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These combi~iatiori sl~owed liiorc cnllua cu~iipored to tlir ?.4.S-l'-co1itai1ii1iy media where the embryos originated liul~i tl~ callus III~IS~CII~. TIic iii~~iibcr ~I'expla~~ts respo~idi~~p was s~m~lar but tllc aicrage I~LIIII~C~ ut'e~~ibryu~ per exl>Ialit \\.;IS cu~iil);~~.;~~i\eI! Io\ver tl1i111 the abo\e-nient1u11cd JELl aeiies ol'~~icd~;~. Tlic c~iib~qus :~l>l>c;~rcd I~~rgcr tliii~~ tlic UII~S III JEM scrle, ;III~ tiley l;~cbctl ;illy atalhs. Fig. 4.2 sllu\rs 111~luction ol' e~~~bryos fro111 111;lturc e111bri.o ;ial:, cul)l;~~ir. Ucs~ rcsl)o~idlng COII~~III~I~IUII \\,IS ?O 11hl 2,411 w~tli 2 1lM kkieti~i tli;ir ilidi~ced 9.3 c~iibryos per c\pl;iiit No sccontl;~ry enlbryos ~ c ubscrvetl ~ e e\en VII prolonged CLIIILISC ul' tlic c\pl;i~it\ 111 11ic b~~~lie 1iiec11111ii \\li~Ic tl~c 111d11ccd c~iibryos gfiidunlly entered c;~lli~s pli;isc V;il.~oua otllcr ~ hpl~~iti s~lcli :la 1);1r1:, o1'111:1lurc e~iibryu ~1x15 (~>lu~iii~l~, radicle. side ;irlns ;lntl ~ii~tltllc purrion). Icaflcl>, stelii aeglnclits ;uid root acg~ilcl~t\ wcrc ci~ll~~rctl on bcst CII~~S)O~~IIIC COIII~II~~IIIUII ui'JLhl20 rIi;~t CUIIL;I~I~C~ I0 piVl ?.1,5-T ;III~ 2 11M ~IIIC~III, I'hc results ;ire allo\\li 111 rlic 1;lblc 3.5. I'lu~nc~le iiolil 1ii;iture c~iibryo ;IXIS ;111tlcaflel:, g;~vc bcst c~iibryi~ ~iiJui.t~o~i li.cij~lcllc) ol'70 ;IIILI JO'% I~II~SCII\CI~. All IC UI~ICI CSPI;II~~~ CKCCP~ r.id~clc ti.uni c~iibryu ,isls sliu\rccl luglicr iiccltlc~icy of II~~LICII~II \+liere ;I t~~iic tlepc~idc~~t ~nducuo~i \ids obhcric~l. 111 ge~icl.;il, clilbryu ~~~~luctioli \V,I~ observed u~tliin 4 to 5 weeks and s~icli phcnornc~lui~ \\;I, obser\cd ii~tli iiinlllre enibryu ;ix~s dc~.~vcd cxpla~its and le:111c1h IIo\+cYcI.. at~111 ii~icl 1.001 \egIIiCliti sllu\\i'd III~IICIIUI~ 011 ;I 1)ruIu11gcd CUI~LI~C on tlic :,;i~iic 1nedi~1111 Iur ;~buut X lo 9 \\ccks. Tlic cnibryo\ ;~ppcared a~ni~l:ir tu tllc ones on cliibryo nsls and leaflet explnl~ls. Tile epldernial I;iyer ol' SI~III and root scgmcnts broke opcn into crevasses and embryos apl)c:~rcd orig~~ii~ti~ig fro111 tlic ililicr tissue.
'I'lle c.;pesllilclith 011 slluol osg;llloycnc>lb wusc Il~u;~tlly d~\~dcd illlo two types namely dlrvct ;111ti ~nillrcct ~nle~liods B;ISIII~ on lliis co~iccpt tllc expl;ll~ts used were 6 c;lt;lgo~i~etillto two groups. I, lixl)l;i~i~s tli;~~ g:l\e tl~rec~ slloot org;~nogenes~s und 2. Euplants that gave liltllrect slioo~ org;inopenesls. E.\pla~ll\ 1i;iving prc-cx~stllig ~nicsistenis or any traces ol'mes~s~cms come u~lticr lis,~ gloul) i111i1 llic olhcri like Ic;~ilc~~, Ic;ll'basc, stel11 segments, ep~cotyl, liypocutyl, root segnients n11d root tip tllel were 11o1 ;lssociated w~tll any mrl.istemat~c tissue conles ulidcr tllc seco~id clabi. Tile ubscrvnt~o~is slioued that ~ndirect
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
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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
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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
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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 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
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
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1 APPENDIX 1. hIURASRICE AND SI(SO)
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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
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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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