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either side of'bws;tl portiol~ oillle espl;~~~t bur d ~d nur elongste well. AM4 cnplant that can bc cons~dcred as itic:ll tbr gene IV,III~(~.I CK~~~IIIICIIIS yve risc to lllultiplc s11oots 3s nllllt~ple cl~iatcra \rl~cre tllc hiloot.; ;II.C lbr~~~sd 1rci111 TIII 1p0r11011~ 011 tl~c si+ollc~~ 1p;ltt of' ,~k~llilr! 11icr1>lt111. I:IC 1.5 ~IIu\\~ regi'~icr,~l~~~g IIIIIIII~>IC ~IIUI)I\ litllli 4\11 ~\~)I,IIII \+l~cre III:III~ ~Iloutb U~I~IIIJIC~ li.0111 tllc ~;Is;II I~U~~IUII:, ui 1I1c ~CIIII)\C~ SI~~I~II h~ld\ I'r~lu~~gcd CLIIILIIC ul' ill1 1llc ;~bu~c-~~~c~~tiu~~ccl L'\~~:IIII> 011 sl~oul ~t~d~~eliti!> III~~IIIIN 1~h111tetl ill L'\~~ 11er C\JI~~IIII \\~lli C:ICI sl~uut but1 ll~\ 1118 tlic ab1111) tu tlc\clul) 111tu CUIII~~~CIS ~)I:IIII 4.1.2.2 Elol~g;~tiul~ ul'tlle slloot buds: I'lilllt gro\\tll r~g~ll~ltorh \bere elll~lloyccl :It IU\V cullcclltr,ltloll:, 10 lllcrc~lhe tI1c ~nunlbcr ut'sllooti elo~~giling ~ C cul,l;1111 I ('l:~blc 4 13) Ijhl'. 2-11], ~IIICIIII ;III~ GAi kcre IC key COIII~)~~I~CII~> 1;ir b11iiot C~OII~;IIICIII, Tllc rlluul cl~~lg~ltio~l 111eil1,1 H~I.C I; IC~ iis C'I:L scrio koln \\111cli Cl:l.? iuln;lllrlllg 5 11hl 2-11' ,111d 2 lib1 ktiieull ;111tl CI:L? collrallllng 2 11hl GAI sl~u\ved beat I.~LLI~I~ by C~OII~:I~III~ 7 3 :III~ X 7 SIIOUI I.CI;I)~CIIVCI~ (1.1~ 4.6 C). EIollg;~tion 011 ~~IIC~III \+,IS iu1111d IU be I~~IIIIIII;I~, E~UII~,I~IUII W:I\ ill50 tcated 011 basal MS, bur ll~c rcsillts were 11ot s~~coui.ng~~~g irlic~i cu~lil);~rcd tu tlic ullicr II.U;I~IIICIIIS. IIowcv~~, prolo~lgcd cul~ure UII MS ioultcd III ille ruuul~g 01.5 lu 10% ul'tlic clo~igallilg shoola. All lilt tested conib111,111o11 sliu\rcd elollg;~t~un uS0.1 to 3 sl~uuls per uxpl,lnl 111 the illirlul step (elong;ll~on I), k111cli al~u\vcd ;I lei~gtl~ ul' 5 tu X c111 111 anutller I\+O wceks. I(ca1 of the explat11 bas placed 011 fl-csli clo~ig;itiun II~C~ILIII~. Ucat results IVC~C ubralncd ~vl1e11 tile elollgatlon \+us done \\~rli CkL? 111 the clulig,it~o~~ I ;III~ CLL7 111 tlic I;ltcr stilgca. The sllool buds gro\~ing 111 cluster'; \+ere cultrlrcd 111 CEL7 fur two to tlirec pas\:~gcs ;11 IO to 12
day inlervals. CAI naa fourid lo be ploy~l~g cruc~al role ill tile clur~p;~t~on 1 phi~sc where its application showed lncre;~sed internodal Ic~~ptli illid bctlc~ Icat' ~~~orliliulupy. Tile slioots cluriil;ited In cocI1 p;ias,l$e \vcrc tr~~~ist>rrctl lu tile louti~~p III~~IILIIII. I cupu\ud to tllc ~uutlng incd~;~ dr;~b~lcally di'crc;~actl the rootilig Srequency. Late roots rnent~oncd abobe, sliowcd \low growth illid they ~uritil~r~ed several root hairs. Sliools, wll~ch did 11o1 lout, ~ crc;~rrlcd lo the P!I;I~C 2 III HIIICII they were pulsc trc;~tetl \v~lli 100 lihl IUA li~r 2 to 3 scc. I
Page 1:
GENETIC TRANSFORMATION OF Cicer ari
Page 4 and 5:
Certificate Ger/$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
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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
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
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 114 and 115: egeneratloll \!;IS dli'licull ;~nti
Page 116 and 117: decreased a[ cllkalinc pH \+li~le 1
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
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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