The Agriculturists 10(2): 109-119 (2012) ISSN-1729-5211

A Scientific Journal of Krishi Foundation Indexed Journal

Impact of Trichoderma-enriched Biofertilizer on the Growth and Yield of

Mustard (Brassica rapa L.) and Tomato (Solanum lycopersicon Mill.)

Md. Manjurul Haque1, G. N. M. Ilias2 and A. H. Molla1*

1
Department of Bioenvironmental Science, Faculty of Agriculture, Bangabandhu Sheikh Mujibur
Rahman Agricultural University, Gazipur-1706, Bangladesh
2
Natore Development Society (NDS), Natore and M/S RASH Agro Enterprise, Natore-6400,
Bangladesh

*Corresponding author and Email: [email protected]

Received: 04 July 2012 Accepted: 15 November 2012

Abstract

Biofertilizer plays a significant role in crop cultivation with reduced the chemical fertilizer use. Three
Trichoderma-enriched biofertilizers were evaluated in mustard and tomato cultivation at field
condition. Sole application of biofertilizers didn’t show remarkable contribution but all Trichoderma-
enriched biofertilizers when supplemented with N fertilizer significantly boosted up the growth and
yield of mustard and tomato. Application of 50% N fertilizer along with 50% Trichoderma-enriched
biofertilizers augmented 108 and 203% yields over control both in mustard and tomato, respectively
which were 81.90 and 61.82% in mustard and tomato at standard doses of Nitrogen, Phosphorus and
Potassium fertilizers. The present results suggest that Trichoderma-enriched biofertilizer could save at
least 50% N fertilizer uses for mustard and tomato and could reduce excessive uses of NPK for crop
cultivation.

Keywords: Trichoderma, biofertilizer, crop growth, yield, biomass, tomato, mustard

1. Introduction biological characteristics of the soil, which in

turns, influence growth and increase plant
There is an agreement that among all the production (Molla et al., 2005; Meunchang et
nutrients amendments applied to soil, N-fertilizer al., 2006). In recent years, biofertilizers the
have by far the most important effects in terms products containing living cells of different types
of crop yield enhancement. There is increased of microorganisms are also used in the integrated
emphasis on the environmental quality due to nutrient supply system. Biofertilizers can convert
continuous use of chemical fertilizers. The nutritionally important elements from
integrated nutrient management i.e. combined unavailable to available form through biological
use of chemical fertilizers with organic materials processes leading to crop yields (Hegde et al.,
such as animal manures, crop residues, green 1999). Many researchers studied the role of
manure and composts are alternatives and are organic manures, which was incorporated with
characterized by reduced input of chemical biofertilizer as stimulating the plant growth and
fertilizers. Organic manures are well established yield of vegetables (Shaheen et al., 2007).
to be useful in fertilization of plants due to thair Several studies have also shown that the use of
beneficial effect on the physical, chemical and compost in agriculture is beneficial to soil, crops
110 Haque et al. /The Agriculturists 10(2): 109-119 (2012)

and environment (Rodd et al., 2002; Rahman et tomato (variety- Rattan) 1 month old, uniform
al., 2006). Moreover, compost acts as a long- and healthy single seedling was transplanted in
term reserve and slow-release sources of major each pit on 3rd December. Seedlings of tomato
nutrient like N, P and K (Sullivan et al., 2002). were grown in raised seed bed (5 g seeds were
sown in seed bed of 3 m x 1 m) of sandy loam
Trichoderma harzianum, a filamentous fungus is soil with addition of well decomposed compost
used as a successful biological control agent to (10 kg/bed). The unit plot size for both crops was
control different soil borne plant pathogens such 2.5 m × 2.0 m and spacing was 50 cm × 50 cm
as Pythium spp. Rhizoctonia solani, Fusarium for tomato. For mustard, line spacing of 30 cm
spp., Sclerotium rolfsii etc (Harman et al., 2004). and plant to plant distance of 4-5 cm were
It was demonstrated that T. harzianum induced maintained after thinning.
defense responses and systemic resistance in
addition to control of plant pathogens (Alfano et 2.2. Source of Trichoderma-enriched
al., 2007). Moreover, several species of biofertilizer (BioF)
Trichoderma promoted growth and development Three biofertilizers enriched with Trichoderma
of seedlings of vegetable and non-vegetable harzianum were used in the present experiment.
crops (Bal and Altintas, 2006; Bal and Altintas, These were: (a) BioF/compost (Trichoderma-
2008). Furthermore, cucumber, bell pepper and enriched organic compost i.e. household kitchen
strawberry yields were increased significantly by wastes were composted with Trichoderma
application of Trichoderma spp. (Elad et al., harzianum); (b) BioF/pellets (T. harzianum
2006). Efficient use of Trichoderma-enriched coated wheat grains i.e. T. harzianum was
biofertilizer may increase yield, reduce the uses allowed to grow over broken wheat grains); and
of N fertilizers, reduce soil borne pathogens and (c) BioF/suspension (Broth of T. harzianum i.e.
improve soil health. The objectives of the present fungal spores-suspension were grown in liquid
study were to investigate the impact of media, contained (g/l) KNO3 1.0, KH2PO 4 0.5,
Trichoderma-enriched biofertilizer on yield and MgSO4 7H2O 0.25, glucose 34, trace element
yield contributing characters and extent of FeCl3 0.5-1.0 and pH of 6.5 was maintained).
reduction of N-fertilizers use alone or in Five pieces of 8 mm sized agar block of
combinations in tomato (Lycopersicon Trichoderma inocula were taken into 500 ml
esculentum Mill.) and mustard (Brassica rapa prior autoclaved (15 min at 121 0C) aforesaid
L.) cultivation under field condition. media in 1000 ml Erlenmeyer flask from 4 days
old culture grown in PDA (3.9% potato dextrose
2. Materials and Methods agar) media. Then the flasks were incubated at
150 rpm in orbital shaker for 7 days at room
2.1. Experimental site, seed sowing and temperature. Afterward the fungal suspension
seedling transplantation was filtered by 12.5 cm Whatman # 1filter paper.
The experiments were carried out at the The obtained fungal spores culture was used as
Agricultural Farm of the Bangabandhu Sheikh BioF/suspension (contained 3.5-6.15 × 10 6
Mujibur Rahman Agricultural University spores/ml approximately).
(BSMRAU), Gazipur, Bangladesh during winter
season (Nov. 2008 to Feb 2009). The soil of the 2.3. Fertilizer application and intercultural
experimental site was analyzed for N based on operations
Micro-Kjeldahl method (Bremner, 1960), for P Fertilizers were applied as per the soil analysis
by description of Olsen et al., (1965) and for K results for the respective experimental field. The
based on procedure described by Jackson (1973). doses of urea, triple super phosphate (TSP) and
Mustard (variety- BARI mustard 16) seeds were muriate of potash (MP) were 135, 54 and 5
sown in line on 1st December. Conversely, for g/plot respectively, for tomato. The equivalent
Trichoderma-enriched biofertilizer 111

doses for mustard were 83, 36 and 5 g/plot. The plant (g) and yield per plot (g) after cleaning and
total amount of TSP and MP and 1/3rd of urea drying.
were applied at final land preparation. The
remaining urea was applied in two equal splits 2.5. Experimental design and statistical
i.e. half at 30 days after transplanting/seed analysis
sowing and half at 60 days after sowing for Experiments were conducted in a randomized
mustard seeds and 75 days after transplanting for complete block design (RCBD) with three
tomato seedlings. Biofertilizers such as replications and eight treatments. The assigned
BioF/compost, BioF/pellets and BioF/suspension treatments were T1 - control (without
were applied @ 618 g/plot, 5 g/plant and 5 biofertilizer and chemical fertilizers), T2 - NPK
ml/plant, respectively. The BioF/compost and fertilizer, T3 – BioF/compost (618 g/plot), T4 –
BioF/pellets were applied to the soil immediately BioF/pellets (5 g/plant for tomato and 25 g/row
before sowing of mustard seeds and for mustard), T5 – BioF/suspension (5 ml/plant
transplanting of the tomato seedlings. In case of for tomato and 25 ml/row for mustard), T6 - 50%
mustard, BioF/suspension @ 25 ml was applied N-fertilizer + 50% BioF/compost, T7 - 50% N
in each row at 2-3 days after seedling emergence fertilizer + 50% BioF/pellets and T8 – 50% N
with water by hand sprayer. On the other hand, fertilizer + 50% BioF/suspension. Analysis of
BioF/pellets @ 25 g were dropped in each row variance and comparison of means were
before the sowing of seeds of mustard. In case of calculated separately using a statistical package
tomato, roots of the seedling were immersed in MSTAT-C (MSU, 1989). The treatment means
BioF/suspension and kept for approximately 30 were compared by the least significance
min before transplanting. Five ml of difference (LSD) test. Based on LSD values
BioF/suspension was applied in each pit different treatment means were separated from
immediately after the seedlings were each other by different letters for easy
transplanted. The intercultural operations viz. interpretation.
gap filling, thinning (only for mustard), weeding,
stalking, pruning, irrigation etc were done as 3. Results and Discussion
needed.
3.1. Impact of Trichoderma-enriched
2.4. Parameters considered for data biofertilizer (BioF) on vegetative growth
collection
Trichoderma-enriched biofertilizer alone did not
Four tomato and 10 mustard plants were increase plant height, number of branches and
randomly selected from each plot in such a way number of leaves per plant in mustard compared
that the border effect was avoided. Data were to NPK fertilizer i.e. T2 (Table 1). However
collected for both tomato and mustard on plant application of Trichoderma-enriched
height (cm), branches and leaves number per biofertilizers along with N-fertilizer (except T8)
plant, root and shoot dry matter at 50% expedited those characters in mustard. The
flowering stage (oven dried at 70°C for 3 days), tallest plant (76.07 cm) was obtained in T2
clusters number per plant and number of flowers treated plots, which was significantly (P≤0.01)
per cluster counted during full bloom. In different from T1, T3, T4, T5 and T8. The shortest
addition, in case of tomato, data were also plant (32.97 cm) was recorded in T4, which was
collected for number of fruits per plant, weight identical to that in T5. As in case of plant height,
of individual fruit (g), and weight of fruits per almost similar observations were recorded for
plant (kg) at harvest. In case of mustard, branch and leaf number per plant of mustard.
additional data were collected on number of
siliqua per plant, length of siliqua (cm), number The impact scenario of Trichoderma-enriched
of seeds per siliqua after harvest, seed yield per biofertilizer in tomato was to some extent
112 Haque et al. /The Agriculturists 10(2): 109-119 (2012)

different from the mustard (Table 1). The plant resulted in the lower number of flowers, number
height of tomato was statistically identical in all of siliqua and length of siliqua compared to T2
treatments except T5, which produced the lowest (Table 2). In case of seed number, the sole
(46.50 cm) plant height. Trichoderma-enriched application of Trichoderma-enriched
biofertilizer alone such as T3 and T4 treatments biofertilizers provided almost similar result with
significantly increased the number of branches T2. Conversely, the numbers of flower and
per plant in tomato but they were very similar to siliqua were significantly boosted up by
the treatments of T2, T6 and T7. Alternatively, the supplementation of N fertilizer with
number of leaves per plant was not increased by Trichodemra-enriched biofertilizers, but the
the sole application of Trichoderma-enriched siliqua length and seed number did not respond
biofertilizer such as the treatments T3, T4 and T5. in the same way.
However, combined application (i.e.
supplemented with N) of Trichoderma-enriched In case of tomato, the supplemented N fertilizer
biofertilizer such as the treatments T6, T7 and T8 with Trichoderma-enriched biofertilizers
increased the number of leaves per plant increased the fruit number and flower cluster
significantly (P≤0.01). The lowest number of (except T6) significantly compared to sole
leaves per plant (26.13) was observed in T4, application of biofertilizers, but the results were
which was not different from T1 and T5. similar to T2 (Table 2). However, the flower
number per cluster did not respond significantly
The leaves number of both crops was by combined treatments (i.e. T6-T8) compared to
significantly lower in sole application of sole application of biofertilizer treatments.
biofertilizers (i.e. T3, T4 and T5) and control. But
when N fertilizer was supplemented with The poorest performance of both crops (mustard
Trichoderma-enriched biofertilizers, it and tomato) was observed in treatment T5 but
significantly increased all parameters of mustard supplementation of N fertilizer to it i.e. treatment
but only leaf number of tomato. Moreover, the T8 expedited the reproductive growth and yield
supplemented N fertilizer with Trichoderma- of tomato significantly and not of mustard (Table
enriched biofertilizers did not create any impact 2). On the contrary, the highest performance was
on plant height and branch number of tomato, noted in most of the parameters at T7 of both
except in the treatment T8. Perhaps, crops but poor performance was noticed in most
supplementation of N fertilizer enhanced the cases at T4 of mustard. The results showed that
proliferation and growth of fungal microbes. The the supplemented N fertilizer with biofertilizers
increased population of Trichoderma might play (i.e. 50% N + 50% BioF) significantly increased
positive role to make available the plant nutrients the number of flower per cluster, number of
in rhizosphere zone that increased growth and siliqua per plant of mustard and flower cluster
development of mustard and tomato. Plant and fruit number per plant of tomato, did not
growth stimulation by Trichoderma spp. and show any encouraging impact on siliqua length
other microbes has been reported in several and seed number of mustard and flower number
crops such as bean (Inbar et al., 1994), cucumber of tomato (Table 2). Perhaps, these characters of
(Kleifeld and Chet, 1992), maize (Björkman et mustard and tomato are more genetically
al., 1994), pea (Naseby et al., 2000) and tomato controlled i.e. poorly responsive. It has been
(Ozbay et al., 2004). reported that the strains of different Trichoderma
species showed variable responses in cucumber,
3.2. Response of Trichoderma-enriched loofah and bitter gourd (Lo and Lin, 2002).
biofertilizer on yield contributing Besides, it was reported that application of
characters Trichoderma spp dramatically increased the
number of fruits per plant in pepper and tomato
The sole application of Trichoderma-enriched grown in greenhouse than untreated control
biofertilizer (i.e. T3, T4 and T5) in mustard (Vinale et al., 2008).
 Trichoderma-enriched biofertilizer
Table 1. Effect of Trichoderma-enriched biofertilizer (BioF) alone or in combination with chemical fertilizers on the growth of mustard and tomato

Mustard Tomato
Treatments
Plant height Branch Leaf Plant height Branch Leaf
(cm) (no/plant) (no/plant) (cm) (no/plant) (no/plant)
T1 (Control) 49.07 d 2.6 c 9.06 c 60.42 a 7.41 b 27.07 d

T2 (NPK fertilizers only) 76.07 a 5.9 a 25.20 a 64.58 a 7.83 ab 61.40 bc

T3 (BioF/compost) 60.33 bcd 3.2 bc 9.73 c 66.00 a 9.25 ab 55.60 c

T4 (BioF/pellets) 32.97 e 1.8 c 6.40 c 59.42 a 7.75 ab 26.13 d

T5 (BioF/suspension) 34.40 e 2.5 c 7.93 c 46.50 b 4.00 c 29.30 d

T6 (50% N + 50% BioF/compost) 73.03 ab 5.4 a 21.40 ab 66.17 a 8.25 ab 69.73 ab

T7 (50% N + 50% BioF/pellets) 64.33 abc 5. 6 a 24.97 a 71.17 a 9.92 a 62.67 bc

T8 (50% N + 50% BioF/suspension) 52.90 cd 5.2 ab 13.67 bc 70.42 a 8.50 ab 73.77 a

CV (%) 10.61 20.46 24.56 8.14 12.26 7.76

LSD (P≤0.01) 14.02 1.97 8.66 12.24 2.3 9.39

In column common letters are not significantly differed at LSD (P≤0.01) level.

113
 114
Table 2. Effect of Trichoderma-enriched biofertilizer (BioF) alone or in combination with chemical fertilizers on yield contributing characters of
mustard and tomato

Mustard Tomato

Treatments Flower Flower Fruit

Flower Siliqua Siliqua Seed
cluster (per
(per cluster) (no/plant) length (cm) (no/ siliqua) (no/cluster) (no/plant)
plant)

T1 (Control) 3.08 c 31.20 bc 4.36 c 10.53 c 9.83 cd 5.45 bc 14.83 c

T2 (NPK fertilizers only) 8.08 a 55.53 ab 6.21 a 16.20 ab 19.22 ab 7.10 ab 36.83 a

T3 (BioF/compost) 5.25 bc 32.80 bc 5.82 ab 14.27 abc 21.67 a 7.00 ab 25.60 b

T4 (BioF/pellets) 3.08 c 29.20 bc 4.71 bc 15.67 abc 13.33 bc 6.17 abc 20.50 bc

Haque et al. /The Agriculturists 10(2): 109-119 (2012)

T5 (BioF/suspension) 5.58 b 24.07 c 4.56 bc 12.40 bc 4.75 d 4.67 c 13.70 c

T6 (50% N + 50% BioF/compost) 5.83 ab 55.80 ab 5.54 abc 16.47 ab 16.42 abc 6.45 ab 35.97 a

T7 (50% N + 50% BioF/pellets) 6.08 ab 62.40 a 5.80 ab 19.07 a 21.00 a 7.75 a 41.03 a

T8 (50% N + 50% BioF/suspension) 6.25 ab 43.07 abc 5.06 abc 13.00 bc 18.73 ab 6.33 abc 39.43 a

CV (%) 18.85 29.62 10.80 14.70 17.97 11.67 13.04

LSD (P≤0.01) 2.43 29.50 1.35 5.15 6.69 1.76 8.86

In column common letters are not significantly differed at LSD (P≤0.01) level.
Trichoderma-enriched biofertilizer 115

3.3. Impact of Trichoderma-enriched Generally, the NO3- form of N fertilizer couldn’t

biofertilizers on yield of mustard and retain long time to the rhizosphere zone of plants
tomato and therefore, split application of N fertilizer was
suggested at different stages of crop growth.
The highest seed yield (1.577 g) per plant of Conversely, the organic amendments and
mustard was found in T6, which was followed by microbes played a roles to slow by release the
T2, T3, T5, T7 and T8 treatments (Table 3). plant nutrients. The microbial population in soil
Similarly, the seed yield per plot of mustard was releases some exudates (organic substances),
also recorded the highest in T6, which was increases nutrient uptake through enhanced root
similar to T2, T3, T5 and T7. The highest yield growth or promotes availability of necessary
promotion (108.36 %) was attained in treatment nutrients and solubilize a number of poorly
T6 and the second highest was 81.90 in T2 soluble nutrients, such as Mn 4+, Fe3+ and Cu2+ etc
treatment (Table 3). These results implied that (Kleifeld and Chet, 1992; Altomare et al., 1999).
the treatment T6 could save at least 50% of N Therefore, in present study the combined
fertilizer. treatments enhanced efficient utilization of
nutrients than the others which expedited crop
Fruit yield of tomato per plant varied growth and yield.
significantly among the treatments (Table 3).
The maximum individual fruit weight was 3.4. Impact on dry matter production
obtained in T8, which did not differ significantly
from treatment T7 but was identical to T2. The Influence of different forms of Trichoderma-
minimum individual fruit weight was recorded in
enriched biofertilizers alone or in combination
control treatment (T1). The highest fruit yield per
plant (1.67 kg) was recorded in T8, which was with N fertilizer on dry matter production of
significantly (P≤0.01) different from with rest of mustard was also noticed in the present study
the treatments. N fertilizer supplemented (Table 4). The highest root (0.66 g/plant) and
treatments T6 and T7 gene the second highest shoot (6.83 g/plant) dry matter weight of mustard
yield, which was significantly lower than the T8 was found in T6 treatment. The lowest root dry
treatment, but significantly higher than the other matter (0.11 g) was recorded in T1, which was
treatments including T2. The lowest fruit yield followed by T4 treatment. Shoot dry weight
per plant (0.55 kg) was found in the control followed the similar trend as root dry weight.
treatment. Yield increase of 203% was recorded There clearly illustrated that dry matter
in the treatment T8, whereas 125 and 156% yield
production was boosted up by the
increase were observed in T6 and T7 treatments,
respectively over control. The yield increase of supplementation of partial N fertilizer to each
61.82% was observed in T2 over control biofertilizer application. The scenario of root and
However, 56, 31 and 9% were observed in sole shoot dry weight of tomato also followed the
biofertilizers i.e. T3, T4 and T5 treatments, similar profile as mustard (Table 4). Similar
respectively over the control. The yields were trend of results was noticed in corn shoot dry
significantly influenced by the combined weight using T. harzianum (Björkman et al.,
treatments i.e. T6-T8 by supplementation of N 1994). Increased root dry weight by application
fertilizer to the biofertilizers. These results of Trichoderma-enriched biofertilizer might be
implied that yield of tomato was significantly one of the important factors for enhancing
increased when Trichoderma-enriched
growth and yield of mustard and tomato.
biofertilizers were combined with chemical N
fertilizer (Table 3). The combined treatment Furthermore, the increased root biomass could
gave significantly higher yield of tomato than the increase the chance for acquisition of nutrients
treatment T2 i.e. standard dose of NPK fertilizer. by exploitation of more volume of soils.
 116
Table 3. Effect of Trichoderma-enriched biofertilizer (BioF) alone or in combination with chemical fertilizers on yield of mustard and tomato

Mustard Tomato

Treatments
Seed yield Yield Percent yield Individual fruit Fruit yield Percent yield
(g/plant) (g/plot) increased wt.(g) (kg/plant) increased

T1 (Control) 0.529 b 69.16 cd - 62.27 d 0.55 d -

T2 (NPK fertilizers only) 1.303 ab 125.80 ab 81.90 82.00 ab 0.89 c 61.82

T3 (BioF/compost) 0.913 ab 99.05 abc 43.21 71.87 c 0.86 c 56.36

T4 (BioF/pellets) 0.533 b 71.36 cd 3.19 71.03 c 0.72 cd 30.90

Haque et al. /The Agriculturists 10(2): 109-119 (2012)

T5 (BioF/suspension) 1.213 ab 101.54 abc 46.82 72.50 c 0.60 d 9.09

T6 (50% N + 50% BioF/compost) 1.577 a 144.1 a 108.36 74.43 bc 1.24 b 125.45

T7 (50% N + 50% BioF/pellets) 1.280 ab 101.1 abc 46.18 86.57 a 1.41 b 156.36

T8 (50% N + 50% BioF/suspension) 0.773 ab 79.27 bcd 14.18 87.17 a 1.67 a 203.63

CV (%) 33.98 24.04 - 4.49 9.0 -

LSD (P≤0.01) 0.88 51.16 - 8.14 0.21 -

In column common letters are not significantly differed at LSD (P≤0.01) level.
 Trichoderma-enriched biofertilizer
Table 4. Dry matter production of mustard and tomato as influenced by Trichoderma-enriched bioferlizer (BioF) alone or in combination
with chemical fertilizers

Treatments Mustard Tomato

Root weight/plant (g) Shoot weight/plant (g) Root weight/plant (g) Shoot weight/plant (g)

T1 (Control) 0.11 e 2.09 e 0.11 g 2.07 d

T2 (NPK fertilizers only) 0.34 c 5.02 c 0.38 d 4.36 c

T3 (BioF/compost) 0.27 cd 3.68 d 0.32 de 3.96 c

T4 (BioF/pellets) 0.14 e 2.45 e 0.28 ef 2.40 d

T5 (BioF/suspension) 0.19 de 2.05 e 0.21 f 2.54 d

T6 (50% N + 50% BioF/compost) 0.66 a 6.83 a 0.69 a 6.57 a

T7 (50% N + 50% BioF/pellets) 0.58 ab 5.70 b 0.59 b 5.42 b

T8 (50% N + 50% BioF/suspension) 0.54 b 5.26 bc 0.48 c 5.08 b

CV (%) 11.78 5.86 7.86 5.37

LSD (P≤0.01) 0.106 0.579 0.075 0.517

In column common letters are not significantly differed at LSD (P≤0.01) level.

117
118 Haque et al. /The Agriculturists 10(2): 109-119 (2012)

In general, the application of biofertilizers in and M/S Ismotara Begum, Proprietor, M/S
different form enhanced dry matter production of RASA Agro-Enterprise, Natore, Bangladesh for
mustard and tomato. The dry matter production providing three types of Trichoderma-enriched
was boosted up while biofertilizers were biofertilizers.
supplemented with N fertilizer. The growth,
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