photon flux
density (APPFD), measured on day 35 after initiation of the 12-h photoperiod. Each datum
represents one fan leaf from a single plant. .................................................................................. 35

Figure 2.6. Sketches of Cannabis sativa L. ‘Stillwater’ plants grown under low (A) and high (B)
photosynthetic photon flux density (APPFD), 9 weeks after initiation of 12-h photoperiod
(illustrated by Victoria Rodriguez Morrison). .............................................................................. 36

Figure 2.7. The relationship between average apical photosynthetic photon flux density (APPFD)
applied during the flowering stage (81 days) and inflorescence dry weight (A), harvest index
(total inflorescence dry weight / total aboveground dry weight) (B), and apical inflorescence
density (based on fresh weight) (C) of Cannabis sativa L. ‘Stillwater’. Each datum is a single
plant............................................................................................................................................... 38

Figure 3.1. Relative spectral photon flux distribution of (A) Pro-650 (Lumigrow) LED fixtures
and (B) UV LED fixtures. ............................................................................................................. 54
 viii

Figure 3.2. (A) The side view and (B) top view of Cannabis sativa L. plants in week 2 after the
initiation of the UV treatments. ‘Low Tide’ under (1) minimum and (2) maximum UV exposure
and ‘Breaking Wave’ under (3) minimum and (4) maximum UV exposure levels. The black scale
bar at the lower right of each image is 5.0 cm. ............................................................................. 62

Figure 3.3. (A) ‘Low Tide’ (LT) and (B) ‘Breaking Wave’ (BW) Cannabis sativa L. stigmas
under minimum UV exposure levels and (C) LT and (D) BW under maximum UV exposure
levels, in week 3 after the initiation of the UV treatments. The white scale bar at the lower right
of (C) applies to (A), and at the lower right of (D) applies to (B). Both scale bars are 1.0 cm. ... 63

Figure 3.4. (A) ‘Low Tide’ and (B) ‘Breaking Wave’ Cannabis sativa L. plants demonstrating
(from left to right) minimum, low, moderate, and high UV exposure levels. The images were
taken in week 3 after the initiation of the UV treatments. The black scale bar at the lower right of
each image is 5.0 cm. .................................................................................................................... 64

Figure 3.5. (A) Adaxial and (B) abaxial sides of youngest, fully-expanded Cannabis sativa L. fan
leaves of ‘Low Tide’ (top row in each image) and ‘Breaking Wave’ (bottom row in each image)
demonstrating UV induced leaf morphology effects with increasing UV-PFDs. Leaves from
plants under minimum UV exposure are on the left, moderate UV exposure in the middle, and
high UV exposure on the right. Scans were taken in week 5 after the initiation of UV treatments.
The black scale bar at the lower right of each image is 2.0 cm. ................................................... 65

Figure 3.6. Gross plant morphology of (A) ‘Low Tide’ and (B) ‘Breaking Wave’ Cannabis
sativa L. plants grown under (from left to right) minimum, low, moderate, and high UV exposure
levels. Images were taken just prior to harvest (i.e., 9 weeks after the initiation of UV
treatments). Note the white spots (powdery mildew) on the adaxial sides of leaves on the far-left
plants. The black scale bar at the upper left of each image is 5.0 cm. .......................................... 68

Figure 3.7. The apical inflorescences of (A) ‘Low Tide’ and (B) ‘Breaking Wave’ Cannabis
sativa L. plants grown under (from left to right) minimum, low, moderate, and high UV exposure
levels. Images were taken at harvest (i.e., 9 weeks after the initiation of UV treatments). The
black scale bar at the upper left of each image is 2.0 cm.............................................................. 70
 ix

LIST OF SYMBOLS, ABBREVIATIONS OR NOMENCLATURE

APPFD: average PPFD at the plant apex integrated over time

Asat: light-saturated NCER

BSWF: biological spectral weighting function

CB: culture basin

CBD: cannabidiol

CBG: cannabigerol

CBGA: cannabigerolic acid

CBN: cannabinol

CCI: chlorophyll content index

COP1: constitutively photomorphogenic 1

DLI: daily light integral

DW: dry weight

Fv/Fm: variable to maximum chlorophyll fluorescence

FW: fresh weight

g: gram

HPS: high pressure sodium

HY5: elongated hypocotyl 5

LAI: leaf area index

LED: light emitting diode

LI: light intensity

LNCER: NCER at LPPFD

LPPFD: localized PPFD at the measured leaf

LRC: light response curve

 x

LSP: light saturation point

m: metre

mg: milligram

NCER: net CO2 exchange rate

PAR: photosynthetically active radiation

PFD: photon flux density

PPFD: photosynthetic photon flux

QY: maximum quantum yield

RH: relative humidity

ROS: reactive oxygen species

SD: standard deviation of the mean

SE: standard error of the mean

SLW: specific leaf weight

TCBD: total equivalent cannabidiol

TCBG: total equivalent cannabigerol

TLI: total light integral

TΔ9-THC: total equivalent Δ9-tetrahydrocannabinol

UDL: under detection limit

UV: ultraviolet

UVA: ultraviolet-A

UVB: ultraviolet-B

UVBE: biologically effective ultraviolet

UVC: ultraviolet-C

UVR8: UV resistant phytoreceptor-8

 xi

Δ9-THC: Δ9-tetrahydrocannabinol

Δ9-THCA: Δ9-tetrahydrocannabinolic acid

 CHAPTER ONE
INTRODUCTION

1.1 INTRODUCTION TO CANNABIS

Cannabis (Cannabis sativa L.) is an annual herbaceous species in the Cannabaceae family, which
also includes hops (Humulus spp.) (Russo, 2019). Cannabis is primarily a dioecious species (i.e.,
male and female reproductive organs usually develop on separate plants), but hermaphrodism
can occur under stressful conditions (Clarke and Merlin, 2016; Larsson and Lagerås, 2015; Punja
and Holmes, 2020). Sexual dimorphism occurs late in cannabis plant development, where female
plants are differentiated from male plants at the onset of flowering (Cristiana Moliterni et al.,
2004; ElSohly et al., 2017). The growth cycle of cultivated cannabis typically consists of three
stages: propagation, vegetative growth, and flowering. Cannabis is typically a short-day plant;
where the vegetative stage is maintained under long days (e.g., 16-h to 18-h days), and the
flowering stage is initiated under short days when a critical uninterrupted dark period per day is
met, with commercial producers typically using a 12-h light and 12-h dark regime (Potter, 2014).
There is the exception of some cultivars that flower depending on plant age rather than the
number of hours of uninterrupted dark exposure, which are labelled as day-neutral or auto-
flowering, but they are not yet extensively used commercially (Clarke and Merlin, 2016).

Unseeded female cannabis flowers are typically cultivated due to their higher cannabinoid
concentration than seeded flowers, vegetative tissues or male flowers (Potter et al., 2018). Each
enlarging female inflorescence consists of bracts that are covered in multicellular, secretory
glandular trichomes that accumulate a sticky resin containing a mixture of secondary metabolites
(Farag and Kayser, 2015; Small, 2017). These secondary metabolites include aromatic terpenes
and a class of terpenophenolic compounds called phytocannabinoids (ElSohly et al., 2017). In
humans, phytocannabinoids bind to endocannabinoid system receptors, specifically the
cannabinoid receptor 1 and cannabinoid receptor 2 (Pertwee, 1997). These receptors allow
cannabinoids to induce psychoactive and therapeutic effects in humans (Gonçalves et al., 2019).
The term phytocannabinoid is used to characterize cannabinoids that originate from plants, as
1
opposed to endocannabinoids which originate from the mammalian brain (Maroon and Bost,
2018). Given the botanical focus of this thesis, phytocannabinoids will be referred to as
cannabinoids hereafter. The dominant cannabinoids in mature cannabis inflorescence tissues are
Δ9-tetrahydrocannabidiolic acid (Δ9-THCA), cannabidiolic acid (CBDA), and cannabigerolic
acid (CBGA), which are converted to the more medicinally relevant compounds, Δ9-
tetrahydrocannabidiol (Δ9-THC), cannabidiol (CBD) and cannabigerol (CBG), through
decarboxylation (Eichler et al., 2012; Zou and Kumar, 2018). Medicinally relevant cannabinoids
can be psychoactive (e.g., Δ9-THC), or non-psychoactive [e.g., CBD, CBG, cannabichromene
(CBC)] (ElSohly et al., 2017; Flores-Sanchez and Verpoorte, 2008). Cannabinoid receptor 1 was
discovered in 1988, followed by cannabinoid receptor 2 in 1993 (Pertwee, 2006). Δ9-THC exerts
its analgesic and psychoactive effects through interacting with cannabinoid receptors, especially
cannabinoid receptor 1 (Gonçalves et al., 2019). The cannabinoids in cannabis inflorescences are
therefore valuable to the medicinal market.

There has been some controversy over differentiating between the subspecies of cannabis;
Cannabis sativa sp. sativa and Cannabis sativa sp. indica (McPartland, 2018), both genetically
and morphologically. McPartland (2018) argues that the interbreeding and hybridization between
subspecies renders the distinction between them irrelevant. Conversely, it is more relevant for
producers and consumers to distinguish cannabis by chemotypes as per their cannabinoid
content, ratio of Δ9-THC to CBD in the inflorescence (i.e., chemotypes) or anticipated product
end-use. Chemotype I has a Δ9-THC to CBD ratio >1, chemotype II has and intermediate Δ9-
THC to CBD ratio ≈1, whereas chemotype III contains has a Δ9-THC to CBD ratio <1 (Small
and Beckstead, 1973). Cannabis product end-uses are usually categorized by either fiber-type or
drug-type. Fiber-type cannabis is cultivated for the edible seeds and oil, and/or for the bast-fibers
used to make textiles (Behr et al., 2016; Clarke and Merlin, 2016), whereas drug-type cannabis is
often cultivated for the unfertilized female flowers containing psychoactive Δ 9-THC, which can
induce relaxation and euphoria (Clarke and Merlin, 2016). In recent years, research has
elucidated the therapeutic benefits of CBD, and all three chemotypes are now cultivated for the
medicinal and recreational use of cannabinoids in the unfertilized female flower (Lewis et al.,
2018). Thus, all three chemotypes could be considered “drug-types”.

2
1.2 HISTORY OF CANNABIS
1.2.1 MEDICINAL USES THROUGH HISTORY

Humans have a long history with cannabis, with cultivation dating back millennia (Clarke and
Merlin, 2016). Pollen fossils indicate that cannabis may have originated in the northeastern
Tibetan Plateau 19.6 million years ago, and also may have dispersed to Europe over a million
years ago (McPartland, 2018). The earliest record of cannabis use dates back to the 28th century
BC, in the Shennong Ben Cao Jing, one of the oldest books of Chinese medicine compiling oral
traditions (Russo, 2007). There is reference to the medicinal use of cannabis throughout history,
including in Ancient Egypt, Greece, and the Roman Empire (Russo, 2007).

1.2.2 PROHIBITION

Cannabis was used as a medicinal drug in the United States pharmacopeia before it was
prohibited in the early 20th century (Valdez and Kaplan, 2019). In 1923, cannabis drug use and
possession became illegal in Canada under the Opium and Narcotic Drug Act (Graham, 2004),
but it was not commonly used. There were only 25 convictions for cannabis possession in
Canada between 1930 and 1946, but after a cultural shift often attributed to the hippie
movement, there were 12,000 convictions in 1972 alone (Kenny and Nolin, 2003). The cannabis
market quickly dominated the illicit drug market in North America and worldwide, in terms of
number of consumers and amount of production [United Nations Office on Drugs and Crime
(UNODC), 2009; 2019]. However, the lack of reliable data and scientific research on the
production and use of cannabis complicates the accurate estimation of its prevalence (UNODC,
2009). In 2007, the majority of the world's cannabis seizures occurred in Mexico (37%) followed
by the United States (26%) (UNODC, 2009). Due to the legal restrictions on cannabis in Canada
and much of the world, there has been very limited scientific research on both cannabis
cultivation and medicinal cannabis use throughout the 20th century (Graham, 2004; Magagnini et
al., 2018).

3
1.2.3 LEGALIZATION AND USAGE

In 2002, Canada became the first country to introduce government-regulated access to cannabis
for a list of medicinal purposes with the Marijuana Medical Access Regulations (Government of
Canada, 2016; Graham, 2004). Subsequently in 2013, the Government of Canada implemented
the Marijuana for Medical Purposes and Regulations, which provided the commercial industry
rules and regulations for production and distribution of dried cannabis inflorescences
(Government of Canada, 2016). Following that, the Government of Canada permitted licensed
producers to sell cannabis oil and allowed authorized users to possess other forms of cannabis in
2015 (Government of Canada, 2016). The Marijuana for Medical Purposes and Regulations was
updated and renamed in 2016 to the Access to Cannabis for Medical Purposes Regulations,
which allowed authorized cannabis users to possess 4 cannabis plants in order to produce their
own medicine (Government of Canada, 2016). As of October 17 2018, the production and sale of
recreational cannabis became legal in Canada under the Cannabis Act (Government of Canada,
2019a). The objectives of this legislation were to reduce cannabis use in youth (under the age of
18), prevent organized crime from profiting from the sale and distribution of cannabis, and to
protect public health and safety by allowing Canadian adults to access regulated, legal cannabis
(Government of Canada, 2019b). Several other countries have decriminalized medicinal and
recreational cannabis use including Uruguay, Georgia, Jamaica, the Netherlands, Israel and even
some US states (Cannigma, 2019; Leggett, 2006; UNODC, 2019).

The legal cannabis market faces the economic challenge of competing with illicit trade
(UNODC, 2019). When governments decide on the tax rates for cannabis products, one
consideration is keep the cost low enough to displace the illegal cannabis market and prevent
organized crime from profiting (UNODC, 2019). In 2019 about 40% of cannabis consumers
obtained cannabis from an illegal supplier (Rotermann, 2020). Despite revenue lost to criminals
involved with the illicit trade of cannabis, the legal cannabis industry benefits the economies of
countries where it has become legal by creating jobs. The legal cannabis industry provided
211,000 Americans with jobs in 2019 (Barcott and Whitney, 2019).

4
The legalization of cannabis across North America has been accompanied with a resurgence of
interest in its medicinal properties. Recent research on cannabinoids has elucidated their
analgesic, anti-inflammatory, appetite stimulating and anti-emetic properties have the potential to
treat a number of ailments including pain, nausea, depression and alleviation of symptoms of
HIV/AIDS, Parkinson's and Huntington's disease, and cancer, to name a few (Guindon and
Hohmann, 2009; Pacher et al., 2006; Slatkin, 2007; Viveros et al., 2005). In the Americas,
cannabis use has increased from 42 million people (i.e., 7% of the population aged 15-65) in
2007 to 57 million people (i.e., 8.4% of the population aged 15-65) in 2017 (UNODC, 2019). In
Canada alone, it was reported in 2015 that 14.7% of the Canadian population aged 15 and older
used cannabis at least once in the past year (Government of Canada, 2015). As cannabis usage
continues to increase with worldwide legalization, cannabis cultivation methods must improve to
keep up with the increasing demand.

1.3 CANNABIS CULTIVATION

1.3.1 CURRENT PRACTICES

Cannabis is produced in the majority of countries around the world, unlike other plant-based
drug production that are often only produced in a select number of countries (UNODC, 2019).
Cannabis is cultivated either in outdoor fields or in controlled environments, such as indoors or
in greenhouses (Chandra et al., 2017; Leggett, 2006; Potter, 2014). According to qualitative data
reported by Member States to the UNODC (2019), both outdoor and controlled-environment
cannabis cultivation increased globally from 2013 to 2017, although controlled-environment
cultivation appears to have a larger increase than outdoor. The increase in controlled-
environment cannabis cultivation is closely associated with an increase in Δ 9-THC content on the
market (UNODC, 2019). The advantage of indoor cultivation (vs. greenhouse or outdoor) is that
it affords complete control over the lighting environment including the manipulation of light
intensity (LI), spectrum or “quality”, as well as photoperiod. Cannabis growers alter the
photoperiod to control the growing cycle, specifically to maintain the vegetative stage or trigger
flowering (Leggett, 2006; UNODC, 2019). Controlled-environment cultivation allows growers to
yield more crops per year than outdoor production in northern climates (ElSohly et al., 2017;
Leggett, 2006). Controlled-environment cannabis cultivation allows for sinsemilla production,
5
which originates from the Spanish term “sin semilla,” meaning without seeds. In other words,
controlled environments protect against pollination from male plants, except in the event of
stress-induced hermaphrodism (Punja and Holmes, 2020). In sinsemilla cannabis production,
female plants are mainly propagated asexually using uniform vegetative cuttings (Chandra et al.,
2017; Clarke and Merlin, 2016), often grown in soilless production systems (Farag and Kayser,
2015; Leggett, 2006). Sinsemilla production results in consistent, unfertilized inflorescences with
higher Δ9-THC concentrations as compared to fertilized inflorescences (Chandra et al., 2017;
Clarke and Merlin, 2016; Leggett, 2006; UNODC, 2019), presumably due to carbon costs of
seed rather than glandular trichome production. Clarke and Merlin (2016) indicate that sinsemilla
growers typically look for six economically desirable traits in their plants: 1) high dry biomass
yields, 2) high proportion of inflorescences compared to stems and leaves, 3) many large
glandular trichomes, 4) high total cannabinoid content in inflorescences, 5) reproducible profile
of cannabinoids, and 6) desirable aromatic terpenes. These traits are not only genetically selected
for, but they may also be impacted by the plant’s growing environment and horticultural
management strategies. Of the environmental parameters in a plant’s growing environment, light
may be the most influential.

1.4 LIGHT INTENSITY IN CANNABIS PRODUCTION

The plasticity of morphological and physiological traits in plants is determined by an equilibrium
between endogenous growth processes and exogenous environmental influences (Barthélémy
and Caraglio, 2007; Jansen et al., 2017). LI and spectrum modulate photosynthetic activity and
photomorphogenic signals, which initiate processes such as cell division and elongation,
directional growth and branching, all of which contribute to plant vegetative growth and
development (Huché-Thélier et al., 2016). In indoor cannabis production, LI is a controlled
environmental input that has a major impact on plant photosynthesis, growth and yield.
Understanding the morphological, physiological and yield-related impacts of LI on cannabis
plants through the flowering stage will assist cannabis growers in determining optimal LI for
their production. In temperate environments, e.g., regions such as Ontario, the low natural LI and
shorter photoperiods (i.e., more than 12-hr nights) than those required in the winter months is not
optimal for cannabis production. As a result, either supplemental lighting in greenhouses or sole-
6
source lighting in indoor production is required winter production. The energy costs related to
lighting (and heat and ventilation associated) in indoor cannabis production make up about 60%
of the total energy consumed by cannabis growers, meaning that lighting is one of the most
expensive environmental inputs (Mills, 2012). Since light is a valuable resource for cannabis
production, the selected LI for production should be chosen to maximize yield in proportion to
energy use. However, there is a lack of scientifically-validated information in the literature on
cannabis’ response to LI in indoor production.

It is well-known that a plant’s photosynthesis, growth and yield (e.g., total aboveground
biomass) respond proportionally to photosynthetically active radiation (PAR) at lower light
levels, followed by a logarithmic phase (gradual decreasing gains in productivity) up to maxima
LI (light saturation point; LSP) whereby further increases of PAR do not result in additional
photosynthesis and biomass production and other plant growth indices (Lobo et al. 2013). For
example, the meta-analysis by Poorter et al. (2019) demonstrates that total dry biomass saturates
in both woody and herbaceous species as function of LI, with high plasticity. Photosynthesis is
often measured as a Net CO2 Exchange Rate (NCER), and is often referred to in the literature as
CO2 assimilation or “A” (Zheng et al., 2006; Bernacchi et al., 2003; Ainsworth and Rogers,
2007). The NCER at the LSP is called the light-saturated NCER, which often referred to as
“Asat” in the literature (Bernacchi et al., 2003; Ainsworth and Rogers, 2007). The LI resulting in
the highest NCER, growth or biomass in proportion to the light energy input is optimal for
production. In this light response curve (LRC), the “optimal” LI is at the end of the linear
increase, before the LSP. The availability of PAR is often a limiting factor (i.e., in the linear
portion of the LRC) for yield in indoor production. Growing crops with insufficient light (i.e.,
below “optimal,” as defined here) limits the yield potential, which in turn wastes the other
production inputs including labour, water, nutrients and electricity. Choosing a LI for production
that is above optimal results in diminished yield returns in proportion to the energy input, thereby
increasing costs without yield gain. The optimum LI may also depend on other factors in the
production environment (including the production goals of the grower), since lighting is only one
of the input costs for production. Overall, LRCs are a useful tool to identify saturating LIs, but
there are no yield LRCs for cannabis growers to use for optimizing their light intensity.

7
1.4.1 EFFECTS OF LIGHT INTENSITY ON CANNABIS PHOTOSYNTHESIS,
GROWTH AND YIELD

Plants in nature are restricted to their permanent locations – i.e., they are sessile and must
acclimate to their surrounding light environment to maintain photosynthesis, growth and yield.
Leaves contain light-activated proteins called photoreceptors that perceive LI, light quality and
light duration and subsequently signal changes to gene expression (Casal, 2013; Thoma et al.,
2020). High-light grown plants typically have higher total vegetative dry mass and more
branching than low-light grown plants (Poorter et al., 2019). The lighting environment in which
an individual leaf is acclimated also has a substantial impact on its morphology (e.g., high-light
grown leaves often have thicker leaves with smaller leaf area) and physiology (e.g., high-light
grown leaves often have higher LSPs compared to low-light grown leaves) (Murchie et al., 2002;
Walker et al., 1989). Consequently, the varying LI within a canopy (i.e., upper canopy leaves
compared to lower canopy leaves) results in leaves with varying morphology and physiology
(Bauerle et al., 2020; Campbell et al., 1992; Namdar et al., 2018). Although plant LRCs all
follow a similar asymptotic trend, specific LRC equations are dependent on several factors, such
as plant species, leaf age and the growing environment in which the plant was acclimated (e.g.,
temperature, CO2 concentration, vapour pressure deficit, and the lighting environment).

Chandra et al. (2008) is a frequently cited study in relation to canopy-level lighting strategies for
optimal cannabis growth (Jaeger, 2019), yield (Coco for Cannabis, n.d.; Mammoth Lighting,
n.d.) and productivity (Downer, 2018; Royal Queen Seeds, 2019). Chandra et al. (2008) found
that Asat in a Mexican variety of cannabis was 24.6 μmol·m−2·s−1 at the estimated LSP of 1500
μmol·m−2·s−1 based on LRCs for single-leaf in the upper canopy; however, the LI that the leaves
were acclimated to and the leaves' ages are unknown. Photosynthetic responses do not directly
predict yield response, especially when important context (e.g., leaf age and light history) is
unreported (Sadras and Richards, 2014).

Although an outcome of photosynthesis in a leaf is CO2 assimilation, there are many factors
within a canopy to consider before correlating leaf-level photosynthesis with aboveground
biomass yield. Leaves in the upper canopy are exposed to higher LI than the lower canopy, so
their NCER, Asat and LSPs are typically higher by comparison (Bauerle et al., 2020; Murchie et
8
al., 2002; Pettersen et al., 2010). Even within the same plant, leaves with higher vertical leaf
positions are younger and have higher Asat (i.e., higher CO2 assimilation capacity) (Bauerle et al.,
2020; Murchie et al., 2002), higher nitrogen and carbon (% dry weight) and higher chlorophyll
content per area (μg·cm–2) compared to lower leaf positions (Gara et al., 2018). Additionally,
respiration must be factored in to comprehend the net CO2 assimilation in a whole plant. Leaf
respiration tends to be more pronounced in leaves that undergo high rates of photosynthesis
compared to lower rates of photosynthesis, therefore, similar to the variability of photosynthesis
throughout a canopy, respiration varies with vertical leaf position (i.e., a leaf at the top of the
plant having the highest Asat is associated with the highest light-dependent and -independent
respiration) (Weerasinghe et al., 2014). Additionally, the canopy does not absorb all available
light; some light is reflected and the transmittance through the canopy depends on the leaf area
index [LAI; m2(leaf)/m2(ground)] and the leaf angle relative to incoming light (Posada et al., 2012).
Since light absorption varies drastically throughout the canopy, leaf photosynthetic responses
vary throughout the canopy as well.

Increased LAI can increase the amount of CO2 assimilation within a plant, but depending on the
level of LAI increase, there could be too much shade within the canopy and the cost of increased
leaf area no longer outweighs the benefit (Peng, 2000). The aforementioned complexity of
varying morphology and physiology means that several parameters must be considered to
accurately understand whole-canopy photosynthesis, and single-leaf gas exchange measurements
from only the upper canopy leaves are not indicative of whole-canopy photosynthesis. Due to the
many factors to consider when relating leaf-level photosynthesis to whole-canopy yield
responses to light, the impact of LI on cannabis yield must be related to the actual yield (i.e.,
cannabis inflorescence weight) to accurately model the relationship. In other terms, a yield LRC
for cannabis will be useful for cannabis growers to understand the relationship between canopy-
level LI and yield.

Some studies have established that cannabis yields are greater at higher LIs relative to low LIs.
For example, Potter and Duncombe (2012) grew cannabis plants under high pressure sodium
(HPS) lamps with varying canopy-level PPFDs during the flowering stage and found that
increasing PPFD from 400 to 900 μmol·m−2·s−1 increased yield an average of 1.3 times higher,
9
across seven cultivars, with no LI treatment effects on floral cannabinoid concentrations.
Vanhove et al. (2011) found that cannabis yields were 1.3 to 3.1 times higher (depending on
cultivar) when plants were grown under approximately 1000 μmol·m−2·s−1 compared to
approximately 450 μmol·m−2·s−1 during the flowering stage. Although these studies provide
some insight into specific yield increase scenarios, the response of cannabis yield to a wide range
of LIs is necessary to create a model for cannabis inflorescence yield response to increasing LI
(i.e., a yield LRC). Eaves et al. (2020) found that cannabis yield increased linearly as PPFD
increased from approximately 500 to 1500 μmol·m−2·s−1 (i.e., yield was on average 2.5 times
higher) during the flowering stage, although there were no data between 1000 and 1500
μmol·m−2·s−1, and PPFD data were reported based on the height the plants were expected to be at
harvest rather than their actual lighting environments in each LI treatment. These studies support
the contention that cannabis has very high saturating PPFD on a yield basis relative to other
crops. Currently, there is a lack of peer-reviewed literature for commercial growers to refer to
when deciding on optimal LI for production (Backer et al., 2019; Eichhorn Bilodeau et al.,
2019). Therefore, studies elucidating cannabis inflorescence yield response to increasing LI (i.e.,
yield LRCs) are required to add to the current literature and guide commercial growers to make
informed decisions on optimal LI for cannabis production.

1.5 ULTRAVIOLET RADIATION IN PLANT PRODUCTION

Ultraviolet (UV) radiation in the solar spectrum is divided into three wavelength ranges: UVA
(315 to 400 nm), UVB (280 to 315 nm), and UVC (100 to 280 nm). The ozone layer of Earth’s
atmosphere absorbs the harmful wavelengths of UVC and some wavelengths in the UVB band
but allows UVA and some UVB wavelengths to reach ground level (De Gruijl and Van der
Leun, 2000). The ozone layer began thinning in the 1970s, allowing some of the shorter, more
harmful UV wavelengths to reach ground level. Upon the discovery of ozone depletion,
researchers sought to identify the risks that shorter wavelength UV radiation might have on life
on Earth. While the stratospheric ozone layer has recovered in most locations, the research
efforts identifying the effects of UV on agricultural plants uncovered its potential benefits. Using
horticultural science to test for optimal UV treatments in different species, UV radiation can be
used to manipulate crops to attain desirable traits.
10
Photomorphogenic responses to UV radiation are achieved through the activation of gene
expression (Jenkins, 2017), such as that of the UV resistant phytoreceptor-8 (UVR8) (Huché-
Thélier et al., 2016; Yin and Ulm, 2017) or through UV-induced oxidative damage (Tossi et al.,
2019). UV radiation activates UVR8, allowing it to bind to the protein called constitutively
photomorphogenic 1 (COP1), which initiates UV signalling (Huché-Thélier et al., 2016; Yin and
Ulm, 2017). The UV dependent interaction between UVR8 and COP1 is required for the
expression of ELONGATED HYPOCOTYL 5 (HY5) transcription factor (Jenkins, 2017). UV
radiation regulates the expression of many genes through UVR8-independent pathways as well.
UVR8-independent pathways may be triggered under more severe UV treatments (i.e., long-term
exposure, exposure to short wavelength radiation such as UVC) that induce oxidative damage.
Oxidative damage includes DNA mutagenesis through the formation of a dimer that inhibits
transcription and replication (Tossi et al., 2019), or through disruption resulting from UV-
induced reactive oxygen species (ROS) (Czégény et al., 2016). Overall, UV radiation can
modulate the expression of hundreds of genes, leading differential expression of plant
metabolism, morphology and physiology (Jenkins, 2017).

Biological responses to UV radiation are more sensitive to the shorter-wavelength (e.g., UVC)
radiation within the UV waveband (i.e., more energetic UV spectra) than to the longer
wavelength radiation (e.g., UVA) (Flint and Caldwell, 2003). Since the biological effects of UV
radiation are highly wavelength specific, many studies express UV treatments by multiplying
spectral irradiance at each wavelength by a Biological Spectral Weighting Function (BSWF).
Applying a BSWF specifically for plant growth responses to UV radiation treatments provides a
more accurate depiction of how the spectra will affect plant growth (Flint and Caldwell, 2003).
Although UV is only present in sunlight in small quantities relative to PAR, its shorter
wavelengths are disproportionately effective in plant response, which is accounted for with the
BSWF. Even after weighting each wavelength, the severity of the UV radiation treatment applied
to a crop is dependent on the intensity, the photoperiod (i.e., hours of UV radiation per day) and
number of days of exposure throughout the crop life cycle. Long-term UV exposure can cause
plant stress, whereas short-term exposure can induce minimal stress that elicits a beneficial
outcome to the organism (Robson et al., 2019; Wargent and Jordan, 2013). Under stronger

11
exposure levels, UV radiation can induce damage to the cellular DNA of a plant, leading to
genome instability and abnormalities in plant development (Friedberg, 2002; Manova and
Gruszka, 2015). DNA damage can be induced directly from the absorption of UV photons or by
the UV-driven production of ROS, which disrupts the balance between ROS production and
ROS scavenging, ultimately leading to oxidative stress (Robson et al., 2019). This oxidative
stress can damage DNA, lipids and proteins. On the other hand, other minor stress responses may
include the UV stimulated expression of genes involved in flavonoid biosynthesis and
antioxidant activity, which may be desirable secondary metabolites in crop production (Robson
et al., 2019). Since the shorter wavelengths in the UV waveband have stronger biological effects
(Flint and Caldwell, 2003), and amount of time of treatment impacts the biological effects of UV
radiation, it is important to consider not only intensity, but also wavelength and time when
evaluating the UV radiation treatments as reported in the literature. However, there is a lack of
consistency with the use of BSWFs in the current literature relating to plant responses to UV
radiation, where recent studies often refer to BSWF by Flint and Caldwell (2003) and older
studies refer to the BSWF by Caldwell (1971). There is also a lack of consistency when reporting
of UV intensity, photoperiod, and wavelength (Huché-Thelier et al., 2016). Therefore, caution is
required when analyzing data on plant responses to UV radiation, and it may be most relevant to
compare the current literature in relative terms rather than absolute UV treatments.

Although there is a large body of literature focused on the effects of UV radiation on agronomic
and horticultural crops, the UV exposure conditions are highly variable between research groups.
Experimental designs typically include versions of 1) low compared to high UV radiation
treatments using UV lamps, or 2) UV radiation exclusion treatments compared to ambient
sunlight UV exposure. Many of these experiments have variable UV to PAR ratios, and use UV
lamps that may contain shorter or longer wavelengths than reported (e.g., UVC or UVA). Some
studies are conducted using sunlight as the source of PAR, while others use various indoor
production lighting methods, with variable spectrums. There are often large differences between
plant UV response in controlled indoor environments compared to outdoor field environments
(Robson et al., 2019). Plants exhibit higher sensitivity to UV radiation as UV:PAR increases
(Behn et al., 2010; Dou et al., 2018), and other spectra present within the treatments can

12
influence the plant sensitivity to UV radiation as well (Palma et al., 2021). Often, the units of
reported UV treatments between studies are highly variable (Huché-Thélier et al., 2016). Given
the wide range of variability between UV exposure conditions, the effects of UV radiation on
plant growth, yield, physiology, and secondary metabolites are highly variable as well.

Despite the variability in the effects of UV radiation on plant morphology and physiology, there
has been a well-established understanding of the “UVB phenotype” (Robson et al., 2019; Jansen
et al., 2017). UVB exposure has been shown to decrease plant stem length in a variety of
monocotyledons and dicotyledons (Barnes et al., 1990; Liu et al., 2013) and increase number of
branches (e.g., in dicotyledons such as bean and rose) and tillers (e.g., in monocotyledons such
as oat and wheat) (Barnes et al., 1990; Torre et al., 2012) leading to reduced internode lengths
and an overall more compact phenotype. Leaf area is highly affected by UVB exposure with
decreases at higher vs. lower levels of UVB in wheat and wild oat, rapeseed, four cultivars of
cucumber, three cultivars of rose and two cultivars of soya (Barnes et al., 1990; Cen and
Bornman, 1993; Krizek et al., 1997; Terfa et al., 2014; Zhang et al., 2014). Other commonly
reported impacts of UVB exposure include increased specific leaf weight (SLW; g·cm–2), a
proxy for leaf thickness (Cen and Bornman, 1993; Zhang et al., 2014), a reduction in total leaf
number (Krizek et al., 1997), a reduction in petiole length (Krizek et al., 1997; Zhang et al.,
2014), and changes in leaf shape (i.e., leaf length:width) (Hectors et al., 2010; Klem et al., 2012;
Robson and Aphalo, 2012). Less frequently reported responses to UVB exposure include
epicuticular wax accumulation, which may decrease UVB penetration by reflectance (Cen and
Bornman, 1993) and leaf epinasty, which is when tissue between leaf venation concaves away
from incident light (Fierro et al., 2015; Jansen, 2002).

Some studies have reported bumpy leaf surfaces and necrotic spots on leaves that were exposed
to UVB (Klem et al., 2012; Torre et al., 2012). In Lactuca sativa L. and Cucumis sativus L.
excluded from UV exposure, higher total dry weight (DW) relative to the UVB exposure
treatment has been reported (Krizek et al., 1997). However, morphological responses to UVB,
such as reduced plant height and increased branching, can occur without alterations to carbon
assimilation and total shoot DW (Barnes et al., 1990). UVB exposure induces early flowering in
poinsettia (Torre et al., 2012), but Setaria and Amaranthus species exposed to UVB
13
demonstrated increases in biomass allocation to main shoot reproductive tissues (Barnes et al.,
1990). UVB radiation also has the potential to reduce disease incidence for cannabis diseases
such as powdery mildew (Austin and Wilcox, 2012; Demkura and Ballaré, 2012). Although there
are many studies reporting the effects of UVB on a variety of species, there are many potential
impacts of UVB on plant morphology, which may be variable depending on the UVB exposure
conditions (e.g., UVB lamps versus sunlight exposure), treatments (e.g., hours per day and days
per growing cycle).

UVB exposure in many plants can impact leaf stress and physiology. Gas exchange parameters
(such as NCER and Asat) are often lower in leaves that are exposed to supplemental UVB (Dou et
al., 2018; Hoffmann et al., 2015; Pacher et al., 2006; Yang et al., 2008; Zhang et al., 2014),
which could indicate damage to the photosystems. Hoffman et al. (2015) also found that
supplemental UVB radiation induced stress in pepper leaves demonstrated by a decline in dark-
adapted chlorophyll fluorescence (Fv/Fm). Chlorophyll content has been found to either increase
or decrease in a wide range of other dicotyledons and monocotyledons in response to UVB
radiation (Neugart and Schreiner, 2018). Plants demonstrate a higher sensitivity to UVB under
higher UV:PAR with respect to total chlorophyll content (Chl a + b; µg cm−2), Asat and Fv/Fm
(Klem et al., 2012). Such variations in physiological and morphological responses to UVB may
be attributed to the highly variable UVB exposure conditions and differences in UV:PAR. For
cannabis production, understanding the effects of UVB exposure on whole plant morphology and
leaf physiology is relevant to production when it comes to establishing a UVB exposure damage
threshold.

1.5.1 EFFECTS OF ULTRAVIOLET RADIATION ON CANNABIS PHOTOSYNTHESIS,

GROWTH AND SECONDARY METABOLITES

The application of UV radiation during cultivation is an area of interest in the cannabis industry
for its purported potential to increase cannabinoid concentration in mature female cannabis
inflorescences tissues (hereafter, inflorescences). It has been shown that cannabis plants with
higher Δ9-THC to CBD ratios typically originated from equatorial regions (i.e., latitudes between
the equator and 30°N or S, whereas plants with low Δ9-THC and high CBD concentrations
typically originate from latitudes north of 30°N or south of 30°S) (Small & Beckstead, 1973;
14
Small & Cronquist, 1976). Pate (1983) proposed a protective function model suggesting that Δ9-
THC protects cannabis tissue (i.e., leaves and inflorescences) from UV radiation. Factors
proposed to contribute to this hypothetical model include: high Δ9-THC production in plants
grown in areas with high UV irradiation (e.g., high altitudes, low latitudes), the UV absorbing
properties of Δ9-THC, and high Δ9-THC production in the female floral tissues which are
responsible for reproductive success and must endure longer UV exposure than the male flowers.
The findings by Pate (1983) showed that Δ9-THC concentration in cannabis inflorescences is
higher in plants with origins that have high UV exposure [in watt-sec·cm–2, 302.5-312 nm,
according to the world distribution map of UV exposure by Schulze and Grafe (1969)] and
conversely there is a negative correlation between CBD concentration and high UV exposure.

UV application may be a useful tool to increase Δ9-THC concentration in indoor production of

modern drug-type cannabis genotypes. Horticultural researchers have evaluated plant responses
to UV in indoor cannabis production settings. Early controlled-environment studies postulate
there is a potential for UV radiation to increase Δ9-THC concentration in cannabis leaf and
inflorescence tissues (Fairbairn and Liebmann, 1974; Lydon et al., 1987), however the
concentration of Δ9-THC in typical inflorescences has increased over time where modern
cannabis genotypes have ≈10× higher Δ9-THC concentration in inflorescence tissue compared to
the older genotypes used in older studies (Dujourdy and Besacier, 2017). It is possible that
modern cannabis genotypes are near their maximum genetic ability to produce Δ9-THC,
impeding their ability to further increase inflorescence Δ9-THC concentration in response to UV
radiation, relative to older genotypes. Giupponi et al. (2020) found that CBD-dominant cannabis
inflorescences exposed to high UV (i.e., higher altitudes) had greater CBD and terpene
concentrations compared to plants exposed to low UV (i.e., lower altitudes), suggesting that Δ9-
THC may not be the only cannabinoid upregulated in response to UV radiation. Cannabinoid
concentration has also been shown to vary with temperature conditions (Bazzaz et al., 1975),
sub-optimal nutrient availability (Caplan et al., 2017; Haney and Kutscheid, 1973; Yep et al.,
2020) and drought stress (Caplan et al., 2019; Haney and Kutscheid, 1973; Latta and Eaton,
1975). In general, there is potential for beneficial responses as a result of environmental changes
in even in modern cannabis genotypes.

15
Currently, there are a few studies that demonstrate the potential to increase cannabinoid yields in
cannabis inflorescences with the application of UV radiation during production (Giupponi et al.,
2020; Lydon et al., 1987; Marti et al., 2014). However, there are no reliable studies
demonstrating an increase in cannabis inflorescence cannabinoid concentration with
reproducible, clearly defined UV treatments (i.e., hours of application, days of application,
intensity in either radiant flux or photon flux units, peak wavelengths) that are relevant to
modern cannabis production (i.e., using supplemental UV lamps in an indoor production facility
and modern cannabis genotypes). There are no studies in the current literature that determine the
effect of UV radiation on cultivars with balanced Δ9-THC:CBD, which would indicate
differences between cannabinoid responses.

16
1.6 THESIS STRUCTURE AND OBJECTIVES
Considering the limited scientific research on LI and UV radiation in cannabis production and
the economic and medicinal importance of this crop, the goal of this thesis was to determine the
effects of these treatments on cannabis photosynthesis, growth, inflorescence yield, and
cannabinoid concentrations. Determining how cannabis responds to various lighting strategies in
the flowering stage will provide insight into the optimization of cannabis production.

The specific objectives were to develop descriptive models can be used widely in the Cannabis
industry and future research relating:

1. LI and cannabis leaf-level photosynthesis, and inflorescence yield and quality.

2. UV exposure levels and cannabis morphology, physiology, and inflorescence yield and
quality via photoprotection mechanism.

Note: Chapter 2 and 3 follow the Frontiers’ style guidelines and have been submitted to
Frontiers in Plant Science, under the research topic entitled ‘Smoke and Mirrors: Reflections on
Improving Cannabis Production and Investigating Medical Potential’. Chapter 2 has been
accepted and published:

Rodriguez-Morrison, V., Llewellyn, D. and Zheng, Y. (2021). Cannabis yield, potency, and
leaf photosynthesis respond differently to increasing light levels in an indoor
environment. Front. Plant Sci. 12:646020. doi: 10.3389/fpls.2021.646020.

17
 CHAPTER TWO
CANNABIS YIELD, QUALITY, AND LEAF PHOTOSYNTHESIS
RESPOND DIFFERENTLY TO INCREASING LIGHT LEVELS IN AN
INDOOR ENVIRONMENT
ABSTRACT
Since the recent legalization of medical and recreational use of cannabis in many regions
worldwide, there has been high demand for research to improve yield and quality. With the
paucity of scientific literature on the topic, this study investigated the relationships between LI
and photosynthesis, inflorescence yield, and inflorescence quality of cannabis grown in an indoor
environment. After growing vegetatively for 2 weeks under a canopy-level PPFD of ≈425
μmol·m–2·s–1 and an 18-h light/6-h dark photoperiod, plants were grown for 12 weeks in a 12-h
light/12-h dark “flowering” photoperiod under canopy-level PPFDs ranging from 120 to 1800
μmol·m–2·s–1 provided by LEDs. Leaf LRCs varied both with localized (i.e., leaf-level) PPFD
and temporally, throughout the flowering cycle. Therefore, it was concluded that the leaf light
response is not a reliable predictor of whole-plant responses to LI, particularly crop yield. This
may be especially evident given that dry inflorescence yield increased linearly with increasing
canopy-level PPFD up to 1800 μmol·m–2·s–1, while leaf-level photosynthesis saturated well
below 1800 μmol·m–2·s–1. The density of the apical inflorescence and harvest index also
increased linearly with increasing LI, resulting in higher-quality marketable tissues and less
superfluous tissue to dispose of. There were no LI treatment effects on cannabinoid
concentration, while there were minor LI treatment effects on terpene concentration. Commercial
cannabis growers can use these light response models to determine the optimum LI for their
production environment to achieve the best economic return; balancing input costs with the
commercial value of their cannabis products.

2.1 INTRODUCTION
Drug-type cannabis (i.e., genotypes grown for their high cannabinoid content; hereafter,
cannabis) is often produced indoors to allow complete control of environmental conditions,
which is important for producing consistent medicinal plants and products (UNODC, 2019;

18
Zheng, 2020). Total reliance on electrical lighting for plant production gives growers the
capability to manipulate crop morphology, yield, and quality using light. However, lighting-
related costs comprise ≈60% of total energy used for indoor cannabis production (Evergreen
Economics, 2016; Mills, 2012); making crop lighting one of the most substantial input costs for
growing cannabis indoors. With recent nationwide legalization in Canada (among many other
regions worldwide), energy demand for indoor cannabis production is expected to increase
rapidly as the industry intensifies production to address rising demand (Sen and Wyonch, 2018).

There are many factors that govern the cost of producing PAR for indoor cannabis production.
These factors include: the capital and maintenance costs of lighting fixtures and related
infrastructure, efficiency of converting electricity into PAR [usually referred to as PAR efficacy;
in units of µmol(PAR)·J–1], management of excess heat and humidity, and uniformity of PAR
distribution within the plant canopy. The most common lighting technologies used for indoor
cannabis production are high intensity discharge (e.g., HPS) and LED (Evergreen Economics,
2016; Mills, 2012). These technologies have widely varying spectrum, distribution, PAR
efficacy, and capital costs. However, regardless of the lighting technology used, the dominant
factor that regulates the cost of crop lighting is the target canopy-level LI.

One common precept in controlled-environment agriculture production is that crop yield

responds proportionally to increasing LI; i.e., the so-called “1% rule” whereby 1% more PAR
equals 1% greater yield (Marcelis et al., 2006). On a per-leaf basis, this principle is clearly
limited to lower light intensities, since light use efficiency [i.e., maximum quantum yield; QY,
μmol(CO2)·μmol–1(PAR)] of all photosynthetic tissues begins to decline at LI well below their
LSPs (Posada et al., 2012). However, in indoor-grown cannabis, it is conceivable that whole-
plant photosynthesis will be maximized when LI at the upper canopy leaves are near their LSP.
This is partly attributable to the inter-canopy attenuation of PAR from self-shading; allowing
lower-canopy foliage to function within the range of LIs where their respective light use
efficiencies are optimized (Terashima and Hikosaka, 1995). This may be especially relevant to
indoor production, where relatively small changes in distance from the light source can impart
substantial differences in foliar LI (Niinemets and Keenan, 2012). Further, distinguished from

19
many other indoor-grown crops, cannabis foliage appears to tolerate very high LI, even when
exposed to PPFDs that are much higher than what they have been acclimated to (Chandra et al.,
2015).

There is a paucity of peer-reviewed studies that have related LI to cannabis cannabinoid

concentration and yield (e.g., mass of dry, mature inflorescence per unit area and time). Perhaps
the most referenced studies report aspects of single-leaf photosynthesis of several cultivars and
under various PPFD, CO2 concentration, and temperature regimes (Chandra et al., 2011; 2015;
Lydon et al., 1987). These works have demonstrated that cannabis leaves have very high
photosynthetic capacity. However, they have limited use in modeling whole canopy
photosynthesis or predicting yield because single-leaf photosynthesis is highly variable;
depending on many factors during plant growth such as: leaf age, their localized growing
environments (e.g., temperature, CO2, and lighting history), and ontogenetic stage (Bauerle et al.,
2020; Carvalho et al., 2015; Murchie et al., 2002; Zheng et al., 2006). While lighting vendors
have long relied on cannabis leaf photosynthesis studies to sell more light fixtures to cannabis
growers, their models are only tangentially related to whole-canopy photosynthesis, growth, and
ultimately yield (Kirschbaum, 2011).

Some forensic studies have utilized various methods to develop models to estimate crop yield
from illicit indoor cannabis production (Backer et al., 2019; Potter and Duncombe, 2012; Toonen
et al., 2006; Vanhove et al., 2011). These models use an array of input parameters (e.g., planting
density, growing area, crop nutrition factors) but, rely on “installed wattage” (i.e., W·m–2) as a
proxy for LI. It is notable that reporting yield as g·W–1 (i.e., g·m–2 / W·m–2) overlooks the
instantaneous time factor inherent in power units (i.e., W = J·s –1). A more appropriate yield
metric would also account for the length of the total lighting time throughout the production
period (i.e., h·d–1 × d), thus factoring out the time units resulting in yield per unit energy input
(e.g., g·kWh–1). Furthermore, area-integrated power does not directly correlate to the canopy-
level light environment due to a myriad of unknowns, such as hang height, light distribution, and
fixture efficacy. It is therefore impossible to accurately ascertain canopy-level LI in these
models. Eaves et al. (2020) reported linear relationships between canopy-level LI (up to 1500

20
µmol·m–2·s–1) and yield; however, they had only one LI treatment above 1000 µmol·m–2·s–1.
Furthermore, they reported substantial inter-repetition variability in their yield models, which
indicates that factors other than LI may have limited crop productivity in some circumstances.
While methodological deficiencies in these studies may limit the confident quantitative
extrapolation of their results to production environments, it is striking that none of these studies
reported evidence of saturation of inflorescence yield at very high LI.

These studies all demonstrate the exceptionally high capacity that cannabis has for converting
PAR into biomass. However, there are also clear knowledge gaps in cannabis’ photosynthesis
and yield responses to increasing LI. In addition, cannabis products are very high-value
commodities relative to other crops grown in indoor environments. This means that producers
may be willing to accept substantially higher lighting-related input costs in order to promote
higher yields in limited growing areas. However, maximizing yield regardless of cost is not a
feasible business model for most cannabis producers; rather there is a trade-off between input
costs and crop productivity by selecting the optimum canopy-level LI (among other inputs) that
will maximize net profits. Further complicating matters, producers must balance fixed costs
which do not vary with crop productivity (such as property tax, lease rates, building security, and
maintenance, etc.) and variable costs (such as the aforementioned lighting-related costs among
other crop inputs) which can have dramatic impacts on crop productivity and yield (Vanhove et
al., 2014). Since indoor crop lighting is a compromise between input costs and crop productivity,
it is critical for growers to select the optimum LI for their respective production environment and
business models.

The objectives of this study were to establish the relationships between canopy-level LI, leaf-
level photosynthesis, and yield and quality of drug-type cannabis. We investigated how plant
growth stage and localized foliar PPFD (LPPFD; i.e., instantaneous PPFD at leaf-level) affected
photosynthetic parameters and leaf morphology, and how growing cannabis at average canopy-
level PPFDs (APPFD; i.e., lighting history) ranging from 120 to 1800 µmol·m–2·s–1 affected
plant morphology, yield, and quality of mature marketable inflorescence. The results of this
study will assist the indoor cannabis industry to determine how much PAR cannabis growers

21
should be providing to the crop canopy in order to maximize profits while minimizing energy
use within their specific production scenarios.

2.2 MATERIALS AND METHODS

The trial area consisted of 2 adjacent deep-water culture basins (CB) located in an indoor
cannabis production facility in southern Ontario, Canada. Each CB (14.6 × 2.4 m) consisted of
24 parallel polystyrene rafts (0.6 × 2.4 m), each containing holes for 16 plant pots, oriented in 2
rows with 30-cm spacing both within- and between-rows. This spacing provided for 384 plants
to be evenly spaced within each CB, at a density of 0.09 m2/plant.

Figure 2.1. Relative spectral photon flux distribution of Pro-650 (Lumigrow) LED fixtures.

Above each CB were 3 racks of LED fixtures (Pro-650; Lumigrow, Emeryville, CA, USA), with
each rack consisting 2 rows of 4 fixtures each; arranged such that all 24 fixtures were uniformly-

22
spaced (1.2 m apart, on-center) relative to each other and centered over the footprint of the CB.
Each rack of fixtures was height-adjustable via a system of pulleys and cables, such that the
hang-height of the 8 fixtures in each rack could be adjusted in unison. Each fixture contained
dimmable spectrum channels for blue (B, peak 455 nm), white (broad-spectrum 5000K) and red
(R, peak 660 nm) which could be individually controlled, wirelessly, through Lumigrow’s
SmartPAR software. The photon flux ratio of B (400-500 nm), green (G, 500-600 nm), and R
(600-700 nm) was B18:G5:R77. Relative spectral photon flux distribution (Figure 2.1) was
measured using a radiometrically calibrated spectrometer (UV-VIS Flame-S-XR; Ocean Optics,
Dunedin, FL, USA) coupled to a CC3 cosine-corrector attached to a 1.9 m × 400 µm UV-Vis
optical fiber.

2.2.1 EXPERIMENTAL DESIGN

The experiment was conducted using a gradient design, whereby plants grown in a common
environment were exposed to a broad range of canopy-level PPFDs with a high level of spatial
variability across the CB. Individual plants were assigned APPFD levels based on rigorous
spatial and temporal evaluations of LI (explained below). Gradient designs can outperform
traditional “treatment × replication” experimental designs when evaluating plants’ responses to a
continuous variable such as LI (Kreyling et al., 2018). Despite the fact they are arduous to setup
and monitor, gradient designs have been successfully used to establish LI effects within other
controlled-environment production scenarios (Bredmose, 1993, 1994; Jones-Baumgardt et al.,
2019).

23
Figure 2.2. Schematic of a single light rack (8 LED fixtures, in magenta) above one third of a
deep-water culture basins (CB). The entire growing area consists of 6 of these light racks. Within
each light rack, each of the 8 target PPFD levels (i.e., the “treatments”) were randomly assigned
to one fixture (i.e., plot). This resulted in a randomized complete block type of experimental
layout, comprised of 8 treatments × 6 replications. However, each treatment plant (in blue) was
assigned an average photosynthetic photon flux density (APPFD) as LI treatment levels,
reflecting the average canopy-level light intensity measured throughout the trial. The APPFD
levels were used as the independent variable in subsequent analyses of plant growth, physiology
and harvest metrics. Each plot was surrounded by non-treatment plants (diagonal lines) to ensure
uniform growing environment and normal planting density.

24
To facilitate setup, the experiment was initially arranged as a randomized complete block type of
design with 6 blocks (i.e., 3 blocks per CB) of 8 PPFD target levels: 200, 400, 600, 800, 1000,
1200, l400, and 1600 μmol·m–2·s–1. Each block consisted of a single rack of LED fixtures, with
the PPFD target levels randomly assigned to individual fixtures (i.e., plots) within each rack. The
two plants located most directly below each fixture were assessed experimentally (Figure 2.2).
PPFD was measured at the apex of each plant using a portable spectroradiometer (LI-180; LI-
COR Biosciences, Lincoln, NE, USA). The initial hang height of each rack was determined by
the maximum height whereby approximately 1600 μmol·m–2·s–1 could be achieved at the apical
meristem of the tallest plant in the highest LI plot. The other treatment levels were subsequently
achieved through dimming; targeting the prescribed PPFD at the apical meristem of the tallest
plant in each plot while maintaining a uniform photon flux ratio of B18:G5:R77 in the entire CB.
Plant height and apical meristematic PPFD were measured twice weekly until vegetative growth
ceased (five weeks after the start of the 12-h photoperiod), and weekly thereafter until harvest.
The prescribed intensity levels in each block were reset each time plant height was measured,
first by raising the rack of fixtures to achieve the target PPFD at the apical meristem of the tallest
plant in the 1600 μmol·m–2·s–1 plot and then adjusting the intensity settings of the remaining
plots accordingly. The trial ran from the beginning of the flowering stage (i.e., when the 12-h
flowering photoperiod was initiated) until harvest, for a total of 81 days (nearly 12 weeks).
While the underlying experimental arrangement was based on a RCBD organization, all analyses
were performed as regressions with LI as the continuous, independent variable.

2.2.2 PPFD LEVELS

Although the prescribed target PPFD levels were maintained at the apical meristem position for
the tallest plant within each plot on regular intervals, these values were not accurate proxies for
the actual PPFD intensity dynamics experienced by each plant throughout the trial due to
variability in individual plant height (on intra- and inter-plot bases), growth rates, and the lengths
of the time periods between PPFD measurements. To account for these temporal dynamics in
apical meristematic PPFD, total light integrals (TLIs, mol·m–2) were calculated for each plant
over the total production time and then back-calculated to APPFD or daily light integral (DLI,
mol·m–2·d–1). The TLIs were based on the product of the PPFD level measured at the start and
25
end of each measurement interval and the length of time the lights were on during each
measurement interval. These interim light integrals were then aggregated to form a TLI for each
plant and divided by the total production time in seconds (i.e., the product of the daily
photoperiod and the number of days). The resulting APPFD levels were then used as the
independent variable (i.e., X-axis) in regressions of LI vs. various growth, yield and quality
parameters. TLI can also be used in yield evaluations whereby the relationship between yield and
TLI becomes a direct measure of production efficacy on a quantum basis (e.g., g·mol–1). This
relationship can be converted to an energy-basis (g·kWh–1), if the fixture efficacy (μmol·J–1) and
spatial distribution efficiency (i.e., proportion of photon output from fixtures that reach the target
growing area) are known.

2.2.3 PLANT CULTURE

Cuttings were taken from mother plants of the ‘Stillwater’ cultivar, a CBD-dominant cultivar, on
August 1 and 15 2019. Cuttings were rooted in stone wool cubes under 100 μmol·m–2·s–1 of
fluorescent light for 14 d and then transplanted into a peat-based medium in 1-gallon plastic pots
and grown under ≈425 μmol·m–2·s–1 of LED light, comprised of a mixture of Pro-325
(Lumigrow) and generic phosphor-converted white LEDs (unbranded) for an additional 14 d.
The apical meristems were removed (i.e., “topped”) from the first batch of clones, 10 d after
transplant, and the second batch were not topped. Propagation and vegetative growth phases both
had 18-h photoperiods. The first CB (CB1) was populated from the first batch of clones on
August 20 2019 and the second CB (CB2) was populated from the second batch of clones on
September 12 2019. In each case, 48 uniform and representative plants were selected from the
larger populations of clones and placed in the plots to be evaluated experimentally. In CB1, the
experimental plants initially had either 9 or 10 nodes and ranged in height (from growing
medium surface to shoot apex) from 34 to 48 cm. In CB2 the experimental plants initially had
either 12 or 13 nodes and ranged in height from 41 to 65 cm. Once the plants were moved to the
CBs, the daily photoperiod switched to 12 h, from 06:30 HR to 18:30 HR.

Plant husbandry followed the cultivator’s standard operating procedures except for the
differences in canopy-level PPFD. Canopy-level air temperature, relative humidity (RH), and

26
CO2 concentration were monitored at 600-s intervals throughout the trial with a data logger
(Green Eye model 7788; AZ Instrument Corporation, Taiwan). Throughout the experiment, the
air temperature, RH, and CO2 concentrations were (mean ± SD) 25.3 ± 0.4°C, 60.5 ± 4.8%, and
437 ± 39 ppm during the day (i.e., lights on) and 25.2 ± 0.3°C, 53.1 ± 3.3%, and 479 ± 42 ppm
during the night. A common nutrient solution was circulated through both CBs. The nutrient
concentrations in the aquaponic solution supplying both CBs were sampled weekly (for 11
weeks) and analyzed at an independent laboratory (A&L Canada; London, ON, Canada). The
nutrient element concentrations (mg·L–1) in the aquaponic system were averaged across the
weekly sampling (mean ± SD): 170 ± 22 Ca, 86 ± 8.2 S, 75 ± 15 N, 57 ± 5 Mg, 32 ± 4 P, 23 ± 8
K, 250 ± 32 Cl, 0.27 ± 0.1 Fe, 0.18 ± 0.07 Zn, 0.050 ± 0.02 Mn, 0.031 ± 0.006 B, and 0.028 ±
0.004 Cu. Mo was reported as below detection limit (i.e., <0.02 mg·L–1) throughout the trial. The
concentrations (mg·L–1) of non-essential nutrient elements were 170 ± 18 Na and 6.7 ± 0.7 Si.
The aquaponic solution was aerated with an oxygen concentrator and the pH and EC were 6.75 ±
0.2 and 1.77 ± 0.15 mS·cm–1, respectively.

2.2.4 LEAF PHOTOSYNTHESIS

Quantifications of leaf-level gas exchange of leaflets on the youngest, fully-expanded fan leaves
were performed on 64 plants (32 plants per CB) each, in weeks 1, 5, and 9 after the initiation of
the 12-h photoperiod using a portable photosynthesis machine (LI-6400XT; LI-COR
Biosciences, Lincoln, NE, USA), equipped with the B and R LED light source (6400-02B; LI-
COR Biosciences). The Light Curve Auto-Response subroutine was used to measure NCER
[μmol(CO2)·m–2·s–1] at PPFD levels of: 2000, 1600, 1400, 1200, 1000, 800, 600, 400, 200, 150,
100, 75, 50, 25, and 0 µmol·m–2·s–1. Since cannabis leaves are palmately compound, the middle
leaflet was used to cover the 6 cm2 chamber of the LI-COR 6400XT for photosynthesis
measurements, because the middle leaflet is the largest. Occasionally, two leaflets of the same
leaf were inserted to ensure the whole chamber was covered, similar to the methods used for oak
seedlings (Goodman et al., 2007). Leaflets were exposed to 2000 µmol·m–2·s–1 for 180 s prior to
starting each light response curve (LRC) and then progressed sequentially from highest to lowest
PPFD to ensure stomatal opening was not a limitation of photosynthesis (Singsaas et al., 2001).
The leaf chamber setpoints were 26.7°C (block temperature), 400 ppm CO 2, and 500 µmol·s–1
27
airflow. The localized PPFD (LPPFD) at each leaflet was measured immediately prior to the LRC
measurement using the LI-180. Asat [μmol(CO2)·m–2·s–1], localized NCER (LNCER), maximum
QY [μmol(CO2)·μmol–1(PAR)], and LSP [μmol(PAR)·m–2·s–1] were determined for each measured
leaflet using Prism (Version 6.01; GraphPad Software, San Diego, CA, USA) with the
asymptotic LRC model: y = a + b·e(c·x) (Delgado et al., 1993) where y, x, a, and e represent
NCER, PPFD, Asat, and Euler’s number, respectively. The LNCER of each leaflet was calculated
by substituting the measured LPPFD into its respective LRC model. The QY was calculated as
the slope of the linear portion of the LRC (i.e., at PPFD ≤200 μmol·m–2·s–1). The LSP is defined
as the PPFD level where increasing LI no longer invokes a significant increase in NCER. The
LSP for each LRC was determined using the methods described by Lobo et al. (2013) by
evaluating the change in NCER (ΔNCER) over 50 μmol(PAR)·m–2·s–1 increments, continuously
along the LRC, until the ΔNCER reached a threshold value, which was determined from the
prescribed measurement conditions and performance specifications of the LI-6400XT. Briefly,
the minimum significant difference in CO2 concentration between sample and reference
measurements is 0.4 ppm (LI-COR Biosciences, 2012). Therefore, given the setup parameters of
the leaf chamber, a ΔNCER of ≤0.33 μmol(CO2)·m–2·s–1 over a 50 µmol(PAR)·m–2·s–1 increment
indicated the LSP.

The ratio of variable to maximum fluorescence (Fv/Fm) emitted from photosystem II in dark-
acclimated leaves exposed to a light-saturating pulse is an indicator of maximum quantum yield
of photosystem II photochemistry (Murchie and Lawson, 2013). Immediately after each LRC,
the leaflet was dark acclimated for ≈900 s and then Fv/Fm was measured with a fluorometer
(FluorPen FP 100; Drasov, Czech Republic). Chlorophyll content index (CCI) was measured on
three fan leaflets from leaves at the bottom and top of each plant in weeks 1, 5, and 9 using a
chlorophyll meter (CCM-200; Opti-Sciences, Hudson, NH, USA). The CCI measurements from
upper and lower tissues, respectively, were averaged on a per-plant basis for each measurement
period.

28
2.2.5 LEAF MORPHOLOGY

On day 35 of the experiment, one leaf from each plant was removed from node 13 (counting
upwards from the lowest node) in CB1 and node 15 from CB2, ensuring that the excised leaves
developed under their respective LPPFD. A digital image of each leaf was taken using a scanner
(CanoScan LiDE 25; Canon Canada Inc., Brampton, ON, Canada) at 600 dpi resolution and then
the leaves were oven-dried (Isotemp Oven Model 655G; Fisher Scientific, East Lyme, CT,
USA), singly, to constant weight at 65°C. The images were processed using ImageJ 1.42
software (National Institute of Health; https://imagej.nih.gov/ij/download.html) to determine leaf
area (LA). The DW of scanned leaves were measured using an analytical balance
(MS304TS/A00; Mettler-Toledo, Columbus, OH, USA). Specific leaf weight (SLW; g·m–2) was
determined using the following formula: DW / LA. See Appendix A for plant growth
methodology, and Appendix B for additional visual observations.

2.2.6 YIELD AND QUALITY

After 81 d of the experiment, the stems of each plant were cut at substrate level and the
aboveground biomass of each plant was separated into three parts: apical inflorescence,
remaining inflorescence, and stems and leaves (i.e., non-marketable biomass), and weighed using
a digital scale (Scout SPX2201; OHAUS Corporation, Parsippany, NJ, USA). Since the plants
from CB2 had the apical meristem removed, the inflorescence from the tallest side branch was
considered the apical inflorescence. The length (L) and circumference (C; measured at the
midpoint) of each apical inflorescence were also measured. Assuming a cylindrical shape, the
density of the apical inflorescence (g·cm–3) was calculated using the formula: apical
inflorescence density = fresh weight/{π·[C/(2·π )2]·L}. The apical inflorescences from 22
representative plants (selected to ensure evenly distributed APPFD) from CB1 were air dried at
15°C and 40% RH for 10 d until they reached marketable weight (i.e., average moisture content
of ≈11%), determined using a moisture content analyzer (HC-103 Halogen Moisture Analyzer;
Mettler-Toledo, Columbus, OH, USA). This ensured that the apical inflorescence tissues selected
for analysis of secondary metabolites followed the cultivator’s typical post-harvest treatment.
The apical inflorescences from CB1 were homogenized on a per-plant basis and ≈2-g sub-
samples from each plant was processed by an independent laboratory (RPC Science &
29
Engineering; Fredericton, NB, Canada) for concentration [mg·g–1(DW) using solvent extraction
followed by ultra-high-performance liquid chromatography with variable wavelength detection
(HPLC-VWD) for cannabinoids and gas chromatography with mass spectrometry detection (GC-
MSD) for terpenes]. Total equivalent ∆9-THC, CBD, and CBG concentrations were determined
by assuming complete carboxylation of the acid-forms of the respective cannabinoids, whose
concentrations were adjusted by factoring out the acid-moiety from the molecular weight of each
compound [e.g., total ∆9-THC = (∆9-THCA × 0.877) + ∆9-THC]. The separated aboveground
tissues from 16 representative plants in each CB were oven-dried (Isotemp Oven Model 655G) to
constant weight at 65°C to determine LI treatment effects on moisture content, which were then
used to determine DW of all harvested materials. The harvest index was calculated as the ratio of
total inflorescence DW (hereafter, yield) to the total aboveground DW, on a per-plant basis.

2.2.7 DATA PROCESSING AND ANALYSIS

Although the initial experimental setup was arranged as a randomized complete block type of
design, blocking played no part in the statistical analysis of the results. Data were analyzed for
random (i.e., blocks) and fixed (i.e., APPFD) effects, which revealed that the variation attributed
to the random effects were not significantly different from 0. On per-CB and per-week bases,
each model from the leaf photosynthesis measurements (i.e., Asat, LSP, LNCER, and QY) were
subjected to non-linear regression using the PROC NLMIXED procedure (SAS Studio Release
3.8; SAS Institute Inc., Cary, NC), with the LPPFD of each measured leaf as the independent
variable, to determine the best-fit models after outliers were removed. In each case, best-fit
models were selected based on the lowest value for the Akaike information criterion (AICc). If
there were no LI treatment effects on a given parameter, then means (± SD) were calculated.
Best-fit models for Fv/Fm and CCI were similarly determined, using LPPFD and APPFD (from
the start of the trial up to the time of measurement), respectively, as the independent variable. On
a per-week basis, Asat, LSP, LNCER, QY, Fv/Fm, and CCI data from CB1 and CB2 were pooled
if the 95% confidence intervals (95% CI) of each element of the respective best-fit models for
the two CBs overlapped, and best-fit models for pooled datasets were then recalculated. The
PROC GLIMMIX Tukey-Kramer test was used (P ≤0.05) on means to determine if there were
differences between the measurement periods (i.e., weeks). If there were any measurement
30
period effects on any element in the models, then weekly models for the respective parameters
were reported.

Computed parameters from single-time measurements (SLW, apical inflorescence density, yield,
and harvest index) were grouped per CB, using the APPFD (at the time of measurement) to
define each datapoint within each CB and PROC NLMIXED was used to evaluate the best fit
model for each parameter using the AICc. Parameter means were computed (on per-CB bases)
when there were no LI treatment effects. If there were LI treatment effects on a given parameter,
datasets from CB1 and CB2 were pooled if the 95% CI of each element of the respective best-fit
models for the two CBs overlapped and best-fit models for pooled datasets were then
recalculated. For parameters with no LI treatment effects, differences between CBs were
evaluated using the 95% CIs of their respective means. For a given parameter, if the 95% CIs the
parameter means for the 2 CBs overlapped, then the data were pooled, and new parameter means
were calculated and presented. Cannabinoids and terpenes from CB1 were modeled, with APPFD
as the independent variable, using PROC NLMIXED to evaluate the best-fit model for each
parameter using the AICc. Best-fit models or parameter means were reported.

2.3 RESULTS
No CB effects were found in any leaf photosynthesis, leaf morphology, and postharvest
parameters; therefore, CB1 and CB2 data were pooled for the development of all models except
secondary metabolites, which were only measured in CB1. In contrast, many of the parameters
that were repeated over time (i.e., in weeks 1, 5, and 9 of the experiment) showed differences
between weeks; whereby the different weeks were modeled separately. Note also that the week-
over-week ranges of LPPFD varied as the plants progressed through their ontogeny, since self-
shading from upper tissues resulted in decreases in maximum LPPFD of leaves selected for
photosynthesis measurements. Nevertheless, a consistent range of APPFDs was maintained
throughout the trial.

31
Figure 2.3. Typical light response curves [net CO2 exchange rate (NCER) response to light
intensity] of two youngest fully-expanded fan leaves of Cannabis sativa L. ‘Stillwater’ grown
under either low or high localized photosynthetic photon flux densities (LPPFD). The low and
high LPPFD were 91 and 1238 μmol·m–2·s–1, respectively. Measurements were made during
week 5 after the initiation of the 12-h photoperiod.

32
Figure 2.4. The light-saturated net CO2 exchange rate (Asat) (A), the light saturation point (LSP)
(B), the localized net CO2 exchange rate (LNCER) (C), and the Fv/Fm (D) of the youngest fully-
expanded fan leaves of Cannabis sativa L. ‘Stillwater’ at the localized photosynthetic photon
flux densities (LPPFD) that the respective leaves were growing under when the measurements
were made, during weeks 1, 5, and 9 after initiation of the 12-h photoperiod. Each datum is a
single plant. Regression lines are presented when P ≤0.05.

2.3.1 LEAF PHOTOSYNTHESIS

Leaf light response curves constructed under different LI and at different growth stages (week 1,
5, and 9) generally demonstrated the trends that the Asat and LSP were higher for plants grown
under high vs. low LPPFD (Figures 2.3, 2.4A-B), especially after the plants had acclimated to
their new lighting environments (i.e., weeks 5 and 9). There were no LPPFD effects on Asat in
week 1, with a mean (± SE, n = 52) of 23.9 ± 0.90 μmol(CO2)·m–2·s–1 (Figure 2.4A). The Asat in
weeks 5 and 9 (Figure 2.4A) and LSP in weeks 1, 5, and 9 (Figure 2.4B) increased linearly with
increasing LPPFD. At low LPPFD, the highest LSP was in week 1. The slopes of the A sat and
LSP models were similar in weeks 5 and 9, but the Y-intercepts for both parameters were
33
approximately twice as high in week 5 vs. week 9. LNCER increased linearly with increasing
LPPFD in weeks 1, 5, and 9 (Figure 2.4C) with the steepest and shallowest slopes apparent at
weeks 5 and 1, respectively. The LNCER model in week 9 had a substantially lower Y-intercept
than the other two weeks. As evidenced by the projected intersection of the A sat and LNCER
models in week 5 (i.e., at LPPFD of 1532 μmol·m–2·s–1), the maximum LPPFD in week 5 (i.e.,
1370 μmol·m–2·s–1) was nearly sufficient to saturate the photosynthetic apparatus at the top of
the canopy. There were no LPPFD effects on QY, but the mean QY in weeks 1 and 5 were higher
than week 9. The mean (± SE) QY were 0.066 ± 0.0013 (n = 54), 0.068 ± 0.0005 (n = 60), and
0.058 ± 0.0008 (n = 63) μmol(CO2)· μmol–1(PAR) in weeks 1, 5, and 9 respectively. The Fv/Fm
decreased linearly with increasing LPPFD in all three measurement periods (Figure 2.4D).

34
Figure 2.5. The specific leaf weight (SLW; on a dry weight basis) of young, fully-expanded
Cannabis sativa L. ‘Stillwater’ leaves in response to the average photosynthetic photon flux
density (APPFD), measured on day 35 after initiation of the 12-h photoperiod. Each datum
represents one fan leaf from a single plant.

35
Figure 2.6. Sketches of Cannabis sativa L. ‘Stillwater’ plants grown under low (A) and high (B)
photosynthetic photon flux density (APPFD), 9 weeks after initiation of 12-h photoperiod
(illustrated by Victoria Rodriguez Morrison).

2.3.2 CHLOROPHYLL CONTENT INDEX AND PLANT MORPHOLOGY

There were no LI treatment effects on CCI either at the top or bottom of the canopy. However
within in each week, the upper canopy CCI were higher than the lower canopy. Additionally, the
CCI in the upper and lower canopy was higher in week 1 vs. weeks 5 and 9. The CCI was
assessed for 91 plants per week. The mean CCI (± SE) were 67.1 ± 0.80, 55.8 ± 2.2, and 52.0 ±
2.1 in the upper canopy and 46.3 ± 1.1, 31.1 ± 0.86, and 31.5 ± 1.1 in the lower canopy, in weeks
1, 5, and 9 respectively. The SLW increased linearly from 35.4 to 58.1 g·m–2 as APPFD
(calculated based on the respective plants’ accumulated PAR exposures up to day 35 of the
flowering stage) increased from 130 to 1990 μmol·m–2·s–1 (Figure 2.5). Plants grown under low

36
vs. high APPFD were generally shorter and wider, with thinner stems, larger leaves, and fewer,
smaller inflorescences (Figure 2.6).

37
Figure 2.7. The relationship between average apical photosynthetic photon flux density
(APPFD) applied during the flowering stage (81 days) and inflorescence dry weight (A), harvest
index (total inflorescence dry weight / total aboveground dry weight) (B), and apical

38
inflorescence density (based on fresh weight) (C) of Cannabis sativa L. ‘Stillwater’. Each datum
is a single plant.

Table 2.1. Cannabinoid concentration in apical inflorescences of Cannabis sativa L. ‘Stillwater’.

Concentration (mg·g–1 of
Cannabinoid
inflorescence dry weight)
Δ-9-tetrahydrocannabinol (Δ9-THC) UDLz
Δ-9-tetrahydrocannabinolic acid (Δ9-THCA) 12.9y ± 0.03
Total equivalent Δ9-tetrahydrocannabinol (TΔ9-THC) 11.3 ± 0.02
Cannabidiol (CBD) 5.53 ± 0.01
Cannabidiolic acid (CBDA) 214 ± 0.4
Total equivalent cannabidiol (TCBD) 193 ± 0.4
Cannabigerol (CBG) UDL
Cannabigerolic acid (CBGA) 4.76 ± 0.01
Total equivalent cannabigerol (TCBG) 4.45 ± 0.009
Cannabinol (CBN) UDL
zunder detection limit of 0.5 mg·g–1 of inflorescence dry weight
ydata are means ± SE of 22 representative inflorescences across all of the APPFDs in CB1

39
Table 2.2. The relationships between average photosynthetic photon flux density (APPFD)
applied during the flowering stage (81 days) and terpene concentration in apical inflorescences
of myrcene, limonene and total terpenes, and the mean concentration for terpenes with no
APPFD treatment effects, of Cannabis sativa L. ‘Stillwater’.

Terpene concentration
(mg·g–1 of inflorescence dry weight)
Terpene
Mean z Regression equation y R2
Total terpenes Y = 0.00230 X + 8.57 0.320
Myrcene Y = 0.00142 X + 2.34 0.464
Limonene Y = 0.000326 X + 1.01 0.246
z
Alpha pinene 0.16 ± 0.01
Beta pinene 0.22 ± 0.01
Terpinolene UDLx
Linalool 0.53 ± 0.01
Terpineol 0.32 ± 0.02
Caryophyllene 2.9 ± 0.2
Humulene 0.65 ± 0.04
3-carene UDL
Cis-ocimene UDL
Eucalyptol UDL
Trans-ocimene UDL
Fenchol 0.22 ± 0.01
Borneol 0.03 ± 0.01
Valencene UDL
Cis-nerolidol UDL
Trans-nerolidol UDL
Guaiol UDL
Alpha-bisabolol 0.38 ± 0.03
Sabinene UDL
z
when there were no APPFD treatment effects on terpene concentration, the means ± SE of 22
representative inflorescences across all of the PPFDs in CB1 are presented
ylinear regression models for the APPFD treatment effects on terpene concentration when P

≤0.05
xunder detection limit of 0.5 mg·g–1 of inflorescence dry weight

40
2.3.3 YIELD AND QUALITY

Cannabis yield was 4.5 times higher for plants grown at APPFD of 1800 μmol·m–2·s–1 relative to
those grown at 120 μmol·m–2·s–1 (Figure 2.7A). Note that yields in the present study are true
oven-DWs. Since fresh cannabis inflorescences are typically dried to 10 to 15% moisture
content to achieve optimum marketable quality (Leggett, 2006), yields in the present study can
be easily adjusted upwards to be comparable any desirable moisture level (e.g., by multiplying
the DW by the FW:DW to determine the additional FW). The harvest index was 1.3 times higher
as APPFD increased from 120 to 1800 μmol·m–2·s–1 (Figure 2.7B). The apical inflorescence
density increased 1.3 times as APPFD increased from 120 to 1800 μmol·m–2·s–1 (Figure 2.7C).

Cannabidiolic acid (CBDA) was the dominant cannabinoid in the dried inflorescences; however,
there were no APPFD treatment effects on the concentration of any of the measured cannabinoids
(Table 2.1). Due to linear increases in inflorescence yield with increasing LI, cannabinoid yield
(g·m–2) increased by 4.5 times as APPFD increased from 120 to 1800 μmol·m–2·s–1. Myrcene,
limonene, and caryophyllene were the dominant terpenes in the harvested inflorescences (Table
2.2). The concentration of total terpenes, myrcene, and limonene were 1.4, 2.0 and 1.5 times
higher at APPFD of 1800 μmol·m–2·s–1 relative to those detected in plants grown at 120 μmol·m–
2·s–1. There were no APPFD effects on the concentration of the other individual inflorescence
terpenes.

2.4 DISCUSSION
2.4.1 CANNABIS INFLORESCENCE YIELD IS PROPORTIONAL TO LIGHT
INTENSITY

It was predicted that cannabis yield would exhibit a saturating response to increasing LI, thereby
signifying an optimum LI range for indoor cannabis production. However, the yield results of
this experiment demonstrated cannabis’ immense plasticity for exploiting the incident lighting
environment by efficiently increasing marketable biomass up to extremely high – for indoor
production – LIs (Figure 2.7A). Even under ambient CO2, the linear increases in yield indicated
that the availability of PAR photons was still limiting whole-canopy photosynthesis at APPFD
levels as high as ≈1800 μmol·m–2·s–1 (i.e., DLI ≈78 mol·m–2·d–1). These results were generally

41
consistent with the trends of other studies reporting linear cannabis yield responses to LI (Eaves
et al., 2020; Potter and Duncombe, 2012; Vanhove et al., 2011), although there is considerable
variability in both relative and absolute yield responses to LI in these prior works. The present
study covered a broader range of LI, and with much higher granularity, compared with other
similar studies.

The lack of a saturating yield response at such high LI is an important distinction between
cannabis and other crops grown in controlled environments (Beaman et al., 2009; Fernandes et
al., 2013; Oh et al., 2009; Faust, 2003). This also means that the selection of an “optimum” LI
for indoor cannabis production can be made somewhat independently from its yield response to
LI. Effectively, within the range of practical indoor PPFD levels – the more light that is
provided, the proportionally higher the increase in yield will be. Therefore, the question of the
optimum LI may be reduced to more practical functions of economics and infrastructure
limitations: basically, how much lighting capacity can a grower afford to install and run? This
becomes a trade-off between fixed costs which are relatively unaffected by yield and profit (e.g.,
building lease/ownership costs including property tax, licensing, and administration) and variable
costs such as crop inputs (e.g., fertilizer, electricity for lighting) and labor. Variable costs will
obviously increase with higher LI but the fixed costs, on a per unit DW basis, should decrease
concomitantly with increasing yield (Vanhove et al., 2014). Every production facility will have a
unique optimum balance between facility costs and yield; but the yield results in the present
study can help cannabis cultivators ascertain the most suitable LI target for their individual
circumstances. Readers should be mindful that this study reports yield parameters as true DWs;
marketable yield can be easily determined by factoring back in the desirable moisture content of
the inflorescence. For example, for a 400 g·m–2 of dry yield, the corresponding marketable yield
would be 444 g·m–2 at 10% moisture content [i.e., 400 g·m–2 + (400 g·m–2 × 10% FW / 90%
DW)].

It is also important to appreciate that PPFD, which represents an instantaneous LI level, does not
provide a complete accounting of the total photon flux incident on the crop canopy throughout
the entire production cycle. While this LI metric is ubiquitous in the horticulture industry and

42
may be most broadly relatable to prior works, there is value in relating yield to the total photon
flux received by the crop. Historically, this has been done by relating yield to installed wattage
on per area bases, resulting in g·W–1 metric (Potter and Duncombe, 2012), which can be more
fittingly converted to yield per unit electrical energy input (g·kWh–1) by factoring in the
photoperiod and length of the production cycle (EMCDDA, 2013). However, since
photosynthesis is considered a quantum phenomenon, crop yield may be more appropriately
related to incident (easily measured) or absorbed photons and integrated over the entire
production cycle (i.e., TLI, mol·m–2), in a yield metric that is analogous to QY: g·mol–1. Unlike
installed wattage, this metric has the advantage of negating the effects of different fixture
efficacy (μmol·J–1), which continues its upward trajectory, especially with LEDs (Kusuma et al.,
2020; Nelson and Bugbee, 2014). The present study did not directly measure lighting-related
energy consumption; however, installed energy flux (kWh·m–2) can be estimated from TLI using
the Lumigrow fixture’s efficacy rating: 1.29 and 1.80 μmol·J–1, from Nelson and Bugbee (2014)
and Radetsky (2018), respectively. Using the average of these values (1.55 μmol·J –1), the
conversion from TLI to energy flux becomes: mol·m–2 × 5.6 = kWh·m–2. At an APPFD of 900
μmol·m–2·s–1 (i.e., TLI of 3149 mol·m–2), the model in Figure 2.7A predicts a yield of 303 g·m–2
which corresponds to an energy use efficacy of 0.54 g·kWh–1. For comparison, doubling the LI
to the highest APPFD used in this trial increases the yield by 70% but results in a ≈15% reduction
in energy use efficacy. It is up to each grower to determine the optimum balance between
variable (e.g., lighting infrastructure and energy costs) and fixed (e.g., production space) costs in
selecting a canopy level LI that will maximize profits.

2.4.2 INCREASING LIGHT INTENSITY ENHANCES INFLORESCENCE QUALITY

Beyond simple yield, increasing LI also raised the harvest quality through higher apical
inflorescence (also called “cola” in the cannabis industry) density – an important parameter for
the whole-bud market – and increased ratios of inflorescence to total aboveground biomass
(Figure 2.7B and 2.7C). The linear increases in harvest index and apical inflorescence density
with increasing LI both indicate shifts in biomass partitioning more in favor of generative
tissues; a common response in herbaceous plants (Poorter et al., 2019) including cannabis
(Hawley et al., 2018; Potter and Duncombe, 2012). The increases in these attributes under high
43
LI may also indirectly facilitate harvesting, as there is correspondingly less unmarketable
biomass to be processed and discarded, which is an especially labour-intensive aspect of
cannabis harvesting.

The terpene composition of the inflorescence – comprised mainly of myrcene, limonene, and
caryophyllene – increased by approximately 25%, as APPFD increased from 120 to 1800
μmol·m–2·s–1 (Table 2.2), which could lead to enhanced aromas and higher quality extracts
(McPartland and Russo, 2001; Nuutinen, 2018). Conversely, total cannabinoid yield increased in
proportion with increasing inflorescence yield since there were no LI treatment effects on
cannabinoid concentration (Table 2.1). Similarly, Potter and Duncombe (2012) and Vanhove et
al. (2011) found no LI treatment effects on cannabinoid concentration (primarily Δ9-THC in
those studies) and attributed increasing cannabinoid yield to enhanced biomass apportioning
towards generative tissues at higher LI. Other studies had contradictory results on the effects of
LI on cannabinoid concentration. Hawley et al. (2018) did not find canopy position effects on Δ9-
THC or CBD levels in a subcanopy lighting (SCL) trial, but they did find slightly higher CBG
concentration in the upper canopy in the control (HPS top-lighting only) and the Red-Green-Blue
SCL treatment, but not in the Red-Blue SCL treatment. While it is not possible to unlink
spectrum from LI in their results, the magnitude of the reported chemical profile differences,
both between canopy positions and between lighting treatments, were relatively minor.
Conversely, Namdar et al. (2018) reported what appeared to be a vertical stratification on
cannabis secondary metabolites, with highest concentrations generally found in the most distal
inflorescences (i.e., closest to the light source, PPFD ≈600 μmol·m–2·s–1). They attributed this
stratification to the localized LI at different branch positions, which were reportedly reduced by
≥60% at lower branches vs. at the plant apex. However, given the lack of LI treatment effects
(over a much broader range of PPFDs) on cannabinoid levels in the present study, it is likely that
other factors were acting on higher-order inflorescences, such as delayed maturation and reduced
biomass allocation, that reduced the concentrations in these tissues (Diggle, 1995; Hemphill et
al., 1980).

44
2.4.3 PLASTICITY OF CANNABIS LEAF MORPHOLOGY AND PHYSIOLOGY
RESPONSES TO LI AND OVER TIME

The objectives of the photosynthesis and leaf morphology investigations in this study were two-
fold: 1) to address the knowledge gap in the relationships between localized cannabis leaf
photosynthesis and yield and 2) observe and report changes in physiology as the plant progresses
through the flowering ontogeny.

General morphological, physiological, and yield responses of plants are well documented across
LI gradients ranging from below compensation point to DLIs beyond 60 mol·m–2·d–1. Recently,
the LI responses of a myriad of plant attributes were compiled across a tremendous range
species, ecotypes and growing environments, and concisely reported them in the excellent review
paper by Poorter et al. (2019). The trends in their LI models align well with primary attributes
measured in the present study, including morphological parameters such as plant height and
internode length, SLW (discussed below), and physiological parameters such as Fv/Fm, LNCER
(i.e., photosynthesis at growth light; Phot/AGL), and Asat (i.e., photosynthesis at saturating light;
Phot/ASL). In general, cannabis photosynthesis and yield responses to localized LI were linear
across the APPFD range of 120 to 1800 μmol·m–2·s–1. Although these results are in agreement
with the contemporary literature on cannabis (Bauerle et al., 2020; Chandra et al., 2008; 2015;
Eaves et al., 2020; Potter and Duncombe, 2012), we also showed substantial chronological
dependencies on leaf photosynthetic indices.

By surveying the photosynthetic parameters of the upper cannabis canopy across a broad range
of LPPFDs and over multiple timepoints during the generative phase, we saw evidence of both
acclimation and early senescence as the crop progressed through its ontogeny. At the beginning
of the trial, the plants were abruptly transitioned from a uniform PPFD (425 μmol·m–2·s–1) and
18-h photoperiod (i.e., 27.5 mol·m–2·d–1) and subjected to a much shorter photoperiod (12-h) and
an enormous range of LI (120 to 1800 μmol·m–2·s–1), resulting in DLIs ranging from 5.2 to 78
mol·m–2·d–1. Furthermore, on a DLI-basis, approximately 1/3 of the plants were exposed to
lower LIs in the flowering vs. vegetative phase (i.e., APPFD <640 μmol·m–2·s–1). These sudden
transitions in both LI and photoperiod resulted in substantive changes in the plants’ lighting
environment at the start of the trial, stimulating various morphological and physiological
45
adaptations with differing degrees of plasticity. The leaves measured in week 1 developed and
expanded during the prior vegetative phase under a different lighting regimen (LI and
photoperiod). The leaves measured in week 5 were developed under their respective LPPFDs
during a period characterized by slowing vegetative growth and transitioning to flower
development. The leaves measured in week 9 would have also developed under their respective
LPPFDs, but since cannabis vegetative growth greatly diminishes after the first five weeks in 12-
h days (Potter, 2014), these tissues were physiologically much older than the leaves measured in
week 5, with concomitant reductions in photosynthetic capacity (Bauerle et al., 2020;
Bielczynski et al., 2017).

These differences in leaf physiological age, plant ontogeny, and localized lighting environments
during leaf expansion vs. measurement resulted in notable temporal variability in leaf-level LI
responses. In week 1, there were no LI treatment effects on Asat and the slopes of the LSP,
LNCER, and Fv/Fm were shallower in weeks 5 and 9. The comparatively lower LI responses in
week 1 were likely due to the reduced adaptive plasticity that mature foliar tissues have vs.
leaves that developed under a new lighting regime (Sims and Pearcy, 1992). In addition, Y-
intercepts for the Asat, LSP, and LNCER models were higher in week 1 than weeks 5 and 9,
which may be partly due to the higher LI (amplified by the longer photoperiod) that the leaves
developed under, during the latter part of the vegetative phase. Further, the A sat, LSP, and
LNCER models in weeks 5 and 9 have comparable slopes, but there is a vertical translation in the
respective models, resulting week 9 models having substantially lower Y-intercepts (i.e.,
approximately half) for these parameters. The interplay of physiological age of foliage and plant
ontogeny (i.e., onset of senescence) on the diminished photosynthetic capacity of the leaves in
week 9 is unknown, but the dynamic temporal nature of cannabis photosynthesis (during
flowering) is manifest in these models.

Given these impacts of physiological age and light history, we posit that cannabis leaf
photosynthesis cannot be used as a stand-alone gauge for predicting yield. Chandra et al. (2008)
and Chandra et al. (2015) provided insight into the substantial capacity for drug-type strains of
indoor grown cannabis leaves to respond to LI; and the results of these trials are much lauded in
the industry as evidence that maximum photosynthesis and yields will be reached under canopy-
46
level PPFDs of ≈1500 μmol·m–2·s–1. However, the 400 to 500 μmol·m–2·s–1 increments in
LPPFD does not provide sufficient granularity (particularly at low LI) to reliably model the
LRCs, thus no models were provided. Further, the LRCs were made on leaves of varying and
unreported physiological ages, from plants exposed to a vegetative photoperiod (18-h), and
acclimated to unspecified localized LI (a canopy-level PPFD of 700 μmol·m–2·s–1 was indicated
in Chandra et al., 2015). The strong associations between a tissue’s light history and its
photosynthesis responses to LI, demonstrated in this trial and by others (Björkman, 1981),
represent a major shortcoming of using leaf LI response models to infer crop growth and yield.
To illustrate, Figure 2.3 shows LRCs of leaves from a single cultivar, at similar physiological
ages (week 5 after transition to 12-h photoperiod) but acclimated to disparate LPPFDs: 91 and
1238 μmol·m–2·s–1. The relative difference in LNCER at higher LIs (≈50%) between these two
curves is representative of the potential uncertainty due to just one of the uncontrolled
parameters (LNCER) in these prior works. Differing physiological ages of tissues at the time of
measurement may have conferred an even larger degree of uncertainty in the magnitude of leaf
responses to LI (Bauerle et al., 2020) than leaf light history. Consideration must also be given to
the different life stages of a photoperiodic crop (i.e., vegetative vs. generative) and the inherent
impact that day length imbues on the total daily PAR exposure (i.e., DLI) which can correlate
better to crop yield than PPFD. Furthermore, for a given DLI, yields are higher under longer
photoperiod (Vlahos et al., 1991; Zhang et al., 2018), ostensibly due to their relative proximity to
their maximum QY (Ohyama et al., 2005). A final distinction between leaf photosynthesis and
whole plant yield responses to LI is the saturating LI: the LSP for leaf photosynthesis were
substantially lower than the LSP for yield, which remains undefined due to the linearity of the
light response model.

Newly-expanded leaves, especially in herbaceous species, are able to vary their leaf size,
thickness and chlorophyll content in response to LPPFD in order to balance a myriad of factors
such as internal and leaf surface gas exchange (CO2 and H2O), internal architecture of the light-
harvesting complexes, and resistance to photoinhibition (Björkman, 1981). In the present study,
the effects of LI on leaf morphology were only evaluated in week 5, when the crop was still
actively growing vegetative biomass. Reductions in SLW (i.e., increases in specific leaf area,

47
SLA) in response to increasing LI are abundant in the literature (Fernandes et al., 2013; Gratani,
2014; Sims and Pearcy, 1992). In particular, Poorter et al. (2019) reported a saturating response
of SLW [also known as leaf mass (per) area; LMA] to LI across 520 species (36% of which were
herbaceous plants), however much of their data was at DLIs lower than the minimum DLI in the
present study (5.2 mol·m–2·d–1), which affected the shape of their SLW response model to LI.
Across similar DLI ranges, the average increase in SLW across 520 species was 1.7× in Poorter
et al., (2019) vs 1.6× in the present study, indicating that cannabis SLW responses to LI are
consistent with normal trends for this parameter.

The lack of LI treatment effects on CCI are also consistent with other studies that have shown
that area-based chlorophyll content is fairly stable across a broad range of LIs (Poorter et al.,
2019; Björkman, 1981), despite substantial variability in photosynthetic efficiency. However,
since there were LI treatment effects on SLW, chlorophyll content on leaf volume or mass bases
would likely have reduced under higher LI. The positional effects on CCI (i.e., higher in upper
vs. lower canopy) were probably due to the interplay between self-shading and advancing
physiological age of the lower leaves (Bauerle et al., 2020). The temporal effects on CCI, which
was higher in week 1 vs. weeks 5 and 9, in both upper and lower leaves, may have been due to
changes in QY over the life-cycle of the crop. Bugbee and Monje (1992) presented a similar
trend; high QY during the active growth phase of a 60-d crop cycle of wheat, followed by a
reduction in QY at the onset of senescence (i.e., shortly before harvest). The decline in
chlorophyll content in the latter phase of the production cycle probably contributed to the
reductions in the photosynthetic parameters (e.g., A sat, LSP, LNCER) of the tissues measured in
week 9 vs. week 5.

Overall, the impact that increasing LI had on cannabis morphology and yield were captured
holistically in the plant sketches in Figure 2.6, which shows plants grown under higher LIs had
shorter internodes, smaller leaves, and much larger and denser inflorescences (resulting in higher
harvest index), especially at the plant apex, compared to plants grown under lower LIs. Like
many other plant species, we have found that cannabis has immense plasticity to rapidly
acclimate its morphology and physiology, both at leaf- and whole plant-levels, to changes in the

48
growing lighting environment. Therefore, in order reliably predict cannabis growth and yield to
LI, it is necessary to grow plants under a broad range of LIs through their full ontological
development, as was done in this study. Without knowing the respective tissues’ age and light
history, instantaneous light response curves at leaf-, branch-, or even canopy-levels cannot
reliably predict yield.

2.5 CONCLUSIONS
We have shown an immense plasticity for cannabis to respond to increasing LI; in terms of
morphology, physiology (over time), and yield. The temporal dynamics in cannabis leaf
acclimations to LI have also been explored, addressing some knowledge-gaps in relating
cannabis photosynthesis to yield. The results also indicate that the relationship between LI and
cannabis yield does not saturate within the practical limits of LI used in indoor production.
Increasing LI also increased harvest index and the size and density of the apical inflorescence;
both markers for increasing quality. However, there were no and minor LI treatment effects on
the concentrations of cannabinoids and terpenes, respectively. This means that growers may be
able to vastly increase yields by increasing LI but maintain a relatively consistent secondary
metabolite profile in their marketable products. Ultimately, the selection of the economic
optimum canopy-level LI for a given commercial production system depends on many
interrelated factors.

Future research should expand to multiple cultivars of different biotypes. Further, since plant
yield responses to elevated CO2 can mirror the responses to elevated LI, the combined effects of
CO2 and LI should be investigated on cannabis yield with an in-depth cost-benefit analysis of the
optimum combination of these two input parameters.

49
 CHAPTER THREE
CANNABIS INFLORESCENCE YIELD AND CANNABINOID
CONCENTRATION IS NOT IMPROVED WITH LONG-TERM
EXPOSURE TO UV RADIATION
ABSTRACT
It is commonly believed that exposing cannabis plants to UV radiation can enhance Δ9-THC
concentrations in female inflorescences and associated foliar tissues. However, a lack of
published scientific studies has left knowledge-gaps in the effects of UV on cannabis; these must
be elucidated before UV can be utilized as a horticultural management tool in commercial
cannabis production. In this study we investigated the effects of UV exposure level on
photosynthesis, growth, inflorescence yield, and secondary metabolite composition of two
indoor-grown cannabis cultivars: ‘Low Tide’ (LT) and ‘Breaking Wave’ (BW). After growing
vegetatively for 2 weeks under a canopy-level PPFD of ≈225 μmol·m–2·s–1 in an 18-h light/6-h
dark photoperiod, plants were grown for 9 weeks in a 12-h light/12-h dark “flowering”
photoperiod under a canopy-level PPFD of ≈400 µmol·m–2·s–1 and 3.5 h·d–1 of supplemental UV
radiation with photon flux densities (PFD) ranging from 0.01 to 0.8 μmol·m–2·s–1 provided by
LEDs with a peak wavelength of 287 nm (i.e., biologically-effective UV doses of 0.2 to 13 kJ·m–
2·d–1). The severity of UV-induced morphology (e.g., whole-plant size and leaf size reductions,
leaf malformations, and stigma browning) and physiology (e.g., reduced leaf photosynthetic rate
and reduced Fv/Fm) symptoms worsened as UV exposure level increased. Dry inflorescence yield
decreased with increasing UV exposure level in LT, but not in BW. In LT, total equivalent Δ9-
THC and total equivalent CBD concentrations decreased with increasing UV exposure level,
whereas there were no UV treatment effects on total equivalent concentrations of individual
cannabinoids in BW. Total inflorescence terpene concentrations decreased linearly with
increasing UV exposure level in both cultivars however, relative concentrations of individual
terpenes varied by cultivar. The potential for using UV to enhance cannabis quality must still be
confirmed before it can be used as a production tool for modern, indoor-grown cannabis
cultivars.

50
3.1 INTRODUCTION

Cannabis is a short-day plant commonly cultivated for its unique secondary metabolites (e.g.,
cannabinoids) that are used both medicinally and recreationally (Small, 2017). Cannabis is often
grown in controlled-environment facilities that are illuminated solely with electrical lighting to
accommodate its photoperiod specificity and produce uniform plants by maintaining prescribed
environmental parameters. Popular sole-source lighting technologies used in the flowering stage
of cannabis production include HPS and, increasingly, LEDs (Cannabis Business Times, 2020);
both technologies normally have little to no UV radiation in their spectra (Radetsky, 2018). In
the natural environment, cannabis plants are exposed a small fraction of UVB radiation relative
to the amount of PAR in sunlight. However, the higher-energy photons in the UVB vs. PAR
spectra are disproportionately effective in evoking plant responses (Flint and Caldwell, 2003),
including changes in morphology, physiology, and metabolism (Huché-Thélier et al., 2016;
Robson et al., 2019). UVC photons have even higher energy than UVB, but solar UVC is
absorbed by the ozone layer and therefore does not reach the Earth’s surface (McElroy and
Fogal, 2008). While UVC is used to inactivate microorganisms such as waterborne pathogens in
recirculating irrigation systems (Younis et al., 2019), UVC is only rarely directly applied to
foliage – to inactivate foliar pathogens through short-term exposures (Aarrouf and Urban, 2020)
– since UVC can cause tissue damage (Stapleton, 1992).

UV radiation can affect many aspects of plant morphology, physiology, and metabolism
(Jenkins, 2017). Plant responses to UV exposure are either induced through pathways mediated
by UVR8 (a UVB-specific photoreceptor) or by UV-induced oxidative cellular damage,
including to DNA (Czégény et al., 2016, Tossi et al., 2019). Typical plant responses to UV stress
include stunted growth, reduced leaf area, increased leaf thickness (Robson et al., 2019),
epicuticular wax accumulation (Cen and Bornman, 1993), and foliar necrosis (Klem et al., 2012;
Torre et al., 2012). From an ecological standpoint, it has been speculated that Δ9-THC (the most
economically valuable psychoactive cannabinoid) production is upregulated in cannabis tissues
under UV exposure to serve as photoprotection. This concept arose from studies that found
comparatively higher Δ9-THC concentrations in cannabis ecotypes that grow in global regions
with relatively high solar UV exposure, such as at low latitudes and high altitudes (Small and
51
Beckstead, 1973; Pate, 1983). However, despite the focus on Δ9-THC in the literature, other
cannabinoids have similar UV absorbing properties (Hazekamp et al., 2005), which may
challenge an ecological explanation for upregulating Δ9-THC over other cannabinoids.
Preliminary controlled-environment studies that were done about three decades ago alluded to
the potential for UV to increase Δ9-THC concentration in cannabis foliar and floral tissues
(Fairbairn and Liebmann, 1974; Lydon et al., 1987). However, the concentration of Δ9-THC in
inflorescence tissues has increased substantially over the past decades, with contemporary
genotypes having ≈10-fold higher Δ9-THC concentrations than the genotypes used these older
studies (Dujourdy and Besacier, 2017). Therefore, modern cannabis genotypes may function
nearer to their maximum capacity for producing Δ9-THC; which could impede their ability to
further increase Δ9-THC production under UV exposure, relative to older genotypes.

Studies on modern cannabis genotypes have shown that environmental stimuli can modify the
cannabinoid composition. For example, inflorescences in CBD-dominant genotypes had greater
CBD concentrations when grown at high vs. low altitude, which may have been a response to
increased UV exposure at higher elevation (Giupponi et al., 2020). Drought-stress and salt-stress
have also been shown to alter the inflorescence cannabinoid composition in modern genotypes
(Caplan et al., 2019; Yep et al., 2020). Therefore, the potential for UV exposure to provoke
changes in the secondary metabolite composition in inflorescences of modern cannabis
genotypes grown in controlled-environments merits scientific investigation. Evaluating the
effects of UV on modern genotypes with relatively balanced concentrations of Δ9-THC and CBD
[i.e., chemotype II; a cultivar with a ratio of Δ9-THC to CBD of ≈1 (Small and Beckstead, 1973)]
would provide insight into whether UV impacts individual cannabinoids differently.

The objectives of this study were to: 1) characterize morphological and physiological responses
of indoor-grown cannabis to UV exposure, and 2) investigate the relationships between UV
exposure levels during the flowering stage and inflorescence yield and secondary metabolite
composition of cannabis genotypes with disparate growth habits and moderate concentrations of
both Δ9-THC and CBD.

52
3.2 MATERIALS AND METHODS
3.2.1 PLANT CULTURE

Cuttings were taken from mother plants of LT and BW, both of which are classified as
chemotype II. After growing for 13 d under humidity domes and fluorescent light
(F32T8/TL850; Philips, Amsterdam, Netherlands) providing ≈100 µmol·m–2·s–1, rooted cuttings
were transferred to 1-gallon pots containing a peat-based substrate, and placed under LED light
comprised of a mixture of Pro-325 (Lumigrow; Emeryville, CA, USA) and generic (unbranded)
white LEDs providing ≈225 µmol·m–2·s–1, for an additional 9 d. Both the propagation and
vegetative growth stages had 18-h photoperiods. The potted plants were subsequently transferred
to a single deep-water CB, where they were placed in floating polystyrene rafts in an indoor
cannabis production facility in southern Ontario, Canada (described in Chapter 2 of this thesis).
There were 384 evenly-spaced plants in the CB at a density of 0.09 m2/plant. The daily
photoperiod was reduced to 12 h on the day the plants were transferred to the CB.

3.2.2 EXPERIMENTAL SETUP

PAR was supplied by 24 LED fixtures (Pro650; Lumigrow Inc. Emeryville, Ca, USA) arranged
evenly over the CB (i.e., 2 rows of 12 PAR fixtures). The LED composition and spectrum of the
PAR fixtures was described in Chapter 2 and the relative photon flux distribution is provided in
Figure 3.1A. Single UV LED fixtures were centered between adjacent PAR fixtures (within
each row), resulting in 2 rows of 11 UV fixtures. The 22 UV LED fixtures had consistent
spectrum output (peak wavelength of 287 nm; Figure 3.1B), but adjustable intensity (with
analog dimmers). According to the conventional definitions of the different UV wavebands
(described above), the photon flux ratio of UVB to UVC was UVB(93):UVC(7). The UV
treatments (described below) were applied daily, in the last 3.5 hours (from 16:00 HR to 19:30
HR) of the PAR photoperiod (from 07:30 HR to 19:30 HR), from the day that the plants were
transferred to the CB. The plants were exposed to the UV treatments for 60 d (≈9 weeks) and
then harvested.

53
Figure 3.1. Relative spectral photon flux distribution of (A) Pro-650 (Lumigrow) LED fixtures
and (B) UV LED fixtures.

54
For each cultivar, 44 representative uniform plants were selected from the larger populations to
be experimentally evaluated. Plots, each consisting of 4 plants, were arranged where 2 plants
were directly underneath each UV LED fixture, and 2 plants were adjacent. There were 3 UV
LED fixture settings that were randomly assigned (within each cultivar) to each plot: off, half
power, and full power. Within each plot, the 2 plants closer to the UV LED fixture were exposed
to relatively higher UV exposure than the 2 adjacent plants. This configuration allowed for a
wide range of evenly-distributed UV exposure levels; ranging from 0.01 to 0.8 µmol·m–2·s–1.

At the start of the UV treatments, experimental plants of LT all had 7 nodes, with heights (from
the substrate surface to the shoot apex) ranging from 14 to 23 cm. Experimental plants of BW
had 8 nodes, with heights ranging from 14 to 20 cm. Experimental plants were surrounded by
plants of the same cultivar to maintain canopy uniformity. The LT cultivar populated the south
half of the CB, while BW populated the north half.

Average hang height (i.e., distance from the bottom of the fixtures to the top of the canopy) was
maintained at 50.5 cm by adjusting the height of the light racks weekly using a system of pulleys
and cables. Canopy-level PPFD (400 to 700 nm) and UV-PFD (270 to 320 nm) were measured at
the apex of each plant weekly, after the light rack height adjustment, using a PAR meter (LI-180;
LI-COR Biosciences, Lincoln, NE, USA) and a radiometrically-calibrated spectrometer (XR-
Flame-S, Ocean Optics, Dunedin, Florida), respectively. A MS Excel tool developed by Mah et
al. (2019) was used to integrate spectrometer data into UV-PFD, biologically-effective UV-PFD
[UV-PFDBE; Flint and Caldwell (2003)], and daily biologically-effective UV dose (kJ·m–2·d–1)
(Table 3.1). At the end of the trial, average PPFD and average UV-PFD were calculated for each
treatment plant by determining the corresponding TLI (mol·m–2) as described in Chapter 2. The
experiment-wise average (± SE, n = 88) PPFD was 408 ± 6.5 µmol·m–2·s–1. The average UV-
PFD for each plant was used as the independent variable (i.e., X-axis) in regressions of UV
exposure vs. the measured growth, yield and quality parameters.

55
Table 3.1. Range of canopy-level UV photon flux density, UV biologically-effective photon flux
density and daily UV biologically-effective dose from UV LEDs with a peak wavelength of 287
nm and a daily 3.5 h photoperiod.
UV UV photon flux UV biologically effectivez photon Daily UV biologically-
exposure density flux density effective dose
target (µmol·m-2·s-1) (µmol·m-2·s-1) (kJ·m-2·d-1)
Minimum 0.01 0.032 0.16
Low 0.1 0.32 1.6
Medium 0.5 1.6 8.3
Maximum 0.8 2.5 13
z
weighted using the BSWF for Plant Growth by Flint and Caldwell (2003)

Plant husbandry and environmental controls followed the cultivator’s standard operating
procedures except for the UV radiation. The air temperature and relative humidity set points
were: 25°C and 60%. There was no CO2 supplementation, with typical concentrations of ≈400
ppm when lights were on. The aquaponic solution was maintained within normal levels of
nutrient concentrations, pH, electrical conductivity and dissolved oxygen, as described in
Chapter 2.

3.2.3 GROWTH MEASUREMENTS

The number of nodes (i.e., primary branches), height (i.e., length of main stem from substrate
surface to the highest point) and widths (i.e., the widest part and its perpendicular width) of each
experimental plant were measured in week 6. Plant height and widths were used to calculate
growth index [(height × width1 × width2) / 300 (Ruter, 1992)] for each plant.

3.2.4 LEAF CHLOROPHYLL AND FLUORESCENCE MEASUREMENTS

CCI [% transmission at 931 nm / % transmission at 653 nm (Parry et al., 2014)] was measured in
upper and lower canopy leaves in week 3. Triplicate measurements from the center leaflet of the
three youngest fully-expanded fan leaves and from three fan leaves at the bottom of each plant,
were taken using a chlorophyll meter (CCM-200; Opti-Sciences, Hudson, NH, USA). The
triplicate measurements were averaged per plant for the upper and lower canopy leaves,
respectively.

56
The ratio of variable to maximum fluorescence (Fv/Fm) emitted from photosystem II in dark-
acclimated leaves exposed to a light-saturating pulse is an indicator of maximum quantum yield
of photosystem II photochemistry (Murchie and Lawson, 2013). During the first 8.5 h of the
PAR photoperiod (i.e., before daily UV exposure), the middle leaflet of the youngest, fully-
expanded fan leaf from each plant was dark acclimated for 15 min and then Fv/Fm measurements
were taken with a fluorometer (FluorPen FP 100; Drasov, Czech Republic). The Fv/Fm
measurements were done weekly on each cultivar, from the start of the trial until evidence of
stress (i.e., reduction of Fv/Fm with increasing UV-PFD) was seen.

3.2.5 LEAF GAS EXCHANGE MEASUREMENTS, LEAF SIZE AND SPECIFIC LEAF
WEIGHT

Quantifications of leaf gas exchange of the middle leaflet of the youngest, fully-expanded fan
leaf on each plant was performed in week 5 during the first 8.5 h of the PAR photoperiod using a
portable photosynthesis machine (LI-6400XT; LI-COR Biosciences, Lincoln, NE, USA)
equipped with the B and R LED light source (6400-02B; LI-COR Biosciences). In situ NCER
was measured with the leaf chamber environmental conditions set to: PPFD of 500 µmol·m–2·s–
1, block temperature of 26.7°C, CO2 concentration of 400 ppm, and air flow rate of 500 µmol·s–1.
If the leaflet did not cover the entire 6 cm2 chamber, the section of the leaflet that was clamped in
the chamber gasket was marked along the outside of the gasket so that leaf area inside the
chamber could be calculated post hoc (described below). After removing the leaflets from the
leaf chamber, whole leaves were excised from the plant and scanned (CanoScan LiDE 25; Canon
Canada Inc., Brampton, ON, Canada) at 600 dpi resolution. Each leaf was oven dried to constant
weight at 65°C (Isotemp Oven 655G; Fisher Scientific, East Lyme, CT, USA). The scanned
images were processed using ImageJ 1.42 software (National Institute of Health;
https://imagej.nih.gov/ij/download.html) to determine the leaflet area within the gas exchange
chamber and the total individual leaf size. The DW of each scanned leaf was measured using an
analytical balance (MS304TS/A00; Mettler-Toledo, Columbus, OH, USA) to determine SLW
[leaf DW / leaf size (g·m–2)].

57
3.2.6 VISUAL OBSERVATIONS

Weekly observations were performed on each plant to evaluate other visual parameters,
including: upward curling of the leaflet margins, leaf shine, leaf-drop, browning of inflorescence
stigmas, leaf epinasty (i.e., concaved leaf tissue between veins), leaf necrotic patches, and
appearance of powdery mildew on the adaxial sides of the leaves. Except for week 1
observations, which occurred 4 d after the start of the UV treatments, all weekly observations
occurred on 7-d intervals. Leaf-drop was recorded as the occurrence of fallen leaves observed on
each plant’s substrate surface. The absence or presence of each respective parameter was
evaluated for each plant, weekly. While these are observational data, weekly minimum UV-
PFDs where parameters were observed were reported, regardless of whether or not all plants
above these UV levels displayed the observed responses.

At various points throughout the trial, representative photos of each cultivar under different UV
exposure levels were taken with a digital camera (iPhone XR iOS 14.4.1; Cupertino, CA, USA).
In week 2, photos of whole plants exposed to minimum and maximum UV exposure, of each
cultivar, were taken. Photos of whole plants in week 3 and whole plants and apical
inflorescences at harvest (i.e., week 9), exposed to minimum, low, moderate and maximum UV
exposure levels (described in Table 3.1) were taken for each cultivar. Photos of the
inflorescences grown under minimum and maximum exposure levels, of each cultivar, were
taken in week 4. In week 5 [i.e., approximately when vegetative growth in cannabis ceases
(Rodriguez-Morrison et al., 2021)], fully-expanded leaves from plants under minimum,
moderate, and high UV exposure were excised from the plants and scanned (CanoScan LiDE 25)
at 600 dpi resolution. All photos were processed using ImageJ 1.42 software to add scale bars.

3.2.7 YIELD AND QUALITY

The FWs of total inflorescence, stems, and leaves were separately weighed for each plant using a
precision balance (EG 2200-2NM; Kern, Balingen, Germany). The separated aboveground
tissues of all plants were oven-dried at 65°C to constant weight (Isotemp Oven 655G) and the
DW (i.e., yield) of the respective tissues were recorded. The apical inflorescences of 18 plants
from each cultivar that were representative of the entire range of UV-PFD exposure levels were

58
air dried at 15°C and 40% relative humidity for 7 d before 2 g sub-samples from each plant were
submitted to an independent laboratory (RPC Science & Engineering; Fredericton, NB, Canada)
for analysis of concentrations [reported in mg·g–1(DW)] of cannabinoids using ultra-high-
performance liquid chromatography and variable wavelength detection (HPLC-VWD) and
terpenes using gas chromatography and mass spectrometry detection (GC-MSD). Total
equivalent ∆9-THC, CBD, and CBG concentrations were determined by assuming complete
carboxylation of the acid-forms (i.e., ∆9-THCA, CBDA and CBGA) of the respective
cannabinoids, whose concentrations were adjusted by factoring out the acid-moiety from the
molecular weight of each respective compound [e.g., total equivalent ∆9-THC = (∆9-THCA ×
0.877) + ∆9-THC].

3.2.8 STATISTICAL ANALYSIS

The treatments in this experiment were continuous, independent variables based on the
calculated UV-PFDs for each individual plant, each week. On per cultivar bases, the best-fit
models (linear or quadratic) for parameters with continuous dependent variables were selected
based on the lowest value for the Akaike Information Criterion (AICc) using UV-PFD as the
independent variable using the PROC NLMIXED procedure (SAS Studio Release 3.8; SAS
Institute Inc., Cary, NC). Analyses also revealed that each dataset had a normal distribution. For
parameters that were measured prior to harvest, UV exposure was determined based on the
weekly UV measurements made until the parameter was measured. If there were no LI treatment
effects on a given parameter, then parameter means (± SD) were calculated.

3.3 RESULTS
The measured canopy-level range of average UV-PFDs (based on the weekly UV measurements)
was 0.01 to 0.8 µmol·m–2·s–1. The average (± SE) step change, relative to the maximum UV-
PFD, between adjacent UV-PFD levels was 2.3 ± 0.43% for both cultivars. For models presented
below, this range was used to contextualize the results since all measured parameters fit within
this range.

59
3.3.1 UV-INDUCED CANNABIS MORPHOLOGY AND PHYSIOLOGY CHANGES

While each entire plant was observed for UV-induced changes in morphology, the recorded
effects occurred primarily in recently developed tissues. Where UV effects were also seen in
older tissues has been highlighted in the text. The data in Table 3.2 are provided to show the
temporal trends in how the parameters relate to each other and how UV sensitivity increases over
time.

The UV-induced changes in cannabis morphology appeared within the first few days of the
initiation of the UV treatments (i.e., in week 1), where the leaflet margins on leaves that had
developed in the vegetative stage (i.e., prior to the initiation of the UV exposure) curled upwards
under UV-PFDs ≥0.33 and ≥0.37 µmol·m–2·s–1 in the LT and BW cultivars, respectively (Table
3.2). Leaves appeared to accumulate epicuticular wax, as demonstrated by the increase in shiny
appearance of adaxial surfaces, shortly after UV exposure began and persisted henceforth. Leaf
shine also appeared to be more prevalent in plants exposed to higher UV-PFDs, more so in BW
vs. LT. In week 2 there were no changes in the extent of upward curling in the leaves that had
developed during the vegetative stage (i.e., older, mid-canopy leaves) however, newly expanded
leaves did not present this symptom (Figure 3.2). In week 3 (about a week after the first
appearance of inflorescences), stigmas of terminal inflorescences began to turn from white to
brown on LT plants exposed to UV-PFDs ≥0.69 µmol·m–2·s–1 and ≥0.30 µmol·m–2·s–1 in BW
(Figure 3.3, Table 3.2). In week 3, early symptoms of upper leaf epinasty started to appear in
upper canopy leaves of plants grown under the highest UV-PFDs (Figure 3.3); observational
measurements on leaf epinasty were initiated in week 5. Leaf-drop occurred in week 3 on plants
under UV-PFDs ≥0.14 and ≥0.23 µmol·m–2·s–1 in LT and BW, respectively (Table 3.2). Fallen
leaves appeared to be predominantly the same leaves that showed upward curling in week 1.

There were no treatment effects on the CCI of the upper canopy leaves of LT in week 3, but the
CCI of the upper canopy leaves of BW decreased linearly and with a 42% reduction from lowest
to highest UV-PFD. The CCI in the lower canopy leaves decreased linearly with the increase of
UV-PFD in week 3, with 60% and 46% reductions from lowest to highest UV-PFD in LT and
BW, respectively (Table 3.3). In week 4, the minimum UV-PFD at which plants exhibited

60
stigma browning (0.22 and 0.14 µmol·m–2·s–1 in LT and BW, respectively) and leaf-drop (0.013
and 0.050 µmol·m–2·s–1 in LT and BW, respectively) were lower than the previous week (Table
3.2). The onset of UV treatment effects on Fv/Fm were observed in week 4 for BW and in week 5
for LT. Both cultivars presented quadratic relationships between UV-PFD and Fv/Fm (Table 3.3).
However, the Fv/Fm values were within 1% (i.e., low stress) of their respective vertices (both
≈0.79, at 0.084 and 0.12 µmol·m–2·s–1 for LT and BW, respectively) from the lowest UV-PFD
up to ≈0.25 µmol·m–2·s–1, followed by an increasing rate of reduction in Fv/Fm with increasing
UV-PFD thereafter. At the highest UV-PFD, the Fv/Fm values were 10% and 21% lower than the
respective vertices for LT (in week 5) and BW (in week 4), respectively. The severity of UV-
induced epinasty was higher in plants exposed to higher UV-PFDs, demonstrated by images of
whole plants (in week 3) of LT (Figure 3.4A) and BW (Figure 3.4B) and single-leaf scans (in
week 5) of LT and BW (Figure 3.5) grown under various UV exposure levels. In week 5, leaves
(particularly in the upper canopy) showed upward curling under minimum UV-PFDs of 0.16 and
0.34 µmol·m–2·s–1 in LT and BW, respectively (Table 3.2, Figure 3.5). In week 5, brown
stigmas were observed at the minimum UV-PFD in LT and 0.14 µmol·m–2 in BW (Table 3.2).
Leaf-drop was observed at very low UV-PFDs of 0.013 µmol·m–2·s–1 in LT and 0.015 µmol·m–
2·s–1 in BW (Table 3.2).

Leaf epinasty was evident in week 5, predominantly in youngest fully-expanded leaves, exposed
to minimum UV-PFDs of 0.13 µmol·m–2·s–1 in LT and 0.14 µmol·m–2·s–1 in BW (Table 3.2). In
week 5, the size of the youngest fully-expanded leaf on each plant decreased linearly by almost
half in both cultivars, while SLW increased by 27% and 21% in LT and BW under highest vs.
lowest UV-PFD (Table 3.3). In week 5, the in situ NCER of the youngest fully-expanded leaves
decreased 31% and 27% in LT and BW, respectively, at highest vs. lowest UV-PFD (Table 3.3).
Brown stigmas and leaf-drop were observed in all experimental plants starting in week 6 (Table
3.2). Starting in in week 7, upper canopy leaves on a few plants grown under intermediate UV-
PFDs (i.e., ranging from 0.12 to 0.69 µmol·m–2·s–1 in LT) began to show brown (necrotic)
patches (Table 3.2, see Appendix C). The minimum UV-PFDs under which leaf epinasty was
evident were marginally lower in week 7 vs. week 5, and substantially lower in week 8 vs. week
7 (Table 3.2). The prevalence of leaves exhibiting necrotic patches increased in BW in week 8

61
vs. week 7 (Table 3.2). While investigating the UV exposure effects on foliar powdery mildew
was not one of the designed objectives of this study, treatment differences were observed and
recorded. In week 9, powdery mildew was visible on the adaxial leaf surfaces on plants exposed
to lower UV-PFDs but was not observed on any plants exposed to UV-PFDs ≥0.076 µmol·m–2·s–
1 in LT and ≥0.090 µmol·m–2·s–1 in BW (Table 3.2, Figure 3.6).

Figure 3.2. (A) The side view and (B) top view of Cannabis sativa L. plants in week 2 after the
initiation of the UV treatments. ‘Low Tide’ under (1) minimum and (2) maximum UV exposure
and ‘Breaking Wave’ under (3) minimum and (4) maximum UV exposure levels. The black scale
bar at the lower right of each image is 5.0 cm.

62
Figure 3.3. (A) ‘Low Tide’ (LT) and (B) ‘Breaking Wave’ (BW) Cannabis sativa L. stigmas
under minimum UV exposure levels and (C) LT and (D) BW under maximum UV exposure
levels, in week 3 after the initiation of the UV treatments. The white scale bar at the lower right
of (C) applies to (A), and at the lower right of (D) applies to (B). Both scale bars are 1.0 cm.

63
Figure 3.4. (A) ‘Low Tide’ and (B) ‘Breaking Wave’ Cannabis sativa L. plants demonstrating
(from left to right) minimum, low, moderate, and high UV exposure levels. The images were
taken in week 3 after the initiation of the UV treatments. The black scale bar at the lower right of
each image is 5.0 cm.

64
 A

Figure 3.5. (A) Adaxial and (B) abaxial sides of youngest, fully-expanded Cannabis sativa L.
fan leaves of ‘Low Tide’ (top row in each image) and ‘Breaking Wave’ (bottom row in each
image) demonstrating UV induced leaf morphology effects with increasing UV-PFDs. Leaves
from plants under minimum UV exposure are on the left, moderate UV exposure in the middle,
and high UV exposure on the right. Scans were taken in week 5 after the initiation of UV
treatments. The black scale bar at the lower right of each image is 2.0 cm.

65
Table 3.2. Minimum UV-PFD (µmol·m–2·s–1) where symptoms were observed in Cannabis
sativa L. ‘Low Tide’ (LT) and ‘Breaking Wave’ (BW) cultivars (CV) in each week after the
initiation of UV treatments, regardless of whether or not all plants above the minimum UV-PFD
presented the observed symptom.

Leaf symptoms Flower Symptoms

Upward Necrotic Powdery
Weekz CV Epinasty Leaf-drop Stigma browning
curling patches mildew
LT 0.33
1 NIy NI NI NI NI
BW 0.37
LT
2 NIEx NI NI NI NI NI
BW
LT 0.14 0.69
3 NIE NI NI NI
BW 0.23 0.30
LT 0.013 0.22
4 NIE NI NI NI
BW 0.050 0.14
LT 0.16 0.13 0.013 0
5 NI NI
BW 0.34 0.14 0.015 0.14
LT 0.13 0w 0
6 NIE NI NI
BW 0.33 0 0
v
LT 0.10 0 0.12 to 0.69 0
7 NIE NI
BW 0.13 0 0.32 0
LT 0.018 0 0.12 to 0.70 0
8 NIE NI
BW 0.034 0 0.20 to 0.51 0
LT 0 0 to 0.076 0
9 NIE NIE NIE
BW 0 0 to 0.090 0
zexcept for observations in week 1, which occurred 4 d after the start of the UV treatments, all
weekly observations occurred on 7-d intervals
yNI: symptom was not investigated
xNIE: no increase in extent of crop sensitivity to UV exposure level was observed
wzero indicates that the symptom was observed at the lowest UV-PFD
vranges are provided when the symptom was observed in only intermediate UV-PFDs

66
Table 3.3. The effects of UV-PFD (µmol·m–2·s–1) applied during the flowering stage on
physiological, morphological and yield parameters of Cannabis sativa L. ‘Low Tide’ and
‘Breaking Wave’.

Regression equationz, R2 or mean ± SDy

Parameter Unit
‘Low Tide’ ‘Breaking Wave’
CCIx of upper canopy leaves in week – 42.0 ± 8.11 y = –19.0x + 36, 0.28
3
CCI of lower canopy leaves in week – y = –30.6x + 41, 0.39 y = –18.7x + 32, 0.27
3
Net CO2 exchange rate (NCER) in µmol(CO2)·m–2·s–1 y = –7.52x + 19, 0.35 y = –6.78x + 20, 0.32
week 5
Fv/Fm in week 2 – 0.78 ± 0.0269 0.78 ± 0.0338
Fv/Fm in week 3 – 0.80 ± 0.0154 0.77 ± 0.0329
Fv/Fm in week 4 – 0.78 ± 0.0224 y = –0.364x2 + 0.0839x + 0.79,
0.87
Fv/Fm in week 5 – y = –0.152x2 + 0.0255x + 0.79, –
0.41
Individual leaf size in week 5 cm2/leaf y = –15.7x + 28, 0.46 y = –10.4x + 18, 0.45
Specific leaf weight (SLW) in week 5 g(leaf)·m–2(leaf) y = 15.4x + 45, 0.34 y = 12.3x + 46, 0.32
Growth index in week 6 – y = –210x + 275, 0.25 y = –145x + 350, 0.11
Increase in height until week 6 cm y = –13.7x + 35, 0.14 y = –11.9x + 36, 0.13
Increase in number of nodes until – y = –3.42x + 10.0, 0.11 y = –3.15x + 9.7, 0.11
week 6
Inflorescence moisture content % 79.0 ± 1.27 79.3 ± 1.01
Leaf moisture content % 69.5 ± 3.59 72.0 ± 2.31
Stem moisture content % 72.4 ± 2.93 74.5 ± 2.17
Apical inflorescence dry weight g·m–2 y = –55.8x + 57, 0.43 y = –22.5x + 26, 0.45
(DW)
Total inflorescence DW g·m–2 y = –95.6x + 235, 0.11 236 ± 63
Leaf DW g·m–2 y = –26.0x + 111, 0.14 y = –40.5x + 100, 0.28
Stem DW g·m–2 49.2 ± 26.5 46.1 ± 21.6
zparameters with UV treatment effects (P ≤0.05) are presented as linear or quadratic models and
the R2
ythe means ± SD are presented for parameters without UV treatment effects
xchlorophyll content index

3.3.2 GROWTH RESPONSES TO UV

Exposure to UV radiation suppressed plant growth, which was recorded after the cessation of the
majority of vegetative growth (i.e., week 6). Growth indices were 61% and 33% lower in plants
grown under the highest vs. lowest UV-PFDs in LT and BW, respectively (Table 3.3, Figure
3.6). Increases in height were 31% and 26% lower in plants grown under the highest vs. lowest
UV-PFDs in LT and BW, respectively. Increases in numbers of nodes were 27% and 26% lower
in plants grown under the highest vs. lowest UV-PFDs in LT and BW, respectively (Table 3.3).

67
There were no UV treatment effects on moisture content of aboveground tissues in either cultivar
(Table 3.3).

Figure 3.6. Gross plant morphology of (A) ‘Low Tide’ and (B) ‘Breaking Wave’ Cannabis
sativa L. plants grown under (from left to right) minimum, low, moderate, and high UV exposure
levels. Images were taken just prior to harvest (i.e., 9 weeks after the initiation of UV
treatments). Note the white spots (powdery mildew) on the adaxial sides of leaves on the far-left
plants. The black scale bar at the upper left of each image is 5.0 cm.

68
3.3.3 RESPONSES OF INFLORESCENCE YIELD, QUALITY AND CANNABINOID
AND TERPENE CONCENTRATIONS TO UV

The most discernable UV exposure effects on inflorescences were the differences in size (Figure
3.7) of the apical inflorescences. The apical inflorescence DW was reduced linearly by 78% and
69% in LT and BW, respectively, from the lowest to highest UV-PFDs. However, the reduction
in apical inflorescence DW under increasing UV exposure only translated to reductions in total
inflorescence DW in LT, which was 32% lower under the highest vs. lowest UV-PFDs.
Approximately 60% of the difference in the total LT inflorescence DW in lowest vs. highest UV
exposure levels arose from the decreases in the DW of apical inflorescences. The leaf DW were
19% and 32% lower under highest vs. lowest UV-PFD in LT and BW, respectively. There were
no UV treatment effects on stem DW.

The effects of UV exposure on the apical inflorescence secondary metabolite composition varied
between the two cultivars. In LT, the concentrations of Δ9-THCA, CBDA and CBGA decreased
linearly by 15%, 21% and 31%, respectively, as UV-PFD increased from lowest to highest
(Table 3.4); with concomitant reductions in the total equivalent concentrations of these
cannabinoids. However, there were no UV treatment effects on Δ9-THC, CBD, CBG or
cannabinol (CBN) concentrations, or the ratio of total equivalent Δ9-THC to total equivalent
CBD (hereafter, Δ9-THC:CBD) in LT. As UV-PFD increased from lowest to highest, the
concentrations of myrcene, limonene, fenchol all decreased in LT, resulting in a combined 41%
decrease in the total concentration of terpenes. In BW, the concentration of Δ9-THC in the apical
inflorescences was 1.6 times higher under highest vs. lowest UV-PFD, while there were no UV
treatment effects on the concentrations of the other cannabinoids. However, the Δ9-THC:CBD
was 1.1 times higher in BW under highest vs. lowest UV-PFD. The concentration of myrcene
and linalool in BW decreased while caryophyllene and guaiol concentrations increased, with
increasing UV-PFD, resulting in a combined 24% decrease in the total concentration of terpenes
at the highest vs. lowest UV-PFD.

69
Figure 3.7. The apical inflorescences of (A) ‘Low Tide’ and (B) ‘Breaking Wave’ Cannabis
sativa L. plants grown under (from left to right) minimum, low, moderate, and high UV exposure
levels. Images were taken at harvest (i.e., 9 weeks after the initiation of UV treatments). The
black scale bar at the upper left of each image is 2.0 cm.

70
Table 3.4. The effects of UV-PFD (µmol·m–2·s–1) applied during the flowering stage on
cannabinoid and terpene concentrations (mg·g–1) in the apical inflorescence of Cannabis sativa
L. ‘Low Tide’ and ‘Breaking Wave’.
Regression equationz, R2 or mean ± SDy
Parameter (mg·g–1)
‘Low Tide’ ‘Breaking Wave’
Δ9-tetrahydrocannabinol (Δ9-THC) 1.66 ± 0.347 y = 1.02x + 1.35, 0.327
Δ9-THC acid (Δ9-THCA) y = –15.9x + 84.3, 0.294 73.8 ± 12.2
Total equivalent Δ9-THC (TΔ9-THC) y = –13.5x+ 75.5, 0.282 66.5 ± 11.1
Cannabidiol (CBD) 1.38 ± 0.447 1.47 ± 0.338
CBD acid (CBDA) y = –33.9x + 130, 0.428 93.8 ± 13.8
Total equivalent CBD (TCBD) y = –29.3x + 115, 0.408 83.7 ± 12.2
Cannabigerol (CBG) 0.657 ± 0.275 1.27 ± 0.217
CBG acid (CBGA) y = –3.31x + 8.59, 0.576 6.04 ± 1.07
Total equivalent CBG (TCBG) y = –2.93x+ 8.21, 0.435 6.85 ± 1.07
Total equivalent Δ9-THC: total equivalent CBD 0.678 ± 0.0387 y = 0.0980x + 0.755, 0.262
Cannabinol (CBN) UDLx UDL
Alpha pinene UDL 0.214 ± 0.0593
Beta pinene 0.235 ± 0.0672 0.440 ± 0.124
Myrcene y = –4.16x + 6.21, 0.376 y = –2.47x + 3.45, 0.363
Limonene y = –0.788x + 1.26, 0.341 1.57 ± 0.423
Linalool 0.274 ± 0.0703 y = –0.147x + 0.222, 0.528
Terpineol 0.254 ± 0.0655 0.379 ± 0.108
Caryophyllene 2.42 ± 0.746 y = 0.520x + 1.13, 0.354
Humulene 0.892 ± 0.324 0.403 ± 0.0842
Fenchol y = –0.118x + 0.219, 0.321 0.257 ± 0.0653
Guaiol 0.801 ± 0.100 y = 0.251x + 0.457, 0.313
Alpha-bisabolol 0.677 ± 0.181 0.355 ± 0.104
Total terpenes y = –7.25x + 14.0, 0.383 y = –2.72x + 9.21, 0.222
z
cannabinoids and terpenes with UV treatment effects (P ≤0.05) are presented as equations and
R2
ythe means ± SD are presented for cannabinoids and terpenes without UV treatment effects
xunder detection limit of 0.5 mg·g–1 of inflorescence DW

3.4 DISCUSSION
Both LT and BW cultivars would be categorized as chemotype II because they have relatively
balanced total ∆9-THC and total CBD concentrations (Small and Beckstead, 1973). However,
they demonstrated disparate morphological attributes: LT had a relatively compact phenotype
with wide leaflets and dense branching and BW had a relatively spindly phenotype with narrow
leaflets and sparse branching. Each cultivar responded to UV exposure with different magnitudes
of severity, but in the majority of the parameters that had UV treatment effects, increasing UV
exposure resulted in distress responses [i.e., damage to plant growth and health following a
strong stress event (Hideg et al., 2013)] that are generally unfavorable for commercial cannabis
production.
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3.4.1 UV RADIATION ALTERS CANNABIS MORPHOLOGY AND PHYSIOLOGY

Cannabis leaf morphology demonstrated substantial plasticity in response to UV radiation

exposure throughout the 9-week flowering stage. The first observed morphological response to
UV was upward curling of leaflet margins during the first week of treatment in plants exposed
UV-PFDs as low as 0.33 µmol·m–2·s–1 in LT and 0.37 µmol·m–2·s–1 in BW. Upward leaf curling
was most evident under higher UV-PFDs, and it occurred primarily on the youngest leaves (i.e.,
that developed just prior to UV exposure). Conversely, leaves that expanded in subsequent
weeks (i.e., under UV exposure) exhibited more typical UV-induced morphology responses such
as reduced leaf size and increased SLW, which were more prominent in plants grown under
higher UV exposure levels (Searles et al., 2001; Zlatev et al., 2012). Further, by week 5, leaves
of both cultivars exhibited similar responses of increasing epinasty with increasing UV-PFD.
Leaves in BW demonstrated more plasticity in terms of epinasty than LT (Figure 3.5), which
may be acclimations for leaf protection by reducing foliar exposure to UV stress (Wilson, 1998;
Fierro et al., 2015). The apparent increase in leaf shine shortly after the initiation of UV
exposures indicates an accumulation of epicuticular wax, which is a common response to UV
exposure in other crops (Steinmüller and Tevini, 1985; Cen and Bornman, 1993; Fukuda et al.,
2008; Valenta et al., 2020). Since epicuticular wax may reflect UV radiation (Cen and Bornman,
1993; Valenta et al., 2020), upregulating epicuticular wax production may reduce the potential
for damage to the photosynthetic machinery.

UV radiation accelerated plant senescence symptoms (i.e., symptoms associated with

deterioration with age), including signs of foliar stress. Female inflorescence maturation can be
characterized by carpel swelling and the transition from white stigmas to a reddish-brown colour
in the final days before harvest (Punja and Holmes, 2020). Although the number of days until the
appearance of inflorescences after moving to the 12-h photoperiod was unaffected by UV
exposure (data not shown), plants exposed to higher UV-PFDs exhibited earlier stigma browning
without carpel swelling (Figure 3.3). It is unknown if premature stigma senescence has any
knock-on effects on other inflorescence development parameters, such as production of
secondary metabolites. However, since stigma browning was observed in different weeks

72
depending on UV exposure levels, this attribute could not be used reliably to determine the
“optimum harvest maturity”.

Foliar chlorophyll content is often negatively correlated to UV exposure level (Neugart and
Schreiner, 2018), which could have negative effects on photosynthesis (Singsaas et al., 2004).
While increasing UV-PFD decreased CCI in the upper canopy leaves of BW and there were no
UV treatment effects on CCI in LT, CCI is an area-based metric that is not directly indicative of
leaf chlorophyll concentration on a biomass basis (i.e., mg·g–1). Chlorophyll concentration in the
upper canopy leaves in both cultivars would probably have been reduced under increasing UV-
PFD due to the trends of increasing SLW (i.e., leaf thickness) under higher UV exposure.
Reduced lower canopy CCI was manifested through higher leaf chlorosis (e.g., Figure 3.6) and
earlier leaf-drop, a phenomenon also seen in sweet basil exposed to UVB radiation (Dou et al.,
2019). Nitrogen from lower canopy leaves is normally remobilized to more active upper canopy
foliage (Havé et al., 2017) as plants age; this appeared to be accelerated by UV exposure given
the reduction in CCI. Foliar necrosis is also a commonly-observed symptom of UV damage in
many species (Maffei and Scannerini, 2000; Zhao et al., 2003; Dou et al., 2019). While the
severity of most of the observed UV stress responses increased with increasing UV exposure,
necrotic patches were observed on upper canopy leaves exposed to intermediate UV-PFDs
(primarily in LT) in the latter weeks of the trial. The acclimation (e.g., epinasty, curling, small
size) of leaves exposed to the highest UV-PFDs may have mitigated foliar necrosis, while the
leaves grown under intermediate UV-PFDs may not have been sufficiently acclimated for long-
term UV exposure. Upper canopy leaves (in week 5 for LT and week 4 for BW) exposed UV-
PFDs <0.25 µmol·m–2·s–1 had Fv/Fm values of ≈0.8, which is normal for unstressed leaves
(Björkman and Demmig, 1987). The reduction in Fv/Fm at higher UV exposure levels in both
cultivars indicates that UV radiation may have induced a stress response in the photosynthetic
machinery of upper canopy leaves.

Lydon et al. (1987) reported no UV treatment effects on measured cannabis morphology and
physiology parameters, which is in stark contrast to the copious morphological and physiological
UV-induced stress responses (outlined above) observed in the present study. While the reported
maximum doses were similar in both studies, the plants in Lydon et al. (1987) may have
73
experienced lower than reported doses due to rapid aging of the cellulose acetate filters (used to
eliminate UVA and PAR wavelengths from their UV spectrum treatments) when exposed to
UVB (Middleton and Teramura, 1993), resulting in substantial reductions in UVB transmissivity.
Additionally, their plants grew for several months under greenhouse conditions (likely including
some UV) prior to exposure to UV treatments, whereas there was no UV exposure in the light
history of the young vegetative plants prior to initiation of the UV treatments in the present
study. Therefore, light history (e.g., spectrum and intensity) and plant age may affect plant
responses to UV stress. Evidently, the plants in the present study were subjected to more
efficacious UV radiation treatments than in Lydon et al. (1987).

3.4.2 UV RADIATION SUPPRESSES CANNABIS GROWTH AND YIELD

Increasing UV radiation exposure suppressed overall plant growth (e.g., height, number of
nodes, plant size) in both cultivars; however, these UV exposure responses were generally more
severe in LT than BW. The growth reductions in both cultivars could be partially attributed to
the UV-induced alterations in leaf morphology that impede biomass accumulation. Reduced
aboveground biomass and lower yields are common effects of exposure to UV radiation
(Teramura et al., 1990; Fiscus and Booker, 1995; Caldwell et al., 2003; Liu et al., 2005).
Reduced leaf area is a typical response to light stress [e.g., high intensity (Poorter et al., 2019) or
UV radiation (Wargent and Jordan, 2013)]. In the present study, individual leaf size and total leaf
biomass reduced with increasing UV exposure, limiting area available for light capture which
reduces a plant’s capacity to convert PAR into biomass (Zlatev et al., 2012). In addition to
morphological changes that impede biomass accumulation, area-based NCER was reduced as
UV exposure increased over the range of UV-PFDs evaluated in both cultivars, which could
further reduce yields under higher UV exposure levels (Kakani et al., 2003).

Total inflorescence DW and the proportion of that DW which is comprised of apical tissues are
two major considerations for commercial cannabis production. The apical proportion may be of
particular interest since these tissues are normally considered premium quality due to their
relatively large size and higher cannabinoid concentrations compared to higher-order (i.e., on
lower branches) inflorescences (Namdar et al., 2018). Despite the UV-induced limitations to

74
foliar biomass accumulation seen in both cultivars, increasing UV exposure only reduced total
inflorescence yield in LT. Within this context, the various growth habits of common indoor-
grown cannabis cultivars may influence their yield responses to UV stress. In the present study,
BW and LT had disparate whole-plant reproductive macro-morphology (i.e., the distribution of
inflorescence biomass within the canopy) under normal indoor conditions. For example, under
minimum UV exposure, the apical inflorescence comprised 24% of the total inflorescence DW in
LT compared to only 11% in BW. Apparently, growth habit may have predisposed BW’s
mitigation of UV-induced yield reductions by partitioning relatively more inflorescence biomass
to positions farther away (i.e., more protected) from the UV source. However, while this may be
a self-protective response to reduce UV exposure to reproductively important (from an
ecological sense) tissues, it also came at commercially-objectionable reductions in inflorescence
quality, such as visually unappealing morphology (Figure 3.7).

To prevent UV-induced yield losses, such as are reported in the present study, it is conceivable
that cannabis plants could be exposed to UV only after the majority of vegetative growth has
completed [i.e., a few weeks after the visual appearance of inflorescences (Potter, 2014)]. This
strategy may minimize UV-induced leaf alterations that could inhibit biomass accumulation.

3.4.3 UV RADIATION ALTERS THE SECONDARY METABOLITE COMPOSITION OF

CANNABIS INFLORESCENCES

The most economically relevant cannabinoids (e.g., Δ9-THC and CBD) are predominantly found
in their acid forms in mature female inflorescence tissues, which are converted to the
psychoactive and medicinal compounds through decarboxylation (Eichler et al., 2012; Zou and
Kumar, 2018). The decarboxylated compounds also exist in relatively low quantities in the fresh
inflorescences, for example, while Δ9-THC concentration increased in BW with increasing UV-
PFD, the concentration was a relatively small proportion of the total equivalent Δ 9-THC;
maximized at 3.3% at the highest UV-PFD. Further, Δ9-THC naturally converts to CBN,
particularly under postharvest conditions and in the presence of oxygen and light. Since CBN
was undetectable in the inflorescences, this is an indicator that the crop was not past peak
maturity (in terms of cannabinoid concentrations) at the time of harvest (Russo, 2007; Aizpurua-
Olaizola et al., 2016).
75
The present study found that there were no UV-induced enhancements to total equivalent
concentrations of Δ9-THC, CBD and CBG. These results are consistent with a recent study that
found no UV treatment effects on Δ9-THC content in a Δ9-THC-dominant cultivar (Llewellyn et
al. 2021), but contrast with studies on older genotypes (Pate, 1983; Lydon et al., 1987). For
example, by exposing greenhouse-grown cannabis to UVBE (based on Caldwell, 1971) doses up
to 13.4 kJ·m–2·d–1, Lydon et al. (1987) found Δ9-THC concentration increased linearly by 28%
under the highest dose vs. control (i.e., 32 vs. 25 mg·g–1). These contrasting results may be due
to the disparate growing conditions (both before and during UV exposure), plant age at the time
of UV exposure and the relative magnitude of cannabinoid concentrations. While the
proportional increases in Δ9-THC content (28%) presented in Lydon et al. (1987) appeared to be
substantial, the magnitude of their increase (e.g., 7 mg·g–1) is probably inconsequential in the
context of cannabinoid composition in modern genotypes which can have Δ9-THC
concentrations that exceed 200 mg·g–1 (Dujourdy and Besacier, 2017).

Pate (1983) reported an increased Δ9-THC:CBD under UV exposure, which suggests that the
production of Δ9-THC may be upregulated and CBD downregulated as temporal adaptations
(i.e., over multiple generations) to the localized environment. However, the results of the present
study do not support this trend, at least as a short-term acclimation response to UV stress.
Additionally, de Meijer et al. (2003) showed that cannabinoid profiles are largely genetically
predetermined, (e.g., a CBD-dominant cultivar is lacking the genetic predisposition to generate
abundant Δ9-THC). This could support the contention that the upregulation of Δ9-THC under UV
stress may be an adaptive response (i.e., over generations) rather than an acclimation response
(i.e., during a single production cycle). Over the past few decades, there have been radical
increases in inflorescence cannabinoid concentrations, which is often attributed to intensive
breeding programs (Chouvy, 2015; Dujourdy and Besacier, 2017; Aliferis and Bernard-Perron,
2020) and the “sinsemilla” cultivation method that eliminates seeds and chiefly produces high
potency female inflorescences (ElSohly et al., 2016). Thus, these factors may have a larger
impact on cannabis inflorescence cannabinoid composition than environmental factors such as
the UV exposure treatments in the present study.

76
While cannabinoids are the primary psychoactive and medicinal compounds in cannabis
inflorescences, volatile terpenes are also economically valuable; both for the aromas that
influence consumer preference and potential medicinal properties (Nuutinen, 2018; Booth and
Bohlmann, 2019). UV radiation equivocally altered the terpene composition, with disparate
responses within and between cultivars. However, total terpene concentrations in both cultivars
decreased linearly with increasing UV exposure, which could depreciate the quality of aromas
and extracts (McPartland and Russo, 2001; Nuutinen, 2018).

While UV exposure did not result in economically relevant increases in cannabinoid or terpene
concentrations in cannabis inflorescences under the conditions of the present study, UV radiation
has been shown to increase concentrations of UV-absorbing secondary metabolites (e.g.,
flavonoids and phenolic compounds) in many species (Huché-Thélier et al., 2016; Robson et al.,
2019), including economically important essential oil producing crops (Schreiner et al., 2012;
Neugart and Schreiner, 2018). However, UV-induced increases in secondary metabolite
concentrations are often concurrent with biomass reductions (Fiscus and Booker, 1995; Caldwell
et al., 2003). This paradox must be evaluated when considering the use UV radiation to
manipulate secondary metabolite concentrations in indoor cannabis production, since the
simultaneous yield reduction may offset any improvements in secondary metabolite composition.

Compared to the UV spectra employed in most other studies, the biologically effective doses in
the present study were dramatically higher for a given photon flux density due to the very short
peak wavelength of the UV LEDs. It is possible that the alternate UV treatment protocols, such
as have been used in other studies, may have more positive results in cannabis production; for
example: longer wavelength, less energetic spectra (Hikosara et al., 2010) and shorter-term (e.g.,
proximal to harvest maturity) exposure (Johnson et al., 1999; Martínez-Lüscher et al., 2013;
Huarancca Reyes et al., 2018; Dou et al., 2019).

3.4.4 IMPLICATIONS OF UV IN INDOOR CANNABIS PRODUCTION AND FUTURE

RESEARCH DIRECTIONS

This study provided insight into the sensitivity of cannabis to relatively short-wavelength UVB
radiation (including a small proportion of UVC) and long-term UV exposure. Increasing UV

77
exposure levels generally had negative impacts on cannabis plant growth, inflorescence yield,
quality, and secondary metabolite composition. The plants exhibited primarily distress responses
to UV radiation, even at low exposure levels. No amount of UV exposure resulted in substantial
increases of cannabinoid concentrations. Perhaps the UV exposure treatments protocol in the
present study (i.e., short-wavelength and long-term exposure) were sufficiently stressful that any
potential eustress responses [i.e., a positive response to mild stress (Hideg et al., 2013)] to
increasing UV intensity were overwhelmed by more abject distress responses.

Many studies have investigated the effects of stratospheric ozone depletion on plant exposure to
UV radiation (Searles et al., 2001; Caldwell et al., 2003) either through ecological or controlled-
environment type research. The ratios between UV and PAR (hereafter, UV:PAR) in controlled-
environment type investigations tend to be much higher than in the solar spectrum in terrestrial
ecosystems (Robson et al., 2019), where plants may exhibit higher sensitivity to UV radiation
including increased secondary metabolite accumulation and reduced photosynthesis and growth
relative to lower UV:PAR responses (Behn et al., 2010; Dou et al., 2019). The other spectra
within the lighting environment have also been shown to influence plant sensitivity to UV
radiation, including biomass accumulation (Palma et al., 2021) and some spectra have even been
shown to counteract UVB-induced damage (Krizek, 2004). Conceivably, through serendipity,
researchers have discovered potential horticultural benefits for providing unnaturally stressful
UV exposure conditions which can enhance pertinent traits in economically relevant crops,
including increasing secondary metabolite concentrations (Huché-Thélier et al., 2016).

Any horticultural UV treatment protocol is comprised of a myriad of factors including: time of

application in the plants’ life cycle, number of days of application, number of hours per day
(including pulsed methods), time of day with respect to the PAR photoperiod, spectrum, and
intensity. While none of the UV exposure levels in the present study would have been
commercially beneficial, results from studies in other species (Huché-Thélier et al., 2016;
Neugart and Schreiner, 2018; Robson et al., 2019; Höll et al., 2019) indicate a strong potential
for there being UV treatment protocols – as yet unidentified through rigorous scientific
investigation and reporting – that could enhance secondary metabolite concentrations in
cannabis.
78
There are other horticulturally relevant reasons, other than enhancing secondary metabolite
concentrations, for having UV wavelengths present in the radiation spectrum in a cannabis
production environment, such as pest management. In the present study there were visible
reductions in powdery mildew on the adaxial foliar surfaces of plants exposed to UV-PFDs
≥0.10 µmol·m–2·s–1, with concomitant reductions in inflorescence yield of only 2-3% in LT (no
yield reductions in BW) at this threshold UV exposure level. UV exposure has also been shown
to reduce powdery mildew in other crops such as grape (Austin and Wilcox, 2012) and
strawberry and rosemary (Suthaparan et al., 2016). UV exposure may also improve plant
resistance to insect herbivory through changes in plant secondary metabolite composition that
affect plant-insect interactions (Demkura et al., 2010).

When making the decision to utilize UV wavelengths (as with any production technology) in
indoor cannabis production, the positive crop outcomes must outweigh factors related to the cost
of deploying the technology including infrastructure and energy costs, fixture lifespan, and
health risks that UV radiation could pose to employees. While UVB LEDs in particular (Kusuma
et al., 2020) and UV lighting technologies in general are less energy efficient than modern
horticultural PAR fixtures (Nelson and Bugbee, 2014; Radetsky, 2018), fluence rates in the UV
spectrum are typically many times lower than the PAR spectrum. Typical functional lifespans of
UVB LEDs are currently much lower (Kebbi et al., 2020) than common horticultural LEDs
(Kusuma et al., 2020); potentially leading to relatively rapid degradation in fluence rates over
time. Given that plant responses in the present study were closely tied to the UV exposure level,
fixture degradation could lead to inconsistencies between sequential crops, which is an important
parameter in the indoor cannabis production industry. Future research could seek to achieve UV
application protocols that promote eustress responses in cannabis secondary metabolite
concentrations while minimizing distress responses (e.g., yield reductions) by using less
energetic UV spectra or shorter-term exposure than were used in the present study. Future
research could also investigate how UV affects cannabis plants grown under different lighting
histories, and seek to determine the ideal developmental stage for UV exposure to achieve the
desired effects in both yield and quality.

79
3.5 CONCLUSIONS
Long-term exposure of various intensities of relatively short-wavelength UV radiation had
generally negative impacts on cannabis growth, inflorescence yield, and inflorescence quality.
By studying two cultivars with similar cannabinoid profiles, we found some differences in
phenotypic plasticity in the temporal dynamics in morphology, physiology, yield, and quality
responses to UV exposure level. Importantly, as it was applied in this study, UV radiation had
substantially reduced yield in one cultivar and had no commercially relevant benefits to
inflorescence secondary metabolite composition. Therefore, potential for UV radiation to
enhance cannabinoid concentrations must still be confirmed before UV can be used as a tool in
cannabis production.

80
 CHAPTER FOUR
GENERAL DISCUSSION AND CONCLUSIONS

Light is a crucial resource for plant growth. It is well known that the manipulation of light
environment (e.g., LI and spectrum) can substantially alter plant development. In indoor plant
production, LI and spectrum can be manipulated to achieve desired plant traits. Determining
optimal LI for indoor cannabis production is of particular interest, since the costs relating to light
energy make up a large component of the total energy consumption. However, there was no
reliable published research demonstrating the relationship between LI and cannabis floral yield
to provide guidance for growers to optimize LI energy cost and yield return. Cannabis is unique
in that it is cultivated not only for its floral yield, but also for the medicinal and recreational
secondary metabolites (e.g., cannabinoids and terpenes) in the inflorescences. These two factors
must be optimized to improve cannabis quality and profitability. Prior work suggests that
wavelengths in the PAR spectrum may influence the secondary metabolite profile in cannabis.
Early studies have alluded to the potential for increasing cannabinoid concentrations with UV
radiation, which has made the UV spectrum a particular topic of current interest in the cannabis
industry. However, this theory has yet to be validated with modern production systems and
modern genotypes that have relevant baseline cannabinoid concentrations. The objectives of this
thesis were to 1) establish the relationships between light intensity and cannabis leaf-level
photosynthesis, and inflorescence yield and quality, and 2) establish the relationships between
UV exposure levels and cannabis morphology, physiology, and inflorescence yield and quality.

To achieve the first objective, the impact of a refined range of LIs (from 120 to 1800 µmol·m–
2·s–1 testing the lower and upper limits of practical LIs used in indoor crop production) on
cannabis leaf-level photosynthesis, and inflorescence yield and quality was evaluated. Likewise,
to achieve the second objective, the impact of various UV exposure levels of short-wavelength
UV radiation (applied during the flowering stage) on morphology, physiology, and inflorescence
yield and quality of two cultivars (LT and BW) was evaluated. The findings demonstrated the
extraordinary plasticity of cannabis’ physiological, morphological and yield responses to

81
increasing LI and UV radiation. In general, the effects of LI and UV radiation, as they were
applied in this thesis, had drastically contrasting effects on cannabis plant health. Increasing LI in
the cannabis growing environment during the flowering stage had an overwhelmingly positive
impact on the cannabis plant, notably shown through the linear inflorescence yield increase up to
1800 μmol·m–2·s–1. In other words, the relationship between LI and cannabis yield did not
saturate within the practical limits of LI used in indoor production. A general trend of stress was
observed in the UV response of both cannabis cultivars, despite differences in phenotypic
plasticity of crop morphology, physiology, yield and quality. Importantly, as it was applied in
this study, UV radiation had no commercially relevant benefits to inflorescence cannabinoid
content, and the total terpene concentrations decreased linearly over the range of UV-PFDs
evaluated. Additionally, yield in LT decreased linearly as UV-PFD increased from the lowest to
highest UV-PFD. The general trends of plant stress could be a result of the minimal UVC
exposure, and the long-term exposure starting during vegetative development. Therefore,
potential for UV radiation to enhance cannabinoid concentrations must still be validated before
UV can be used as a tool in cannabis production.

There were no LI treatment effects on inflorescence cannabinoid concentrations therefore

commercial cannabis growers can modify their LIs without altering the cannabinoid profile of
their product. The quality of the inflorescences improved with increasing LI (in addition to the
yield increases), including denser, larger apical inflorescences with increased concentrations of
aromatic terpenes. Contrarily, UV radiation reduced total terpene concentrations and the size and
proportion of apical inflorescences relative to other inflorescences on the plant in both cultivars.
Cannabis growth (i.e., height increases and increases in number of nodes) was generally
suppressed by UV radiation and increased by LI. The SLW (i.e., leaf thickness) increased
linearly both as LI and UV exposure level increased over the ranges evaluated. Leaves that
developed under UV radiation had abnormal morphology including leaf epinasty, upward curling
of leaflet margins, and asymmetry. The severity of these symptoms worsened as UV exposure
level increased.

On the physiological level, although plants exposed to high LIs showed some signs of light stress
in the upper canopy leaves (i.e., through linear decreases in Fv/Fm), the Asat and in situ NCER
82
increased with increasing canopy-level LI. The findings also demonstrated that leaf-level
photosynthetic responses to LI vary substantially with leaf age and light history. Therefore, leaf-
and plant-level photosynthetic responses cannot reliably predict cannabis yield responses to LI.
Increasing cannabis exposure to UV radiation decreased NCER and induced physiological (i.e.,
linear decrease in Fv/Fm over the range of UV PFDs evaluated) and visible (necrotic spots on
leaves) signs of stress. Exposure to UV radiation also accelerated the severity of symptoms
related to plant senescence (e.g., lower canopy leaf-drop and stigma browning). Cannabis plants
have a very capacity to utilize high photon densities (i.e., high LI) demonstrated through
increased photosynthesis and concomitant yield increases. On the contrary, highly energetic
photons (i.e., UV spectrum) trigger morphological and physiological stress in cannabis plants
through UVR8 specific and non-specific pathways (Tossi et al., 2019).

This study has provided the first reliable models for lighting strategies in the flowering stage for
indoor cannabis production. As evidenced through the research in this thesis, high LIs are
beneficial for increasing cannabis yields, but further evaluations are required to establish a
beneficial method of UV application. This research will assist growers in making informed
decisions about the optimum LI to use for their specific production systems. Ultimately, the
selection of the economic optimum canopy-level LI for a given commercial production system
depends on many interrelated factors, including the infrastructure limitations of a particular
cultivation operation. However, particularly since this is the forefront of cannabis horticulture
research, many research questions remain. Since plant yield responses to elevated CO2 can
mirror the responses to elevated LI, the combined effects of CO2 and LI should be investigated
on cannabis yield with an in-depth cost-benefit analysis of the optimum combination of these
two input parameters. The present study evaluated the effects of LI from red and blue LEDs.
Since it is well documented that different light spectra impact plant development differently,
further research is required to understand the impacts of LI of other spectra (e.g., full spectrum)
on cannabis morphology, physiology, and inflorescence yield and quality. Previous studies have
also shown that the PAR spectra influences plant responses to UV radiation, and that plants may
be less sensitive to UV radiation when paired with high PAR intensities. Further investigations

83
are required to understand the interactive effects of UV and PAR radiation (e.g., spectrum and
intensity) on cannabis responses to UV treatments.

Although the cannabis plants in the present study demonstrated stress responses to even the low
UV exposure levels, given that there are various possible UV treatments (e.g., spectrum,
intensity and temporal application), other UV application methods can be explored. Future
research could seek to achieve a method to promote eustress responses in cannabis secondary
metabolite concentration by using less energetic UV spectra than the present study. The plants in
the present study were exposed to UV radiation for 60 days, including during some of the
vegetative development, which could have been a sufficient amount of time to induce the
observed distress responses. Given that studies on other essential-oil producing species have seen
increases in secondary metabolite concentrations with shorter-term UV exposure, future research
could also investigate how UV affects cannabis plants with different lighting histories, and the
ideal development stage for UV exposure to achieve the desired effects in both yield and quality.
This study evaluated the effects of LI on a CBD-dominant cultivar, and the effects of UV
radiation on two balanced Δ9-THC to CBD cultivars with disparate morphology. Future research
on cannabis photobiology should also expand to evaluate multiple cultivars of all chemotypes.

84
 REFERENCES
Aarrouf, J., Urban, L. (2020). Flashes of UV-C light: An innovative method for stimulating plant
defences. PLoS One. 15:7. doi: 10.1371/journal.pone.0235918.

Ainsworth, E.A., Rogers, A. (2007). The response of photosynthesis and stomatal conductance to
rising [CO2]: mechanisms and environmental interactions. Plant Cell Environ. 30(3):258–
270. doi: 10.1111/j.1365-3040.2007.01641.x. PMID: 17263773.

Aizpurua-Olaizola, O., Soydaner, U., Öztürk, E., Schibano, D., Simsir, Y., Navarro, P.,
Etxebarria, N., Usobiaga, A. (2016). Evolution of the cannabinoid and terpene content
during the growth of Cannabis sativa plants from different chemotypes. J. Nat. Prod.
79:324–331. doi: 10.1021/acs.jnatprod.5b00949.

Aliferis, K.A., Bernard-Perron, D. (2020). Cannabinomics: Application of metabolomics in

cannabis (Cannabis sativa L.) research and development. Front. Plant Sci. 11:554.
doi: 10.3389/fpls.2020.00554.

Austin, C.N., Wilcox, W.F. (2012). Effects of sunlight exposure on grapevine powdery mildew
development. Phytopathology. 102:857–866. doi: 10.1094/PHYTO-07-11-0205.

Barcott, B., Whitney, B. (2019). Special report: Cannabis jobs count. Leafly.

Barnes, P.W., Flint, S.D., Caldwell, M.M. (1990). Morphological responses of crop and weed
species of different growth forms to ultraviolet-B radiation. Am. J. Bot. 77:1354–1360.
doi: 10.2307/2444596.

Barthélémy, D., Caraglio, Y. (2007). Plant architecture: A dynamic, multilevel and

comprehensive approach to plant form, structure and ontogeny. Ann. Bot. 99:375–407.
doi: 10.1093/aob/mcl260.

Backer, R., Schwinghamer, T., Rosenbaum, P., McCarty, V., Eichhorn Bilodeau, S., Lyu, D.,
Ahmed, M.B., Robinson, G., Lefsrud, M., Wilkins, O., Smith, D.L. (2019). Closing the
yield gap for cannabis: A meta-analysis of factors determining cannabis yield. Front.
Plant Sci. 10:495. doi: 10.3389/fpls.2019.00495.

Bauerle, W.L., McCullough, C., Iversen, M., Hazlett, M. (2020). Leaf age and position effects on
quantum yield and photosynthetic capacity in hemp crowns. Plants 9:271. doi:
10.3390/plants9020271.

Bazzaz, F.A., Dusek, D., Seigler, D.S., Haney, A.W. (1975). Photosynthesis and cannabinoid
content of temperate and tropical populations of Cannabis sativa. Biochem. Syst. Ecol.
3:15–18. doi: 10.1016/0305-1978(75)90036-8.

85
Beaman, A.R., Gladon, R.J., Schrader, J.A. (2009). Sweet basil requires an irradiance of 500
μmol·m-2·s-1 for greatest edible biomass production. HortScience 44:64–67. doi:
10.21273/HORTSCI.44.1.64.

Behn, H., Albert, A., Marx, F., Noga, G., Ulbrich, A. (2010). Ultraviolet-B and
photosynthetically active radiation interactively affect yield and pattern of monoterpenes
in leaves of peppermint (Mentha x piperita L.). J. Agric. Food Chem. 58:7361–7367. doi:
10.1021/jf9046072.

Behr, M., Legay, S., Žižková, E., Motyka, V., Dobrev, P.I., Hausman, J.F., Lutts, S., Guerriero,
G. (2016). Studying secondary growth and bast fiber development: The hemp hypocotyl
peeks behind the wall. Front. Plant Sci. 7:1733. doi: 10.3389/fpls.2016.01733.

Bernacchi, C.J., Calfapietra, C., Davey, P.A., Witting, V.E., Scarascia-Mugnozza, G.E., Raines,
C.A., Long, S.P. (2003). Blackwell Publishing Ltd. Photosynthesis and stomatal
conductance responses of poplars to free-air CO2 enrichment (PopFACE) during the first
growth cycle and immediately following coppice. New Phytol. 159: 609–621. doi:
10.1046/j.1469-8137.2003.00850.x

Bielczynski, L.W., Łaçki, M.K., Hoefnagels, I., Gambin, A., Croce, R. (2017). Leaf and plant
age affects photosynthetic performance and photoprotective capacity. Plant Physiol.
175:1634–1648. doi: 10.1104/pp.17.00904.

Björkman, O. (1981). "Responses to different quantum flux densities," in: Encyclopedia of Plant
Physiology: New Series: Vol. 12A, Physiological Plant Ecology 1, eds. L. Lange, P.S.
Nobel, C.B. Osmond, and H. Ziegler (Heidelberg, Berlin: Springer-Verlag), 57-107.

Booth, J.K., Bohlmann, J. (2019). Terpenes in Cannabis sativa – From plant genome to humans.
Plant Sci. 284:67–72. doi: 10.1016/j.plantsci.2019.03.022.

Bredmose, N. (1993). Effects of year-round supplementary lighting on shoot development,

flowering and quality of two glasshouse rose cultivars. Sci. Hortic. 54:69–85. doi:
10.1016/0304- 4238(93)90084-4.

Bredmose, N. (1994). Biological efficiency of supplementary lighting on cut roses the year
round. Sci. Hortic. 59:75–82. doi: 10.1016/0304-4238(94)90094-9.

Bugbee, B., Monje, O. (1992). The limits of crop productivity. Bioscience. 42:494–502. doi:
10.2307/1311879.

Caldwell, M.M. (1971). Solar UV irradiation and the growth and development of higher plants.
Photophysiology 6:131–177. doi: 10.1016/b978-0-12-282606-1.50010-6.

Caldwell, M.M., Ballaré, C.L., Bornman, J.F., Flint, S.D., Björn, L.O., Teramura, A.H.,

86
 Kulandaiveli, G., Tevini, M. (2003). Terrestrial ecosystems, increased solar ultraviolet
radiation and interactions with other climatic change factors. Photochem. Photobiol. Sci.
2:29–38. doi: 10.1039/b211159b.

Campbell, R.J., Marini, R.P., Birch, J.B. (1992). Canopy position affects light response curves
for gas exchange characteristics of apple spur leaves. J. Am. Soc. Hortic. Sci. 117:467–
472. doi: doi.org/10.21273/JASHS.117.3.467

Cannabis Business Times. (2021). 2020 State of the industry report. (2020) Valley View, OH:
GIE Media, Inc.

Cannigma. (2019). Cannabis regulation around the world. The Cannigma.

Caplan, D., Dixon, M., Zheng, Y. (2017). Optimal rate of organic fertilizer during the flowering
stage for cannabis grown in two coir-based substrates. HortScience 52:1796–1803. doi:
10.21273/HORTSCI12401-17.

Caplan, D., Dixon, M., Zheng, Y. (2019). Increasing inflorescence dry weight and cannabinoid
content in medical cannabis using controlled drought stress. HortScience 54:964–969.
doi: 10.21273/HORTSCI13510-18.

Carlini, E.A., Leite, J.R., Tannhauser, M., Berardi, A.C. (1973). Cannabidiol and Cannabis
sativa extract protect mice and rats against convulsive agents. J. Pharm. Pharmacol.
25:664–665. doi: 10.1111/j.2042-7158.1973.tb10660.x.

Carvalho, F.E.L., Ware, M.A., Ruban, A. V. (2015). Quantifying the dynamics of light tolerance
in Arabidopsis plants during ontogenesis. Plant Cell Environ. 38:2603–2617. doi:
10.1111/pce.12574.

Casal, J.J. (2013). Photoreceptor signaling networks in plant responses to shade. Annu. Rev.
Plant Biol. 64:403–427. doi: 10.1146/annurev-arplant-050312-120221.

Cen, Y., Bornman, J.F. (1993). The effect of exposure to enhanced UV‐B radiation on the
penetration of monochromatic and polychromatic UV‐B radiation in leaves of Brassica
napus. Physiol. Plant. 87:249–255. doi: 10.1111/j.1399-3054.1993.tb01727.x.

Chandra, S., Lata, H., Khan, I.A., Elsohly, M.A. (2008). Photosynthetic response of Cannabis
sativa L. to variations in photosynthetic photon flux densities, temperature and CO2
conditions. Physiol. Mol. Biol. Plants 14:299–306. doi: 10.1007/s12298-008-0027-x.

Chandra, S., Lata, H., Khan, I.A., ElSohly, M.A. (2011). Photosynthetic response of Cannabis
sativa L., an important medicinal plant, to elevated levels of CO2. Physiol. Mol. Biol.
Plants 17:291–295. doi: 10.1007/s12298-011-0066-6.

87
Chandra, S., Lata, H., Mehmedic, Z., Khan, I.A., ElSohly, M.A. (2015). Light dependence of
photosynthesis and water vapor exchange characteristics in different high Δ9-THC
yielding varieties of Cannabis sativa L. J. Appl. Res. Med. Aromat. Plants 2:39–47. doi:
10.1016/J.JARMAP.2015.03.002.

Chandra, S., Lata, H., ElSohly, M.A., Walker, L.A., Potter, D. (2017). Cannabis cultivation:
Methodological issues for obtaining medical-grade product. Epilepsy Behav. 70:302–
312. doi: 10.1016/j.yebeh.2016.11.029.

Chouvy, P.-A. (2015). The supply of hashish to Europe report prepared for the EMCDDA. Paris,
France.

Clarke, R.C., Merlin, M.D. (2016). Cannabis domestication, breeding history, present-day
genetic diversity, and future prospects. CRC. Crit. Rev. Plant Sci. 35:293–327. doi:
10.1080/07352689.2016.1267498.

Coco for Cannabis. (n.d.). How much light (ppf) do you need for indoor cannabis?. Coco
Cannabis. Retrieved from: https://www.cocoforcannabis.com/how-much-light-ppf-do-
you-need-for-indoor-cannabis/.

Cristiana Moliterni, V.M., Cattivelli, L., Ranalli, P., Mandolino, G. (2004). The sexual
differentiation of Cannabis sativa L.: A morphological and molecular study. Euphytica
140:95–106. doi: 10.1007/s10681-004-4758-7

Czégény, G., Mátai, A., Hideg, É. (2016). UV-B effects on leaves-Oxidative stress and
acclimation in controlled environments. Plant Sci. 248:57–63. doi:
10.1016/j.plantsci.2016.04.013.

Delgado, E., Parry, M.A.J., Lawlor, D.W., Keys, A.J., Medrano, H. (1993). Photosynthesis,
ribulose-1,5-bisphosphate carboxylase and leaf characteristics of Nicotiana tabacum L.
genotypes selected by survival at low CO2 concentrations. J. Exp. Bot. 44:1–7. doi:
10.1093/jxb/44.1.1.

Demkura, P. V., Ballaré, C.L. (2012). UVR8 mediates UV-B-induced Arabidopsis defense
responses against Botrytis cinerea by controlling sinapate accumulation. Mol. Plant.
5:642–652. doi: 10.1093/MP/SSS025.

Diggle, P.K. (1995). Architectural effects and the interpretation of patterns of fruit and seed
development. Annu. Rev. Ecol. Syst. 26:531–552. doi:
10.1146/annurev.es.26.110195.002531.

Dou, H., Niu, G., Gu, M., Joseph G., M. (2018). Responses of sweet basil to different daily light
integrals in photosynthesis, morphology, yield, and nutritional quality. HortScience.
53:496–503.

88
Downer, S. (2018). The ultimate lighting guide for cannabis cultivation. Medium. Retrieved
from: https://medium.com/@sabinedowner/the-ultimate-lighting-guide-for-cannabis-
cultivation-cc60b22df835.

Dujourdy, L., Besacier, F. (2017). A study of cannabis potency in France over a 25 years period
(1992–2016). Forensic Sci. Int. 272:72–80. doi: 10.1016/j.forsciint.2017.01.007.

Eaves, J., Eaves, S., Morphy, C., Murray, C. (2020). The relationship between light intensity,
cannabis yields, and profitability. Agron. J. 112:1466–1470. doi: 10.2139/ssrn.3310456.

Eichhorn Bilodeau, S., Wu, B.-S., Rufyikiri, A.-S., MacPherson, S., Lefsrud, M. (2019). An
update on plant photobiology and implications for cannabis production. Front. Plant Sci.
doi: 10.3389/fpls.2019.00296.

Eichler, M., Spinedi, L., Unfer-Grauwiler, S., Bodmer, M., Surber, C., Luedi, M., Drewe, J.
(2012). Heat exposure of cannabis sativa extracts affects the pharmacokinetic and
metabolic profile in healthy male subjects. Planta Med. 78:686–691. doi: 10.1055/s-
0031-1298334.

ElSohly, M.A., Mehmedic, Z., Foster, S., Gon, C., Chandra, S., Church, J.C. (2016). Changes in
cannabis potency over the last 2 decades (1995-2014): Analysis of current data in the
United States. Biol. Psychiatry 79:613–619. doi: 10.1016/j.biopsych.2016.01.004.

ElSohly, M.A., Radwan, M.M., Gul, W., Chandra, S., Galal, A. (2017). Phytochemistry of
Cannabis sativa L. Phytocannabinoids. 103:1-36. doi: 10.1007/978-3-319-45541-9_1.
Springer, Cham.

European Monitoring Centre for Drugs and Drug Addiction. (2013). Cannabis Production and
Markets In Europe. Lisbon, Portugal: EMCDDA Insights, No. 12.

Evergreen Economics. (2016). Cannabis Agriculture Energy Demand Study Final Report. San
Diego, CA: San Diego Gas & Electric Company.

Fairbairn, J.W., Liebmann, J.A. (1974). The cannabinoid content of Cannabis sativa L. grown in
England. J. Pharm. Pharmacol. 26:413–419. doi: 10.1111/j.2042-7158.1974.tb09306.x.

Faust, J.E. (2003). "Light," in Ball Redbook, ed. D. Hamrick (Batabia, IL: Ball Publishing), 71–
84.

Farag, S., Kayser, O. (2015). Cannabinoids production by hairy root cultures of Cannabis sativa
L. SciRes. Am. J. Plant Sci. 6:1874–1884. doi: 10.4236/ajps.2015.611188.

Fernandes, V.F., de Almeida, L.B., da Feijó, E.V.R.S., da Silva, D.C., de Oliveira, R.A., Mielke,
M.S., do Costa, L.C.B. (2013). Light intensity on growth, leaf micromorphology and

89
 essential oil production of Ocimum gratissimum. Brazilian J. Pharmacogn. 23:419–424.
doi: 10.1590/S0102-695X2013005000041.

Fierro, A.C., Leroux, O., De Coninck, B., Cammue, B.P.A., Marchal, K., Prinsen, E., Van Der
Straeten, D., Vandenbussche, F. (2015). Ultraviolet-B radiation stimulates downward leaf
curling in Arabidopsis thaliana. Plant Physiol. Biochem. 93:9–17. doi:
10.1016/j.plaphy.2014.12.012.

Fiscus, E.L., Booker, F.L. (1995). Is increased UV-B a threat to crop photosynthesis and
productivity?. Photosynth. Res. 43:81–92. doi: 10.1007/BF00042965.

Flint, S.D., Caldwell, M.M. (2003). A biological spectral weighting function for ozone depletion
research with higher plants. Physiol. Plant. 117:137–144. doi: 10.1034/j.1399-
3054.2003.1170117.x.

Flores-Sanchez, I.J., Verpoorte, R. (2008). PKS activities and biosynthesis of cannabinoids and
flavonoids in Cannabis sativa L. plants. Plant Cell Physiol. 49:1767–1782. doi:
10.1093/pcp/pcn150.

Friedberg, E.C. (2002). The intersection between the birth of molecular biology and the
discovery of DNA repair. DNA Repair. 1:855–867. doi: 10.1016/S1568-7864(02)00112-
X.

Fukuda, S., Satoh, A., Kasahara, H., Matsuyama, H., Takeuchi, Y. (2008). Effects of ultraviolet-
B irradiation on the cuticular wax of cucumber (Cucumis sativus) cotyledons. J. Plant
Res. 121:179–189. doi: 10.1007/s10265-007-0143-7.

Gara, T., Darvishzadeh, R., Skidmore, A., Wang, T. (2018). Impact of vertical canopy position
on leaf spectral properties and traits across multiple species. Remote Sens. 10:346. doi:
10.3390/rs10020346.

Giupponi, L., Leoni, V., Pavlovic, R., Giorgi, A. (2020). Influence of altitude on phytochemical
composition of hemp inflorescence: A metabolomic approach. Molecules 25:1381. doi:
10.3390/molecules25061381.

Gonçalves, J., Rosado, T., Soares, S., Simão, A., Caramelo, D., Luís, Â., Fernández, N., Barroso,
M., Gallardo, E., Duarte, A. (2019). Cannabis and its secondary metabolites: Their use as
therapeutic drugs, toxicological aspects, and analytical determination. Medicines 6:31.
doi: 10.3390/medicines6010031.

Goodman, R.C., Apostol, K.G., Jacobs, D.F., Wilson, B.C. (2007). Comparing cold-stored and
freshly lifted water oak (Quercus nigra) seedlings based on physiological parameters.
National Proceedings, Forest and Conservation Nursery Associations, 2006. Fort Collins,
CO.

90
Government of Canada. (2015). Canadian tobacco alcohol and drugs (CTADS): 2015 summary.
Retrieved from: https://www.canada.ca/en/health-canada/services/canadian-tobacco-
alcohol-drugs-survey/2015-summary.html.

Government of Canada. (2016). Understanding the new Access to cannabis for medical purposes
regulations. Retrieved from: https://www.canada.ca/en/health-
canada/services/publications/drugs-health-products/understanding-new-access-to-
cannabis-for-medical-purposes-regulations.html.

Government of Canada. (2019a). Cannabis legalization and regulation. Department of Justice.

Retrieved from: https://www.justice.gc.ca/eng/cj-jp/cannabis/.

Government of Canada. (2019b). What you need to know about cannabis. Health Canada.
Retrieved from:
https://www.canada.ca/en/services/health/campaigns/cannabis/canadians.html.

Graham, S.D. (2004). Medical marijuana: Canada’s regulations, pharmacology, and social
policy. Can. Pharm. J. 137:23–27. doi: 10.1177/171516350413700104

Gratani, L. (2014). Plant phenotypic plasticity in response to environmental factors. Adv. Bot.
2014:1–17. doi: 10.1155/2014/208747.

De Gruijl, F.R., Van der Leun, J.C. (2000). Environment and health: 3. Ozone depletion and
ultraviolet radiation. CMAJ. 163:851–855.

Guindon, J., Hohmann, A.G. (2009). The endocannabinoid system and pain. CNS Neurol.
Disord. Drug Targets 8:403–21. doi: 10.2174/187152709789824660.

Haney, A., Kutscheid, B.B. (1973). Quantitative variation in the chemical constituents of
marihuana from stands of naturalized Cannabis sativa L. in East-Central Illinois. Econ.
Bot. 27:193–203. doi: 10.1007/BF02872989.

Havé, M., Marmagne, A., Chardon, F., Masclaux-Daubresse, C. (2017). Nitrogen remobilization
during leaf senescence: Lessons from Arabidopsis to crops. J. Exp. Bot. 68:2513–2529.
doi: 10.1093/jxb/erw365.

Hawley, D., Graham, T., Stasiak, M., Dixon, M. (2018). Improving cannabis bud quality and
yield with subcanopy lighting. HortScience 53:1593–1599. doi: 10.21273/hortsci13173-
18.

Hazekamp, A., Peltenburg, A., Verpoorte, R., Giroud, C. (2005). Chromatographic and
spectroscopic data of cannabinoids from Cannabis sativa L. J. Liq. Chromatogr. Relat.
Technol. 28:2361–2382. doi: 10.1080/10826070500187558.

91
Hectors, K., Jacques, E., Prinsen, E., Guisez, Y., Verbelen, J.P., Jansen, M.A.K., Vissenberg, K.
(2010). UV radiation reduces epidermal cell expansion in leaves of Arabidopsis thaliana.
J. Exp. Bot. 61:4339–4349. doi: 10.1093/jxb/erq235.

Hemphill, J.K., Turner, J.C., Mahlberg, P.G. (1980). Cannabinoid content of individual plant
organs from different geographical strains of Cannabis sativa L. J. Nat. Prod. 43:112–
122. doi: 10.1021/np50007a009.

Hideg, É., Jansen, M.A.K., Strid, Å. (2013). UV-B exposure, ROS, and stress: Inseparable
companions or loosely linked associates?. Trends Plant Sci. 18:107–115. doi:
10.1016/j.tplants.2012.09.003.

Hikosara, S., Ito, K., Goto, E. (2010). Effects of ultraviolet light on growth, essential oil
concentration, and total antioxidant capacity of Japanese mint. Environ. Control Biol.
48:185–190. doi: 10.2525/ecb.48.185.

Hoffmann, A.M., Noga, G., Hunsche, M. (2015). High blue light improves acclimation and
photosynthetic recovery of pepper plants exposed to UV stress. Environ. Exp. Bot.
109:254–263. doi: 10.1016/j.envexpbot.2014.06.017.

Höll, J., Lindner, S., Walter, H., Joshi, D., Poschet, G., Pfleger, S., Ziegler, T., Hell, R., Bogs, J.,
Rausch, T. (2019). Impact of pulsed UV-B stress exposure on plant performance: How
recovery periods stimulate secondary metabolism while reducing adaptive growth
attenuation. Plant Cell Environ. 42:801–814. doi: 10.1111/pce.13409.

Huarancca Reyes, T., Scartazza, A., Castagna, A., Cosio, E.G., Ranieri, A., Guglielminetti, L.
(2018). Physiological effects of short acute UVB treatments in Chenopodium quinoa
Willd. Sci. Rep. 8:371. doi: 10.1038/s41598-017-18710-2.

Huché-Thélier, L., Crespel, L., Gourrierec, J. Le, Morel, P., Sakr, S., Leduc, N. (2016). Light
signaling and plant responses to blue and UV radiations: Perspectives for applications in
horticulture. Environ. Exp. Bot. 121:22–38. doi: 10.1016/j.envexpbot.2015.06.009.

Jaeger, K. (2019). Study reveals little-known trick to make marijuana plants grow more bud for
less. Marijuana Moment. Retrieved from: https://www.marijuanamoment.net/study-
reveals-little-known-trick-to-make-marijuana-plants-grow-more-bud-for-less/.

Jansen, M.A.K. (2002). Ultraviolet-B radiation effects on plants: Induction of morphogenic

responses. Physiol. Plant. 116:423–429. doi: 10.1034/j.1399-3054.2002.1160319.x.

Jansen, M.A.K., Klem, K., Robson, T.M., Urban, O. (2017). "UV-B induced morphological
changes - an enigma," in UV-B radiation and plant life: molecular biology to ecology, ed.
B. Jordan. doi: 10.1079/9781780648590.0000.

92
Jenkins, G.I. (2017). Photomorphogenic responses to ultraviolet-B light. Plant Cell Environ.
40:2544–2557. doi: 10.1111/pce.12934.

Johnson, C.B., Kirby, J., Naxakis, G., Pearson, S. (1999). Substantial UV-B-mediated induction
of essential oils in sweet basil (Ocimum basilicum L.). Phytochemistry 51:507–510. doi:
10.1016/S0031-9422(98)00767-5.

Jones-Baumgardt, C., Llewellyn, D., Ying, Q., Zheng, Y. (2019). Intensity of sole-source light-
emitting diodes affects growth, yield, and quality of Brassicaceae microgreens.
HortScience. 54:1168–1174. doi: 10.21273/HORTSCI13788-18.

Kakani, V.G., Reddy, K.R., Zhao, D., Sailaja, K. (2003). Field crop responses to ultraviolet-B
radiation: A review. Agric. For. Meteorol. 120:191–218. doi:
10.1016/j.agrformet.2003.08.015.

Kebbi, Y., Muhammad, A.I., Sant’Ana, A.S., do Prado-Silva, L., Liu, D., Ding, T. (2020).
Recent advances on the application of UV-LED technology for microbial inactivation:
Progress and mechanism. Compr. Rev. Food Sci. Food Saf. 19:3501–3527. doi:
10.1111/1541- 4337.12645.

Kenny, C., Nolin, P.C. (2003). Cannabis: Report of the senate special committee on illegal
drugs. Toronto, ON: University of Toronto Press.

Kirschbaum, M.U.F. (2011). Does enhanced photosynthesis enhance growth? Lessons learned
from CO2 enrichment studies. Plant Physiol. 155:117–124. doi: 10.1104/pp.110.166819.

Klem, K., Ač, A., Holub, P., Kováč, D., Špunda, V., Robson, T.M., Urban, O. (2012). Interactive
effects of PAR and UV radiation on the physiology, morphology and leaf optical
properties of two barley varieties. Environ. Exp. Bot. 75:52–64. doi:
10.1016/j.envexpbot.2011.08.008.

Kreyling, J., Schweiger, A.H., Bahn, M., Ineson, P., Migliavacca, M., Morel-Journel, T.,
Christiansen, J.R., Schtickzelle, N., Larsen, K.S. (2018). To replicate, or not to replicate -
that is the question: How to tackle nonlinear responses in ecological experiments. Ecol.
Lett. 21:1629–1638. doi: 10.1111/ele.13134.

Krizek, D.T., Mirecki, R.M., Britz, S.J. (1997). Inhibitory effects of ambient levels of solar UV-
A and UV-B radiation on growth of cucumber. Physiol. Plant. 100:886–893. doi:
10.1111/j.1399-3054.1997.tb00014.x.

Krizek, D.T. (2004). Influence of PAR and UV-A in determining plant sensitivity and
photomorphogenic responses to UV-B radiation. Photochem. Photobiol. 79:315. doi:
10.1562/2004-01-27-ir.1.

93
Kusuma, P., Pattison, P.M., Bugbee, B. (2020). From physics to fixtures to food: Current and
potential LED efficacy. Hortic. Res. 7:56. doi: 10.1038/s41438-020-0283-7.

Larsson, M., Lagerås, P. (2015). New evidence on the introduction, cultivation and processing of
hemp (Cannabis sativa L.) in southern Sweden. Environ. Archaeol. 20:111–119. doi:
10.1179/1749631414Y.0000000029.

Latta, R.P., Eaton, B.J. (1975). Seasonal fluctuations in cannabinoid content of Kansas
marijuana. Econ. Bot. 29:153–163. doi: 10.1007/BF02863315.

Leggett, T. (2006). A review of the world cannabis situation. Bull. Narc. 58:1–155.

Lewis, M.A., Russo, E.B., Smith, K.M. (2018). Pharmacological foundations of cannabis
chemovars. Planta Med. 84:225–233. doi: 10.1055/s-0043-122240.

LI-COR Biosciences. (2012). Using the LI-6400 / LI-6400XT Portable Photosynthesis System
Version 6. Lincoln, NB: LI-COR Inc

Liu, L.X., Xu, S.M., Woo, K.C. (2005). Solar UV-B radiation on growth, photosynthesis and the
xanthophyll cycle in tropical acacias and eucalyptus. Environ. Exp. Bot. 54:121–130. doi:
10.1016/j.envexpbot.2004.06.006.

Liu, B., Liu, X. bing, Li, Y.S., Herbert, S.J. (2013). Effects of enhanced UV-B radiation on seed
growth characteristics and yield components in soybean. F. Crop. Res. 154:158–163. doi:
10.1016/j.fcr.2013.08.006.

Llewellyn, D., Golem, S., Foley, E., Dinka, S., Maxwell, A., Jones, P., Zheng, Y. (2021).
Cannabis yield increased proportionally with light intensity, but additional ultraviolet
radiation did not affect yield or cannabinoid content. Preprints. doi:
10.20944/preprints202103.0327.v1.

Lobo, F. A., de Barros, M.P., Dalmagro, H.J., Dalmolin, Â.C., Pereira, W.E., de Souza, É.C.,
Vourlitis, G.L., Rodríguez Ortíz, C.E. (2013). Fitting net photosynthetic light-response
curves with Microsoft Excel - a critical look at the models. Photosynthetica. 51:445–456.
doi: 10.1007/s11099-013-0045-y.

Lydon, J., Teramura, A.H., Coffman, C.B. (1987). UV-B radiation effects on photosynthesis,
growth and cannabinoid production of two Cannabis sativa chemotypes. Photochem.
Photobiol. 46:201–206. doi: 10.1111/j.1751-1097.1987.tb04757.x.

Maffei, M., Scannerini, S. (2000). UV-B effect on photomorphogenesis and essential oil
composition in peppermint (Mentha piwperita L.). J. Essent. Oil Res. 12:523–529. doi:
10.1080/10412905.2000.9712150.

94
Magagnini, G., Grassi, G., Kotiranta, S. (2018). The effect of light spectrum on the morphology
and cannabinoid content of Cannabis sativa L. Med. Cannabis Cannabinoids 1:19–27.
doi: 10.1159/000489030.

Mah, J.J., Llewellyn, D., Zheng, Y. (2019). Protocol for converting spectrometer radiometric
data to photon flux units [Microsoft Excel Spreadsheet]. Guelph, University of Guelph.
Available from: http://www.ces.uoguelph.ca/TechNotes.shtml.

Mammoth Lighting. (n.d.). Optimal light intensity. Mammoth Light. Retrieved from:
https://mammothlighting.com/pages/photosynthetic-response-of-cannabis-sativa-l.

Manova, V., Gruszka, D. (2015). DNA damage and repair in plants: From models to crops.
Front. Plant Sci. 6:885. doi: 10.3389/fpls.2015.00885.

Marcelis, L.F.M., Broekhuijsen, A.G.M., Meinen, E., Nijs, E.M.F.M., Raaphorst, M.G.M.
(2006). Quantification of the growth response to light quantity of greenhouse grown
crops. Acta Hortic. 711:97–103. doi: 10.17660/actahortic.2006.711.9.

Martínez-Lüscher, J., Morales, F., Delrot, S., Sánchez-Díaz, M., Gomés, E., Aguirreolea, J.,
Pascual, I. (2013). Short- and long-term physiological responses of grapevine leaves to
UV-B radiation. Plant Sci. 213:114–122. doi: 10.1016/j.plantsci.2013.08.010.

Maroon, J., Bost, J. (2018). Review of the neurological benefits of phytocannabinoids. Surg.
Neurol. Int. 9:91. doi: 10.4103/sni.sni_45_18.

Marti, G., Schnee, S., Andrey, Y., Simoes-Pires, C., Carrupt, P.A., Wolfender, J.L., Gindro, K.
(2014). Study of leaf metabolome modifications induced by UV-C radiations in
representative vitis, cissus and cannabis species by LC-MS based metabolomics and
antioxidant assays. Molecules 19:14004–14021. doi: 10.3390/molecules190914004.

McElroy, C.T., Fogal, P.F. (2008). Ozone: From discovery to protection. Atmos. Ocean. 46:1–
13. doi: 10.3137/ao.460101.

McPartland, J.M., Russo, E.B. (2001). Cannabis and Cannabis extracts: Greater than the sum of
their parts?. Cannabis Ther. 1:103–132. doi: 10.1300/J175v01n03_08.

McPartland, J.M. (2018). Cannabis systematics at the levels of family, genus, and species.
Cannabis Cannabinoid Res. 3:203–212. doi: 10.1089/can.2018.0039.

De Meijer, E.P.M., Bagatta, M., Carboni, A., Crucitti, P., Moliterni, V.M.C., Ranalli, P.,
Mandolino, G. (2003). The inheritance of chemical phenotype in Cannabis sativa L.
Genetics. 163:335–346. doi: 10.1093/genetics/163.1.335.

Middleton, E.M., Teramura, A.H. (1993). Potential Errors in the use of cellulose acetate and

95
 Mylar filters in UV‐B radiation studies. Photochem. Photobiol. 57:744–751. doi:
10.1111/j.1751-1097.1993.tb02948.x.

Mills, E. (2012). The carbon footprint of indoor Cannabis production. Energy Policy 46:58–67.
doi: 10.1016/J.ENPOL.2012.03.023.

Murchie, E., Hubbart, S., Chen, Y., Peng, S., Horton, P. (2002). Acclimation of rice
photosynthesis to irradiance under field conditions. Plant Physiol. 130:1999–2010. doi:
10.1104/pp.011098.

Murchie, E., Lawson, T. (2013). Chlorophyll fluorescence analysis: A guide to good practice and
understanding some new applications. J. Exp. Bot. 64:3983–3998. doi:
10.1093/jxb/ert208.

Namdar, D., Mazuz, M., Ion, A., Koltai, H. (2018). Variation in the compositions of cannabinoid
and terpenoids in Cannabis sativa derived from inflorescence position along the stem and
extraction methods. Ind. Crops Prod. 113:376–382. doi: 10.1016/j.indcrop.2018.01.060.

Nelson, J.A., Bugbee, B. (2014). Economic analysis of greenhouse lighting: Light emitting
diodes vs. high intensity discharge fixtures. PLoS One. 9. doi:
10.1371/journal.pone.0099010.

Neugart, S., Schreiner, M. (2018). UVB and UVA as eustressors in horticultural and agricultural
crops. Sci. Hortic. 234:370–381. doi: 10.1016/j.scienta.2018.02.021.

Niinemets, Ü., Keenan, T.F. (2012). Measures of light in studies on light-driven plant plasticity
in artificial environments. Front. Plant Sci. 3:156. doi: 10.3389/fpls.2012.00156.

Nuutinen, T. (2018). Medicinal properties of terpenes found in Cannabis sativa and Humulus
lupulus. Eur. J. Med. Chem. 157:198–228. doi: 10.1016/j.ejmech.2018.07.076.

Oh, W., Cheon, H., Kim, K.S., Runkle, E.S. (2009). Photosynthetic daily light integral influences
flowering time and crop characteristics of Cyclamen persicum. HortScience. 44:341–344.
doi: 10.21273/HORTSCI.44.2.341.

Ohyama, K., Manabe, K., Omura, Y., Kozai, T., Kubota, C. (2005). Potential use of a 24-hour
photoperiod (continuous light) with alternating air temperature for production of tomato
plug transplants in a closed system. HortScience. 40:374–377. doi:
10.21273/HORTSCI.40.2.374.

Pacher, P., Bátkai, S., Kunos, G. (2006). The endocannabinoid system as an emerging target of
pharmacotherapy. Pharmacol. Rev. 58:389–462. doi: 10.1124/pr.58.3.2.

Palma, C.F.F., Castro-Alves, V., Morales, L.O., Rosenqvist, E., Ottosen, C.-O., Strid, Å. (2021).

96
 Spectral composition of light affects sensitivity to UV-B and photoinhibition in
cucumber. Front. Plant Sci. doi: 10.3389/fpls.2020.610011.

Parry, C., Blonquist, J.M., Bugbee, B. (2014). In situ measurement of leaf chlorophyll
concentration: Analysis of the optical/absolute relationship. Plant, Cell Environ.
37:2508–2520. doi: 10.1111/pce.12324.

Pate, D.W. (1983). Possible Role of Ultraviolet Radiation in Evolution of Cannabis Chemotypes.
Econ. Bot. 37:396–405.

Peng, S. (2000). Single-leaf and canopy photosynthesis of rice. Studies in Plant Science. 7:213–
228. doi.org/10.1016/S0928-3420(00)80017-8

Pertwee, R.G. (1997). Pharmacology of cannabinoid CB1 and CB2 receptors. Pharmacol. Ther.
74:129–180. doi: 10.1016/S0163-7258(97)82001-3.

Pertwee, R.G. (2006). Cannabinoid pharmacology: The first 66 years. Br. J. Pharmacol.
147:S163. doi: 10.1038/sj.bjp.0706406.

Pettersen, R., Torre, S., Gislerød, H. (2010). Effects of leaf aging and light duration on
photosynthetic characteristics in a cucumber canopy. Sci. Hortic. 125:82–87. doi:
10.1016/J.SCIENTA.2010.02.016.

Poorter, H., Niinemets, Ü., Ntagkas, N., Siebenkäs, A., Mäenpää, M., Matsubara, S., Pons, T.
(2019). A meta‐analysis of plant responses to light intensity for 70 traits ranging from
molecules to whole plant performance. New Phytol. 223:1073–1105. doi:
10.1111/nph.15754.

Posada, J.M., Sievänen, R., Messier, C., Perttunen, J., Nikinmaa, E., Lechowicz, M.J. (2012).
Contributions of leaf photosynthetic capacity, leaf angle and self-shading to the
maximization of net photosynthesis in Acer saccharum: a modelling assessment. Ann.
Bot. 110:731–741. doi: 10.1093/aob/mcs106.

Potter, D.J. (2014). A review of the cultivation and processing of cannabis (Cannabis sativa L.)
for production of prescription medicines in the UK. Drug Test. Anal. 6:31–38. doi:
10.1002/dta.1531.

Potter, D.J., Duncombe, P. (2012). The effect of electrical lighting power and irradiance on
indoor-grown cannabis potency and yield. J. Forensic Sci. 57:618–622. doi:
10.1111/j.1556-4029.2011.02024.x.
Potter, D.J., Hammond, K., Tuffnell, S., Walker, C., Di Forti, M. (2018). Potency of Δ 9-
tetrahydrocannabinol and other cannabinoids in cannabis in England in 2016:
Implications for public health and pharmacology. Drug Test. Anal. 10:628–635. doi:
10.1002/dta.2368.

97
Punja Z. K., Holmes, J. E. (2020). Hermaphrodism in marijuana (Cannabis sativa L.)
inflorescences – Impact on floral morphology, seed formation, progeny sex ratios, and
genetic variation. Front. Plant Sci. 11:718. doi: 10.3389/fpls.2020.00718

Radetsky, L.C. (2018). LED and HID Horticultural Luminaire Testing Report Prepared For
Lighting Energy Alliance Members and Natural Resources Canada. Troy, NY:
Rensselaer Polytechnic Institute.

Robson, T.M., Aphalo, P.J. (2012). Species-specific effect of UV-B radiation on the temporal
pattern of leaf growth. Physiol. Plant. 144:146–160. doi: 10.1111/j.1399-
3054.2011.01546.x.

Robson, T.M., Aphalo, P.J., Banaś, A.K., Barnes, P.W., Brelsford, C.C., Jenkins, G.I.,
Kotilainen, T.K., Łabuz, J., Martínez-Abaigar, J., Morales, L.O., Neugart, S., Pieristè,
M., Rai, N., Vandenbussche, F., Jansen, M.A.K. (2019). A perspective on ecologically
relevant plant-UV research and its practical application. Photochem. Photobiol. Sci.
18:970–988. doi: 10.1039/c8pp00526e.

Rodriguez-Morrison, V., Llewellyn, D., Zheng, Y. (2021). Cannabis yield, potency, and leaf
photosynthesis respond differently to increasing light levels in an indoor environment 2.
Preprints. doi: 10.20944/preprints202101.0163.v1.

Rotermann, M. (2020). What has changed since cannabis was legalized?. Stat. Canada 31:11–20.
doi: 10.25318/82-003-x202000200002-eng.

Royal Queen Seeds. (2019). The indoor marijuana grower’s guide to artificial lights. Royal
Queen Seeds. Retrieved from: https://www.royalqueenseeds.com/blog-cannabis-
cultivation-tips-how-to-set-up-indoor-grow-lights-n670.

Russo, E.B. (2007). History of cannabis and its preparations in saga, science, and sobriquet.
Chem. Biodivers. 4:1614–1648. doi: 10.1002/cbdv.200790144.

Russo, E.B. (2019). The case for the entourage effect and conventional breeding of clinical
cannabis: No “strain,” no gain. Front. Plant Sci. 9:1969. doi: 10.3389/fpls.2018.01969.

Ruter, J.M. (1992). Influence of source, rate, and method of applicating controlled release
fertilizer on nutrient release and growth of “Savannah” holly. Fertil. Res. 32:101–106.
doi: 10.1007/BF01054399.

Sadras, V.O., Richards, R.A. (2014). Improvement of crop yield in dry environments:
Benchmarks, levels of organisation and the role of nitrogen. J. Exp. Bot. 65:1981–1995.
doi: 10.1093/jxb/eru061.

98
Schreiner, M., Mewis, I., Huyskens-Keil, S., Jansen, M.A.K., Zrenner, R., Winkler, J.B.,
O’Brien, N., Krumbein, A. (2012). UV-B-induced secondary plant metabolites - Potential
benefits for plant and human health. CRC. Crit. Rev. Plant Sci. 31:229–240. doi:
10.1080/07352689.2012.664979.

Schulze, R., and Grafe K. (1969). "Consideration of sky ultraviolet in the measurement of solar
ultraviolet radiation," in The Biological Effects of Ultraviolet Radiation, ed. Urbach, F.,
Pergamon Press, New York.

Searles, P.S., Flint, S.D., Caldwell, M.M. (2001). A meta-analysis of plant field studies
simulating stratospheric ozone depletion. Oecologia. 127:1–10. doi:
10.1007/s004420000592.

Sen, A., Wyonch, R. (2018). Cannabis Countdown: Estimating the Size Of Illegal Markets and
Lost Tax Revenue Post-Legalization. Toronto, ON: C.D. Howe Institute.

Sims, D.A., Pearcy, R.W. (1992). Response of leaf anatomy and photosynthetic capacity in
Alocasia macrorrhiza (Araceae) to a transfer from low to high light. Am. J. Bot. 79:449-
455. doi: 10.2307/2445158.

Singsaas, E.L., Ort, D.R., DeLucia, E.H. (2001). Variation in measured values of photosynthetic
quantum yield in ecophysiological studies. Oecologia 128:15–23. doi:
10.1007/s004420000624.

Singsaas, E.L., Ort, D.R., Delucia, E.H. (2004). Elevated CO2 effects on mesophyll conductance
and its consequences for interpreting photosynthetic physiology. Plant, Cell Environ.
27:41–50. doi: 10.1046/j.0016-8025.2003.01123.x.

Slatkin, N.E. (2007). Cannabinoids in the treatment of chemotherapy-induced nausea and

vomiting: beyond prevention of acute emesis. J. Support. Oncol. 5:1–9.

Small, E. (2017). Cannabis: A complete guide. CRC Press, Boca Raton, FL.

Small, E., Beckstead, H.D. (1973). Cannabinoid phenotypes in Cannabis sativa. Nature.
245:147–148. doi: 10.1038/245147a0.

Small, E., Cronquist, A. (1976). A Practical and Natural Taxonomy for Cannabis. Taxon.
25:405–435.

Stapleton, A.E. (1992). Ultraviolet radiation and plants: Burning questions. Plant Cell. 4:1353–
1358. doi: 10.1105/tpc.4.11.1353

Steinmüller, D., Tevini, M. (1985). Action of ultraviolet radiation (UV-B) upon cuticular waxes
in some crop plants. Planta. 164:557–564. doi: 10.1007/BF00395975.

99
Suthaparan, A., Solhaug, K.A., Bjugstad, N., Gislerød, H.R., Gadoury, D.M., Stensvand, A.
(2016). Suppression of powdery mildews by UV-B: Application frequency and timing,
dose, reflectance, and automation. Plant Dis. 100:1643–1650. doi: 10.1094/PDIS-12-15-
1440-RE.

Teramura, A.H., Sullivan, J.H., Lydon, J. (1990). Effects of UV‐B radiation on soybean yield
and seed quality: a 6‐year field study. Physiol. Plant. 80:5–11. doi: 10.1111/j.1399-
3054.1990.tb04367.x.

Terashima, I., Hikosaka, K. (1995). Comparative ecophysiology of leaf and canopy

photosynthesis. Plant. Cell Environ. 18:1111–1128. doi: 10.1111/j.1365-
3040.1995.tb00623.x.

Terfa, M.T., Roro, A.G., Olsen, J.E., Torre, S. (2014). Effects of UV radiation on growth and
postharvest characteristics of three pot rose cultivars grown at different altitudes. Sci.
Hortic. 178:184–191. doi: 10.1016/j.scienta.2014.08.021.

Thoma, F., Somborn-Schulz, A., Schlehuber, D., Keuter, V., Deerberg, G. (2020). Effects of
light on secondary metabolites in selected leafy greens: A review. Front. Plant Sci.
11:497. doi: 10.3389/fpls.2020.00497.

Toonen, M., Ribot, S., Thissen, J. (2006). Yield of illicit indoor cannabis cultivation in the
Netherlands. J. Forensic Sci. 51:1050–1054. doi: 10.1111/j.1556-4029.2006.00228.x.

Torre, S., Roro, A.G., Bengtsson, S., Mortensen, L.M., Solhaug, K.A., Gislerød, H.R., Olsen,
J.E. (2012). Control of plant morphology by UV-B and UV-B-temperature interactions.
Acta Hortic. 956:207–214. doi: 10.17660/ActaHortic.2012.956.22.

Tossi, V.E., Regalado, J.J., Iannicelli, J., Laino, L.E., Burrieza, H.P., Escandón, A.S., Pitta-
Álvarez, S.I. (2019). Beyond arabidopsis: Differential UV-B response mediated by
UVR8 in diverse species. Front. Plant Sci. 10:780. doi: 10.3389/fpls.2019.00780.

United Nations Office on Drugs and Crime (UNODC). (2009). World Drug Report 2009. New
York, NY: United Nations.

United Nations Office on Drugs and Crime (UNODC). (2019). World Drug Report 2019. New
York, NY: United Nations.

Valdez, A., Kaplan, C.D. (2019). Deconstructing US marijuana prohibition policies in the early
twentieth century. Aztlan A J. Chicano Stud. 44:111–140.

Valenta, K., Dimac-Stohl, K., Baines, F., Smith, T., Piotrowski, G., Hill, N., Kuppler, J., Nevo,
O. (2020). Ultraviolet radiation changes plant color. BMC Plant Biol. 20:253. doi:

100
 10.1186/s12870-020-02471-8.

Vanhove, W., Van Damme, P., Meert, N. (2011). Factors determining yield and quality of illicit
indoor cannabis (Cannabis spp.) production. Forensic Sci. Int. 212:158–163. doi:
10.1016/J.FORSCIINT.2011.06.006.

Vanhove, W., Surmont, T., Van Damme, P., De Ruyver, B. (2014). Filling in the blanks. An
estimation of illicit cannabis growers’ profits in Belgium. Int. J. Drug Policy 25:436–443.
doi: 10.1016/j.drugpo.2014.01.020.

Viveros, M., Marco, E., File, S. (2005). Endocannabinoid system and stress and anxiety
responses. Pharmacol. Biochem. Behav. 81:331–342. doi: 10.1016/j.pbb.2005.01.029.

Vlahos, J.C., Heuvelink, E., Martakis, G.F.P. (1991). A growth analysis study of three
Achimenes cultivars grown under three light regimes. Sci. Hortic. 46:275–282. doi:
10.1016/0304- 4238(91)90050-9.

Walker, D., Jarvis, P., Farquhar, G., Leverenz, J. (1989). Automated measurement of leaf
photosynthetic O2 evolution as a function of photon flux density. Philos. Trans. R. Soc.
Lond. B. Biol. Sci. 323:313–326.

Wargent, J.J., Jordan, B.R. (2013). From ozone depletion to agriculture: Understanding the role
of UV radiation in sustainable crop production. New Phytol. 197:1058–1076. doi:
10.1111/nph.12132.

Weerasinghe, L.K., Creek, D., Crous, K.Y., Xiang, S., Liddell, M.J., Turnbull, M.H., Atkin, O.K.
(2014). Canopy position affects the relationships between leaf respiration and associated
traits in a tropical rainforest in Far North Queensland. Tree Physiol. 34:564–584. doi:
10.1093/treephys/tpu016.

Wilson, M.I. (1998). PhotomorphologicaI and Photochernical Effects of UV-B Radiation on

Brassica napus (L.) and Arabidopsis thaliana (L.) Heynh: Morphological, Cellular and
Structural Biological Changes. [Thesis]

Yang, Y., Yao, Y., He, H. (2008). Influence of ambient and enhanced ultraviolet-B radiation on
the plant growth and physiological properties in two contrasting populations of
Hippophae rhamnoides. J. Plant Res. 121:377–385. doi: 10.1007/s10265-008-0163-y.

Yep, B., Gale, N. V., Zheng, Y. (2020). Aquaponic and hydroponic solutions modulate NaCl-
induced stress in drug-type Cannabis sativa L. Front. Plant Sci. 11:1–14. doi:
10.3389/fpls.2020.01169.

Yin, R., Ulm, R. (2017). How plants cope with UV-B: From perception to response. Curr. Opin.
Plant Biol. 37:42–48. doi: 10.1016/j.pbi.2017.03.013.

101
Younis, B.A., Mahoney, L., Schweigkofler, W., Suslow, K. (2019). Inactivation of plant
pathogens in irrigation water runoff using a novel UV disinfection system. Eur. J. Plant
Pathol. 153:907–914. doi: 10.1007/s10658-018-01608-8.

Zhang, L., Allen, L.H., Vaughan, M.M., Hauser, B.A., Boote, K.J. (2014). Solar ultraviolet
radiation exclusion increases soybean internode lengths and plant height. Agric. For.
Meteorol. 184:170–178. doi: 10.1016/j.agrformet.2013.09.011.

Zhang, X., He, D., Niu, G., Yan, Z., Song, J. (2018). Effects of environment lighting on the
growth, photosynthesis, and quality of hydroponic lettuce in a plant factory. Int. J. Agric.
Biol. Eng. 11:33–40. doi: 10.25165/j.ijabe.20181102.3420.

Zhao, D., Reddy, K.R., Kakani, V.G., Read, J.J., Sullivan, J.H. (2003). Growth and physiological
responses of cotton (Gossypium hirsutum L.) to elevated carbon dioxide and ultraviolet-B
radiation under controlled environmental conditions. Plant, Cell Environ. 26:771–782.
doi: 10.1046/j.1365-3040.2003.01019.x.

Zheng, Y. (2020). Soilless production of drug-type Cannabis sativa. Acta Hortic. (In press.).

Zheng, Y., Blom, T., Dixon, M. (2006). Moving lamps increase leaf photosynthetic capacity but
not the growth of potted gerbera. Sci. Hortic. 107:380–385. doi:
10.1016/j.scienta.2005.09.004.

Zlatev, Z.S., Lidon, F., Kaimakanova, M. (2012). Plant physiological responses to UV-B
radiation. Emirates J. Food Agric. 24:481–501. doi: 10.9755/ejfa.v24i6.481501.

Zou, S., Kumar, U. (2018). Cannabinoid receptors and the endocannabinoid system: Signaling
and function in the central nervous system. Int. J. Mol. Sci. 19. doi:
10.3390/ijms19030833.

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 APPENDIX A

SUPPLEMENTARY INFORMATION: PLANT GROWTH RESPONSE TO

INCREASING LI
METHOD

The number of nodes and main stem length (i.e., from the substrate surface to the tip of the
tallest shoot, hereafter, height) were measured twice weekly until day 42, when vegetative
growth had ceased.

RESULTS

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Supplementary Figure 1. The accumulated increase in Cannabis sativa L. ‘Stillwater’ (A) main
stem length (cm) and (B) number of nodes from day 0 (i.e., day from initiation of 12-h flowering
photoperiod) over time. Each datum is the mean ± SE increase of main stem length from day 0 of
all plants from that particular day (CB1 n = 43, CB2 n = 48).

Supplementary Figure 2. Cannabis sativa L. ‘Stillwater’ (A) main stem length (cm) and (B)
number of nodes on day 42 of the flowering stage in response to average photosynthetic photon
flux density (PPFD) calculated based on the respective plants’ PAR accumulated exposures up to
day 42 of the flowering stage. Each datum is a single plant.

Around day 42 after the initiation of the 12-h photoperiod, plant the plants ceased growing
vegetatively (Supplementary Figure 1A, B). After the cessation of vegetative growth (i.e., on
day 42), height increased linearly from 98.2 to 118 cm (i.e., 1.2 times higher) in CB2, but height
in CB1 was not affected with mean ± SD of 105 ± 70 (Supplementary Figure 2A), as APPFD
increased from 130 to 1930 μmol·m−2·s−1 (calculated based on the respective plants’
accumulated PAR exposures up to day 42 of the flowering stage). Across the same increasing

104
range of APPFD, number of nodes increased from 20 to 24 (i.e., 1.2 times higher) and 23 to 26
(i.e., 1.1 times higher) in CB1 and CB2, respectively (Supplementary Figure 2B). The
“topping” technique from plants in the vegetative stage lowered absolute main stem length and
number of nodes.

DISCUSSION

After the initiation of the 12-h photoperiod, plants continued to grow their vegetative organs
until around day 40. The present study found that “untopped” plants responded to increasing
APPFD with increased height. In contrast, topped plants did not demonstrate significant increases
in height, likely due to the lack of apical dominance which could evenly distribute the height
increases between to the two nodes at the top of the plant and quell the overall height increase
(Barbier et al., 2017). Nevertheless, increasing APPFD increased the number of nodes in plants
whether they were topped or not. Tall plants can be difficult to manage and harvest, hence
growers must consider the potential height increase when increasing their canopy-level LI, which
depends on the use of the topping technique.

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 APPENDIX B

SUPPLEMENTARY INFORMATION: PHOTOBLEACHING AT HIGH LI

METHODS

Photobleaching was recorded based on the visible observation, including a measurement of

PPFD directly at the location where the phenomenon occurred.

RESULTS

Supplementary Figure 3. Image of inflorescence photobleaching 70 d after the initiation of 12-

h photoperiod.

Photobleaching occurred on the apical inflorescences of treatment plants under average PPFDs
ranging from 1040 to 1850 μmol·m−2·s−1 (Supplementary Figure 3).

DISCUSSION

When light intensity exceeds a tissue’s photoprotective capacity for long-term periods, damage
occurs such as photobleaching (i.e., the photodestruction of chlorophyll) (Björkman and
Demmig, 1987; Havaux and Niyogi, 1999; Henley et al., 1991; Mooney et al., 1974). However,
the photobleaching that occurred in this trial was visible only on the top of apical inflorescences,
which are not the primary photosynthetic tissue. Additionally, photobleaching was only observed

106
on certain plants, not all the plants under high LIs. Further research is required to understand this
phenomenon.

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 APPENDIX C

Supplementary Figure 4. Necrotic patches on ‘Breaking Wave’ (left) and ‘Low Tide’ (right)
Cannabis sativa L. leaves 9 weeks after the initiation of UV treatments. The black scale bar at
the lower right is 2.0 cm.

108