Journal of Essential Oil Bearing Plants

ISSN: 0972-060X (Print) 0976-5026 (Online) Journal homepage: https://www.tandfonline.com/loi/teop20

Influence of Harvesting Time on Essential Oil

Content, Chemical Composition and Pesticidal
Activity of Artemisia maritima Growing Wild in the
Cold Desert Region of Western Himalayas

Swati Walia, Arti Rana, Ashok Singh, Mohit Sharma, S.G. Eswara Reddy &
Rakesh Kumar

To cite this article: Swati Walia, Arti Rana, Ashok Singh, Mohit Sharma, S.G. Eswara Reddy
& Rakesh Kumar (2019) Influence of Harvesting Time on Essential Oil Content, Chemical
Composition and Pesticidal Activity of Artemisia�maritima Growing Wild in the Cold Desert
Region of Western Himalayas, Journal of Essential Oil Bearing Plants, 22:2, 396-407, DOI:
10.1080/0972060X.2019.1610077

To link to this article: https://doi.org/10.1080/0972060X.2019.1610077

Published online: 22 May 2019.

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 TEOP 22 (2) 2019 pp 396 - 407 396
ISSN Print: 0972-060X
ISSN Online: 0976-5026

Influence of Harvesting Time on Essential Oil Content, Chemical

Composition and Pesticidal Activity of Artemisia maritima Growing
Wild in the Cold Desert Region of Western Himalayas

Swati Walia 1,2, Arti Rana 2, Ashok Singh 4, Mohit Sharma 1,3
,
S.G. Eswara Reddy 1,2* and Rakesh Kumar 1,2*

Academy of Scientific and Innovative Research

1

Agrotechnology of Medicinal, Aromatic and Commercially Important Plants Division

2

3
Natural Product Chemistry and Process Development Division
4
High Altitude Biology Division, CSIR-Institute of Himalayan Bioresource Technology,
(Council of Scientific and Industrial Research), Post Box No. 6, Palampur 176 061 (H.P.), India
Received 02 November 2018; accepted in revised form 16 April 2019

Abstract: Essential oil (EO) of Artemisia maritima plants growing naturally in the cold desert region
has been extracted by hydro distillation at different growth stages during 2015-16. The constituents of the EO
have been characterized by gas chromatography (GC) and GC-mass spectrometry. Nineteen compounds were
identified. EO content and composition vary throughout the vegetation period of the plant. The best time for
higher EO content (0.45±0.15 %) is August, while for higher 1,8-cineole (59.24±1.63 %) is during October. EO of
A. maritima was characterized by high content of 1,8-cineole, bornyl acetate, terpinen-4-ol, camphor, sabinene
and germacrene D which varied from 90.54±7.76 to 92.41±6.50 % of the total oil content. The EO of A. maritima
was evaluated for their acaricidal activity against two-spotted spider mite (Tetranychus urticae) (Acarina:
Tetranychidae). EO reported excellent fumigant activity against T. urticae with LC50 value of 323.86, 27.05, 15.48
and 12.15 mgL-1 after 1, 2, 3 and 4 h of treatment, respectively as compared to positive control i.e. Propargite 57
% EC (755.72 mgL-1). In repellent activity, 10000 mgL-1 showed significantly higher repellency (95.83±1.67 %) to
adult T. urticae and was at par with positive control i.e. Azadirachtin 0.15 % EC (96.67±3.33 %) and 8000 mgL-1
(91.67±2.20 %) as compared to other concentrations studied.

Key words: Artemisia maritima, harvesting time, 1,8-cineole, Tetranychus urticae, fumigant.

Introduction mintic and very effective against round worms

Artemisia maritima L. is an aromatic perennial (Ascaris spp.). Distinctive parts of the plant have
herb commonly known as sea wormwood (fam- been used for medicinal purposes; leaves for
ily Asteraceae). It is distributed in the western asthma, stomach torment (antispasmodic) and
Himalayas at altitudes of 7000-9000 feet and CNS ailments, while seeds are used as an appe-
grows abundantly in north-west Kashmir, dry tem- tizer, expectorant, aphrodisiac, ophthalmia, a cure
perate region of Himachal Pradesh 1 and to a for indigestion, diarrhoea, inflammation, a heart
certain extent in Chamoli district of Garhwal stimulant and tonic 3. The essential oil of A. mar-
Himalayas (Uttarakhand) 2. The plant is commer- itima shows anti-infective, antibacterial, antifun-
cially utilized in India for santonin which is anthel- gal, antispasmodic, antimalarial and bronchodila-

*Corresponding authors (S.G. Eswara Reddy; Rakesh Kumar)

E-mail: < [email protected]; [email protected] > © 2019, Har Krishan Bhalla & Sons
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 397
tory activities 1,4. The chemical composition of its and dropping of leaves 14,15. Tetranychus urticae
essential oil has been extensively studied. The was developed resistance to different acaricides
main compounds of the essential oil of A. mar- due to the non-judicious application and short life
itima are 1-8 cineole, camphor, borneol, bornyl cycle. The EOs reported toxicity, antifeedant, re-
acetate and germacrene-D 5. However, the con- pellent and ovicidal activities against different
tent of essential oils and its constituents reported pests 16,17 including T. urticae 18-20. Although, aca-
varying significantly depending upon variations in ricidal properties of different EOs have been re-
climatic and agricultural conditions, growing sea- ported on T. urticae, but no report is available for
son, growth/harvesting stage, plant parts, cultiva- the EO of A. maritima. Therefore, the present
tion practices and origin 6. experiment was planned to study the fumigant and
The chemical composition of Artemisia spp. repellent activity of A. maritima against T. urticae.
from different origins show a dominant presence The population of A. maritima is abundantly
of 1,8-cineole, α-thujone, β-thujone, davanone, available in the barren land of Lahaul-Spiti and
artemisian ketone, β-caryophyllene or caryo- Kinnaur district of Himachal Pradesh between
phyllene oxide, 1,8-cineole, camphor, and borneol 2700 m and 3400 m but the local people are not
are the major compounds present in essential oil making its effective use due to lack of knowledge
A. maritima 7-9. The variation in the occurrence 1
. Though local people use raw Artemisia plants
of certain compounds in a plant is a function of for insect repellency in the fields but they are un-
any one of, or a combination of, three factors: aware of its insecticidal property which can be of
genetically determined properties, the age of the great use in industries and can lead to good eco-
plant, and the environment. Ideally, an experiment nomic returns. Many studies have been conducted
evaluating the relative importance of these fac- to determine essential oil content and composi-
tors should be designed with randomized blocks tion of A. maritima, but there is no information
and plants of equal age, collected by careful sam- regarding its harvesting time and insecticidal ac-
pling in a random and systematic manner from tivity of its essential oil. Hence this study was
the natural populations. In several species of conducted to find out the effect of harvesting time
Lamiaceae, at flowering, essential oil is at the high- on essential oil content and compounds of A. mar-
est level, while in other species, flowering has a itima from its natural population grown in cold
lower influence 10. The present investigation deals desert region of western Himalayas to obtain the
with the variations of yield and composition of the highest essential oil yield and to study its insecti-
essential oil obtained at different growth times by cidal properties for industrial use.
plants collected from the same stand to limit the
influence of the factors previously cited. Materials and methods
Botanical formulations have received much im- Experimental site
portance in organic farming/sustainable manage- The experiment was performed at the research
ment of insect pests due to low toxicity to natural center of CSIR-Institute of Himalayan Biore-
enemies of insects and non-target organisms. Due source Technology, Palampur (HP) India, Centre
to multiple modes of action, plant products are for High Altitude Biology, CeHAB situated at
gaining more attention as an alternative method Tandi, Keylong Distt. Lahaul and Spiti, HP (3450
of control. Two-spotted spider mite (Tetranychus m amsl, 32°33’58.8" N, 76°58’21.8" E), during
urticae Koch) (Acarina: Tetranychidae) is an 2015-2016. Weather data (maximum and minimum
important pest of different cultivated crop plants temperature, rainfall, soil temperature, and rela-
grown under protected and field conditions caus- tive humidity) were obtained from weather tower
ing losses in most years. Adults and nymphs suck situated in CeHAB Ribling farm, Lahaul and Spiti,
the sap from the lower surface of leaves which HP (Figure 1). Weekly average data was recorded
results in yellowing and discoloration 11-13. In se- during different harvesting time which was pooled
vere incidence, the mites construct the spider webs for 2015-16. In 2015-2016, the maximum tempera-
on the leaves and stem which results in drying ture ranged from 17.34 to 21.46°C, whereas mini-
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 398
mum temperature ranged from 4.02 to 11.09°C gen with 1.05 mL min-1 flow rate. Oven tempera-
during the harvesting period. Likewise, average ture was programmed at 70°C fo 4 min and then
soil temperature ranged from 15.16 to 18.51°C, increased by 4°C min-1 to 220°C, with maintained
mean relative humidity ranged between 38.56 to temperature of injector (240°C) and detector
68.80 % and 62.23 mm total rainfall was received (250°C).
during the experimental period. Gas chromatography-mass spectrometry analy-
sis was carried out by Shimadzu QP2010 GC-
Plant material MS with AOC-5000 Auto injector and ZB- 5MS
Fresh plant material of A. maritima was col- capillary column (30 m x 0.25 mm, 0.25 μm film
lected from Lahaul valley (latitude 32o.57 1 N and thickness) in which nitrogen with 1.05 mL min-1
longitude 77o.04 1 E) of Himachal Pradesh (In- flow rate was used as carrier gas. Sample injec-
dia). The plant material was authenticated, and a tion of 2 μL with split ratio of 10 and mass scan
voucher specimen was deposited in the herbarium 40-800 amu. Injector and ion source temperature
(PLP 17794) of CSIR-IHBT, Palampur. were maintained at 240°C and 250°C, respectively.
At initial oven temperature used was 70°C for 4
Experimental details min with a rising rate of 4°C min-1 upto 220°C
A. maritima fresh plant samples were harvested and then held isothermal at 220°C for 5 min 22.
from natural population during the fourth week, in Compounds were identified on the basis of reten-
the afternoon, in different times of the growing tion indices (calculated using homologous series
phase of the plant at the monthly interval i.e. from of n-alkanes from C8 to C24) and peak area chro-
June to October (during 2015 and 2016). In each matogram. Components of oil were then identi-
month 500 g of fresh aerial biomass viz. stem and fied by comparing retention indices (RI) values,
leaves were used for distillation. Material was mass spectra and NIST (National Institute of
chopped and hydro-distillated in Clevenger appa- Standards and Technology) library 23,24.
ratus for four hours. Three replicates were used
for evaluating essential oil content and two repli- Test mites
cates for the composition of the oil. The experi- Tetranychus urticae used for the study was
ment was conducted in a randomized complete collected from the field and reared on Phaseolus
block design. Essential oil content was measured vulgaris at 25±2°C, 60±5 % relative humidity and
as mL and % ratios (v/w), as per procedure given L16:D8 h of photoperiod under laboratory for more
in European Pharmacopoeia 21. Extracted oil than 30 generations.
samples were dried over anhydrous sodium sul-
phate, and stored at 4°C until analysis. Essential Acaricidal activities of essential oils against
oil chemical composition was analyzed in Natural T. urticae
Product Chemistry and Process Development di- Fumigant toxicity
vision of the Institute. Fumigant toxicity of the EO of A. maritima
against adult T. urticae was evaluated as per the
Identification of compounds through Gas method described by Mozaffari et al. 18 with little
Chromatography (GC) and Gas Chromato- modification by Reddy et al. 17. Five concentra-
graphy-Mass Spectrometry (GCMS) tions of essential oil collected in the month of
Essential oil composition of A. maritima was August (100, 50, 40, 20 and 10 mg L–1 volume by
analyzed by Shimadzu GC-2010 gas chromato- air) were prepared.
graph (Shimadzu, Tokyo, Japan) equipped with P. vulgaris leaf discs (3 cm diameter) were
flame ionized detector (FID) and ZB- 5MS (5 % placed in Petri dishes (60 ml capacity) lined with
diphenyl, 95 % methyl polysiloxane stationary water-saturated cotton wool. With a fine camel-
phase) capillary column (30 m x 0.25 mm, 0.25 hair brush, 10 adult mites were released on the
μm film thickness) from SGE International, surface of the discs. Each concentration was rep-
Ringwood, Australia. Carrier gas used was nitro- licated thrice. Filter paper disks (Whatman No.
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 399
1) cut into 2 cm in diameter and placed on the top design used, and treatment means were compared
of petri dishes with cello tape and impregnated at P < 0.05 level of significance. The fumigant
with five concentrations of EO. The commercially toxicity experiment was repeated for three times
available acaricide Propargite 57 % EC (Omite®) and mean data were used for analysis. Fumigant
available in the market at recommended dose was toxicity data from all bioassays were corrected
used for the control of T. urticae and other mite for mortality in the control using Abbott formula
species was used as a positive control for com- 26
. The median lethal concentration (LC50), me-
parison. Observations on number of dead mites dian lethal time (LT50) and their corresponding 95
were recorded at hourly interval up to 4 hours. % confidence intervals, Chi-square and regres-
sion coefficients were determined following Probit
Repellent activity analysis 27 using statistical software, version 16.
Repellent activity of EOs was tested as per the The data on repellence was subjected to one way
method followed by Kogan & Goeden 25 with little ANOVA using statistical software, version 16 and
modification by Reddy et al. 15. Briefly, 100 mg means were compared by Duncan Multiple Range
of EO was dissolved in 10 mL of distilled water Test (DMRT).
containing 0.05 % Triton-X100 LR spreader and
sonicated for complete dissolution. Five concen- Results and discussion
trations of test solutions (10000, 8000, 4000, 2000 Essential oil content
and 1000 mgL-1) were prepared from stock solu- The fresh samples of A. maritima on
tions from the higher concentration and poured in hydrodistillation yielded pale yellow oil with pleas-
glass petri dishes. Fresh P. vulgaris leaf discs (3 ant odor. Essential oil content varied from 0.25 to
cm diameter) were prepared from P. vulgaris 0.45 % on the weight of fresh leaf material in the
leaves. Half of the leaf disk was immersed into a season that lasts from June to October. Signifi-
test solution for five seconds, and after drying, cantly higher essential oil content was obtained in
the other half of the disc was immersed in water, August as compared to other months. This may
which served as control. After drying, leaf discs be because of the reason that flowering in A.
were placed in Petri dishes lined with water-satu- maritima occurs during July-August, also these
rated cotton wool. With a fine camel-hair brush, months recorded maximum temperature, rainfall
10 adult mites were released on the leaf discs. and relative humidity (Figure 1) which do not pro-
There were seven treatments and each treatment duce sufficient IAA for cell division and leads to
was replicated three times. The commercially flower formation 28, due to which highest essen-
available neem based formulations i.e. Neem Baan tial oil content is reported during this period. Kaul
(Azadirachtin 0.15 % EC at 5 ml L-1) and neem et al. 29 also reported low oil content in summer
oil at 5 ml L-1 available in the market at recom- when the day temperature cross 40°C and high
mended dose for the control of T. urticae and oil content during moderate rainfall with high rela-
other mite species was used as a positive control tive humidity in rose-scented geranium. The
for comparison. Observations on the number of present result showed that flowering stage i.e.,
mites present in each half of the leaf disc was during July-August is best suitable harvesting stage
counted after 1 h. The percent repellence was for A. maritima, a further delay of harvest de-
calculated according to the following formula: creased the essential oil content of A. maritima.
% Repellency = (C-T/N)*100; Where, T = num- An earlier study in Garhwal region of Uttara-
ber of mites in the treatment, C = number of mites khand, also reported higher essential oil content
in the control, N = Number of mites used in the in Artemisia when harvested during the third
test. week of August 30. Likewise, Sharma et al. 5 in
A. maritima reported 0.33 % essential oil content
Statistical analysis when harvested in the month of July in the Cold
Statistical analysis of data was done as per the Desert region of Himachal Pradesh. However,
standard analysis of variance technique for the Sah et al. 9 reported opposite results with higher
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 400

Figure 1. Monthly mean weather data (temperature, rainfall , and relative humidity)
during the harvesting season at Lahaul &Spiti, India. RH, Relative humidity
(0.50 %) essential oil content in A. maritima with or camphor (44.3-44.4 %) as major constituents
late harvesting during the month of November. 32
. Sah et al. 9 identified chrysanthenone (25.7 %)
as the main compound in A. maritima EO. These
Essential oil composition data illustrate the existence of different
Fifteen components were determined and iden- chemotypes that may be due to differences in lo-
tified by GC and GC-MS, representing about 77.43 cation, soil type, altitude, and physiological stage
to 88.12 % of the essential oil of A. maritima of development. The variation in certain com-
(Table 1). Comparison of the analytical data of pounds in a plant is due to genetic makeup, the
the oils revealed marked differences in qualita- age of the plant and the environment.
tive and quantitative composition. The major con- Among different harvesting time, harvesting in
stituents of the essential oil were 1,8-cineole, the month of October recorded a significantly
bornyl acetate, terpinen-4-ol, camphor, sabinene, higher percentage of 1,8-cineole which was 28.47
germacrene D, etc. The essential oil of A. mar- % higher than August month which recorded low-
itima possessed the highest amount of 1,8-cin- est (Figure 2). 1,8-cineole was reported maximum
eole during all the harvesting stages. These re- in October followed by July>June>
sults are in accordance with the reports of Jaitak September>August. Each compound had a unique
et al. 31 and Sharma et al. 5 on A. maritima. The profile for accumulation as a function of tempera-
altitudinal impact was exhibited by an investiga- ture and precipitation 33. As October month re-
tion of Jaitak et al. 31 where plants collected from corded lowest temperature and rainfall (Figure 1),
different regions of western Himalaya revealed so it can be assumed that maximum accumula-
distinctive percentage of 1,8-cineole with 23.8 % tion of 1,8-cineole occurred at low mean tempera-
from Pooh area (2500 m), 37.3 % from Rhongtong tures (between 10-18°C), and at low precipita-
pass (3500 m) and 44.2 % from Lahaul & Spiti tion level. Significantly higher percentage of Cam-
(4500 m) district of HP. Generally, the composi- phene and bicyclogermacrene were also reported
tion of A. maritima essential oil varied between in the month of October and lowest in June. Bornyl
investigations published in the literature. Some acetate and Terpinen-4-ol was found significantly
older publications reported α-thujone (63.3 %) 7 higher in the month of June, which was 67.95 %
 Table 1. Effect of different harvesting times on essential oil content and
composition (v/w %) of A. maritima (pooled data of 2015 and 2016)
Total

Borneol

Linalool
Camphor

Sabinene

Camphene
1,8-Cineole

β-Myrecene
Phellandrene
Terpinen-4-ol
Germacrene D

Bornyl acetate
Sabinyl acetate

Santolina triene
Bicyclogermacrene

Essential oil (v/w %)

Chrysanthenyl acetate

Litt. RI 908 953 976 991 1031 1033 1098 1143 1165 1177 1262 1285 1291 1480 1494
Exp. RI 902 950 973 990 1024 1033 1098 1146 1161 1180 1256 1283 1286 1479 1493
June 0.37c 0.69c 0.87de 2.68a 1.57cde 1.68bcd 41.28bc 4.31a 1.78d 0.99cde 14.20a 2.03a 13.47a 0.58cde 1.14abc 0.33cd 88.12ab
July 0.41b 4.88a 0.99d 2.14bc 2.72bc 2.53ab 43.09b 0.84bcd 3.87b 1.11cd 11.39b 1.04bc 8.10d 2.05a 0.95bcd 0.38c 86.39abc
August 0.45a 0.44d 2.16ab 2.52b 2.18bcd 2.82a 38.87de 0.86bc 5.52a 1.28bc 9.71bcd 1.19b 13.01abc 1.23bc 1.44a 0.65b 84.25abd
e
September 0.33d 3.87ab 1.34c 0.81e 5.81a 1.33ce 39.02cd 1.23b 3.43bc 3.66a 11.16bc 1.19b 13.15ab 1.47ab 0.39 0.07e 88.14a
ab
October 0.25e 0.00e 2.44a 1.29d 3.12b 2.05bc 49.94a 0.52bcde 1.75de 2.68ab 9.69bcde 0.90cd 8.02de 0.63cd 1.36 1.11a 77.43e
SEm ± 0.004 0.49 0.18 0.20 0.53 0.31 0.75 0.28 0.38 0.49 0.89 0.17 1.40 0.26 0.16 0.09 2.26
Swati Walia et al., / TEOP 22 (2) 2019 396 - 407

LSD (<0.05) 0.013 1.46 0.54 0.59 1.58 0.92 2.23 0.84 1.13 1.46 2.63 0.52 4.16 0.77 0.46 0.25 6.73

RI, retention indices; SEm±, Standard Error Mean; LSD, Least Significant Difference
401
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 402

Intensity

min.
Figure 2. GC chromatograms of essential oil of
Artemisia maritima harvested in the month of October
and 46.54 % higher, respectively, as compared to Valeriana officinalis, major compounds viz.
October which recorded the lowest value. A sig- valerenal, valerenic acid, and α-humulene con-
nificantly higher value of Camphor and germa- tents increased with the age of plants. Since 1,8-
crene D was reported in the month of August, cineole is the major and most important constitu-
which further decreases with delaying time and ent of A. maritima essential oil, so to get its maxi-
reported 215.42 % and 269.23 % higher as com- mum percentage plant should be harvested in
pared to October and September, respectively. October.
Sabinene, linalool and chrysanthenyl acetate were
reported significantly higher in June which de- Acaricidal activity of EO of A. maritima
creases with delaying harvesting time or say ma- Fumigant toxicity of EO of A. maritima against
turity, while the opposite was seen in case of T. urticae
Borneol and β-myrcene which significantly in- The fumigant toxicity of EO of A. maritima
creased in September with the lowest value in against adults of T. urticae for dose-response bio-
the month of June. Other compounds of essential assay to calculate the LC50 and LT50 to kill 50 %
oil did not show any consistent trend over the har- of the test insect relative to control values at dif-
vesting time. Previous studies have shown that ferent hours after treatment was presented in
changes in the oil composition as a result of growth Table 2 and Table 3. Results showed that, all the
development are dependent on the species and concentrations of EO showed toxicity to T.
the nature of compounds. Some compounds are urticae after treatment. LC50 values of EO at 1,
increased with the age of plants while some de- 2, 3 and 4 h after treatment was 323.86, 27.05,
creases. Chauhan et al. 34 found that thymol re- 15.48 and 12.15 mgL-1, respectively. Similarly, LT
corded its maximum amount in Origanum vulgare 50 to kill 50 % of test mite at 100, 50, 40, 20 and
at the late vegetative stage, while for γ-terpinene, 10 mgL-1 was 1.35, 1.58, 1.61, 2.71 and 3.94 h,
the maximum % was recorded at the reproduc- respectively as compared to positive control i.e.
tive stages (flower initiating, full blooming, and fruit Propargite 57 % EC (755.72 mgL-1). The toxicity
set). Letchamo et al. 35 also reported that in of A. maritima oil against T. urticae varies with
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 403
Table 2. LC50 values for the fumigant toxicity of essential
oil of A. maritima against T. urticae (HAT)

Time (HAT) LC50 (mgL-1) 95 % Cl (mgL-1) Slope ± SE Chi square P value

1h 323.86 96.41-417.50 0.92 ± 0.36 1.02 0.80

2h 27.05 12.72-71.29 0.99 ± 0.31 0.19 0.98
3h 15.48 8.98-21.32 1.71 ± 0.35 1.30 0.73
4h 12.15 7.80-15.90 2.45 ± 0.45 1.35 0.72
Propargite 57 % EC (24 h) 755.72 95.68-1511.05 1.32 ± 0.37 1.24 0.74

HAT: Hours after treatment

Cl: Confidence limits

Table 3. LT50 values for the fumigant toxicity of

essential oil of A.maritima against T. urticae

Concn. LT50 (h) 95 % Cl (h) Slope ± SE Chi square P value

100 mgL-1 1.35 1.06 - 1.60 3.98 ± 0.68 1.61 0.40

50 mgL-1 1.58 1.24 - 1.88 3.44 ± 0.61 0.35 0.84
40 mgL-1 1.61 1.23 - 1.94 3.13 ± 0.59 0.06 0.97
20 mgL-1 2.71 2.02 - 4.28 1.87 ± 0.55 0.27 0.87
10 mgL-1 3.94 2.98 - 7.90 2.17 ± 0.62 1.32 0.52

h- Hours after treatment

Cl- Confidence limits
time after treatment/spray. Initially the mortality In the present study, EO of A. Maritima
will be slow and then gradually increased and showed good fumigant toxicity to adult T. urticae.
constant after some time because these EOs take Our results conform with the results of others who
time to act on insect as compared to synthetic reported the fumigant toxicity of other EOs to T.
pesticides. Similarly, the higher dose of EO took urticae. Piper aduncum and Thuja orientalis
less time to kill the 50 % of mite population as oil showed good fumigant activity to T. urticae
compared to lower concentrations (Table 2). Use 38,39
. In a similar experiment, Eucalyptus
of higher dose of EO (100 mgL-1), the air would codonocarpa, and E. dives oil (10 μl L-1 air)
very probably be saturated with fumes of EO much showed excellent fumigant action against T.
more quickly, thereby attaining more mortality. urticae 40. Similarly, EO of Cuminum cyminum,
Volatile nature of EO plays an important role Syzygium aromaticum, Mentha spicata, Achil-
for the toxicity, however, not essentially all the lea mellifolium and A. filipendulina also reported
constituents of the EO have the same degree of fumigant activity to T. urticae 41-43.
volatility/mode of action. The chemical constitu-
ents’ of EO may vary depending upon genotype, Repellent activity of EO of A. maritima
soil, climatic conditions as well as season and ex- against T. urticae
traction method 36. The toxicity of the EO is also Repellent activity of EO of A. maritima was
varied between plant species, chemical composi- evaluated against T. urticae and results were pre-
tion and proportion of the constituents. The in- sented in Table 4. Among the tested concentra-
secticidal activities of EOs are may be due to the tions, A. maritima at 10000 mgL-1 showed signi-
major chemical constituents but not always 37. ficantly (one way ANOVA; F 6, 20 = 42.74;
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 404
Table 4. Repellent activity of essential oil of A. maritima oil against T. urticae

Concn. (mgL-1) % repellence

10000 95.83 ± 1.67 a

8000 91.67 ± 2.20 a
4000 61.67 ± 3.00 c
2000 47.50 ± 5.00 d
1000 35.83 ± 3.63 e
Azadirachtin 0.15 % EC 96.67 ± 3.33 a
Neem oil 0.50 % 80.00 ± 5.77 b
F value F6, 20 = 42.74; p< 0.0001

Means followed by the same letters within a column do not differ significantly by DMRT
p<0.0001) higher repellency (95.83±1.67 %) to amounts throughout the period under study. The
adult T. urticae and was at par with positive con- plant can be harvested in the month of July to
trol i.e. Azadirachtin 0.15 % EC (96.67±3.33 %) August for getting higher essential oil, however,
and A. maritima oil at 8000 mgL-1 (91.67±2.20 for higher 1,8 cineole the plant is to be harvested
%) followed by 4000 mgL-1 (61.67±3.0 %) and in the month of October. Monoterpene 1,8-cin-
2000 mgL-1 (47.50±5.0 %). Similarly, the other eole, bornyl acetate, terpinen-4-ol, camphor,
positive control (Neem oil 0.50 %) which is used sabinene, and sesquiterpene germacrene D are
for the control of T. urticae in greenhouse and found to be alternatively the main components of
field conditions is inferior to higher concentration the oil: monoterpene before and after the flower-
of A. maritima oil. Most of the EO and its com- ing and sesquiterpene before flowering. The EO
pounds presently used as repellents. The monot- of A. maritima showed excellent fumigant and
erpenes (eg., limonene, cineole, eugenol, citronel- repellent activity to T. urticae as compared to
lal, etc.) present in the EOs are may be mainly positive controls i.e. Propargite, Azadirachtin/
responsible for repellent activity of T. urticae. In neem oil. Production of any botanical formulation
the present study, EO of A. maritima showed requires continuous availability of plant material
excellent repellent activity to T. urticae. Present on a commercial scale. The population of A. mar-
results are in agreement with the reports of other itima is abundantly available in barren land but
researchers who reported repellent activity to T. the local people are not making its effective use
urticae. The EO from Protium bahianum Daly due to lack of knowledge of processing techniques
44
, Piper aduncum L. 38, Callistemon sieberi, E. and processing equipment. The standardization of
bicostata, E. ovata, E. sideroxylm, E. manni- process technology for the extraction of essential
fera, E. dives, E. elata, E. condonocarpa, oil from plant parts and crude extracts of A. mar-
Kunzea ericoides, Melaleuca armillaris and M. itima should be further evaluated against target
fulgens 19, M. pulegium 18 , Elettaria carda- insect pests in the field. Based on field bio-effi-
momum L 45, Lavandula x intermedia Emeric cacy data and feasibility, the botanical formula-
ex Loisel, Hyssopus officinalis 46, S. aromaticum tion may be recommended for the commercial
and M. Spicata 41 showed promising repellent application in the greenhouse/field conditions for
activity to T. urticae. the target pests.

Conclusion Acknowledgments
In general, it can be concluded that harvesting The authors are grateful to the Director, CSIR-
time improves the essential oil content and oil com- IHBT, Palampur for necessary facilities during
position of A. maritima as components of the es- the course of study. We also thank Mr. Shiv Kumar,
sential oil show fluctuations in their relative Senior Technical Officer for providing technical
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 405
support during the work. Authors are also grate- nology, GOI, New Delhi, and Council of Scien-
ful to Dr. Bikram Singh, Chief Scientist (Retd.), tific and Industrial Research (CSIR) is also ac-
NPC&PD division for help in identification of knowledged. IHBT communication number for
essential oil compounds. Financial grant received this article is 4228.
from SERB, Department of Science and Tech-

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