EFFECT OF TRICHODERMA BIOFERTILIZER ON

Vigna radiate

PROJECT REPORT

SUBMITTED FOR THE DEGREE OF MASTER OF SCIENCE

IN BIOTECHNOLOGY

SUBMITTED BY SUPERVISED BY
Ms. AYUSHI NEGI Dr. POONAM RANI

DEPTARMENT OF BIOTECHNOLOGY MEERUT

INSTITUTION OF ENGINEERING AND TECHNOLOGY
MEERUT
(2019-2021)

1
 Candidate’s Declaration

I hereby certify that the work which is being presented in this project report entitled EFFECT
OF TRICHODERMA BIOFERTILIZER ON Vigna radiatafor fulfillment of the requirement
of the degree of Master of Science in Biotechnology submitted in the Department of
Biotechnology, Meerut Institute of Engineering & Technology, Meerut, (U.P.), India. This is
an authentic record of my own work carried out during the final yearof M.Sc. degree, under
the supervision of Dr. Poonam Rani, Assistant Professor, Department of Biotechnology,
M.I.E.T. Meerut.

Name Roll number Signature

AYUSHI NEGI 190979227004

This is to certify that the above statement the candidate is correct to the best of my
knowledge. I wish her for her future endeavor.

Dr. Poonam Rani

Assistant Professor
Department of Biotechnology
M.I.E.T., Meerut- 250002

2
 ACKNOWLEDGMENT

In the accomplishment of this project successfully, many people have best owned upon me
their blessing and the heart pledged support, this time I am utilizing to thank all the people
who have been concerned with this project.

Primarily I would thank to god for being able to complete this project with success. Then I
owe my deep gratitude to my project guide Dr. Poonam Rani, whose valuable guidance has
been the ones that helped me patch this project and make it full proof success. Her
suggestions and her instructions have served as the major contribution towards the
completion of the project.

I am thankful and fortunate enough to get constant encouragement, support and guidance of
my family, my colleagues, friends and all teaching and non-teaching staff of the department
which helped me in successful completion of my project.

AYUSHI NEGI

3
 ABSTRACT

Microorganism such as bacteria, fungus, are mainly considered as pathogens but some
beneficial fungus such as Trichoderma viride can be used as plant growth promoter. T. viride
powder in market is sell as bio fertilizer. The present study is conducted to observe effect of
Trichoderma viridebio fertilizer on plant growth and development. For this soil was collected
in bulk from CCS Biotechnology herbal garden, MIET, Meerut and processed for
experimental purpose. Then three treatments C,T5,T10 and T15 in triplicates were prepared.
Plant species Vigna radiate was selected for experiment as it can easily be grown and
maintained. Pre-treated seeds of V. radiate were sown in different treatments and seed
germination percentage was calculated along with shoot height, root length and vigour
indexafter seed germination where parameters except seed germination percentage were
measured and calculated on 3rd,5th and &7th day. Results showed that seed germination
percentage, shoot height, root length and vigour index were high in T10 treatment. On the
basis of results it can be concluded that 10 gm T.viride in 1 Kg soil is better for the seed
germination and growth parameters of Vigna radiate.

Key words:-Trichoderma viride, Vigna radiate, seed germination, seed vigour index,
biofertilizer.

4
 CONTENTS

ACKNOWLEDGEMENT
ABSTRACT
CONTANTS
CHAPTER
1. INTRODUCTION.
1.1- Introduction of Trichoderma.
1.2- Ecology of pulses.
1.3- Ecology of green gram.
1.4- Aims and objective of use of Trichoderma
1.5-Work Plan.

2. REVIEW AND LITERATURE.

3. MATERIAL AND METHOD.
3.1- Soil collection and processing.
3.2- Plant material.
3.3- Seed germination experiment.
3.4- Ecology of plant samples.
3.4.1- vignaradiata
3.5- pH of the soil.
3.6- Seed germination percentage.
3.7- Seed vigour index.
3.8- Plant Attributes (Shoot height and Root length)
3.9- Statistical Analysis.

4: OBSERVATION
4.1 Table no.1. Seed germination percentage, seedling shoot height, seedling root length and
seed vigour in different treatment in Vigna radiate.
4.2 Table no.2. pH of the Soil treatments.

5
5: RESULT AND DISCUSSION
5.1. Soil analysis and parameters.
CONCLUSION
SUMMARY
REFERENCE

6
 INTRODUCTION

1.1 INTRODUCTION OF TRICHODERMA:

Trichoderma - A bio-control agent for management Soil borne diseases. Trichoderma is a
very important and effective biological mean for plant diseases, who restrict all the diseases,
especially the soil borne diseases. Trichoderma is a free-living fungus which is easily found
in soil and root ecosystem. It is highly interactive in root, soil, and leaves environment. It
restrict the growth of pathogens, who disturb the plant growth and soil nutrition. (Gary E.
Harman, Charles R. Howell, Ada Viterbo, Ilan Chet and Matteo Lorito, 2004)

Scientific classification
Kingdom Fungi
Division Ascomycota
Class Sordariomycetes
Order Hypocreales
Family Hypocreaceae
Genus Trichoderma
Species T.viride

Binomial name: Trichoderma viride

The biostimulant effect of Trichoderma spp. on horticultural crops are highly variable. Thus,
practical use of Trichoderma sp. requires feasible formulated products and suitable substrates.
Trichoderma viride work as a fertilizer. Nowadays, there is a growing interest in the use of
beneficial microorganisms in agriculture, and some of them are being commercialized as
biopesticidal and biofertilizer formulations (Zilli et al., 2019). Fungi of the genus
Trichoderma are among the most studied, due to their multiple effects on plants, acting as
disease biocontrol agents (Harman, 2011) on the growth and development of plants and, also,
for their ability to induce defense responses against phytopathogens, insect damage and
abiotic stress (Vitti et al., 2015) Basically Trichoderma works as a fertilizers and that helps

7
in the growth On plants Plant species Vigna radiata was chosen for the experimental purpose
because the plant species are easy to grow , generally used by human beings for daily
domestic purposes and saw the effect of Trichoderma on that. Growth note in alternative days
and check them in daily bases and saw the effect of different-different amount of
Trichoderma’s effect. The effect of Trichoderma spp. In promoting plant development can be
observed in the germination of the seeds, rooting, sprouting of cuttings, growth of branches,
increase of leaf area,accumulation of organic matter.

Benefits of Trichoderma:

Disease control: Trichoderma is a biocontrol agent and used forsoil borne diseases. It works
successfully against the pathogenic fungi belonging to various genera, viz. Fusarium.(Rakesh
kumar singh, 2010) Plant growth promoter: Trichoderma also work as a fertilizerwho is
enhance the plant growth. Plant grow very rapidly, with the leaves and shoot height and root
length, which increases the plant ability to resist drought. (Rakesh kumar singh, 2010)

Bioremediation: Bioremediation is a process where biological organisms are used to remove

harmful environmental pollutant bymetabolic process. Trichoderma strains play an important
role in the process of bioremediation of soil that are contaminated with pesticides and
herbicides. Trichoderma have an ability to restrict the growth of insecticides:
Organophosphates and carbonates. (N. Rana singh, A. Saurabh and M. Nedunchezhiyan,
2006)

Bio fertilizer: Trichoderma species can easily improve the plant health even in the absence of
harmful pathogens. Fungal can easily grow in soil and make such useful environment and
work as a fertilizer. (Helga George, 2019) Antibiotic production: Trichoderma produce the
chemicals that are toxic to the fungi. Some compounds are volatile and travel through the air.
(Helga George, 2019) Trichoderma is most useful and important for all types of plant and
vegetables growth such as cotton, tomato, potato, pulses, sunflower, ginger, turmeric, tea,
pepper, red gram, black gram, green gram etc.Trichoderma is also used for seed treatment.

8
 1.2: ECOLOGY OF PULSES:

In this process we did comparison of two pulses, to see the impact on Trichoderma on the
plant growth.

1.3: ECOLOGY OF GREEN GRAM (MUNG BEAN or MOONG)

Botanical name: Vigna radiata

Family: Leguminoseae
Origin: India and Central Asia

Fig.1. Vigna radiata

Area and Distribution:

Green gram is cultivated since ancient time and India is the largest producer of this legume.
Green gram is cultivated in the countries of India, Burma,Srilanka, Pakistan, China, Fiji and
Africa. In India almost all the state produce Green gram in large quantity. It is grown in huge
amount. In India some states production rate is very high and those states are: Odisha,
Maharashtra, Andhra Pradesh, Madhya Pradesh, Gujrat, Rajasthan and Bihar. (Pallvi G)

9
Importance of Green gram:

Green gram is also known as Mung bean or moong dal. Green gram is a richest
source of protein(25%). Mung dal consume in various way as a Dal, as a sprouts and also
make a good or healthy sweet dish.(netmeds 2020)

Green gram is also used as green manure crop, and most important it have an ability to fix the
atmospheric nitrogen, and also help in preventing soil erosion. It is a short duration crop, and
it can be use as a cattle food, and various health benefit features in green gram.Mung dal is
one of the richest source of plant based protein and in mung dal have 211 calories and
14.2gm of protein. Without increasing the fat, mung dal help to increase the protein level in
the body.(netmeds 2020)

Mung dal also help to reduce the cholesterol level, reduce the symptoms of constipation and
reduce the risk of developing heart diseases. Mung dal also take as a sprouts forms, sprouts
changing the nutrition level and more beneficial for our health. (netmeds 2020)

Nutrition
Calories – 212
Fat - 0.8 grams
Protein - 14.2 grams
Carbs - 38.7 grams
Fibre - 15.4 grams
Folate (B9) - 80% of the Reference Daily Intake (RDI)
Manganese - 30% of the RDI
Magnesium - 24% of the RDI
Vitamin B1 - 22% of the RDI
Phosphorus - 20% of the RDI
Iron - 16% of the RDI
Copper - 16% of the RDI
Potassium - 15% of the RDI
Zinc - 11% of the RDI
* Source as Per USDA (netmeds 2020)

10
Green gram is powerhouse of antioxides, and also very beneficial for our health and help to
fight against all the diseases, rich source of protein, iron etc. Green gram also increase the
immunity and help to protect from all diseases.
Control Blood pressure: High Blood Pressure is a risk factor for heart diseases and stroke
which are a leading cause of mortality. The richness of potassium and magnesium and fibre
in green gram to regulate blood.
Cognitive health: Green gram is full overpower with iron and protein which helps in carrying
oxygen and nutrition in the blood to all the vital organ and tissues.
For Weight Loss: Green gram is a rich source of all the powerful nutrition and also help to
manage fat in our body. Green gram is also use as a diet food, and for the healthy diet moong
dal is also take as a sprouts and as a healthy saladin our diet.
Green gram is a full of nutrition values like vitamins, protein, magnesium, fibre etc. Adding
mung bean is a simplest and convenient way to maintain overall health and when consumed
as a part of a balanced diet it may help to prevent obesity, lowers cholesterol regulates blood
pressure and blood sugar levels, lowers inflammation and prevents cancer. This versatile dal
melds well with all the ingredients and can be prepared in a multitude of ways, add them in
your daily diet.

1.4- Aims and objective of present work

1. To observe effect of Trichoderma powder on pH.

2. To observe the effect of Trichoderma fertilizer on seed germination percentage
3. To observe the effect of Trichoderma fertilizer on plant attributes i.e. shoot height,
root length.
4. To observe the effect of Trichoderma fertilizer on vigour index.

11
1.5: Work plan

1. Collected bulk fertile soil sample from the MIET herbal Garden.
2. Prepared soil to make treatments (triplicate) for experimental purpose.
3. Collected plant species (botanical name) seed from the Amazone.com .
4. Procured Trichoderma fertilizer from the Amazone.com for present study.
5. Soil bed was prepared as control and treatments.
6. Seeds were pre-treated and sown.
7. After 5 days radicles and plumes were visible
8. After Seed germination seedlings were uprooted and saplings growth were recorded
on 3rd ,5th and 7th day
9. Experiment was set up in random design.
10. Raw data was compiled and standard analysis was performed.

12
 2: REVIEW AND LITERATURE

Beneficial bacteria and fungi are used to prevent diseases and enhance plant growth since
ancient time and are considered as plant growth promoting microbes because they are found
in association with the roots of various plant species (Kloepper et al., 1991;Sajjad et al.,
2001; Shanmugaiah 2005, 2007). These microbes may occupy the rhizosphere, root or leaf
surfaces or may colonize intracellular spaces and vascular tissues inside the plant (Preston,
2004).

They encompasses symbionts relationship, protect their host against various aggressions,
determine the ecological success of plants. They may modify plant communities to improve
the inventory of diversity and functions of in situ plant-associated microorganisms (Selosse et
al., 2004). Tricoderma sp. showed effective result in plant growth promotion and disease
control in a wide range of crops (Manjula et al.,2002).

Trichoderma also use as a bio pesticides which help to protect the various crop plants from
the pest. Application of Trichoderma formulates with strain mixture perform better than the
individual strain. (Sanjeev kumar, et al, 2014). Now day rice cultivation faces many
environmental problems caused by the chemical bio fertilizer. Trichoderma used in the rice
field and observed that the increases the nutrition and soil fertility, which help in the plant
growth. (SanjitDebnathet al 2020).

Trichoderma have a high biotechnological value, with several biological control agents.
Trichoderma work as a biocontrol agents as well, they have other beneficial effects on plants
and promote the plant growth.(Rosa Hermosa et al.,2013). Trichoderma helps in the plant
growth and i.e.

also known as growth promoters. (Robert hill et al 2014).Trichoderma has an ability to act as
biocontrol agent against the plant pathogens. Trichoderma spp. are also used as a pathogens
control and increase the efficiency of crops. (Gary E Harman, 2006).

Trichoderma is the most commonly isolated soil fungi, which help in the plant growth and
protect the plants and contain the pathogens population under the different soil condition.
(Sheridan L Woo et al 2008).

13
Trichoderma species are effective in the control of soil/seed borne fungal diseases in different
crop plants (Kubicek et al., 2001). Trichoderma sp. and other beneficial root fungi also
improve the plant growth and productivity (Balasubramanian, 2003).

Trichoderma sp. has been reported beneficial for many crops such as beans (Phaseolus
vulgaris) cucumber (Cucumissativus), pepper (Capsicum annum), carnation (Dianthus
carophyllus), maize (Zea mays), and wheat (Tritichumaestivum) (Lo and Lin, 2002).

Pseudomonas species which mixed with Trichoderma are effective root colonizers and
biocontrol agents, the production of antibiotics and other antifungal metabolites including
antibiotics, hydrogen cyanide and siderophores (O'Sullivan and O'Gara, 1992). They are
Increased dry weight and plant height were recorded with Pseudomonas sp.MML2212 and
Pseudomonasfluorescens on rice and green gram when compared with the control
(Mathivanan et al., 2005, Shanmugaiah et al., 2005, 2008).

To overcome the problem by help of the beneficial microbes in a biocontrol prepration to

combine the diseases suppressive activity. (Meyer & Roberts, 2002). Considerable evidence
has been collected in recent years to support and identify the benefits associated with the use
of endophytic bacteria in crop protection (Siddiqui &Ehteshamul-Haque, 2001; Hallmann et
al., 1995; 1997, 1998; Tariq et al., 2009).

These include phosphate solubilization activity (Altomare et al., 1999), indole acetic acid
production (Tariq et al., 2009) and production of a siderophore (Leong, 1986). Among the
endophytic plant growth promoting bacteria, Pseudomonas also have has been shown to
improve plant growth and it is known to synthesize growth stimulating plant hormones
(Ehteshamul-Haque et al., 2007; Tariq et al., 2009).

Similarly Trichoderma spp., are free living fungi that are common in soil and root ecosystem
are well known for their biocontrol potential (Ehteshamul-Haque&Ghaffar, 1992; Qureshi et
al., 2012; 1990; Woo et al., 2006). supplemented with trimethoprim (Gould et al., 1985).

Nowadays microbial inoculants have been used as an alternative technique of commercially

available chemicals to promote growth of both bulb and non bulb plants (Pareek et al.
2017).plant nutrition due to the uptake of Pand other elements by extra radical mycorrhizal
hyphae(Abdelhameed and Metwally 2019)

A large number of antibiotics such as polyketides, trichodermin, peptaiboils, trichodermol

and steroids are produced by the Trichoderma species which encourage plant growth

14
(Harman et al. 2012). Calvet et al. (1993) has reported a synergistic effect. T. viride and their
combination increase the area in onion leaf with AM. Our findings are comparable to the
results of Mwangi et al. (2009). Some reported that co-inoculation of T. harzianum and AM
fungi increased plant heights and shoot and root dry weights of tomato seedlings and tea
cutings.

Rasool et al., (2011) reported that the increasing in seedling growth and nutrient uptake in
tomato by Trichoderma isolates Bhattari and Hess (1993) found that the increased yield
response in cultivation of spring wheat (Triticumastivum L.) with Azospirillium spp. of
Nepalese origin. Yadav et al., (2000) also found that Azotobactor increases yield and nitrogen
economy in wheat under field condition.

Over all the treatments with Azotobacterchroococcum + Pseudomonasstriata +

Trichodermais in concordance to find the highest growth rate of length and diameter of
seedlings. Mishustin and Shilnikova (1969), Azcon and Barea (1975), Trappe (1977), Lin et
al., (1987) and Barbara and Wong (1989). Bagyaraj and Manjunath (1980) , Smith (1980) ,
Maronek et al., (1981), Mishra and Kapoor (1986) and Verma (1993) have also reported that
the association of microorganism like Azotobacterchroococcum with Pseudomonasstriata,
Azotobacterchroococcum with Trichodermaviride were found beneficial in the availability of
nutrients.

Azotobacterchroococcum + Pseudomonasstriata + Trichodermaviride. Sundara Rao (1968)

and Dobreiner and Day (1974) have also reported the accumulation of more nutrients in
leaves due to the metabolization and assimilation through roots with the help of
microorganisms. Trichodermaspp. have been extensively studied as potential biocontrol
agents (e.g. Lynch 1990; Papavizas 1992). Trichodermaspp. can stimulate the growth of a
number of vegetable and bedding plant crops (e.g. Baker 1988; Lynch et al. 1991a,b ).

Lynch et al. (1991a,b) investigated the effect of Trichoderma on the growth of lettuce, and its
ability to control damping-off diseases caused by Rhizoctoniasolani and Pythiumultimum.
The direct effect of Trichoderma strains on lettuce establishment and growth in the absence
of pathogens. To reduced the emergence time of seedlings compared to the controls the
fungal treatments help. From their results, and those of Ousley et al. (1994) ,

Specific Trichodermastrains have the potential to consistently increase plant growth ( Lynch
et al. 1991a ). Ousley et al. (1993) investigated the effect of autoclaving plant growth
promoting Trichoderma isolates on the germination and growth of lettuce. Some strains

15
turned out to be good plant growth promoters.Trichoderma is a balance between growth
inhibiting and growth promoting properties and autoclaving some strains alters this balance.
Some commercial preparations of Trichoderma are available for controlling plant diseases
( Coley-Smith et al. 1991 ),. The use of biocontrol and plant growth promoting strains of
Trichoderma may be a useful asset in

Mungbean (Vignaradiata) is a key legume crop grown predominantly in South and Southeast
Asia. Biotic and abiotic stresses cause significant yield reduction in mungbean, and among
these, fungal diseases are particularly important. Although disease management practices,
including physical, chemical, and biological methods have been researched and described in
the literature, few of these are available or have been used by growers. Here we review the
economic impact, and sustainable management options for the soil-borne and foliar fungal
diseases of mungbean as well as major challenges to manage these diseases. Potential use of
all possible components of integrated management practices including host resistance,
fungicides, biocontrol agents, natural plant products, and cultural practices etc. are discussed.
Major diseases include powdery mildew, anthracnose, Cercospora leaf spot, Fusarium wilt,
Rhizoctonia root rot and web blight, Macrophomina charcoal rot/dry root rot and blight.
Review of the literature indicated an absence of resistance to Rhizoctonia root rot, little
sources of resistance for dry root rot and anthracnose. Major resistant genes (R genes) and
quantitative trait loci (QTL were identified for powdery mildew and Cercospora leaf spot,
which may be potentially used in Marker assisted selection (MAS).

Although the mechanisms of induced systemic resistance (ISR) by biocontrol agents have
been studied with Macrophomina blight, there is little information on the mechanisms and
use of systemic acquired resistance (SAR) in managing fungal diseases of mungbean. Several
studies targeted exploiting biological control for soil-borne root rot diseases. Botanical
products, such as plant extracts, are also found effective to manage root and foliar diseases.
However, many of these studies were limited to laboratory and/or green house experiments.
Thus, long-term field studies are required for further exploitation of biological methods and
commercial applications.

Mungbean mainly is grown in rain-fed climates and variability in climate such as elevated
temperature and CO2 within the rain-fed ecologies leads to varying intensities of biotic stress
(Chakraborty et al., 2000; Sharma et al.,2007) which cause significant loss in production.
Foliar and root rot fungal diseases are major production constraints in South Asia and South

16
East Asia. Charcoal rot/dry root (Macrophominaphaseolina) and
Rhizoctoniaroot(Rhizoctoniasolani) are economically important soil-borne vascular diseases,
causing wilt and root rot complex (Alam et al., 1985; Iqbal and Mukhtar, 2014).

Among majorsoil-borne vascular diseases, dry root rot and wilt is a major concern sincethe
pathogen affects the plant during all growth stages and subsequently causes significant yield
loss. Yield loss due to dry root rot was reported to be 11% in Northern India (Kaushik et al.,
1987) and up to 44% in Pakistan (Bashir and Malik, 1988).

Dry root rot was also reported first time in Shanxi province of China in 2010 (Zhang et al.,
2011). Mungbeanplants with wilt and root rot symptoms, with incidence of 80–90% in
susceptible genotypes,was also reported in 1979 in southwest Ontario (Anderson, 1985).
Anthracnose (Colletotrichumlindemuthianum or C. truncatum or C.gloeosporioides) (Shen et
al., 2010), Cercospora leaf spot (Cercosporacruenta or C.canescens or C. kikuchii or
C.caracallae) (Joshi et al., 2006), and powdery mildew (Figure 1h) (Erysiphepolygoni or
Podosphaerafusca) (Ryley et al., 2010), Macrophominablight (M. phaseolina) and web blight
(R. solani) (Alam et al.,1985; Iqbal and Mukhtar, 2014) are the major foliar diseases of
mungbeanas causing yield loss ranging 20–60% in different continents.

A wide range of yield losses (23–96%) due to Cercospora leaf spot was reported from field
trials conducted at different states of India (Kaur, 2007; Chand et al.,2012; Bhat et al., 2014)
and up to 61% from Pakistan (Iqbal et al., 1995).The impact of powdery mildew on
mungbean also was reported from different countries. Yield loss due to powdery mildew was
reported up to 21% in the Philippines (Quebral and Cowell, 1978), up to 40% in Australia
(Kelly et al., 2017), and from 20 to 100% in different regions of India. Yield losses from
powdery mildew was reported 35% from Gujarat, western India (Khunti et al., 2002), 20–
40% from Chhattisgarh, central-eastern India (Khare et al., 1998) and 20–40% in
Maharashtra, western-central India (Mandhare and Suryawanshi, 2008), and from 9 to 50% in
Uttarakhand and Uttar Pradesh of Northern India (Pandey et al., 2009).

Reddy et al. (1994b) also reported 100% loss from Maharashtra State, India due to powdery
mildew diseases at seedling stage. A wide range of yield losses (24–67%) due to anthracnose
disease was estimated from several mungbean growing areas in India (Deeksha and Tripathi,
2002; Kulkarni, 2009; Shukla et al., 2014). Alternaria leaf spot (Alternariaalternata) is also
reported in South Asia, but economic impact is minor i.e., only 10% loss reported from
Jammu and Kashmir, India (Maheshwari and Krishna, 2013).

17
Web blight has been a problem for several decades in Pakistan (Alam et al., 1985) and in
India, where it was reported from diverse geographical areas including Kanpur and Uttar
Pradesh, northern India (Dwivedi and Saksena, 1974), Punjab, northwest India (Bains et al.,
1988), Madhya Pradesh, central India (Tiwari and Khare, 1998), Rajasthan (south India),
Bihar, Haryana, and Himanchal Pradesh (northern India) (Anonymous, 2014). About 30–40%
of yield loss due to web blight was reported from Rajasthan (Anonymous, 2000) and 20–40%
seedling mortality due to Rhizoctonia infection was reported from Jabalpur, central India
(Tiwari, 1993).

There is a little information available regarding the dynamics in the prevalence and incidence
of diseases in mungbean at temporal and spatial scale. Fusarium wilt caused by
Fusariumoxysporum and/or F. solani was a minor disease of mungbean in Australia.
However, the incidence and severity of the disease has increased substantially in recent years
and yield losses of up to 80% were reported in the susceptible mungbean cultivars (Kelly,
2017).

Outbreaks and spread of diseases were reported in other legume crops, such as soybean
(Sconyers et al., 2006) and chickpea (Sharma and Ghosh, 2017). For example, Asian soybean
rust (Phakosporapachyrhizi) was a problem in Asia and South America, but then spread
rapidly across eight states of southeastern United States within a few years of first detection
in Louisiana in 2004 (Sconyers et al., 2006).

It was speculated that an extreme weather event (hurricane) was responsible for the
introduction and spread of the Asian soybean rust. Sharma et al. (2015) also reviewed dry
root rot (Macrophominaphaseolina) as an emerging disease of chickpea in semiarid tropic
region and disease intensity has been increased in a past decade. They further speculated that
changes in weather pattern, such as high temperature and drought stress during reproductive
stages of the chickpea increased the dry root rot intensity. Climate change could have positive
or negative or neutral impact on the dynamics of crop diseases depending on the types of
crops, diseases or geographical regions (Luck et al., 2011).

Climate change would increase average global temperature, CO2 level, and cause more
extreme rain/drought events (Meehlet al., 2005). Severity of some diseases, such as brown
spot of soybean (Septoriaglycines) and sheath blight of rice (Rhizoctoniasolani) increased
with elevated levels of CO2 (Kobayashi et al.,2006; Eastburn et al., 2010), whereas variable
results were reported for powdery mildew of wheat and barley (Thompson et al., 1993;

18
Hibberd et al., 1996). In South Asia, spot blotch in wheat (Cochliobolussativus) has increased
substantially in recent years and it is speculated that elevated night temperatures due to
climate change has contributed to this (Sharma et al., 2007).

In Australia, root and crown rot of wheat (Fusarium pseudograminearum) is expected to

increase due to climate change as the disease was high with elevated CO2, temperature, and
drought (Melloy et al.,2010). There is no information available on the impact of climate
change on the dynamics of mungbean diseases. However, based on knowledge of similar
pathogens/diseases in other crops, we can speculate that the pressure of soil-borne diseases
caused by Fusarium, Macrophomina, and Rhizoctonia in mungbean may increase due to
climate change. It is difficult to predict the effect of climate change on foliar diseases since
they are influenced by the combination of temperature, rainfall and relative humidity, which
can't be precisely predicted in most situations of climate change. Use of different cultural
practices and physical methods to eliminate seed- borne pathogens were found effective to
reduce the foliar and root rot diseases of mungbean in fields.

Field sanitation, crop rotation, removal of crop debris and weed hosts in the vicinity of the
crop reduced the Cercospora foliar blight in mungbean (as reviewed in Sharma et al., 2011)
Removing root rot infected mungbean plants reduced sclerotialoads in the field and delayed
sowing and maintaining wider spacing between the plants reduced powdery mildew
incidence (as reviewed in Satyagopal et al., 2014).

Plastic mulching increased sclerotial mortality of M. phaseolina and reduced pathogen

infection (Yaqub and Shahzad, 2009). Mungbean seed treatment with gamma rays (60Cobalt)
for 0–4 min and 90 days of storage had a suppressive effect on root rot fungi (Ikramet al.,
2010). Hot water emersion treatments (55–65°C) were effective to eliminate seed-borne
infection with Colletotrichumacutatum and C. gloeosporioides of mungbean (Lee et al.,
2007).

In South Asia, mungbean is commonly rotated with rice and wheat. It is reported anecdotally
that root rot diseases in mungbean have been increased in South Asia and other Asian
countries due to continuous rotation with rice. Several soil-borne pathogens, such as
Rhizoctonia, Fusarium etc. are common problem in rice and wheat (Kobayashi et al., 2006;
Melloy et al., 2010).

These fungal genera also infect mungbean, but more studies are required to determine if the
same species and strains also infect mungbean. If it is practical, adding diversity in the crop

19
rotations would help for the sustainable management of these soil-borne diseases in
mungbean. Crop diversification and use of diverse cultural practices, such as crop rotation,
plant residue management, adjusting the planting dates etc., are recommended as effective
strategies for managing crop diseases in conditions of climate change (Juroszek and von
Tiedemann, 2011).

Use of host–resistance is an effective, economical, and eco-friendly method for managing

mungbean fungal diseases. In this section, we synthesize the available information regarding
the identification of sources of resistance,available methods for efficient and reliable disease
reaction phenotyping,identification of molecular markers associated with disease resistance
genes and their potential use to improve disease resistance traits in mungbean. Trichoderma
are a very vast and useful group of microorganism which play an important role in plant
growth (Lidia et al., 2014). Plants or Crops losses before the crop harvesting just because of
plant pathogens, such as viruses, bacteria, fungi etc. plant pathogens still controlled by the
different techniques, one of the technique is to use the Trichoderma which can directly kill
the plant pathogens (Roberto N. Silva et al., 2019). Trichoderma spp. widely used in
agriculture application i.e. also known as biological control mechanism .The use of
Trichoderma spp. in agriculture and industry order. (Noor A Badaluddinet al., 2020).

Trichoderma play an important role in the environment, Trichoderma comprise edible and
medicinal mushrooms but also the pathogens of humans. (Lidia Blaszczyket al, 2014). As we
know bio fertilizer may be better option as eco-friendly to maintain the soil fertility. The
Trichoderma help in the vegetative growth of plant species. (Sanjay Mahato and Srijana
Neupane.2017)

Reliable and efficient methods are available to screen for reaction to foliar diseases of
mungbean including Cercospora leaf spot, powdery mildew, and anthracnose. Screening of
these foliar diseases can be successful in natural field conditions where disease pressure is
high, or if artificial inoculation with pathogen spores is available (Iqbal et al., 2004; Yadav et
al., 2014a,b).

For other foliar diseases caused by hemibiotrophic (Cercospora spp.) or necrotrophic

pathogens, disease can also be evaluated in the greenhouse with artificial inoculation. Several

20
disease rating scales/systems were developed to assess foliar diseases of mungbean
(Wongpiyasatid et al.,1999; Khunti et al., 2005; Suryawanshi et al., 2009).

Mungbean germplasm accessions can be screened by inoculating with the pathogen in the
controlled environments. Reliable and efficient methods were developed for
screeningmungbean seedlings against powdery mildew in the greenhouse (Wongpiyasatid et
al., 1999; Kasettranan et al., 2010); and in the laboratory using a detached leaf assay (Reddy
et al., 1987). For the assessment of foliar diseases, both qualitative and quantitative rating
scales were used (Reddy etal., 1994b; Wongpiyasatid et al., 1999; Khunti et al., 2005;
Marappa,2008; Suryawanshi et al., 2009).

Root rot and wilt diseases are sporadic and highly variables due to genotypes × environment
(G × E) interaction, therefore it is very difficult to get consistent results while screening in
natural fields. Therefore, host genotypes are usually screened by inoculation at seedling
stages in controlled environment for soil-borne diseases (M. phaseolina, R. solani, and F.
solani). Different methods such as paper towel (Khan and Shuaib, 2007) and sick pot/field
inoculation methods by inoculating the fungus grown in sorghum or maize grains (Dubey et
al.,2009; Choudhary et al., 2011) were used for the evaluation of root rot disease.Sources of
resistance against powdery mildew, Cercospora leaf spot, anthracnose, Macrophomina blight
and dry root rot have been identified the majority of studies targeted resistance to Cercospora
leaf spot and powdery mildews and were conducted in the field.

There have been fewer studies to identify root rot and anthracnose resistance sources, and
these were conducted in both lab/glasshouse and field experiments. Most of the identified
resistant materials were derived from cultivars/recombinant lines /breeding lines/land races;
however, some were from wild relatives (Marappa, 2008) and mutant lines (Wongpiyasatidet
al., 1999). Since screening trials for resistance against Alternaria leaf spot, anthracnose, and
root rot diseases are limited, more attention is required on these. These resistant lines from
difference sources can be utilized as donors for developing resistant varieties.

21
 3: MATERIAL AND METHOD

3.1: Soil collection and processing

Garden soil was collected from Herbal garden at backyard of Department of Biotechnology
and Microbiology, MIET, Meerut, (U.P), India (Figure 3). Soil samples were collected in
zipped polybags in triplicate, and transferred to the laboratory for analysis. In laboratory soil
samples wereprocessed such as crushed, sieved, and properly mixed and allow to air dried
until constant weight was achieved.Treatments (Tc,T5,T10,T15) were prepared as
control(Tc) where 100 % pure herbal garden soil was taken, treatment(T5) made by adding
5gm Trichoderma fertilizer in 100% pure soil, and T10,T15 treatment contain 10gm and 15
gm Trichoderma fertilizer in 100% pure soil successively. After that control and treated soil
was filled in plastic glasses in equal quantity and rest for some time after sprinkling water on
them to make soil soft and favourable for seed germination.

Figure 2. Map of the study site. (Uttar Pradesh) Figure 3. Map of Department of
Biotechnology Garden (MIET College)

22
 3.2: Plant material:

Seeds of selected plant species (Vigna radiate) and Trichoderma fertilizer were purchased
from Amazone.com.

3.3: Seed germination experiment:

Equal in size and healthy and size Vigna radiate was selected and dipped in water for 24
hours. Seeds were settle down in bottom of beaker were chosen for pot experiment. These
selected seeds were aseptically sown in amended and unamended pots. Each experiment was
performed in the variety of plants. Water spray when required in 24 hours interval starting
from the 3 day of showing up to 7 days and germination percentage was obtained.
Appearance pf plume on the soil surface was considered as germination.

3.4: Ecology of plants samples:

3.4.1: Vigna radiate

Vigna radiate, Moong bean or Green gram of family Fabaceae, is native to subcontinent and
mainly cultivated in India, Pakistan, China, Thailand, Bangladesh. In India almost all the
state produce green gram in large quantity. It is grown in huge amount. In India some states
production rate is very highand those states are: Odisha, Maharashtra, Andhra Pradesh,
Madhya Pradesh, Gujrat, Rajasthan and Bihar etc. It is adapted to wide range of well drained
soils, but it is best suitable for fertile sandy loams. It is upright annual legume ranging in
height from 15 cm to 1m; the average height of mature plant is 0.9m. Junctions of the
branches and stems are stipule. The first flowers appear seven to eight weeks after planting
and the crops reaches maturity in 12 to 14 weeks. Pods are borne at top of plant. Seeds are
green and almostglobular. Pods are covered with long, spreading, deciduous silky hairs
ofpulses food crop plants. Green gram is also known as Mung bean or moong dal. Green
gram is a richest source of protein (25%). Mung dal consume in various way as a Dal, as a
sprouts and also make a good or healthy sweet dish.

23
METHODS

3.5: pH of the Soil.

Soil pH are measured in 1:1 water: soil ratio. The entire process willtake about five minutes
per sample for preparation and analysis. Plus fifteen minutes of shaking time for each batch.
10 g of soil are used for this analysis. One standard check sample is include in each batch.

Principle

Soil pH can be measured at either 1:1 water. Soil ratio According to NAPT, they are very
similar 1:1 soil-water pH = 0.99, soil: water– 0.04.

can be measured in the field and lab testing is pH inherently inaccurate.

Equipment

Hanna HI5522 with appropriate probes

Small beaker with glass stir rod OR Falcon tube with cap.

Reciprocating shaker(if using falcon tube)

Reagents

pH calibration standards( 4.01, 7.01,10.01)

DI Water.

Procedure

Measure 10 grams of soil into the beaker or Falcon tube.

Add 20 ml of DI water.

Stir frequently or place on a shaker for 15 minutes.

Calibrate the pH and according to the instruction on the screen.

24
Insert rinsed PH,temperature probes into the each sample. extent possible, keep the probes
still and not against the wall or bottom the vessels. This may require the tipping of the vessel,
using a vessel that is tall and this, or decanting the supernatant in a test tube. Record results
when the numbers stabilize.When not measuring, make sure that the probes are rinsed and
submerged in 3M KCL. Between measurements, rinse each probe well with DI water and
gently pat dry.

3.6: Seed Germination Percentage

Germination percentage is an estimation of the viability of seeds, calculated as total number

of germinated seeds to the total number of seeds sown expressed in percentage (Rani et al.,
2016).

Germination percentage (%) = Total no. of germinated seeds × 100.

Total no. of seeds

3.7: Seed vigour index (SVI)

This is calculated by determining the germination percentage and seedling length of the same
seed lot. Seed vigour index is calculated by multiplying germination (%) and seedling length
(shoot and root length).The seed lot showing the higher seed vigour index is considered to be
more vigorous (Abdul-Baki and Anderson, 1973.)

Seed Vigour Index = germination percentage x seedling length (shoot height + Root length)

3.8: Plant Attributes (Shoot height and root length):

Plant shoot height and root length were measured with the help of measuring scale (cm).

3.9: Statistical Analysis:

The statistical analyses were conducted to determine mean and standard deviation using MS-
Excel 2007 (Microsoft Inc.).

25
 4. OBSERVATION:

Table:1. Seed germination percentage, seedling shoot height, seedling root length and
seed vigour in different treatment at different interval in Vigna radiate. (Mean ±S.D)

DAYS Parameter Treatments

C T5 T10 T15
Seed Germination 73.00±27.82 99±0.62 100.00±00.16 100.00±05.02
Seedling shoot height (cm) 8.00±0.30 10±0.28 11±0.25 10±0.23
3 rdDay Seedling Root length (cm) 2.9±0.23 3.22±0.25 3.46±0.27 3.6±0.24
Seed Vigour index 280±3.60 600±3.50 670±3.77 500±2.51
Parameter C T5 T10 T15
Seedling shoot height(cm) 9.00±0.50 9.5±0.25 11.25±0.55 10.00±0.45
Seedling Root length (cm) 2.3±0.24 3.1±0.40 3.2±0.5 3.00±0.5
Seed Vigour index 890±0.25 1300±2.60 1400±3.21 1380±2.64
5 th Day

Parameter C T5 T10 T15

Seedling shoot height(cm) 10.00±0.54 10.5±0.26 12.00±0.53 11.50±0.23
Seedling root length (cm) 2.4±0.25 3.00±0.26 4.00±0.27 3.67±0.3
7 th Day Seed Vigour index 900±0.26 1320±2.64 1650±3.21 1400±2.64

Table 2.pHof the Soil treatments (Mean± S.D).

Parameters Treatments

Control T5 T10 T15

pH 6.9±0.12 6.7±0.15 6.7±0.15 6.6±0.15

Fig. 4.pHmeasure of the herbal garden’s soil.

26
Fig.5. Collected bulk fertile soil sample from Fig.6. Soil before Processing.
From herbal garden of MIET.

Fig.7 .Processing of the soil.

27
Fig.8. Seed of vigna radiate

28
5 gm treatment:

Fig.9. Vigna radiate Seed Sown . Fig.10. After 10 days of water

treatment(spray of water to soil moist)

Fig.11. At the start of seed germination Fig.12. Seedling of plant after 15 days.

29
10 gm treatment :

Fig.13. Vigna radiate Seed Sown. Fig.14.After 10 days of water

treatment(spray of water to soil moist)

30
Fig.15. At the start of seed germination. Fig.16. Seedling of plant after 15 days.

31
15 gm treatment :

Fig.17. Vigna radiate Seed Sown Fig.18. After 10 days of water treatment
(spray of water to soil moist)

Fig.19. At the start of seed germination. Fig.20. Seedling of plant after 15 days.

32
 5: RESULT AND DISCUSSION:

5.1 Soil analysis and parameters:- Genrally soil is acidic or alkaline in nature and this can be
measured by the testing their ph value. A ph value is measure of the hydrogen ion
concentration varies over a wide range. When used of Trichoderma viride the nature of soil
was acidic in nature.Trichoderma viride enhance the production of the soil and work also a
pesticide. After using of Trichoderma soil fertility was also increased which help in crop
production.

Seed germination is considerd as first physiological activity where senstivity of plant species
can be observed clearly in respect of their growth rate. Seed germination in all the treatment.
Seed germination percentage in all amendments was observed C (73%),T5(99%), T10(100%)
and T15 with100 % for (Vigna radiata), whereas minimum seed germination percentage was
observed in Vigna radiatain Control (75%) and maximum seed germination percentage is 100
% in T10 and T15. Minimum shoot height in 3rdday of V. radiatais (8.0cm) in Control and
Maximum shoot height is obseved in T10 (11cm), Minimum root length of vigna radiata is
observed in Control (2.9cm) and maximun observed in (3.46 cm) in T10, and Seed vigour
index minimum in control (280) and maximum in T10 (670).

In 5th day observation minimum shoot height of control i.e. (9.0cm) and maximum shoot
height is observe in T10 i.e. (10cm), and minimum root length observed in control i.e.
(2.3cm) and maximum root length obseved in T10 i.e. (3.2cm) and the seed vigour index in
control observed minimum i.e. (890) and maximum in T10 i.e. (1400).

In 7th day observation minimum shoot height of control i.e.(10cm) and maximum obseved
shoot height in T10 i.e.(11.50cm), Minimum root length obseved in control i.e.(2.4cm) and
maximum root length observed in T10 i.e. (4cm), and the seed vigour index minimum
obseved in control i.e. (900) and maximum in T10 i.e. (1650).

M Asaduzzaman et al.,el1986 advocated that Trichoderma species helping seed germination,

here seed germination percentages and the vigour index were significantly affected by the
application of deifferent Trichoderma species. In chilli seeds gave the highest vigour index
values with Trichoderma species. Trichoderma species useful to enhance the germination of
seeds and as well as reduce the delayed germination.(Rajesh R. Waghunde et al.,el2016)find
result that the Trichoderma species help in the agriculture sector and improved the agriculture
production and some modern practices affect the environment as well. Trichoderma various

33
species used as a biocontrol agent. In historical development of Trichoderma species mode of
against different biological agents. Trichoderma spp have good potential in agriculture as
well. They ammend the abiotic stress and improve the physiological response and they
enhance the nitogen use and improve the efficiency of different crop.Trichoderma work as a
fertilizers and it helps to the plant in its growth, different amount of trichoderma works
effectively. It helps plants to his proper growth within the month.

It is known that some Trichoderma strains can stimulate plant growth, atleast in part by
increasing the nutrient uptake and efficiency of nitrogen use(Altomare et al., 1999; Yedidia et
al., 2001). Our data confirm the ability of the commercial biocontrol agent T. viride strain to
increase canopy and stem growth in Moong and indicate that, in this respect, T.viride is more
effective than T.viride strain P1, a fungus mainly used forlaboratory work and not developed
commercially to a significant extent. Moreover, our findings demonstrate that the extent of
growth stimulation is largely dependent on the Moong genotype, suggesting that the response
to Trichodermaviride. is under genetic control. Shoot dry weight was increased in most lines,
except for TA209 with P1 and for the wild accession LA1777 with T22. Interestingly, P1 was
able to reduce the stemlength of LA1777 and had a similar, although not significant, effect on
the other indeterminate line under study, Corbarino.

The importance of the plant genetic background has already been reported for the interaction
between maize and T.viride T22 (Harman et al.,2004b). Commercial trials on several T22-
treated hybrids and inbred lines have revealed the expected yield increases in most genotypes,
with a few actually showing a yield reduction (Harman et al., 2004b). Genetic analysis has
demonstrated that the maize response is largely conditionedby dominant genes (Harman,
2006).

The importance of the Moong genotype for the outcome of the beneficial plant–fungus
interaction was also assessed for root growth promotion, with a significant increase in root
dry weight obtained by treating Corbarino and TA209 with strain P1 and a significant
decrease for M82 and TA209 with strain T22. Trichoderma spp. also modified the root
architecture of Moong in a differential way among the five tested lines. In three of the five
lines, T. atroviride P1 increased root length and reduced lateral development .

Bycontrast, in most cases, root length was unaffected by T. viride T22, whereas lateral
development was stimulated by this fungus .We have not investigated the mechanisms
underlying these differential responses. A recent study on Arabidopsis thaliana suggested that

34
two strains of T. atroviride and T.virens stimulated lateral root development and reduced
primary root length by producing indole-3-acetic acid (IAA) and auxin-like compounds
(Contreras-Cornejo et al., 2009).

Preliminary data indicate that both Trichoderma strains used in this work synthesize IAA
(results not shown), and we have demonstrated previously their ability to release secondary
metabolites with an auxin-like effect on plants (Vinale et al., 2008a, b). Further, Gravel et al.
(2007) suggested that another strain of T. atroviride stimulates Moong root growth in a
‘controlled manner’ by balancing the synthesis and degradation of IAA and or by limiting Et
synthesis through hydrolysis of its precursor molecule 1-aminocyclopropane-1-carboxylic
acid (ACC). However, cytokinins could also be involved, as demonstrated for plant growth-
promoting rhizobacteria (PGPR) (Ortiz-Castro et al., 2009), as the production of cytokinin-
like molecules, e.g. zeatin, and of GA3 or GA3-related gibberellins has been reported as
being possibly correlated with the biofertilization potential of Trichoderma (Benitez et al.,
2004). An antagonistic auxin–cytokinin cross-talk may explain the opposite effects of T.
viride T22 on the shoot and root development of the Moong lines TA209 and, possibly, M82.

Trichoderma to promote a balanced growth of plants depends on a fine regulation of

hormones and hormone-like compounds in the rhizosphere, our data indicate that other
factors, such as the genotype and the physiological status of the plant, or the cultural
conditions (i.e. hydroponics vs.soil or high vs. low fertilizer content), are involved. Beside
the well-characterized direct biocontrol activity of Trichoderma species on soil pathogens, it
has more recently become clear that root colonization by these microorganisms induces plant
resistance to foliar diseases, in the absence of direct contact between Trichoderma and the
pathogen (Bigirimana et al.,1997; Harman et al., 2004a; Shoresh et al., 2010). Our results
confirm that damage from B. cinerea infection on Moong leaves can be limited by
rhizosphere colonization with either T. viride T22 or T. atroviride P1,although the former
species appears to be more effective. However, major differences were also detected among
the five tested Moong lines with regard to the Trichodermainduced tolerance to the pathogen.
Apart from the wild species line LA1777, whose very low susceptibility to B.cinerea was
further boosted by interaction with either Trichoderma strain, lesion expansion was initially
limited by T22 in all tested lines. At later times, T22 reduced significantly the average lesion
area in only TA209 and Corbarino, i.e. the two lines which are the least able to defend
themselves from the pathogen in sterilized soil (Fig. 3b). Corbarino and LA1777, being a
local variety selected by farmers in low-input agricultural conditions and a wild species

35
accession, respectively, might be more ecologically fit and therefore able to benefit the most
from rhizosphere interaction with beneficial microflora. Information about the pedigree of the
seed company- bred line TA209 is too sparse to confirm or disprove this hypothesis.

The susceptible line TA209 also showed alleviation of disease symptoms with T.atroviride
P1, which was not effective for the other S. lycopersicum lines, and even increased the
pathogen susceptibility of M82, a result that is worthy of further investigation. Notably, the
very limited or negative Trichodermainduced responses to B. cinerea of line M82 also
extended to the transcription of defence genes. The ability of different strains of T. viride,
including T22, to significantly restrict the development of B. cinerea symptoms on Moong
has been reported previously (De Meyer et al., 1998; Dik and Elad, 1999; O'Neill et al., 1996;
Seaman et al., 2003).

However, only one study was performed on different Moong varieties (two), but the effect of
the genotypic differences was not investigated (Dik and Elad, 1999). In addition, reports on
thegenotypic variation of the maize response to T. viride T22 did not addressthe variability in
induced plant resistance (Harman, 2006; Harmanet al.,2004b). Our results clearly highlight
the existence of a genetic componentof the plant response to Trichoderma spp. in terms of the
induction ofsystemic resistance. This is in agreement with reports demonstrating that genetic
background affects the response of different cucumber varieties to PGPR species (Liu et al.,
1995), which actually are considered to share with Trichoderma spp. similar mechanisms of
ISR (Harman et al.,2004a).

In order to further study the Trichodermamediated systemic plant increase in pathogen

tolerance, we analysed the transcriptional activation of several defence genes in Moong
leaves: PR1b1, a PR1 gene used as a marker for SAR activation and SA-mediated responses
(Tornero et al., 1997); PR-P2, a PR4 gene mainly induced by SA in Moong (Bertini et
al.,2003; Fiocchetti et al., 2006; Linthorst et al., 1991; Van Kan et al.,1995); PINI and PINII,
which are induced through the JA signal transduction pathway (Doares et al., 1995; Fidantsef
et al., 1999); and TomLoxA and TomLoxC, encoding two enzymes of the lipoxygenase
(LOX) family, which is involved in JA response and defence (Feussner and
Wasternack,2002; Porta and Rocha-Sosa, 2002). Root penetration by Trichoderma is known
to increase the expression of defence-related genes and the production of antimicrobial
compounds, but only transiently (Shoresh et al., 2005; Yedidia et al., 1999, 2003).

36
However, our results demonstrate that rhizosphere colonization by Trichoderma can support
the transcription of some defence-related genes at low but significant levels for a relatively
long period of time, at least in some plant genotype Trichoderma species' interactions. This
effect was particularly strong for PR1b1 and PR-P2 expression, suggesting that the long-term
response to Trichoderma in Moong may involve SA signalling.Similarly, transcriptomic
analysis revealed constitutive PR5 up-regulation in 7-week-old Moong plants sown in T.
hamatum 382-inoculated soil (Alfano et al.,2007).

When plants are challenged with a pathogen soon after the establishment of the interaction
with Trichoderma, they are primed to react more strongly, increasing defence gene
expression and the activity of protective enzymes sooner and to higher levels than in
untreated plants(Shoresh et al.,2005; Yedidia et al., 1999, 2003). Our data demonstrate that
this effect lasts long after the onset of the interaction between Trichoderma and the plant as
reduced lesion size and higher expression levels of defence genes in systemic leaves were
detected in lines LA1777,TA209 and Corbarino. Generally, enhanced accumulation of RNA
concerned PIN and, to some extent, lox genes, suggesting the involvement of an ISR
response 2 months after treatment with the biocontrol agent. Higher induction of PR
proteins/activities by pathogens in plants treated with Trichoderma spp. has been reported in
cucumber and maize (Harman et al., 2004b; Shoresh et al., 2005). However, in this work, the
expression levels of PR1b1 and PR-P2 were higher than in controls on plants treated only
with Trichoderma , but, in most cases, decreased below the control value after subsequent
inoculation with the pathogen .

These data are in agreement with those from proteomic analysis of a three-player interaction
(pathogen, plant and Trichoderma) (Marra et al., 2006), and strongly suggest that the
presence of the biocontrol agent reduces the intensity of some plant responses to pathogen
infection. Concerning the response of lox genes, T. asperellum was reported to up-regulate
lox1 transcription in roots, but not leaves, of cucumber (Shoresh et al., 2005) and of lox2 in
plants of Arabidopsis thaliana (Segarra et al., 2009) soon after treatment. Based on these
results, the authors proposed that Trichoderma-enhanced plant defences depend on JA
signalling and hence the activation of the phenylpropanoid pathway (through activation of
PAL genes transcription), eventually leading to induced accumulation of phytoalexins and
other antimicrobial metabolites (Shoresh et al., 2005; Yedidia et al., 2003).

37
We also found that an increased resistance of the various Moong lines to B.cinerea
corresponded to enhanced expression of lox genes, although the opposite was not always true
. In the case of the PINI and PINII genes, their expression was often induced and maintained
at a level higher than that in controlsin Trichodermatreated and B. cinerea-inoculated Moong,
and generally correlated with increased pathogen tolerance. Several lines of evidence indicate
that the plant defence reaction mediated by Trichoderma involves the JA pathway and
stimulation of Et-responsive genes (Korolev et al., 2008; Shoresh et al., 2005).

However, the reported activation of Pal1 (Shoresh et al., 2005; Yedidia et al., 2003) could
also increase SA biosynthesis. Indeed, Martinez et al. (2001) proposed that the reduction of
powdery mildew symptoms in melon cotyledons by a cellulase from T. longibrachiatum
occurs through two parallelmechanisms: active cellulase induces an oxidative burst and hence
increases PAL activity and the synthesis of SA and phenolic compounds; at the same time, it
induces LOX activity, JA synthesis and Et accumulation, thus additionally stimulating PAL
and the phenylpropanoid metabolism (Martinez et al., 2001).

It has also been suggested that ISR may be mediated by different signalling routes, depending
on the BCA and challenging pathogen (Korolev et al., 2008). In terms of the activation of
different signal transduction pathways, our gene expression data suggest the following
mechanism for the Moong response: (i) in the absence of pathogen infection, a preventative
treatment of the plant with Trichoderma, as typically used in agriculture, induces an increase
in the expression of some PR genes, such as the SAR markers PR1b1 and PR-P2, indicative
of a priming of the defence reaction; (ii) when plants are challenged by B. cinerea,
pretreatment with Trichoderma might mitigate the SA-dependent gene expression; (iii) soon
after infection, expression of PINI, PINII, TomLoxA and TomLoxC is enhanced, possibly as
a consequence of the priming effect, which results in an increased systemic resistance,
probably caused by promotion of the JA-mediated response.

Taken together, our data demonstrate that the plant response to ‘elite’strains of Trichoderma
spp., such as the commercial biocontrol agent T.viride T22 or the widely studied T. atroviride
P1, is affected by plant genetic variability and thus is under genetic control in Solanum
species ofsection Lycopersicon. Genotypic differences in any of the plant components of the
complex cross-talk with Trichoderma can be evoked to explain this effect, including the
genotype ability to attract and sustain root colonization by the fungus, different sensitivities

38
to the effectors produced by the BCA,variability in the perception and signal transduction of
any of the hormones whose concentrations are controlled by Trichoderma spp., and so on. An
explanation of the mechanisms that underlie plant genetic control of theinteraction was
beyond the scope of this work and will require a massive research effort. However, our
findings hold promise that studies aimed at the identification of the major plant genetic
determinants involved in the interaction with Trichoderma spp. will lead to the selection and
breeding of Moong genotypes with an improved capacity to benefit from rhizosphere
colonization by these microorganisms.

39
 CONCLUSION
Effect ofTrichoderma on shoot length and root length of Mungbean (Vignaradiata) was
positively significant. When use of 10 gm Trichoderma in 1 kg soil then we observed that
seed germination percentage, Shoot height, Root length and seed vigour index shows the
satisfactory results, which means 10 gm of Trichoderma in 1kg soil gave the benificial result ,
which enhance the plant growth.Thus it is concluded that Trichoderma is the beneficial fungi
for the growth of the crops and other plants.Hence it is indicated that Trichoderma viride can
be a growth promoter and be used as a biofertilizer, which enhance the plant growth and
protect the plants to the pest.

40
 SUMMARY

Many benificial bacteria and fungi work as a pesticide and bio fertilizer, which enhance the
crop production. one of them is Trichoderma, Trichoderma work as a pesticide, biofertilizer
and this is also help to make the fertilizer from the kitchen waste as well. Vigna radiata which
is very benificial for the health and which also provide the proteins to our body. In the study
of Trichoderma effect on plant, One edible plant species Vigana radiata were choosen for the
experimental purpose as these species are easily to grow and generally used by the human in
their daily diet. This experiment’s objective was Trichoderma effect on plant growth.The
effect of Trichoderma in plant growth was very fined and also enhance the growth of plant,
also ammend the PH value of the soil. Amendment on chemical properties, Seed
germination ,Shoot height, Root length and Seed vigour index of the plant species was
studied.For the present experimental purpose, soil sample was collected from the herbal
garden of the MIET, Meerut. Seeds of plant species were procured from Amazon.com for the
experiment. Also determine the Ph value of the soil by the digital pH meter where observed
that soil was acidic.Calculation of seed germination percentage, Seed vigour index, Shoot
height and Root length, Where seed vigour index was calculated by Baki and Anderson
(1973) mthod.

Plant species (v. radiata) were grown in the treatment (TC, T5, T10, T15), Seed germination
percentage in all amendments where the maximum Seed germination percentage observed in
the T10 and the minimum observed in the Tc.Maximum shoot height observed in the Tc and
the Minimum shhot height obseved in T10. Root length maximum observed in T10 and
Minimum Root length obseved in T10. Plant Vigour Index maximum obseved in the T10
and the Minimum plant vigour index observed in theTC treatment.Trichoderma used for as
plant pramoter and a ferilizer, which enhance the production of crops and also work as a
pesticides..

41
 References:-

1. Adandonon, A. (2000): Damping-off of cowpea in Benin and South Africa M.Sc.

Thesis. University of Pretoria, Pretoria, South Africa.
2. Chet, I., Harman, G. E. and Baker, R. (1981): Trichoderma hamatum : its hypal
interaction with Rhizoctonia solani and Pythium spp. Micro. and Eco., 7: 29-38.
3. Ciccarese, F., Frisullo, S., Amenduni, M. and Cirulli, M. (1992):Biological control of
Sclerotium rolfsii root rot of sugar beet withTrichoderma harzianum. (Eds.): E.C.
Tjamos, G.C. Papavizas and R.J. Cook. pp. 243-248.
4. Cilliers, A. J., Herselman, L. and Pretoius, Z. A. (2000): Geneticvariability within and
among mycelial compatibility groups of Sclerotiumrolfsii in South Africa.
Phytopathol., 90: 1026-1031.
5. Elad, Y., Chet, I. and katan J. (1980): Trichoderma harzianum: A biocontrol agent
effect against Sclerotium rolfsii and Rhizoctonia solani. Phytopathol., 70: 119-121.
6. ISSN 2320-5407 International Journal of Advanced Research (2015),Volume 3, Issue
2, 153-158 157 Gawande, S. and Sharma, P. (2003):Changes in host enzyme activity
due to induction of resistance against downy mildew in cauliflower. Ann. Agric. Res.,
24(2):322-331.
7. Harman G.E. (2006): Overview of mechanisms and uses ofTrichoderma spp.
Phytopathol., 96: 190–194.
8. Harman, G. E. (2000): Myths and dogmas of biocontrol Changes in perceptions
derived from research on Trichoderma harzianum T22. Pl Dis., 84(4): 377-393.
9. Howell, C. R. (2003): Mechanisms employed by Trichoderma spp. In the biological
control of plant diseases: the history and evolution of current concepts. Pl. Dis., 87: 4-
10. ICRISAT, Patancheru, A. P., India:303-347. Jangid, P. K., Mathur, P. N. and M.
Kusum (2004): Role of chitinase and beta -1, 3 glucanase elicitation in the
Trichoderma harzianum induced systemic resistance in capsicum.
10. Indian phytopathol., 57(2): 217-221. Johnson, L. F. and Curl, E. A. (1972):Methods
for research and their ecology of soil borne plant pathogens.Burgess Pub. Com. Mim.,
pp. 247. Karpagavalli, S. and Ramabadan,R. (2001): Effect of fungicides and
Trichoderma species on cellulolytic enzyme production, damping-off incidence and
seedling vigour of tomato. Plant Dis. Res. 16(2): 179-185. Morton, D. J. and Stroube
W.H. (1955): Antagonistic and stimulating effects of soil micro-organism of

42
 Sclerotium. Phytopathol., 45:417-420. Schirmbock, M.,Lorito, M., Hayes., C.K.,
ArisanAtac, I., Scla, F., Harman, G. E. and Kubicek,C. P. (1994): Parallel formation
and synergism of hydrolytic enzymes and peptaibol antibiotics, molecular
mechanisms involved in the antagonistic action of Trichoderma harzianum against
phytopathogenic fungi. Appl. Environ. Microbiol., 60: 4364- 4370.

11. Waghunde R.R., Shelake R.M and Sabalpara A.N (2016) Trichoderma: A significant
fungus for agriculture and environment. African journal of Agricultural
Research.11(22):1952-1965. DOI: 10.5897/AJAR2015.10584
12. Chang YC, Chang YC and Baker(1986) Increased growth of plants in the presence of
the Biological Control Agent Trichoderma harzianum. Plant disease.70(2): 145-148.
DOI: 10.1094/ PD-70-145
13. Asaduzzaman M, Alam MJ, Islam MM (2010) Effect of Trichoderma on seed
germination and seedling parameters of chilli. Journal of science foundation. 8(1-2),
141-150. https://doi.org/10.3329/jsf.v8i-2.14637
14. Yaqub F and Shahzad S(2009) Effect os solar heating by polyethylene mulching on
sclerotial viability and pathogenicity of sclerotium rolfsii on mungbean and sunflower
. Pak. J. Bot., 41(6):3199-3205.
15. Melloy P, Hollaway G, Luck J, Norton R M, Aitken E and Chakraborty S (2010)
Production and fitness of Fusarium pseudograminearum inoculum at elevated carbon
dioxide in FACE. Global change biology 16(12) doi: 10.1111/j.1365-
2486.2010.02178.X
16. Shukla K.K, Newsom RK, Naja M, Singh N et al. (2014) Estimation of the mixing
layer height over a high altitude site in central Himalyan region bt ussing Doppler
Lidar Journal of Atmospheric and Terrestrial physics, 109:48-53
doi:10.1016/j.jastp.2014.01.006
17. Eastburn DM, Degennaro Mm, Delucia EH, Dermody O and Mcelrone A (2010)
Elevated atmospheric carbon dioxide and ozone alter soyabean diseases at SoyFACE.
Global change biology 16(1) doi:10.1111/j.1365-2486.2009.01978.x
18. Sharma M, Ghosh R, Tarafdar A et al. (2019) Exploring the Genetic cipher of
chickpea through identification and multi-environment validation of resistant sources
against fusarium wilt. Front. Sustain. Food syst. (3:78), doi:
10.3389/fsufs.2019.00078

43
19. Vitti A, Sofo A, Scopa A and Nuzzaci M (2015) Sustainable Agriculture practices in
disease defence of traditional crops in southern italy: The case study of tomoto cheery
protected by Trichoderma harzianum T-22 against cucumber mosaic virus (CMV)
The sustainability of agro-food and natural resource system in the mediterranen basin
(9); doi: 10.1007/978-3-319-16357-4_9
20. Dubey SC, Madugula S, and Singh BK(2007) Evalution of Trichoderma species
against Fusarium oxysporum fsp. Ciceris for integrated management of chickpea wilt.
Biological control 40(1):118-127; doi:10.1016/j.biocontrol.2006.06.006
21. Sharma P, Sharma M, Manokaran R and Shanmugam V(2014), Status of Trichoderma
research in india: A review.Indian phytopath. 67(1): 1-19 (2014).
22. Gawade SB, Zanjare SR, Suryawanshi AV and Shelar VR(2009). Efficacy of
bioagents and botanicals on seed mycoflora and seed quality in mungbean.
Agricultural science digest- A research journal 36(1), 30-34, 2016
23. Rasool F, Ahmad F, Ashraf S et al. (2014). Field performace of Trichoderma species
against wilt disease complex of chickpea caused by Fusarium oxysporum f. sp. Ciceri
and Rhizoctonia solani. Turkish journal of agriculture and forestry 38(4), 447-
454,2014. Doi: https://doi.org/10.3389/fpls.2015.00868
24. Siddiqui ZS, Yasmeen R (2017). Physiological responses of crop plants Trichoderma
harzianum in saline environment. Acta Botanica Croatica 76(2), 154-162, 2017.doi:
https://doi.org/10.1515/botcro- 2016-0054
25. Renukadevi P et al. (2016) Exploring the potential of Trichoderma for management of
seed and soil- borne disease of crops Integrated pest management of tropical
vegetable crops. Doi: 10.1007/978-94-024-0924-6_4
26. Blaszczyk L, Lisiecka J et al (2014). Trichoderma spp.- application and prospects for
use in organic farming and industry. Journal of plant protection research 2014;
54(4):309-317, doi: https://doi.org/1o.2478/jppr-2014-0047
27. Silva RN, Monteiro VN, et al., (2019) Trichoderma/ Pathogens/ Plant interaction in
pre harvest food security . Journel hompage: www.elsevier.com/locate/funbio (2019),
doi: https://doi.org/10.1016/j.funbio.2019.06.010
28. Malloy KM, Milling LS. The effectiveness of virtual reality distraction for pain
reduction: a systematic review. Clin Psychol Rev. 2010 dec;30(8):1011-
8.Doi:10.1016/j.cpr.2010.07.001. Epub 2010 jul 13.

44
29. Badaluddin NA, Zin NA. Biological functions of Trichoderma spp. for agriculture
applications. Annals of agriculture sciences.(65) 168-178. Doi:
https://doi.org/10.10.1016/j.aoas.2020.09.003
30. Blaszczyk L, Siwulski M, Sobieralski Ket al (2014). Trichoderma spp. – application
and prospects for use in organic farming and industry. Journal of plant protection
research 54(4), 2014
31. Kumar S, Thakur M and Rani A (2014). Trichoderma: Mass production, formulation,
quality control, delivery and its scope in commercilization in india for the
management of plant diseases. African journal of agriculture research. 9(53), 3838-
3852 (2014). Doi: 10.5897/AJAR2014. 9061
32. Mahato S,and Neupane S (2017). Comprative study of impact of Azotobacter and
Trichoderma with other fertilizers on maize growth. Journal of maize research and
development (2017) 3(1): 1-16. Doi: http://dx.doi.org/10.3126/jmrd.v3il.18915
33. Debnath S, Chakraborty G et al (2020)Potential of Trichoderma species as
biofertilizer and biological control on plant cultivation. Biotecnologia Vegetal 20, 1:1-
16
34. Hermosa R, Rubio MB et al (2013).The contribution of Trichoderma to balancing the
costs of plant and defences. International microbiology (2013) 16:69-80.doi: 10.2436/
20.1501.01.181
35. Hill R, Stewart A (2014). Application of Trichoderma in plant growth
promotion.Biotechnology and Biology of Trichoderma 2014, 415-428. Doi:
https://doi.org /10.1016/B978-0-444-59576-8.00031-X
36. Harman GE (2006). Overview of mechanisms and uses of Trichiderma spp. APS
Publications 2006, 96(2), Doi: https://doi.org/10.1094/PHYTO-96-0190
37. Woo SL, Vinale F, et al (2008). Trichoderma- plant-pathogen interactions, Soil
Biology and Biochemistry 2008, 40(1), 1-10. Doi:
https://doi.org/10.1016/j.soilbio.2007.07.002
38. Harman GE, Howell CR, Viterbo A et al (2004). Trichoderma species- Opportunities,
Avirulent plant symbionts. Doi: 10.1038/nrmicro797
39. Rana singh N, Saurabh A and Nedunchezhiyan M , 2006. Use of Trichoderma in
Disease Management, 0rissa review 2006
40. Kumar RS (2010). Trichoderma: A bio-control agent for management soil born
diseases. Agropedia
41. George H, (2019). Trichoderma improves plant growth and kills fungal pathogens.
45
42. G Pallvi: Greengram:origin, distribution and harvesting,pulses,agronomy Agriculture
in india

46