ELICITATION OF RESISTANCE BY RAW COW MILK AND TRICHODERMA VIRIDE IN

CHILLI AGAINST LEAF CURL DISEASE

Arun Kumar and P. C. MaliCentral Arid Zone Research

Institute, Jodhpur-342003, IndiaEmail: arpurster@gmail.com

Chilli or hot pepper (Capsicum annuum) is an effective medicine for various types of pains in humans

Herpes zoster-related pain, diabetic neuropathy, post- mastectomy pain, headaches, osteoarthritis, and the post-surgical pains in case of cancers

The crop is widely cultivated as vegetable and spice crop in the

irrigated belt of arid and semi-arid areas

CHILLI-THE CROP

Rajasthan is the 6th largest producer in India

Jodhpur region produces 42.7% red chilli on 20,000 ha of land

Jodhpur district alone contributes 20% of total production of red chilli in the State

Continued..

BACKDROPBACKDROP

Popular chilli cvs. Mathania Red, Mehsana, Haripur-Raipur and Mandoria

Large and fleshy fruits, mild pungency and brilliant red colour with rich spicy flavour

Susceptible to fungal and viral diseases along with nematode

DISEASES OF CHILLIDISEASES OF CHILLI•Leaf curl disease

(Begomovirus ChiLCuV, after Senanayake et al. 2007. Plant Pathology 56, 343).

•Die-back (Colletotrichum capsici (Syd.) Butler & Bisby)

•Root Knot nematodes (Medoidogyne incognita )

SYMPTOMSSYMPTOMS• Curling of upper Curling of upper

leaves with leaves with shortened shortened internodesinternodes

• Severely infected Severely infected plants produce plants produce clusters of reduced clusters of reduced sized leaves with sized leaves with fewer flowers and fewer flowers and fruitsfruits

• Small sized fruits Small sized fruits are produced with are produced with deformed seeds deformed seeds

Leaf Curl Disease of chilliLeaf Curl Disease of chilli

Small sized fruits Clusters of reduced size leaves

Year Fruit Yield

(kg ha-1)

Before 1990 600

1991-2008 140

Green Fruit Yield of Chilli

Farmers extensively use a number of insecticides to protect the crop from LCD

Use of pesticides has failed to minimize the losses

Causes environmental pollution, resistance in insect vectors and health hazards

EXISTING MANAGEMENT EXISTING MANAGEMENT PRACTICESPRACTICES

BIO-CONTROL AGENTSBIO-CONTROL AGENTS

• Bio-control agents have emerged as a new method of managing plant diseases

• Different botanicals (Plant spp.), animal products and other materials such as ash, cow urine and dung, fish meal and milk etc.

• Rhizobacteria (PGPR), leaf proteins, proline and Trichoderma spp. reduce diseases in many plant species manipulating host plant's physical and biochemical properties

USE OF MILK USE OF MILK IN PLANT DISEASE IN PLANT DISEASE MANAGEMENTMANAGEMENT

Indigenous ancient tool against plant diseases

Amino acids viz. proline, isoleucein and phenyl alanine in milk induce resistance against plant diseases

Phenolic anti-microbials are also present

Potassium phosphate content strengthens host immune system against diseases

Amino Acid Content of Raw Cow’s Milk *

Amino Acid In RCM (mg-100 ml.)

Amino Acid In RCM (mg-100 ml.)

Glutamic acid 731 Tyrosine 172

Isoleucine 165 Threonine 146

Lysine 268 Methionine 86

Phenylalanine 171 Histidine 84

Hydroxyproline

330 Glycine 66

Aspartic acid 261 Tryptophan 23

Valine 202 Cystine 21

Serine 197 Arginine 18

Leucine 326 Alanine 110

* Payne-Botha, S. and Bigwood, E. J. (1959). Amino-acid content of raw and heat sterilized cow’s milk British J. Nutrition 13: 385-389.

Certain amino acids demonstrated to act as inducers of plant resistance against biotic or abiotic stresses in many host-pathogen systems

Besides inducing resistance these amino acids also enhance growth of plants and act as osmo-protectants

AMINO ACIDS AND PLANT DISEASE PROTECTION

TREATMENT (T 1)Treated chilli seeds with RCM (1:1)

for 24 hrs

Dried in shade

Treated these seeds with Trichoderma viride ( @ 6g-kg seed)

Nursery soil treated with T. viride

(@ 10g2) after mixing with FYM

Seed sowing in nursery

45 DAS roots of plant saplings were dipped in RCM (15% or 1:6) for 20 min. before transplanting

20 DAS

RCM (15%) sprayed on the transplanted plants

FARMER’S PRACTICE(Control)

Most of these practices are chemical sprays with or without chemical seed treatment

Chilli-root-dip treatment with RCM

Trichoderma in nursery soil grows with developing root system of the treated plant and

protects the roots from infection

Treatment*

Incidence (%)

Protection over FP (control)

T-1 19.4 48.4

FP (Control) 37.6 ---

On-farm Management of LCD (Pooled data of 2000-’04)

*T-1: Seed treatment with RCM (1:1) for 24 hrs and T. viride (0.6%). Root dip for 10 min. in RCM (15%). Application of T. viride (10 g-m2) with FYM, and RCM (15%) spray 20 days after transplantation in field.FP: Variety of practices that vary from farmer to farmer. Most of these are non-organic, without any seed treatment.

Average Yield Parameters of Average Yield Parameters of ChilliChilliTrea

tment* 

Plant heigh

t (cm)

Root lengt

h(cm)

No. of

fruits plant-

1

(g)

Fruit size(cm)

Fruit wt.

plant-1

(g)

Yieldplot-1

Kg-1

  

LengtLengthh

WidthWidth

T 1T 1 66.3 20.2 66.4 10.4

4.5 306.2 3.32#

T 2T 2 57.5 13.9 40.1 9.2 3.9 219.4 2.72

*T1. *T1. Seed treatment with RCM (50% dilution with water for 24 h) and Trichoderma viride (0.6%) with soil treatment with T. viride (10g m-2 ) mixed with FYM T2. Farmer’s Practice # more than 0.5 kg ha-1

increase

TC

ON-FARM LCD MANAGEMENT IN CHILLION-FARM LCD MANAGEMENT IN CHILLI

Treatment

Farmer’s Practice

 

Elicitation of Defense Related Enzymes by Amino Acids

Scavenging enzymes –peroxidases (POX) and phenolic generators - phenylalanine ammonia lyase (PAL) are induced in response to both biotic and abiotic stresses (Schneider and Ullrich, 1994; van Loon, 1997)

Treatment

Enzyme Soluble Proteins

(mg g‑1 dry leaf wt.)

PPO( OD

minute‑1 mg‑1

protein)

POX( OD

minute‑1 mg‑1

protein)

PAL(Mole t

cinnamic acid mg

protein-1 h-

1)

T0.0037 0.6718 0.7091 118.7

C0.0020 0.3458 0.7744 90.2

RCM and RCM and T. virideT. viride Induced Induced Biochemical Biochemical Changes Changes in in Leaves of ChilliLeaves of Chilli

New strategy for managing plant diseases

Elicitors are characteristically non-specific

Induced general resistance is effective against a range of pathogens

Previously susceptible plant becomes resistant after application of an inducer

INDUCED RESISTANCEINDUCED RESISTANCE

Contd…

Elicitors enhance resistance response to extrinsic stimulus without altering the genome

Only metabolic changes in plants help in defending against infections

Environment friendly

SYNTHESIS Induced biochemical and molecular

mechanisms of defense responses are generally correlated with enzymatic responses in plants

Peroxidase (POX) is a useful marker of plant development, physiology, infection and stress

POX contributes to resistance by oxidation of phenolic compounds in chilli

POX is a key enzyme in the biosynthesis of lignin that limits the extent of pathogen spread

PAL is critical precursor of a cascade of defense reactions leading to ISR

Particulars Farmers’ Practice(T 2)

Raw Cow’s Milk + T. viride treatment (T1)

CostSeed 375.00 (1.49)* 375.00 (1.52)FYM 1990.74 (7.92) 1990.74 (8.06)Fertilizer 1226.85 (4.88) 1171.71 (4.74)Pesticides 1157.41 (4.61) 281.48 (1.14)Irrigation 14814.26 (58.97) 14880.26 (60.26)Machinery 2078.70 (8.27) 2078.70 (8.42)Labour 3481.48 (13.86) 3916.08 (15.86)Total 25124.44 (100.00) 24693.97 (100.00)

ReturnProduction (q ha-1) 110 138 (25.45)Production (Rs. ha-1) 33000 41400Unit cost of production (Rs. q-1) 228.40 178.94Net return (Rs. ha-1) 7875.56 16706.03B-C ratio 1.31 1.68

Cost and return from chilli crop ( Rs. ha –1)

*Figures within parentheses are percentages to the total

BCAs and Induced

Resistance

LCD of Hot

Pepper

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Conclusion

Results

ChiLCuV induced disease posing hazards in increasing the production

• Susceptible plant turns resistant by inducer application

• Milk and T. viride have primed defense responses

•RCM (1:1 & 1:6) protected chilli from LCD

•Metabolites increased in treated plants

•Defense related enzymes showed increased activity

•RCM &T.v treated seeds and T.v applied nursery soil with RCM sprays of 15%dilutioncapable of stimulating IR in chilli

ENVIRONMENTALPROTECTION

SOCIALRESPONSIBILITY

THE PLEDGETHE PLEDGE

Towards Green Pesticides

SUSTAINABLE PRODUCTION