ISOLATION AND CHARACTERIZATION OF ENTOMOPATHOGENIC ...

More documents

Recommendations

Info

3.6 EVALUATION OF FOOD GRAINS FOR MASS PRODUCTION OF Metarhizium anisopliae (Ma2) AND Verticillium lecanii (Vl1) The crushed grains of sorghum, bajra, navane, maize, rice and wheat with 1 per cent yeast extract were assessed for their suitability as substrates for mass production of M. anisopliae and V. lecanii. In addition to grains, agro waste viz., crushed maize cobs, wheat bran, rice bran, baggase and press mud with and without molasses were also tested. To each of these substrates, sterile distilled water was added in order to bring the moisture content to 50 per cent. After thorough mixing, the bottles were plugged with cotton and autoclaved at 15 psi and 121°C for 30 minutes. Circular agar discs of 5 mm diameter were taken from the eight day old fungal culture grown on SMAY plates. One disc was inoculated to each bottle and mixed with it to disperse the inoculum. The bottles were incubated in BOD incubator at 25±1°C. Four replications were maintained for each treatment. The spores were harvested from sixth day onwards at definite intervals of upto 25 days by sampling 1 g of the digested material. The spore suspension of each sample was made by dispersing the inoculum in 10 ml sterile water blank with one drop of 0.02 per cent Tween-80, serially diluted and the spore count estimated using a haemocytometer. 3.7 GENETIC IMPROVEMENT OF STRAINS The genetic improvement of the fungi was attempted for V. lecanii Vl1 and M. anisopliae Ma1 through mutagenesis using UV radiation. 3.7.1 Mutation using ultra-violet rays Seven days old culture was taken and the conidial suspension prepared in sterile distilled water. The suspension was centrifuged at high speed (10000 rpm) to pellet the conidia. The conidial pellet was washed in 0.1M phosphate buffer (pH 7) and re-suspended in the same buffer. The spore concentration was later adjusted to 10 6 conidia/ml. The spores were then exposed to UV radiation in Petridish for 20 minutes at different heights from UV bulb in the UV cabinet. Later, the surviving population was analyzed by plating the mutagenised culture on SMAY agar. The bank of mutants which had the highest killing percentage were selected for further studies. 3.8 PERSISTENCE OF SPORES OF Metarhizium aniospliae (Ma2) AND Verticillium lecanii (Vl1) IN SOIL AND PHYLLOPLANE Survivability of spores in soil and phylloplane in simulated field conditions (in glasshouse) and field conditions were studied from January 2001 to May 2002. Spores of M. anisopliae and V. lecanii cultured on PDA plates were mixed with soil @ 4 x 10 6 conidia per g of dried and sterilized soil and filled in earthen pots. In addition, the same spore load was sprayed on leaves one set of potted soil with cotton plant of one month old was stored under shade in the laboratory and the other in cage i.e., open to sunlight and rains, simulating field situation. Samples were drawn at monthly intervals from individual pots and composite sample was observed for the presence of viable conidia in soil and phylloplane. Ten gram of soil sample was suspended in 100 ml sterile water containing 0.02 per cent Tween-80 and shaken on orbital shaker (120 rpm) for 2 hours. The suspension was sterially diluted and plated on SMAY medium maintaining four replications per dilution and incubated at room temperature (26±2°C). Colonies of the fungus formed on the plate were counted after 72 h.
Sl. No. Table 2. Details of different pesticides used in the experiments Common name Trade name Group 0.25 R.d 0.5 R.d R.d 2 x R.d 1. Endosulfan 35 EC Endocel Insecticide 0.70ml/l 1.40ml/l 2.80ml/l 5.60ml/l 2. Quinalphos 25 EC Ekalux Insecticide 0.50ml/l 1.00ml/l 2.00ml/l 4.00ml/l 3. Monocrotophos 36 Nuvacron Insecticide 0.62ml/l 1.25ml/l 2.50ml/l 5.00ml/l 4. Chlorpyriphos20EC Nuchlor Insecticide 0.50ml/l 1.00ml/l 2.00ml/l 4.00ml/l 5. Oxydemeton methyl 25 EC Metasystox Insecticide 0.35ml/l 0.70ml/l 1.40ml/l 2.80ml/l 6. Dichlorvos 78 SL Nuvan Insecticide 0.12ml/l 0.25ml/l 0.50ml/l 1.00ml/l 7. Dicofol 20 EC Kelthane Insecticide 0.72ml/l 1.45ml/l 2.50ml/l 5.00ml/l 8. Malathion 50 EC Malathion Insecticide 0.50ml/l 1.00ml/l 2.00ml/l 4.00ml/l 9. Dimethoate 30 EC Rogar Insecticide 4.25ml/l 8.50ml/l 1.70ml/l 3.40ml/l 10. Carbendazim 50WP Bavistin Fungicide 0.25g/l 0.50g/l 1.00g/l 2.00g/l 11. Iprodione Rovrel Fungicide 0.62ml/l 1.25ml/l 2.50ml/l 5.00ml/l 12. Chlorothalonil75WP Kavach Fungicide 0.50g/l 1.00g/l 2.00g/l 4.00g/l 13. Propiconazole25WP Tilt Fungicide 0.25ml/l 0.50ml/l 1.00ml/l 2.00ml/l 14. Mancozeb 75 WP Indofil M- 45 Fungicide 0.50g/l 1.00g/l 2.00g/l 4.00g/l 15. Triadimefon 25 WP Bayleton Fungicide 0.25g/l 0.50g/l 1.00g/l 2.00g/l 16. Hexaconazole Contaf Fungicide 0.25ml/l 0.50ml/l 1.00ml/l 2.00ml/l 17. Wettable sulfer 80% Thiovit Fungicide 1.37g/l 2.75g/l 5.50g/l 11.0g/l 18. Oxyflurfon Goal Weedicide 0.12ml/l 0.25ml/l 0.50ml/l 1.00ml/l 19. Pendimethalin30EC Stamp Weedicide 2.00ml/l 4.00ml/l 8.00ml/l 16.0ml/l 20. Attrazine 50% Atratof Weedicide 0.33g/l 0.66g/l 1.33g/l 2.66g/l 21. Butahclor 5% Machete Weedicide 11.0g/l 22.0g/l 4.00g/l 8.80g/l 22. Fluchloralin 45 EC Basalin Weedicide 0.66ml/l 1.33ml/l 2.67ml/l 5.34ml/l 23. Glyphosate 41 SL Roundup Weedicide 1.45ml/l 2.50ml/l 5.00ml/l 10.0ml/l 24. Alachlor 50 EC Lasso Weedicide 0.69ml/l 1.39ml/l 2.78ml/l 5.56ml/l R.d.: Recommended dose
Page 1 and 2: ISOLATION AND CHARACTERIZATION OF E
Page 3 and 4: Chapter No. I INTRODUCTION C O N T
Page 5 and 6: Contd….. Table No. Title 14. Spor
Page 7 and 8: Contd….. Table No. Title 39. Per
Page 9 and 10: Plate No. LIST OF PLATES Title 1. M
Page 11 and 12: I.INTRODUCTION The increased use of
Page 13 and 14: II. REVIEW OF LITERATURE The variou
Page 15 and 16: The entomopathogenic fungi so far i
Page 17 and 18: Table 1. Contd….. Sl. No. Insect
Page 19 and 20: Milner et al. (2002) studied the ef
Page 21 and 22: 25°C for sporulation and biomass p
Page 23 and 24: Pathogencity of various strains/iso
Page 25 and 26: Gopalkrishnan and Mohan (2000) test
Page 27: necessary serial dilutions under ph
Page 31 and 32: 3.9 EVALUATION OF DIFFERENT FORMULA
Page 33 and 34: Fig. 1. Mycopathogens of insect pes
Page 35 and 36: Table 5. Contd….. Month/s visited
Page 37 and 38: Table 5. Contd….. Month/s visited
Page 39 and 40: Table 5. Contd….. Mycosis noticed
Page 41 and 42: Totally eight isolates of N. rileyi
Page 43 and 44: Metarhizium anisopliae Vertucillium
Page 45 and 46: Ma 1 (Dharwad) Ma2 (Dharwad) Ma 3 (
Page 47 and 48: Table 8. Mortality caused by the is
Page 49 and 50: Helicoverpa armigera Dimond Black M
Page 51 and 52: Plate 5. The moratality of Hlelicov
Page 53 and 54: Table 11. Total biomass production
Page 55 and 56: Table 12. Total biomass production
Page 57 and 58: Table 14. Sporulation of Metarhiziu
Page 59 and 60: Mean spore yield x 10 8 /g 25 20 15
Page 61 and 62: Among the non-grain substrates, the
Page 63 and 64: Mean spore yield x 10 4 /g 35 30 25
Page 65 and 66: Sl. No. Table 19. Effect of fungici
Page 67 and 68: Per cent inhibition 100 90 80 70 60
Page 69 and 70: Sl. No. Table 21. Effect of insecti
Page 71 and 72: Out of nine insecticides, endosulfa
Page 73 and 74: Sl. No. Table 23. Effect of weedici
Page 75 and 76: Days after storage Table 25. Persis
Page 77 and 78: Table 26. Survival of Metarhizium a
Page 79 and 80:
Table 28. Survival of Metarhizium a
Page 81 and 82:
Sl. No. Table 29. Survival of Metar
Page 83 and 84:
Sl. No. Table 30. Survival of Metar
Page 85 and 86:
Per cent germination 90 80 70 60 50
Page 87 and 88:
Sl. No. Table 33. Survival of Verti
Page 89 and 90:
Page 91 and 92:
Page 93 and 94:
4.9 PATHOGENICITY OF Verticillium l
Page 95 and 96:
Sl. No. Table 39. Per cent mortalit
Page 97 and 98:
Culture grown on rice grains Harves
Page 99 and 100:
Tr. No. Table 41. Per cent reductio
Page 101 and 102:
Tr. No. Table 43. Per cent reductio
Page 103 and 104:
The fungal spray at higher dosage (
Page 105 and 106:
Table 46. Effect of wild types and
Page 107 and 108:
The hyphal and conidial characters
Page 109 and 110:
In an earlier experiment, bagasse w
Page 111 and 112:
M. anisopliae exhibited rapid reduc
Page 113 and 114:
higher mortality of aphids. The com
Page 115 and 116:
14. Oil formulations proved better
Page 117 and 118:
CHARNLEY, A.K. AND ST. LEGER, R.J.,
Page 119 and 120:
IGNOFFO, C.M., MARSTON, N.L., HOSTE
Page 121 and 122:
MULLER, E.J., 2000, Control of the
Page 123 and 124:
ST LEGER, R.J., BUTT, T.M., STAPLES
Page 125 and 126:
APPENDIX I Mean monthly weather dat
Page 127 and 128:
Maharashtra Association for the Cul
Page 129:
ISOLATION AND CHARACTERIZATION OF E
show all

ISOLATION AND CHARACTERIZATION OF ENTOMOPATHOGENIC ...

Create successful ePaper yourself

Delete template?

Save as template?