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CERTIFICATE Certified that the entire work embodied in this thesis entitled "Genetic Transformation of Cicer arietinum L. for Insect Resistance" has been carried out by B. Jayanand under my guidance in the Genetic Transformation Laboratory, International Crops Research Institute for the Semi Arid Tropics (ICRISAT), Patancheru, Hyderabad, India and that no part of it has been submitted elsewhere for any degree or diploma. I(\( ,L, ,% ' 1 1 r 1 Dr K. K SHARMA Research Supervisor Hyderabad, Inha January, 2003 DR. KIK4RI '( St-14RM4 Cen-'icT ' . .OlVtO v ICRIS"' . L Andhla FrdJ,sh 502323, INUIA.
Page 1: GENETIC TRANSFORMATION OF Cicer ari
Page 5 and 6: DECLARATION 9 hereby aeclare that t
Page 7 and 8: "I'm convinced biotecllr~ology is g
Page 9 and 10: 2.2.2.1 Organogenesis 2.2 2.2 Sonla
Page 11 and 12: 4.1.2.2 Eloiig;itio~~ ofllie siloor
Page 13 and 14: SUMMARY Agriculture con~ributes maj
Page 15 and 16: adventitious slioot buds. Tliese se
Page 17 and 18: initially maintained in ~onviron""
Page 19 and 20: I~iduct~o~; of e~nbryos from varlou
Page 21 and 22: LIST OF FIGURES Figure 3.1 Diagr:~m
Page 23 and 24: Figure 4.5 A - I; Stages of ~nultip
Page 25 and 26: Figure 4.1 2 A, B Histological sect
Page 27 and 28: Figure 4.18 A, B PCR arnplificatio~
Page 29 and 30: Figure 4.21 A, B Southern blots of
Page 31 and 32: structurd and filnctiolial Impol.ta
Page 33 and 34: dishes for people of the developing
Page 35 and 36: tumefaciens, popularly kl~own as a
Page 37 and 38: Wakhulu, 1994). However, regenerati
Page 39 and 40: agalnsr varlous biotic and ahiotic
Page 41 and 42: equiren~ei~ts (Santbrd, 1993) The o
Page 43 and 44: Agrobactenu~ii (Zaenen et dl, 1974)
Page 45 and 46: ~ - /'- Binury vectors: A binary ve
Page 47 and 48: essential. In practice, temllnator
Page 49 and 50: 2.2.1 Food legumes There are severa
Page 51 and 52: effect of silver nitrate was studie
Page 53 and 54:
from germinated seeds of a multipur
Page 55 and 56:
thidiazuron (TDZ) was studied to in
Page 57 and 58:
protoplasts isolated from mesophyll
Page 59 and 60:
legume was considered to be liighly
Page 61 and 62:
sequentlal subculture on cytokinin
Page 63 and 64:
germinating the seeds of pasture le
Page 65 and 66:
phase (Scarpa et a]., 1992). Direct
Page 67 and 68:
~$2.3 Other metltods Protoplasts we
Page 69 and 70:
organogenesis rather than somatic e
Page 71 and 72:
l~izogeites for the integration of
Page 73 and 74:
natural biological controls with pr
Page 75 and 76:
ased on these reports, an estimate
Page 77 and 78:
Closrridi~r~n (Barloy et al., 1998)
Page 79 and 80:
loss of toxin activity (Uaw~thya et
Page 81 and 82:
point to consider would be the targ
Page 83 and 84:
!Chrirlre e~!ib~~,o rr.ri~: Tl11s c
Page 85 and 86:
Cold shock: Tlir explants benr~ng t
Page 87 and 88:
des~gnated as AB2, !\.as prepared b
Page 89 and 90:
EJJkcr ofpH on 1~111l11ple slloor r
Page 91 and 92:
effect of varlous co~iccntriirio~is
Page 93 and 94:
3.3 Histological studies oa multipl
Page 95 and 96:
lock was fiirtlier trimmed so that
Page 97 and 98:
dose (LD-50). This cxperimel~t \bas
Page 99 and 100:
4, ~IICI~L)I)LII~~IC~C'~ \\ere J~I'
Page 101 and 102:
for about 3 to J Jaya alid \\ere su
Page 103 and 104:
sectior~ :uid c\l~bl~blird pln111s
Page 105 and 106:
SII~)~I~XI~VI~~ \\,la t;~hen (;I~~U
Page 107 and 108:
gclle was probed uirh rlie PCll ti.
Page 109 and 110:
2. The nie~iib~;~~ir n;ir c.~~eli~i
Page 111 and 112:
w~tli kltlctin. More iii~niber ol'c
Page 113 and 114:
'I'lle c.;pesllilclith 011 slluol o
Page 115 and 116:
and klnet111 by using AM! ,mil A>,l
Page 117 and 118:
Sl~uot regeaeruliu~~ ill dil'fercl~
Page 119 and 120:
day inlervals. CAI naa fourid lo be
Page 121 and 122:
co~llbinatio~i \villi rncli otl1c1-
Page 123 and 124:
addition to tile tubes collt:llllll
Page 125 and 126:
varying concenlr;~lions ol'TD% (0.
Page 127 and 128:
oftlie seeds was tiound 10 be lo\~t
Page 129:
~~IIOII~IC DYA ror SOLIIII~SII L~~~
Page 132 and 133:
(Shade et al. 1991: blorru~i ct ol.
Page 134:
selection arid prepordtloii ul' c\l
Page 137 and 138:
appropriate conce~itralio~i of TDZ
Page 139 and 140:
cotyledoli incl~ided. Aalllary ~iii
Page 141 and 142:
terms of leaflel ~norpholugy atid i
Page 143 and 144:
wilting or drylng of tl~e pliints.
Page 145 and 146:
molecul~ir analysis of tlic I'III~~
Page 147 and 148:
with a variable copy 11~11nber CBI
Page 149 and 150:
Explant Preparation -Germmate seeds
Page 151 and 152:
Altieri, M.A., Whitcomb., (1980) We
Page 153 and 154:
Proceedings of rhr Natiorlul Acader
Page 155 and 156:
Biron, D., Vincent, C., Giroux, M..
Page 157 and 158:
Chilton, M.D.. Drummon, M.H., Merlo
Page 159 and 160:
Bucrllirs rhrrrmgre~iesis subsp. Is
Page 161 and 162:
Dillen, W., Clercq, J. de., Goossen
Page 163 and 164:
Cella, R., Falavigna, A., (eds.) Cl
Page 165 and 166:
Gavidia, I., Zaragoza, C., Segura,
Page 167 and 168:
Gulati, A., Jaiwal, P K., (1990) Cu
Page 169 and 170:
Hooykaas, P.J.J., Klapwijk, P.M., N
Page 171 and 172:
~ .- Jones, B., Davey, MR., (1991)
Page 173 and 174:
Khan, M,R.l., L.M., Heath, L.C., Sp
Page 175 and 176:
Last, D.I., Brettel, R.I.S., Chanib
Page 177 and 178:
Malik, K.A., Khan, S.T.A., Saxella,
Page 179 and 180:
Mickel, C.E , Standish, J.. (1947)
Page 181 and 182:
egeneration in excised cotyledon cu
Page 183 and 184:
Parr, J.F., (1974) Effects of pesti
Page 185 and 186:
' Potrykus, I., Spangenberg, G., (1
Page 187 and 188:
Rech, E.L., Golds. T.J., Hammatt. N
Page 189 and 190:
Ryan, Clarence A., (1990) Proteasr
Page 191 and 192:
.'/' ' Sh kar, S., Ram, H. Y. M., (
Page 193 and 194:
Stiff, C.M., Leuba, V.. Sun. 9.. Le
Page 195 and 196:
.-A. Uawithya, P , Tuntitippawan. T
Page 197 and 198:
__-- Wa_a_s5JL(l1)96) "Yes. But Doe
Page 199 and 200:
Zambryski. P., Holsters, bl., Kruge
Page 201:
2. Fixative used for fisntio~~ of t
Page 204 and 205:
Table 4.2 Induction of elnbryos 011
Page 206:
Table 4.4 Induction of embryos 011
Page 209 and 210:
~ Table 4.7 lnductio~~ of multiple
Page 211 and 212:
Table 4.9 Effect of pH of tile clll
Page 213 and 214:
Table 4.11 Effect of age of tllc es
Page 215 and 216:
Table 4.13 Effect of media composit
Page 217 and 218:
'Table 4.15 1.etl1nl dose testing :
Page 220:
I'relii~rutior~ ofi~xillary ~lleris
Page 224 and 225:
Figure 3.4 llestrictio~~ Innp of tl
Page 226:
Itestrictiol~ III;I~) 01' the plasn
Page 230:
Induclio~~ of son~atic cn~bryos fro
Page 234:
Effect uf co~~centrr~tio~t of BAP o
Page 250:
.A cu~irj~lr~t~ scl~clrie 01' rcgel
Page 256:
C!osrr ol)sei~v:~~iuri tlc\~clo~)i~
Page 260:
CliS I~i~locl~cl~tic;~l ;I,s;I) of
Page 264:
l:iyuris 4.18 IY'lt ;~ri~l~Ijl'ic;l
Page 268:
1:igitt.e 4.?O 1)S.t 111.uIile 01'
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