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(Shade et al. 1991: blorru~i ct ol. 20011) II;I~ opcncii 11cr\ \15t;is lor tra~~sli.rrl~lg agronoln~cally 1lnport:lnt tr;l~ts lilto othcr crup pl.iilr\ tu dcvclup cllte c~ilt~\ars (Hirch. 19971. Thc trcliliology c~llpll.~t~c~illy aupport\ lllc ir,l~~sl;.l, III~~.~~.III~II ;III~ c\l)rcsaloll ol' foreign gsries 111 l~etcrologou> orga~iisiiis. I~~IUIIUIIIIC;I~~~ IIII~)O~I,IIII ~CIIC:, 10 1111l1ru\c tll~ crop ylrld qualirativcly ;IS \\,ell as qu:~i~tiiat~\~cIy II:I\~ beell ~;ul.~tcd ;III~I TIOIIVLI 1'1.0111 vartous sourccs. TI~csc 111clu~lc gclics Ibr li~l~giil r~h~st;i~lcc- C~II~III:I~C~ :ICI ~IUC~III;I~CS: blral I.es1st:llii.e- cu;it prutc~l~. rcplic.~>c ;lnd r~bu\~i~lc III~II[)IIIII~ I)~U~CIII\: lpc:,i ~CII~I~III~C- a~iiylasc ~~il~ib~torr, ~irutci~~.i\e i~ili~b~tvir ;IIIJ 111 ~O\III gcilca ,111c1 ~CIIL,~ hr IIII!II~ILIII;I~ improvenicnl- ?S iilbi1111111 ~CIIC B~ulceI~~iuIog) c~~~o~iip,~~~lllg gc11e11c CII~IIICC~III~ 011;'r:, ave~it~es to o ~crco~~~c tlle b;lrr~cr> 111 ~c111e\ III~ I~IJ\IIIILIIII cl~~chl~c;~ ~~I~ILILICIIUII 5.1 'l'issuc r~lllure atlldirs p~.otocol.; aplx,iscd 111 il~ickl)c;i rutlr,lllc cnlbr)o~c~lcs~\ 111:it (lid IIO~ \;I~I>I) Ihc ~ ~ l ~ L l l ~ C 1 ll ~ b l l ~CIICIIL ~ S ll,lll~~~~llll~lll~~ll k,,ll~) lCllUll\ ~11' l(
leatlets (Dilieshkuniar er al.. 1094). %earill ;ind I&\ \\;re ii>cii IU iildiice soiii;nic ciilbryos from inimature uotyletlo~~s \'cry liigli ti.cql~elicy ofpI;i~itlc~~ \\ere regcner;~lcd tio111 tllcse embryos (Hila el al. 1997) Hunc\e~. .ill l / l ~ 6 I)~I~IOL,U~\ ~ re~t~r,~l~~d 111' II~~I~CIIIIY 01 rnaturat~on and regcliclniioli ol' nliole p1~11tb ii.1 \olii,nic c~i~br~o~ei~c~~~ ~p;iili\\i~~, rliis was rc-co11firmt.d 111 our c\perlliitIi1\ A tcr! good ~~.CLIIICIIC! ul' ~II~~~ICIIOII \\;IS obhcr\ed tion1 m;iturc eiiibryo ;I\IS :iiit! Ic;itl~.i c\l)l;iii!s I:~iibryopi'l~li II~\IIC \\.,IS ~hsci\ed to iiiducc bcticr e~iibryugcnc.;ia 1Llc~c1l~l1)ll liarlle ol')utliig lc.~Ile~\ \\.;I\ ;II\ 1'11~ p~ihli~llcd prolocolh were repeatedly ~ricd \rill1 Iirilc ur 110 ~LICCC\A. I'ICIC r~i1111> s~~ggesI 111~11 ~C~CII~~:IIIO protocol vla soiiinlic ei~ibryugciics~h calilior be ;I rcll.ibls t11ic lirr gci~cl~c ~~~iislbrin;lt~oli exper~iiicnts 5.1.2 Ori~g~~~~ogc~~csis: ~!IC~O~)~U~);I~~IIIOII ,iiitI ~~~~I~V'IIIUII \),I tlirtc~ or t11~11rcc1 organogclie>ls 1i;ive beell e\le~~\~\cli ~itlti~cd :III~\CVCI:II 01 ICI~UIIZ \\ere ~p~lbl~~~l~rd. liiclus~o~i of I~~L'~I~ICI~I:IIIC IUIICI ii~lli IC c\I)I,II~I\ \L,I\ liltl~id ti OC cruci;iI for regcncr;ltiuil of slluoi butla .ind i\liulc 1)1;111t\ 111 CIIIC~~)C:I. I.;II.I) rcj)ort\ ~III cli~ckpca sho~ed callu> III~II;II~UI~ ,111tl ~IU\\III \\JIII 11u regciici,~t~
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
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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
Page 28 and 29:
Figure 4.20 A, B DNA profile of gen
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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
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
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ioteclinologicsl i~iiprovemel~t of
Page 64 and 65:
combinat~ons and concentrdtions of
Page 66 and 67:
egenerated via somatic eliibryogene
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egenerat~on from protoplasts and ca
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transformants were identified by sy
Page 72 and 73:
urgent econoniical control measures
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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 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: 5.0 DISCUSSION During tile la31 dcc
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
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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