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egeneratloll \!;IS dli'licull ;~nti could 1101 be ocl~icvctl n.itl~ 111s tcsted 111cdiu. M;lture elnbryo axli, shout tip. ;lxlIl;iry bud, cot)li'llo~i.~ry node .ind d\~li;lr) III~~I\~~III (AM) explnllts sl~u~vcd sl~uut rcge~~cr;~l~u~~ ;IIIIUII~~I \\ IIICII ;1\111;1ry II~~~I\I~III c\p1;111ts ,AM2 :III(I AM4 were ~CICCICJ ,IS [lie best UIICL OL'C F I J ~ 4). Sla~~d:~rdi~:~tio~~ ol' il~~ll~cliu~~ ni~di11111: SI,III~~:I~~I/;IIIUII oi';111 ;I~I)~UPII,IIC I~I~L~I~IIII ii~r III(~LIC~I~~II LII'II~LII~I~)~~ hl~out\lio\\h II~IIIIII)/C hliuut II~~UCIIUII I~UIII III~IIIIIC CIII~~)O iixih by 11511ig DAl> ;IS 111c 1pri11~1p;iI grot\ tli regt~l:i~ur ivlt~lt~~~lc slioul~ urigi~i:itcd o11c al'tcr tllc otl~cr ~~iy~~cliru~ioti~l~ ;it IIIC \IIUUI 111) ;IIICI iixlll;ir) bud rcg1011~ 01' I~I,IIIIIC eillbryo :IXCS. Thc ~spli~~it gre~i I~CC IU ~ULII t1111es III sl/e ;illil Iiicri~tcIii;iLIc IC~IUIIS br,~~~cli~d IU 111uIt1ple \tiout\ 'rllc otllul cullib~~~;~l~u~l~ ~ulll;llllitly 2-11' ;III~~\I\;'+ \4llll Di\l' 111 ~CIICI.;II sI10i~ed ciicuuraylilg results, h1;11iy ~I:IIII gru~,th rcgl~l;i~or\ lilw LC~IIII~, ~ICIU~~IIII, lAA iilld IDA ucse citlicl- i~liIividu,illy or 111 combtnolion \LIIII UAI' sliowcd soinc ~rcdundiint rcsults of lo\+ li.eqiic~~cy 111ul:iplc illout ~~iduct~oil and llrcy were IUI tzibulalcd due to ltlclr ~iiiig~iiticant rcsponic I'ur ilioot i~lduct~o~i Separate crperime~us for ~ hc sla~~d;~rdi~,itiut~ oi'sliout illduct~o~~ mcdlum were done uslng TDZ as pr~~icipal niillt~plc slioot ~nduclng grow111 regulator 111 cun?b~nnt~o~i w~th 2-if'
and klnet111 by using AM! ,mil A>,l4 c\l1l.i11tr Hoth 111c c\;l,l;~~~t\ hliu\\cd lllorc or less s11i11lar ~iuiiibcr ot' ~nilliiplc rIiou~\ per e\l)l.ii~r (l',~blc 4 X i rI)L \\;IS 11act1 ;II various conccntr,itioii, ralig111g liutu 2 lo IOU phi. Tile 111rdln C~III~III:UI~IIS \\ere 11;1111cd 11s JCR acrles \\liicli tilso cu~is~atcd ot'?'i)Z \\nil 2-11' ,111d LIII~IIII. SI;III(I;~ILI~/:~~I~II ot' he I~~C~IIIIII \\;la done hccpll~g tile I,IIU st,i~cs ui ~IUII~:IIIOII ;IIIJ suo1111g 111to ~,o~is~tIcl.;i~~ol~ :is cu~~ci'~itl.;~l~ui~ ,IIIL~ IIIIIC vl cllIl!lrc 01' c\lll;1111\ 011 I'~)/-COIII:IIIIIII$ III~.~I;I r110\v \1g11ilic;1111 ctf'cc~ UII a!~bsci]t~c~~t diuul cIo~ig:it~o~~. IIIC~\ISIUI~ 01' !.il' i111tI L~IICIII~ \\;I< :IISO t'o~111ci easc1111~11 ;IS 111c) \\ere ;IISO IIIcI~I~I~~I 111 ~IIC SIOUI cIoi~g,itio~~ 111c~li~11i1 [SEM) ;I( Iu~~cr ~UII~~III~~~IIUIIS. 11 Iun le\cl IJI'I'DZ (41141, J['l
Page 1:
GENETIC TRANSFORMATION OF Cicer ari
Page 4 and 5:
Certificate Ger/$ed /La/ /Ae eldire
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I deem il as a great pleastire to e
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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
Page 20 and 21:
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
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Figure 4.20 A, B DNA profile of gen
Page 30 and 31:
1'1.0 INTRODUCTION L~fe is the ter~
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them. About 10% of the Earth's land
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observations GkI Shull's experiment
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(Nene and Haware, 1980). The legume
Page 38 and 39:
jf.0 REVIEW OF LITERATURE Large par
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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
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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 82 and 83: 3.1 Pl:~nt 111nteri:il sllrl cultur
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 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
Page 150 and 151: 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
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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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