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Mmrtion and Assessment of DNA Spectrophotometric method Quantification of DNA was done using spectrophotometer (UV- 160 A, Sh~madzu) by diluting 0 5 pl of DNA with 995 p.l of sterile distilled water Concentration of DNA was calculated using the formula At 5015 pl of DNA taken, where At is 260 nm and 280 nm wavelength at which 0 D values were taken DNA concentration was compared with the known concentration of Lambda DNA Agarose Gel Electrophoresis The DNA was also quantified using agrose gel electroporesis Thirty ml of 0 8% Agarose gel (0 24 g Agarose, and 30 ml IX TBE) was prepared by boiling for 5 mln and allowed to cool when the temperature of agarose solut~on reaches 45 C Two pI of Ethidium bromide (2 pg/pl) was added mixed well before the gel was poured into a preset agarose gel unit with two 26-well combs one below each other and allowed to solidify After solidifying, the gel was transferred to gel running unit and tank was filled with 0 5 X TBE and combs were removed The DNA sample (2 pl) was diluted with 2 p1 of orange dye (10 ml 0 5 M EDTA, 1 ml 5 M NaCI, 50 mi glycerol, 39 ml distilled water and sufficient orange dye powder in 100 ml distilled water) and 5 1 of 0 5 X TBE buffer and loaded into the gel and )L uncut DNA (0 5 p1 I pl, 2 PI and 3 p1) is added to I*, 26" 27" and ~2~ wells at 50 ng/@, UX) nglpl, 200 ng/pl, and 400 ngp1 concentration The gel was run at 80 V for 30 min and visualized under a U V Transilluminator for DNA Gel
whoresis method was preferable than spectrophotometric mahod since the DNA vidzed by this method was more pure than the other method AFLP-PCR amplification conditions AFLP core reagent and starter primer kits were purchased from (L~fe Technologies Invitrogen, Gatherburg, and U S A) The analys~s was done as per the manufacturer's protocol with slight modifications Restriction Digestion of Gtnomic DNA Component EcoRVMsel (Invitrogen) Restriction enzyme buffer Genomic DNA Total 1 10 p1 of cocktail was distribrlted in each labeled tube 2 Vortex and briefly centrihged 3 Digestion was canied out in a final voiume of 10 pl at 37 C for 2 h Ligation of Adapters. r Component T4 DNA ligase 10 X buffer Starting cone. I 25 1JIp1 I X Song Initial IUlpI Adapter ligation solution 4 Final conc. I 25 U//pl 10 X 250-600 ~JPI Vol. added per sample (PJ) 1 2 8 I I Volume added Per sample (pl) 1 I
Page 1: EVALUATION AND CHARACTERIZATION OF
Page 4 and 5: Dr. RP Thrkur Senior Scientist (Pla
Page 6 and 7: We certify that the thesis entitled
Page 8 and 9: My docere thanks are also due to Mr
Page 11: About the bost GENERAL INTRODUCTION
Page 14 and 15: Bblogiul control About biocontrol a
Page 16 and 17: af caupatiile biological control ag
Page 18 and 19: MATERIALS AND METHODS (GENERAL) Gen
Page 20 and 21: INTRODUCTION Species of Trrchaferma
Page 22 and 23: REVIEW OF LITERATURE Quantitative e
Page 24 and 25: espectively m vrho On seed coated w
Page 26 and 27: phte The disc of the A. Jaws was pl
Page 29 and 30: Fig 3 h?dlor groundnut growlng d~st
Page 31 and 32: Table 1 Population dynamics of TnWr
Page 33 and 34: Bunker and Kusum Mathur (2001) eval
Page 35 and 36: INTRODUCTION '2bs genus Trichodermo
Page 37 and 38: AFLP with a range of prima pairs d
Page 39 and 40: dried mycdium 0 1 - 0.15% by weight
Page 41 and 42: molearlrr data, morphology, physiol
Page 44 and 45: The spore suspension of Trrchoderma
Page 48 and 49: 1. 5 ul of ligation mixture was add
Page 50 and 51: Thermal cycling for dectivr amplifi
Page 52 and 53: plates the comb was placed in such
Page 54 and 55: 'rn Protocol 'XI2 amplification: Tb
Page 56 and 57: Amplified genomic DNA was restricte
Page 58 and 59: calculated using SIMQUAL program of
Page 60 and 61: 2) T. fertile of second group has t
Page 62 and 63: 8) T. aureoviride had the following
Page 64 and 65: ~roup V ~ncludes three specles of t
Page 66 and 67: Fig. 1 1. UPGMA Dendrogram using mo
Page 68 and 69: AFLP Results: Molecular characteriz
Page 70: viz., 7: longibrachia!um and 7: pse
Page 73 and 74: Fig. 14. Multidimensional scaling o
Page 75 and 76: Pnc@basi~m section. 'h second major
Page 77 and 78: Fig. 16. UPGMA den&- of 25 Trichode
Page 79 and 80: DISCUSSION Taxonomy of Trichodenna
Page 81 and 82: with Trichodenna section distribute
Page 83 and 84: study re selected on the basis of a
Page 85 and 86: mODUCTION Biological control of pat
Page 87 and 88: REMEW OF LITERATURE Chitina5e.s are
Page 89 and 90: chitill, hifiarin and A s of ~athog
Page 91 and 92: ~ntifungd Compounds MATElWLS AND ME
Page 93 and 94: mom temp- for thirty min and a b s
Page 95 and 96: deterioration Before use it was dhe
Page 97 and 98:
pec- model DV-20) at a wavelength o
Page 99 and 100:
sucking gd buffer ~ris - HCI 01 .O
Page 101 and 102:
~cetic acid Water Methodology Prepa
Page 103 and 104:
suining and w i n g of- coomuie Bri
Page 105 and 106:
Fig 18 Effect of volatile antibioti
Page 107 and 108:
Fig. 20. Preliminary screening of T
Page 109 and 110:
On other hend the isolates capable
Page 111 and 112:
Table 5. Effect of Mch0denna volald
Page 113 and 114:
Table 7. Amount of protein prmh.tce
Page 115 and 116:
DISCUSSION hRUoan wnthtion of groun
Page 117 and 118:
(1993) stated Chibl~c V h m I: hani
Page 119 and 120:
Chapter IV Evaluation of selected T
Page 121 and 122:
aspergiUi hss allowed developmat of
Page 123 and 124:
control of PI- diseases by biologic
Page 125 and 126:
01. (1983) compand two rum-tYPe Wut
Page 127 and 128:
esim to A. flavvs -on end assuming
Page 129 and 130:
Methodology Biomass of Trkhoderm is
Page 131 and 132:
preparation of Trkhoderma Inoculum
Page 133 and 134:
~~~ergiUusflovus seed infection rou
Page 135 and 136:
T~~I. I I. Randomization of 8 b-eab
Page 137 and 138:
Fig 23 Blocontrol of A finjrr\ cont
Page 139 and 140:
phosphate buffer dine (PBS), (pH 7.
Page 141 and 142:
The plates were washed in three cha
Page 143 and 144:
Biomass of Trkhoderma isolates: RES
Page 145 and 146:
8 7 2 6 l a 5 cn I 0 1-T I Trichode
Page 147 and 148:
Different isolates of Tricltoderm s
Page 149 and 150:
population dynamics of biocontrol a
Page 151 and 152:
Management of aflatorin contaminati
Page 153 and 154:
~ ~ 17 bpopulation l ~ dynamics of
Page 155 and 156:
Management of aflatoxin contaminati
Page 157 and 158:
Table 19 Population dpImks of bioco
Page 159 and 160:
Management of anatorin contaminatio
Page 161 and 162:
~ ~ b21. l e Population dynamics of
Page 163 and 164:
Effect of ~richoderma SPP. on seed
Page 165 and 166:
Fig 26 Groundnut seed showing seed
Page 167:
During field experiments conducted
Page 170 and 171:
I lo 2 week old pcaMf plmts with fo
Page 172 and 173:
ad~sh was induced by 7: h m m when
Page 174 and 175:
Summary
Page 176 and 177:
Besides mor~holoEYa identification.
Page 178 and 179:
PPseud~koninPi (T 29) produced more
Page 180 and 181:
~~nclusionS isolation --- 212 isola
Page 182 and 183:
,4bdel-satar, M. A., Khalil, M. S.,
Page 184 and 185:
uisset, J. (1992). Trichodemm atrov
Page 186 and 187:
Busby, W. F. J. R.9 and Wogan, G. N
Page 188 and 189:
~~cological Society. 57,2549. Denni
Page 190 and 191:
Grondona, 1.9 Hermosa1 M, R.1 Tejad
Page 192 and 193:
IARC. (1993). ]ARC on the evaluatio
Page 194 and 195:
Kuhh K.3 Lirkfeldf E-9 Samuek, \\..
Page 196 and 197:
~ackenzie, D. W. R. (1998). As~er~i
Page 198 and 199:
Muller, U., and Wolfenbarger. L. (1
Page 200 and 201:
Raguchander, T., Samiap~an$ and Arj
Page 202 and 203:
Sanders, T* ". (19*'). Effect of hu
Page 204 and 205:
Turoczi, G., Fekete, C., Ketenyi, Z
Page 207 and 208:
Identification of Trichoderrna Spec
Page 209 and 210:
conidiophOWS, shape of the phlalide
Page 211 and 212:
(7, longibrochiarunt) showed the ma
Page 213 and 214:
soil and pod ~Mlpling, Soil was sam
Page 215 and 216:
1 3 4 5 AF'LP and SNP D i a ~ oof s
Page 217 and 218:
- indue hat- in plan& (15.57). wa e
Page 219 and 220:
drlxol~tCeIhKdCaI- 111@~d~d3~~iimlc
Page 221 and 222:
-inmfo~~bpoplLtianr@-~wnnmwbrb~r~--
Page 223 and 224:
~ O f ~ ~ m l p t n 2 ) . T h i ~ d
Page 225 and 226:
T~wi*olrtu,w~~pandwithtbt~duuaof~bu
Page 227 and 228:
In this WdY have unable to diffennw
Page 229 and 230:
2. Bhst R V. 1989. Risk to human hc
Page 231 and 232:
1 49. VmthS V. Bhnt. 1998 MWXIN tn
Page 234 and 235:
collecl~on number and gene sequence
Page 236 and 237:
T*BLE4*. mP marken s~gn~ficantly a.
Page 238 and 239:
- 0.00s TI@.' R TI43 R TI70 R 'T~LI
Page 240 and 241:
FIGURE 5. UPGMA dcndrogram of thc A
Page 242:
Allen S.G., Dobrenz Heritability of
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