AGRICULTURAL RESEARCH STATION(Rajasthan Agricultural University)

SRIGANGANAGAR

Development, production and popularization of Development, production and popularization of biocontrol technology for disease control in semi-biocontrol technology for disease control in semi-arid region of Rajasthan, India- arid region of Rajasthan, India- A success storyA success story

R. B. GaurR. B. Gaur and R.N. Sharmaand R.N. Sharma

INTRODUCTIONINTRODUCTION

Indiscriminate use of pesticides against crop diseases has created several problems related with environmental pollution, development of resistance in pathogen and accumulation of harmful residue in soil, water and plants. This has prompted the necessity for development of alternative that could be practical and effective for eco friendly disease management.

Steady efforts were therefore made to develop cost effective, eco friendly, viable bioagent based technology to manage important soil borne plant pathogens such as Phytophthora, Rhizoctonia, Fusarium, Sclerotinia which have detrimental effect on kinnow, cotton, chickpea and mustard crop of economic importance in the region.

Step 1: Establishment of bioagent production unit

* Keeping this in view, a pilot project under a mission mode on “Development, production and demonstration of bio-control agents under integrated pest management” was submitted under the leadership of R.K. Joshi and S. Gangopadhyay.

* A bioagent production unit was finally established in 1995 under the funding (Rs. 33 lac) of DBT, government of India .

* This was the first bioagent based lab in the State.

Step 2: Preliminary studies(i) Collection of bioagent isolates & mass production of TrichodermaPreliminary bioagent based studies(1995-1998) were conducted by Gangopadhyay and Joshi.Initially, different isolates of bioagents were collected from biocontrol labs of Bangalore, Coimbatore and Pantnagar to select the suitable strains for the region. Exotic isolate of Trichoderma harzianum (Th-8) obtained from Pantnagar was used for mass production of bioagent .

Large scale multilocational field demonstrations were conducted in farmers’ fields and Central State Farm, Suratgarh in cotton and chickpea crops in the fields having known history of root rot infestations.

Seed treatment with Trichoderma-SD 4g/kg seed (talc-based formulated product) plus soil application Trichoderma at 5 kg per hectare were found effective in controlling root rot in cotton and chickpea crops (Gangopadhyay and Joshi, 1997). This was the first bioagent based recommendation in the zone.

(ii) Field demonstrations & trials

Step 3: Technology refinement and development(i) Potent bioagent isolate of Trichoderma identified

Work on bioagent based technology refinement was initiated during 1998-1999 by Gaur and Sharma.

Collection of soil samples from the zone -1b (Sriganganagar and Hanumangarh district) of Rajasthan were made to isolate potent Trichoderma strains among 13 local isolates.

Thereby during 1998, the commercial production of T. harzianum (Th-8) in bioagent production lab was substituted with TG-1 strain of T. harzianum.

(Gaur and Sharma, 1998).

(ii) Suitable substrates for Trichoderma multiplication were identified

Following substrates were evaluated

a. Wheat bran + 2% molassesb. Sugarcane bagasse + 2% molassesc. Gram shell d. Paddy huske. Sorghum grains f. Sodium alginate granules impregnated with 2% molasses g. FYM h. Faecal pellets of goat & sheepi. Standard talk based formulation

(Gaur et. al., 2005).

Substrates Sporulation

(x107 cfu/g)

Remarks

Faecal pellets of goat & sheep

70.85 Rapid development of bioagent, dense colony, early profuse sporulation.

Gram shell 69.52 Good growth, early profuse sporulation.

FYM 66.07 Rapid growth of bioagent. Dense colony. Sporulation at moderate stage.

Paddy husk 33.70 Poor uneven growth. Delayed sporulation

Table: Growth and sporulation of T. harzianum on substrateafter 15 days of incubation.

(iii) Shelf-life of talc based formulation of Trichoderma was estimated

viability was estimated (1999-2000) at temp. ranging between 0 to 40 0c during 180 DOSThe initial mean population of Trichoderma on 0 days was 270 x 10 6 cfu/g.After 180 DOS it declined to 20x10 6 cfu/g.Shelf - life of 90 – 105 days at 25 oC was worked out for commercial produce of Trichoderma

(Gaur et al., 2005)

(iv) Delivery system of antagonist to seed & soil was worked outTechnique of soil application of Trichoderma was found significantly superior over seed treatment.

Pre incubation of Trichoderma in FYM 15 days before soil application gave better results than its alone application.

Wheat bran based formulation of Trichoderma gave better control of disease than talc based formulation for soil application.

Trichoderma @ 10 g kg-1 seed gave better control than the lower dose of 4 g kg-1 seed. Technique of seed dip, solid matrix priming did not prove effective.

Seed treatment involving single bioagent with or without spray combination though provided significantly better control of disease in comparison with control but were found inferior when compared with mixture of two different bioagents.

(Gaur et al., 2005)

(v) Delivery system of antagonist for paint formulation was worked out

Field studies were conducted (2004-05 to 2006-07) to findout environmentally accepted paint formulation, alternative to metalaxyl for the control of foot rot disease in kinnow.

Different paint formulations of Trichoderma and Pseudomonas were explored against Phytophthora stem lesions in Kinnow.

Table: Efficacy of different paint formulations against the foot rot disease of kinnow (2004-05 to 2006-07)

Painting formulationsTrunk lesion

recovery

% increase in

yield

Aqueous spore suspension of T. harzianum* followed by painting with linseed oil after 5 days

55.21(48.0) 42.83

Talc based T. harzianum formulation** @ 100 g/l water followed by painting with linseed oil after 5 days

97.33(80.73) 87.93

Painting with talc based T. harzianum formulation** @ 100 g/l water

58.21(49.74) 48.64

Painting with talc based T. harzianum formulation** @ 100 g/l rice starch solution

75.48(60.87) 68.31

Painting with talc based P. fluorescens formulation*** @ 100 g/l water followed by painting with linseed oil after 5 days

98.67(84.58) 93.59

Control (Untreated){-}128.10(4.1

) (-) 78.54

CD at 5%CV (%)

6.477.30

* 5 x109 conidia/ml ;** 2 x 107 cfu/g ;*** 2 x 109 cfu/g

(vi) Biocontrol agents isolated from different production system

Various types of biocontrol agents (mycoparasites) were isolated (2001-02 to 2002-03) from field rhizospheric soil collected from different production system in the region in association with various pathogens Identity was confirmed by Indian Type Culture Collection, IARI, New Delhi and Agharkar Research Institute, Pune.

(Gaur, 2004).

Production system

Bioagent (mycoparasite)*

Khejri + Mung Trichoderma harzianum, T. viridi, Gliocladium virens (3)

Khejri + Moth T. harzianum (1)

Khejri + Guar T.harzianum, T.viridi, G.virens, G.penicelloides (4)

Khejri + Chickpea T.harzianum, T.viridi ,T.koningii, G.virens, G. penicelloides, Chaetomium globosum, Penicillium pinophilum (7)

Pasture + Khejri + chickpea

(0)

Pasture+ Chickpea Penicillium funiculosum (1)

Chickpea + Pasture (0)

Chickpea alone T.koningii (1)

Pasture land (0)

Table: Bio-control agents (mycoparasite) isolated from khejri based production systems (2001-2002 to 2002-2003)

(i)  Khejri based production system

Soil samples collected below khejri (Prosopis cineraria) invariabily yielded higher bioagent population. Among various species of Trichoderma (mycoparasites) isolated, T. harzianum and T. viridi were found to be widely disturbed while T. koningii was found least distributed in soil.

Ber/Khejri + Chickpea production system harboured maximum Mycoparasites.

(Gaur, 2004)

Table 11: Bio-control agents (mycoparasite) isolated from Kinnow based production systems (2001-2002 to 2002-2003).

Production system

Bioagent (mycoparasite)

Pre bearing

Kinnow + Mung Trichoderma harzianum, Aspergillus terreus

Kinnow + Moth T. harzianum

Kinnow + Chickpea T. harzianum

Bearing

Kinnow + Mung

T.harzianum, Aspergillus versicolar, A.flavus ,A. fumigatus , Penicillium

Kinnow + Moth T. harzianum, Aspergillus candidus

Kinnow + Chickpea T.harzianum, T.viridi, T.koningii, T hamatum, Chaetomium globosum, Cladosporium oxysporium

(ii) Kinnow based production system

Bearing Kinnow orchard harboured more bioagents and soil borne pathogens compared to pre bearing orchard. The

Aspergillus group was predominant during kherif season when the mung and moth crop was taken as intercrop.Trichoderma harzianum was found widely distributed while Chaetomium globosum was found least distributed mycoparasites in soil.

(Gaur, 2004).

(vii) Less dry root rot incidence in chickpea under Khejri (Prosopis cineraria):

Studies were conducted at cultivators field. Chickpea crop grown under khejri based production system and as a sole crop was observed during 2001-2002 to 2002-2003. During both the years, 4 fields each measuring 1 ha. was observed for dry root rot incidence. Dry root rot (R. bataticola) incidence was significantly less (22 %) in chickpea under khejri (Prosopis cineraria ) based production system compared to sole crop of chickpea. (Gaur, 2004)

(viii) New consortium formulation of bioagents was developed

Systematic studies in laboratory, cage house and multilocational field trials were conducted (2002-03 to 2005-06) to find out the best consortium formulation of bioagents for controlling Sclerotinia rot (Sclerotinia sclerotiorum) of Indian mustard (Brassica juncea).

Talc based consortium formulation (2 x 108 cfu/gm) of T. hamatum (HP-20) + T. viride (Tv-1) in the ratio of 1 : 1 (w/w) was developed & commercelised.

Gaur and Sharma, 2006

(Sharma et al., 2008).

(ix) New yeast bioagent isolated & identified

An hitherto unreported yeast bioagent Sporidiobolus pararoseus (KFY) was isolated from skin of kinnow fruit.

(x) Bioagents compatibility with fungicides worked out:

The compatibility of potential bioagents with recommended and newly developed fungicides was investigated & tolerance limit worked out.

Metalaxyl, fosetyl-Al, mancozeb, cymoxanil 8% + mancozeb 64% mixture and copper oxychloride fungicides were found compatible with bioagent T. harzianum (TG-1) and tolerance limits (ED50) of these fungicides were >1000 µg/ml.

Metalaxyl proved their compatibility with another potential bioagent T. viride also where even >1000 µg/ml concentration was under safe tolerance limit (ED50). Copper oxychloride, mancozeb, fosetyl-Al and cymoxanil 8% + mancozeb 64% mixture fungicides showed moderate to good compatibility with this bioagent

(Gaur and Sharma, 2010.)

Step 4: Recommendation provided to Package & Practices (2001-2010)

Management of soil borne diseases of chickpea and cotton (Rhizoctonia bataticola), mustard (Sclerotinia sclerotiorum) and kinnow (Phytophthora) was recommended through use of locally developed formulations of Trichoderma ie. Trichoderma SD, Trichoderma mixture.

KINNOWKINNOWPhytophthoraPhytophthora

For integrated disease management of Phytophthora rot of kinnow stem painting with metalaxyl (Ridomil –MZ 72 WP) @ 20 g/l of linseed oil in association with soil drenching with metalaxyl @ 25 gm/tree or Trichoderma harzianum @ 60 gm/ tree in 30-40 litre of water 2 times each during February and August at 15 DAS interval was recommended.

Year: 2002Year: 2002 ((27.08.2002)27.08.2002) Gaur et. al., 2004

Table 1. Integrated management of Phytophthora in kinnow mandarin Table 1. Integrated management of Phytophthora in kinnow mandarin orchardorchard

Treatment* recovery in trunk lesion size

(%)

Fruit /plant**

Yield/

Plant (kg)

Net Monitory

return (Rs. /tree)Pooled

(1999-2001)

Stem painting and soil Stem painting and soil drenching with drenching with metalaxylmetalaxyl

57.5 57.5

(50.0)(50.0)

307.3 307.3

(17.5)(17.5)

47.047.0 135.2135.2

Stem painting with metalaxyl and soil application of Trichoderma 60g/tree

37.0

(37.5)

235.3

(15.4)

34.7 170.7

Control (Diseased) - 60.5

(4.1)

17.3

(4.2)

5.3 32.0

CD (P = 0.05)

CV (%)

3.5

6.9

0.8

3.3

2.8

6.0

-

* Stem painting with metalaxyl (Ridomil MZ 72 WP) @ 20 g/l of linseed oil. ** Soil drenching of fungicide @ 1 g/l of water (25 g/tree)

Tr..No.

1

4

8

KINNOWPhytophthora

Biorational paint formulation against Phytophthora: Stem painting with talc based Pseudomonas fluorescens (2 x 109 cfu/gm) orT. harzianum (2 x107 cfu/gm) @ 100 gm/lit water subsequently followed bypainting with linseed oil after 5 days at two times each during February andAugust gave significant recovery in trunk lesion size of Phytophthora andenhanced fruit yield (Gaur et al., 2010).

Table: Efficacy of different paint formulations against the foot rot disease of kinnow (2004-05 to 2006-07)

Painting formulationsTrunk lesion

recovery

% increase in yield

Ridomil @ 20 g/l linseed oil 100.00(90.00) 98.73

Talc based T. harzianum @ 100 g/l water followed by painting with linseed oil after 5 days

97.33(80.73) 87.93

Talc based T. harzianum @ 100 g/l rice starch solution

75.48(60.87) 68.31

Talc based P. fluorescens @ 100 g/l water followed by painting with linseed oil after 5 days

98.67(84.58) 93.59

Control (Untreated) {-}128.10(4.1) (-) 78.54

CD at 5%CV (%)

6.477.30

T . harzianum @ 2 x 107 cfu/g ; P. fluorescens @ 2 x 109 cfu/g

Kinnow

Citrus cankerFor the control of citrus canker two spray of Streptocycline @ 100 mg (100ppm) in combination with Copper oxychloride @ 2gm/lit.

of water or Pseudomonas fluorescens @ 0.2 per cent at 15 days was recommended.

Year: 2006 (7.3.2006) (Gaur and Sharma, 2010)

Table: 1. Controlling citrus canker through chemicals and biorationals

Treatments Per cent Disease Index

2003 2004 2005 PooledFoliar spray of Streptocyclene @ 0.01% + Bavistin @ 0.1%

22.67

(28.43)

40.30

(39.40)

25.00 (30.00)

29.32

(32.00)

Foliar spray of Streptocyclene @ 0.01% + Copper oxychloride @ 0.2%

17.64

(24.80)

32.10

(34.47)

20.17 (26.63

23.30

(28.63)

Foliar spray of talc based Pseudomonas fluorescens @ 0.2%

21.93

(27.93)

37.50

(37.73)

20.83 (27.10)

26.75

(30.92)

Control 47.00

(43.30)

86.95

(68.93)

72.50 (58.40)

68.82

(56.88)

C D at 5 %

C V ( % )

2.95

5.14

3.30

4.15

1.48

4.86

1.72

4.66

T.No.

2

3

6

11

Kinnow Pre harvest fruit dropping & post harvest fruit rotting

To check pre-harvest fruit dropping and post-harvest fruit rotting three sprays of locally isolated yeast bioagent Sporidiobolus pararoseus @ 109 cfu/ml in association with Gibbrelic acid @ 20 ppm during the month of April, August and September and two alone during March and October were recommended (Gaur et al., 2009).

For the control of dry root rot of cotton seed treatment with For the control of dry root rot of cotton seed treatment with Trichoderma harzianumTrichoderma harzianum @ 10 gm/ kg seed and soil application @ 10 gm/ kg seed and soil application withwithTrichoderma harzianumTrichoderma harzianum @ 10 kg/ ha pre incubated in 200 kg @ 10 kg/ ha pre incubated in 200 kg of FYM for 10-15 days was recommended.of FYM for 10-15 days was recommended.

Year: 2003 (22.03.2003) (Gaur et al., 2005).

COTTON

Table 1. Incidence of root rot (Rhizoctonia bataticola) and germination percentage in cotton under different application manipulation techniques of bio agent (Trichoderma harzianum) under green house conditions

Tr.No.

Treatment Germination (%) Root rot (%)

Pooled

(1999-2001)

Pooled

(1999-2001)1 DST with talc based Trichoderma

harzianum @ 4 g/Kg seed 70.6 (57.2) 40.9 (39.8)

2 DST with talc based T.harzianum @ 10 g/Kg seed

71.1 (57.5) 33.4 (35.3)

8 SA of 10 days pre-incubated wheat bran based T. harzianum (@ 15 g/ 3 kg soil) in FYM (100g) at sowing time

72.8 (58.6) 19.5 (26.2)

19 Control (Only Pathogenic Inoculum) 48.3 (44.0) 61.3 (51.5)

CD at 5%

CV (%)

2.4

2.6

1.4

2.4

Table 2. Biocontrol of root rot (Rhizoctonia bataticola) of desi cotton at cultivator’s fields

Treatments Av. Mortality

(%)

(5 Locations)

Av. Seed Cotton

Yield (Q/ha) (5

Locations)Soil application of 10-15 days pre incubated T. harzianum @ 10 kg ha-1 + 200 kg FYM ha-1

2.4 (8.73) 9.90

Seed treatment with T. harzianum @ 10 g kg-1 seed + soil application of Zinc sulphate @ 25 kg ha-1

15.1(22.83 7.70

Seed treatment with T. harzianum @ 4 g kg-1 seed + soil application of Zinc sulphate @ 25 kg ha-1 (Recommended package)

17.0 (24.32) 7.40

Control 38.1 (38.12) 5.60

CD at 5%

CV (%)

3.1

9.5

1.3

12.6

Tr.No.

1

2

3

4

Table 3. Economics of biological control of cotton root rot (Rhizoctonia bataticola

Treatments Mortality

(%)

Av. Seed

Cotton

Yield (Q/ha)

B : C Ratio

Soil application of 10-15 days pre incubated T. harzianum @ 10 kg ha-1 + 200 kg FYM ha-1

2.4 (8.73) 9.9 4.9 : 1

Seed treatment with T. harzianum @ 10 g kg-1 seed + soil application of Zinc sulphate @ 25 kg ha-1

15.1 (22.83) 7.7 4.1: 1

Seed treatment with T. harzianum @ 4 g kg-1 seed + soil application of Zinc sulphate @ 25 kg ha-1

(Recommended package)

17.0 (24.32) 7.4 4.0 : 1

Control 38.1 (38.1) 5.6 -

CD at 5%

CV (%)

3.1

9.5

1.3

12.6

-

Tr.No.

1

2

3

4

Chickpea

For the control of dry root rot of Chickpea seed treatment with Trichoderma harzianum @ 10 gm/ kg seed and soil application with Trichoderma harzianum @ 10 kg/ ha pre incubated in 200 kg of FYM for 10-15 days was recommended.

Year: 2003 (14.8.2003) Gaur et al., 2005

Table 1. Incidence of root rot (R. bataticola) and germination percentage in chickpea under different application manipulation technique of bio-agent (T. harzianum) under green house conditions (1999-2000 to 2001-2002)

Treatment % Germination % Root rot

Seed pelleting with T. harzianum (5 g talc based/ml) and fresh cow dung (50 g) using 25% jaggery solution

73.7  27.0

Soil application of 10 days pre-incubated wheat bran based T. harzianum (@ 15 g/ 3 kg soil) in FYM (100g) as basal dose at sowing time

84.3 19.0

Soil application of wheat bran-based T. harzianum (@ 15 g/ 3 kg soil) and FYM (100g) mixture as basal dose at sowing time

82.4 24.1

Control (Only Pathogenic Inoculum ) 62.6 79.5

CD (P=0.05) CV (%)

3.53.7

3.75.8

T.N.

4

8

9

13

Table 1. Incidence of root rot (R. bataticola) and germination percentage in chickpea under different application manipulation technique of bio-agent (T. harzianum) under green house conditions (1999-2000 to 2001-2002)

Tr.No. Treatments % Germination % Root rot

1 Dry seed treatment with talc based Trichoderma harzianum @ 4 g/Kg seed

83.5 41.3

2 Dry ST with talk based T.harzianum @ 10 g/Kg seed

84.8 35.2

8 Soil application of 10 days pre-incubated wheat bran based T. harzianum (@ 15 g/ 3 kg soil) in FYM (100g) as basal dose at sowing time

84.3 19.0

13 Control (Only PathogenicInoculum ) 62.6 79.5

CD (P=0.05) CV (%)

3.53.7

3.75.8

Table 2. Biocontrol of root rot of chickpea in Hanumangarh district under unirrigated condition (2002-2003)

Treatments Mortality % Location

Hanumangarh

Yield (Q/ha.)

Location

Hanumangarh

B:C ratio

Soil application of 10-15 days pre- incubated wheat bran based T.harzianum @ 10 kg/ha + 200 kg FYM/ha.

3.3 19.0 10.1: 1

ST with talc based T.harzianum @ 10 gm/kg seed +soil application of Zinc sulphate @ 25 kg/ha.

9.0 14.7 10.7 : 1

ST with Bavistin @ 1.5 gm/kg seed. 25.0 10.1 4.7 : 1

Control 36.0 9.4

CD (P=0.05)

C.V.(%)

2.8

7.3

2.9

13.4

-

T.No

1

2

3

4

Table 3. Biocontrol of root rot of chickpea in Sriganganagar district under irrigated conditions (2002-2003)  

Treatments Av.% Plant mortality

(4 Locations)

Av. Seed yield (Q/ha)

(4 Locations)

B:C Ratio

Soil application of 10-15 days pre-incubated wheat bran based Trichoderma @ 10 kg/ha + 200 kg FYM /ha.

2.5 14.8 5.4:1

ST with talc based T.harzianum @ 10 gm/kg seed + soil application of ZnSO4 @ 25 kg/ha.

19.7 11.9 5.3:1

ST with Bavistin @ 2 gm /kg seed (D.R)**

35.0 9.8 4.7:1

Control 40.2 9.0 -

C.D.at 5%

C.V. (%)

3.3

8.6

1.5

9.7

T.No.

1

2

3

4

Mustard For the control of stem rot of mustard seed treatment with mixture of Trichoderma hamatum and Trichoderma viride (1:1) @ 10 gm/ kg of seed followed by foliar spray of same bioagent combination @ 0.2 per cent at 50 DAS was recommendation.

Year: 2006 (2.9.2006) Gaur et. al.,2010

In vitro antagonistic activity of bioagents on Sclerotinia sclerotiorum

24 native and exotic isolates/strains were tested.10 isolates of Trichoderma: harzianum (Th-1), harzianum (TG-1), harzianum (Th-8), harzianum (HP-22), viride (T v-1), viride (T v-2), viride (HP-21), virens (Tvs), koningii (Tk), hamatum (HP-20)

3 species of Gliocladium: virens (Gv), deliquescens (Gd), penicelloids (RH-12)

3 isolates of Chaetomium: globosum (HP-29), globosum (RH-6), globosum (Cg)

3 species of Penicillium: P. sp. (HP-3), funiculosum (RH-8), pinophilum (JT-3)

5 species of Aspergillus: versicolar (HP-2), fumigatous (HP-5) candidus (HP-12), flavus (HP-17), terreus (HP-19)

On the basis growth inhibition and sclerotia produced in culture 9 bioagent were tentavely selected. These were:

T. viride (T v-1), T. harzianum (Th-1), T. harzianum (TG-1), T. virens (Tvs), G. virens (Gv ), G. deliquescens (Gd), T. hamatum (HP-20), T. viride (HP-21) T. harzianum (HP-22)

Table: Effect of seed coating with mycelial culture of bioagents on Sclerotinia rot of mustard (Pot trial)

S.No. Seeds coated with bioagents

Germination (%)

Mortality (%)at 65 DAS

Vigourindex

1 T.harzianum (TG-1) Local

90.00(75.00) 20.03(26.58) 919

2 Gliocladium virens (GV) Coimbatore

76.67(61.20) 20.08(26.60) 686

3 T.harzianum (HP-22) Local

86.67(68.87) 25.00(30.00) 855

4 T.hamatum (HP-20) Local

81.67(64.67) 27.82(31.80) 725

5 G.deliquescens (Gd)

Banglore

73.33(59.03) 28.17(32.05) 572

6 T.viride-1 (TV-1)

Coimbatore

86.67(68.87) 33.23(35.20 802

7 T. harzianum (TH-1) Coimbatore

86.67(68.87) 33.77(35.15) 796

8 T.virens (TVS)

Banglore

81.67(65.00) 35.5(36.60) 772

25 Control (Only pathogen) 33.33(35.27) 97.34(83.30) 163

  CD at 5%

CV (%)

8.218.90

4.275.91

74.389.66

Table:1.Biological control of Sclerotinia sclerotiorum causing stem rot of mustard through bioagent mixture (Pot).

Tr. No. Germination (%) Av.

(2005 & 06)

Mortality(%)Av.

(2005 & 06)

Shoot length

(cm) Av.(2005 & 06)

Root length (cm) Av.

(2005 & 06)

Vigour index Av. (2005 &

06)

T.hm+

T.vir

88.96

(71.37

8.50

(16.84)

14.41 8.34 2030.50

T. har(TG 1)

+G.vir

88.68

(72.23)

12.27

(20.42)

14.52 8.26 2024.50

T.har

(HP-22)

+T.vir

84.65

(67.72)

13.37

(20.33)

12.63 7.37 1728.50

Control 59.69

(50.61)

76.87

(61.45)

10.36 6.62 1008.00

CD

Cv(%)

5.05

5.37

2.99

7.44

1.33

7.13

0.90

8.21

174.99

7.13

Table 4. Bioefficacy of talc based mixed formulations of bioagents against Sclerotinia rot of mustard under field conditions at ATC, Hanumangarh

Treatment

ST/FS

2004-05 2005-06

Disease

incidence (%)

Yield

(q/ha)

Disease

incidence (%)

Yield

(q/ha)

B: C

ratio

Thm + Tv/

Thm + Tv

11.3

(19.7)9.9

2.2

(8.6)32.5 8.0: 1

TG + Gv/

TG + Gv

11.4

(19.8)9.6

2.7

(9.4)32.2 7.9: 1

Control31.0

(33.8)7.6

10.6

(19.0)27.1 -

CD at 5%

CV (%)

2.1

5.8

1.3

10.0

1.3

6.0

NS

7.1

Note:

Seed treatment with Trichoderma hamatum (HP-20) local + T. viride (TV-1) Coimbatore (1:1) @ 10 g/kg seed followed by foliar spray of bioagent @ 0.2 percent at 50 DAS was found significantly superior in checking the stem rot of mustard. This treatment provided minimum mortality due to stem rot, maximum germination and higher vigour index.

The compatibility of potential bioagents with recommended and newly developed fungicides was investigated & tolerance limit worked out.

Bioagent compatibility

Compatibility of T. harzianum with fungicidesMetalaxyl, fosetyl-Al, mancozeb, cymoxanil 8% + mancozeb 64% mixture and copper oxychloride fungicides were found compatible with bioagent T. harzianum (TG-1) and tolerance limits (ED50) of these fungicides were >1000 µg/ml.

(A) Higher Concentrations of 1000, 500 and 250 ppm1-3. Aliette, 4-6. Apron, 7-9. Bavistin, 10-12. Copper blue, 13-15. Curzate M8, 16-18. Folicure, 19-21. Dithane M-45, 22-24. Insignia, 25-27. Vitavax power, 28. Control

(B) Lower Concentrations of 100, 50 and 25 ppm1-3. Aliette, 4-6. Apron, 7-9. Bavistin, 10-12. Copper blue, 13-15. Curzate M8, 16-18. Folicure, 19-21. Dithane M-45, 22-24. Insignia, 25-27. Vitavax power, 28. Control

Compatibility of T. viride with fungicides

Metalaxyl proved their compatibility with another potential bioagent T. viride also where even >1000 µg/ml concentration was under safe tolerance limit (ED 50). Copper oxychloride, mancozeb, fosetyl-Al and cymoxanil 8% + mancozeb 64% mixture fungicides showed moderate to good compatibility with this bioagent

(A) Higher Concentrations of 1000, 500 and 250 ppm1-3. Aliette, 4-6. Apron, 7-9. Bavistin, 10-12. Copper blue, 13-15. Curzate M8, 16-18. Folicure, 19-21. Dithane M-45, 22-24. Insignia, 25-27. Vitavax power, 28. Control

(B) Lower Concentrations of 100, 50 and 25 ppm1-3. Aliette, 4-6. Apron, 7-9. Bavistin, 10-12. Copper blue, 13-15. Curzate M8, 16-18. Folicure, 19-21. Dithane M-45, 22-24. Insignia, 25-27. Vitavax power, 28. Control

Step 5: Bioagent production:

This centre developed two talc based bioagent formulation of Trichoderma (Trichoderma SD, Trichoderma Mixture) for commercial use against soil borne diseases prevalent in the area.

Trichoderma SD

Trichoderma SD is talc based formulation of T. harzianum (TG-1). Production of this bioagent is being taken up since 1995 in IPM bioagent production unit setup at this centre under the funding of DBT, Bangalore. This formulation of bioagent is recommended for the control of root rot of cotton, gram, Phytophthora of kinnow and other soil borne diseases.

Trichoderma Mixture is talc based formulation of two different strains of Trichoderma evolved during 2006 against Sclerotinia stem rot of mustard (Gaur and Sharma 2006). In this formulation exotic strain of T. viride-1 (Tv-1) from Coimbatore and native strain of T. hamatum (HP-20) is used in the ratio of 1 : 1.

Trichoderma Mixture

During the year in 1995 when the production unit was established, the production of this bioagent was only 16 Kg. However in subsequent years production increased substantially reaching 873 kg in 2009 year. During the period between 1995 and 2009 this unit has produced a total of 8307 kg of Trichoderma SD.

During the period between 2007 and 2009 a total of 1548 kg of Trichoderma Mixture has been produced at this centre.

Trichoderma SD

Trichoderma Mixture

Fig. 2. Commercial sale of Trichoderma spp. by private & govt. agencies in Zone 1b (Sriganganagar and Hanumangarh districts) of Rajasthan

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Hence since establishment of bioagents production unit, a total of 9855 kg of Trichoderma formulations has been produced for commercial sale to the cultivators and different Govt. and semi Govt. agencies.

Step 6: Income generation through technology developed

The use of Trichoderma for the control of soil born diseases, a technology developed at the center and recommended for the zone after duly approval by research and extension wing of university and state, has given substantial source of income generation through direct commercial sale of Trichoderma to the cultivators and other Govt. and semi Govt. agencies by IPM production unit.

Fig. 7. Income generation through biocontrol technology at Research Centre

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The total income through sale of this product was only Rs. 960=00 when the production unit was established in 1995. However this income reached to the level of Rs. 1,10,760=00 (Rupees One lac ten thousand seven hundred sixty) during the period of 2009. Since the establishment of bio agent production unit, this center has already earned approximately Rs. 11,69,340 = 00 (Rupees Eleven lac sixty nine thousand three hundred forty only) during the period between 1995 and 2009 .

Step 7: Process changes and technology impact

Bioagent use technology for the control of soil borne disease was transferred to cultivators systematically through front line, field and IPM based demonstration. Initially during 1996 – 98 technology was popularized through large scale (30-32 ha) IPM based demonstrations on cotton and chickpea and the bioagent was distributed among the farmers as a free incentive. Since establishment of bioagents production unit, a total of 9855 kg of Trichoderma formulations has been produced for commercial sale to the cultivators and different Govt. and semi Govt. agencies.

This successful transfer of bioagent technology has led to generate interest of State (Rajasthan) Agricultural Department in establishing mini bioagent production unit in the zone. A growing number of private companies have now entered in market to meet out the growing demand of these bioagents. The commercial sale through these alternate sources in the zone between 2007-08 to 2009-10 was approximately 87 metric tonnes.

(i) Area covered under the technology by research centre

Bioagent based technology developed at this centre has been accepted by the cultivators of zone 1b comprising Sriganganagar and Hanumangarh district of the Rajasthan. The response was quite encouraging. During the initial year of 1995-96, an approximate area of only 200 hectare was covered under this technology. However after 15 year of journey on ecofriendly road approximately 13,633 hectare area has been covered under this technology through commercial sale of bioagent by this centre.

23 85 204 127 142 318 1278354

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Fig. 4. Area covered through biocontrol technology under different crops by Research Centre

Area covered (ha)

(ii) Area covered under the technology by Govt. & Private agencies

Beside research centre, an area of about 1, 22, 697 hectare has also been covered through commercial sale by Govt. and private agencies. This clearly indicate impact of technology generated by this centre.

3708540821

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Fig. 6. Area covered through biocontrol technology under different crops in Zone 1b of Rajasthan by private & govt. agencies

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This successful transfer of bioagent technology has contributed significantly in managing soil borne diseases of the area and improving soil health.

ReferencesReferences

Gangopadhyay, S. and Joshi, R.K. (1997). Efficacy of Trichoderma in controlling root-rot of cotton and chickpea. Indian Phytopathology, Golden Jubilee Proceeding, pp. 325-326.

Gaur, R. B. (2004). Develop IPM package for the arid-agri-silvi-horti-pastoral system. Final Progress Report (Sept. 2000 – Nov. 2004) under NATP Sub Project submitted to

CAZRI (ICAR). pp. 1-172. Gaur, R.B., Kaul, M.K. Sharma, R.N. and Sharma, R.R. (2004). Integrated disease

management of Phytophthora rot of kinnow. J. Mycol. Pl. Pathol.34 :465-469.

Gaur, R.B. and Sharma, R.N. (2002). Efficacy of mycoparasite for Rhizoctonia root rot in cotton. Indian Phytopath., 55: 368 pp.

Gaur, R.B. and Sharma, R.N. (2002). Studies on antagonism of Trichoderma, Gliocladium and Cheatomium species against Rhizoctonia bataticola of cotton. Indian

Phytopath., 55: 372 pp.

Gaur, R.B. and Sharma, R.N. (2010). Prevalence of canker in kinnow (Citrus deliciosa Ten.) and its management. Indian Phytopath., 63 (2): (Accepted).

Gaur, R.B. and Sharma, R.N. 2010. Biocontrol of root rot in cotton and compatibility of potential bioagents with fungicides. Indian J. Pl. Prot. (Under communication).

Gaur, R. B. and Sharma, R. N. 2006. Biocontrol of Sclerotinia sclerotiorum causing stem rot of mustard. Seasonal Summary Rabi 2005-06, Irrigated North Western Plain Zone–1b, Agric. Res. Station, Sriganganagar. pp: 12-13.

Gaur, R.B., Sharma, R.N., Gautam, V.S. and Dangi, R.P. (2010). Management of Sclerotinia rot of mustard through bioagents. Indian Phytopath., 63 (3): (Accepted).

Gaur, R.B., Sharma, R.N. and Kaul, M. K. (2010). Foot rot incidence in Kinnow (Citrus deliciosa) orchards and its management through biorational paint formulations. Indian Journal of Horticulture (Under communication).

Gaur, R.B., Sharma, R.N. and Kaul, M.K. (2009). Studies on physiological and pathological induced fruit dropping, post-harvest fruit decay and biochemical changes in kinnow fruits (Citrus deliciosa Ten.) in relation to pre- harvest spray scheduling. Final

report of the Project submitted under National Horticulture Mission, Govt. of Rajsthan, Jaipur. pp. 1-73.

Gaur, R.B., Sharma, R.N. and Sharma, R.R. (2005). Shelf-life of talc-based formulation of Trichoderma and soil application for biological control of dry root rot of chickpea. J. Mycol. Pl. Pathol., 35: 380-384.

Gaur, R.B., Sharma, R.N., Sharma, R.R. and Gautam, V.S. (2005). Efficacy of Trichoderma for Rhizoctonia root rot control in chickpea. J. Mycol. Pl. Pathol, 35:

144-150.

Gaur, R.B., Sharma, R.N. and Singh, V. (2005). Manipulations in the mycoparasite application techniques against Rhizoctonia root rot of cotton. Indian Phytopath., 58: 402-409.

Sharma, R.N., Maharshi, R.P. and Gaur, R.B. (2008). Sporidiobolus pararoseus Fell & Tallman – an antagonistic yeast with biocontrol potential. Current Science, 95:

1003- 1004.