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3.1 Pl:~nt 111nteri:il sllrl culture col~ditiul~a: hfatorc hccdh ut'cliiclq~c~~ (('I~~,I,~IIC,/IIIIIIII 1.) CLI~II\,I~ C.235, ,I !\~cIcly gro\\~i ~LIIIIV;IS 111 Il~di;~, ncrc s~~rl';~cc s~cril~~cd NIIII ~(I'?U (1 '\) c111,11iol (or 1 1i1i11 , U loo I~~CI~II~IC ~l~lor~(lc for 10 11111i . ii~i~l r111seci 5 l1111es 111 h~ct.11~ d~>tillcd \v,~ti.r ~IILIS 10 ~U,II\III~ u~erl11g111. The dc-~ii~ted 5ecda \rere Lcpt iir~ gcniii~l,~~~oii 011 hli~~;lzll~gi' ,111d SI\OU;'> (IOO?) I~IC~ILIIII (\IS: we ;~ppe~i(i~\ I), cir 011 ;I II~~L~ILII~: q)ec~licci tiir tllc cyil,~~il \>I~~>\IKIIIOII ('LIIILIIC ~i~ccl~;~ nc~c ohcd ;I, liqul(I or ;IS scln~-,olld Iho\111g 0.8'io (\v \) I)ilku-lj;~cto :I:I :I rcq~~~r~d dilcl 1pII \\;I\ ;iiljusted to 5.8 i~lilcss o~licr\\~sc ~iie~itio~ictl. All llic ~~hsui. cultuscb \\cl.c I~I;IIII~~III~C~ ;II 20ilYi' u~idsr contlnuou:, cool irl111c l~glit ~proi~dcd by lluo~~c~cci~t I;iinp\ (30 11Lln 'S I)
!Chrirlre e~!ib~~,o rr.ri~: Tl11s call be cunsidercd :IS \verc soaked uicril~glit ;iiid tile sscd co;it \\,is ic~ilo\cd IC tile O-do)-oltl accdlilig. Thc seeds i'illo\\ III~ I~IU~I~III~. Tlic eo~yledo~~~ \+ere bplit ope11 JII~ IC CIII~I:)~ ;I\I~\!i~\ ici~~o\c~l c:~~cliill~ ,111cl CLIIILII:~~ 011 11ic ciiibryo ~~~cilictiu~i 1iicd1i1111 (Elhli 111:11 cunsiatcd of I0 11hl 2.4.T-1 :ind 2 1111 killc1111 Synthetic ourci~is. 2.4.5-T and 2,4-0 iirc lllc ba\ic co~iipoi~cii~~ ui' il~e enlbryoge~lic indiiur~uii media. All tile 111cdi:i were re,ted un ~lintilrc cniblyo akis eul>lalil li~r opli~i~izatio~~. The ~~iducrio~i ilicdi;~ were tiiu type!, 01ic 2.4.5-l' based cilitl llie olhcr 2.4-U based. Tllc lornicr was iiariied ;is JEJI scrics niid llic lalcs \\a> ilamcd na JIIM serlcs. 111 JLIhl series tlic 2,4.5-T \+\.as :idded ;it 2. 5, IO ;ind 15 phl :~nd in cach tscntniclil ~ar~able COIICCII~~B~IOI~S or TDZ at 0.5 and 1.0 phi. BAP ;it 0.5 ;!lid 1.0 j1J1, Lcnrlii ;II 0.5. 2 O ,ind 5.0 phl nlld kinetin at
Page 1:
GENETIC TRANSFORMATION OF Cicer ari
Page 4 and 5:
Certificate Ger/$ed /La/ /Ae eldire
Page 6 and 7:
I deem il as a great pleastire to e
Page 8 and 9:
CONTENTS ABBREVIAI'IONS SUMMARY LIS
Page 10 and 11:
3.2.2.3 Shoot elongat~on 3.2.2.4 Ro
Page 12 and 13:
ABBREVIATIONS 2-il' 2-isopci~~cnpl
Page 14 and 15:
frequency was observed wit11 2,4,5-
Page 16 and 17:
in vitro grown plants w;is ocli~eve
Page 18 and 19:
LIST OF TABLES Table 3.1 Pr~~iier c
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Table 4.1 1 Effect ul'age uf lhe ex
Page 22 and 23:
Figure 4.1 A - D inducl~on of sonla
Page 24 and 25:
Figure 4.9 A - C Alternative llleti
Page 26 and 27:
Figure 4.15 A, B Restriction analys
Page 28 and 29:
Figure 4.20 A, B DNA profile of gen
Page 30 and 31:
1'1.0 INTRODUCTION L~fe is the ter~
Page 32 and 33: them. About 10% of the Earth's land
Page 34 and 35: observations GkI Shull's experiment
Page 36 and 37: (Nene and Haware, 1980). The legume
Page 38 and 39: jf.0 REVIEW OF LITERATURE Large par
Page 40 and 41: plants from them enlhuscd scientist
Page 42 and 43: introduction into protoplasts using
Page 44 and 45: evident by 1983 that the Agrobacter
Page 46 and 47: The above promoters were found to b
Page 48 and 49: gained an illstant popi~larity owi~
Page 50 and 51: legume crop species. Cotyledon expl
Page 52 and 53: 1994). Clonal propagation of FI int
Page 54 and 55: were induced to for111 callus when
Page 56 and 57: ' the ernblyos into plantlets on MS
Page 58 and 59: and nprII for selection. PCR and So
Page 60 and 61: Agrobacleriuiii and ill otlier expe
Page 62 and 63: ioteclinologicsl i~iiprovemel~t of
Page 64 and 65: combinat~ons and concentrdtions of
Page 66 and 67: egenerated via somatic eliibryogene
Page 68 and 69: egenerat~on from protoplasts and ca
Page 70 and 71: transformants were identified by sy
Page 72 and 73: urgent econoniical control measures
Page 74 and 75: applications of traps. Technically
Page 76 and 77: in the laboratory and releasing tl~
Page 78 and 79: have been exhaustively reviewd (Dai
Page 80 and 81: larvae of certaln Insects were unab
Page 84 and 85: 0.5. 2.0 and 5.0 pM were added. 111
Page 86 and 87: Lec!/i?is: Young anci juve~iile lea
Page 88 and 89: AM3 explant: This was a by-product
Page 90 and 91: PM) 2nd GAJ (2 5 PM), eltller singl
Page 92 and 93: of the plastic bag was removed rwlc
Page 94 and 95: - . -- laopropyl alcoliol ; Absolul
Page 96 and 97: The sl~des were Illell muiaited and
Page 98 and 99: 6. The bupcniatant \\,I,, t,ikcli i
Page 100 and 101: 5. Microcal~lers were allowed to se
Page 102 and 103: till the growtli of bacterial cells
Page 104 and 105: ;I! 37 'C for 3-24 Iiours 111 d;irL
Page 106 and 107: PCN for 111cl.4 ge!iiJ: I'C'I< tbr
Page 108 and 109: 4. Tliu pel \\as \r;~alicd 3 ti~iir
Page 110 and 111: 4.0 RESULTS $loaf of tllc c\l~cr~~l
Page 112 and 113: These combi~iatiori sl~owed liiorc
Page 114 and 115: egeneratloll \!;IS dli'licull ;~nti
Page 116 and 117: decreased a[ cllkalinc pH \+li~le 1
Page 118 and 119: either side of'bws;tl portiol~ oill
Page 120 and 121: IBA sl~owed bro\\~~i~~g i~lld de:~l
Page 122 and 123: humidity uf 50% 111 the dayrinie a1
Page 124 and 125: 4.2 Histological studies ~II nlultl
Page 126 and 127: sliowetl no liirtlie~ yrowlli Tllc
Page 128 and 129: Conlirmntioll of plllati!e t~.~ll~a
Page 131 and 132: 5.0 DISCUSSION During tile la31 dcc
Page 133 and 134:
leatlets (Dilieshkuniar er al.. 109
Page 136 and 137:
used with an add111ou;ll care :u~d
Page 138 and 139:
tissue to regenerate illto lesser i
Page 140 and 141:
prololigcd culture on TUL sho\\~d 1
Page 142 and 143:
exceed 5 cni Icngtl~. Loiigcr shoot
Page 144 and 145:
stem and lravca Secol~d;lrq ~'IIICI
Page 146 and 147:
arnplitication of the sopccIi\i. 90
Page 148 and 149:
Explant Preparation -Geminate seeds
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6.0 REFERENCES Adkins, A.L.. Godwin
Page 152 and 153:
Avenido, R.A., Hattori, I
Page 154 and 155:
Benfey, P.N., Chua, N.1-I., Ilcgiii
Page 156 and 157:
Cnrvalho. IC1.H C dc , Uui Vat1 LC
Page 158 and 159:
Chu, P.W.G., Andersoli. B.J., Khan.
Page 160 and 161:
Deblaere, R., Reyneerts, A., Hotte,
Page 162 and 163:
Drun~mond, M.H., Gordoil. M.P., Nes
Page 164 and 165:
Foster, J.G., Coulombe, B.A., Van S
Page 166 and 167:
Gill, R., Saxstla, I'.K., (1992) Di
Page 168 and 169:
Hess, D., (1970) Versueche zur tran
Page 170 and 171:
Hopkinson, M.J. ;md Collins, I1.M.,
Page 172 and 173:
Karthikeyan, A.S., Sarina, K.S., Ve
Page 174 and 175:
Krens, F.A., Molendijk, L., L., Wul
Page 176 and 177:
Li, Z.Y., Tanner, G.J., Larkin. P.J
Page 178 and 179:
Maxan~, A.M., Gilbert, W., (1977) A
Page 180 and 181:
field conditions. Procc.rdbigs o/Na
Page 182 and 183:
Ohara, A., Akasako, Y., Daimon, H.,
Page 184 and 185:
llechrnlics sur les prnidilies des
Page 186 and 187:
Rand, G.M. (Ed.) (1995) Ftordir~~rc
Page 188 and 189:
Richardson, M.J., (199 I ) Seed sto
Page 190 and 191:
Sarria, R., Calderon, A., Thro, A.M
Page 192 and 193:
Shri, P.V., Davls, T.M., (1992) Zen
Page 194 and 195:
Tegeder, M. Koli~i, I-I., Nibbbe, M
Page 196 and 197:
Venkatachalam, P., Klshor, P.B.K.,
Page 198 and 199:
Winans, S.C., (1992) Tiyo way chemi
Page 200 and 201:
1 APPENDIX 1. hIURASRICE AND SI(SO)
Page 203 and 204:
Table 4.1 Induction of sorrlalic er
Page 205 and 206:
Table 4.3 111ducti011 of en~bryos O
Page 208 and 209:
'Table 4.6 Dificrent combinations o
Page 210 and 211:
Table 4.8 Effect oI'TDZ, 2-iP :~nd
Page 212 and 213:
Table 4.10 Effect of i~lclusio~~ or
Page 214 and 215:
Table 4.12 Induction of multiple sh
Page 216 and 217:
Table 4.14 Effect of media constitu
Page 218:
Figure 3.1 Dii~gralll~l~i~tic rcpre
Page 222:
Figure 3.3 A - C Diagrn~nn~atic rep
Page 225 and 226:
I . Nhu 1 51 ?C,%- .pal5125 &pa1512
Page 228:
I
Page 232:
Induction of somatic embryos from m
Page 238:
I.iy111. J.5 SI:i;i.\ iil ~ ~ l ~ ~
Page 254:
IIistologic:~l stutlic\ of dcveiopr
Page 258:
Rc\[ricfiot~ ;111;1l)si\ ur ill? ~
Page 262:
I'C'R :~llipLiiir:iriuil 01'700 bl)
Page 266:
Fixi1r.c 17 \r:15 -l)K,.\ ;!lid J"
Page 272:
1:igurr 4.22 Sui~lllcrn :III:II!s~$
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