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4.2 Histological studies ~II nlultlple slloot de\tloplllel~t I'I.~III Ahl.1 espl~l~~t: As described 111 secliull 4.1.2,l. Ah14 elplants alio\\.cd ;~dvcn~~t~uus niul~iple shoot induction ir~lll alloul bud> OII~II~;I~III~ (i.0111 dilji'reiit 11;1r1\ uI' IC ~C~CIICI.,IIIII~ ;I~c;I. Histological slut11i.a ol' ~iitlltiplc slloo~ III~IICIIUII 1j.oi11 ~llebe c\lll;ii~ts \\ere co~iductctl 10 colitirm tile for~ii;lt~oli oT~iit~ltiple ~~dvc~~ti~iou~ ~ilet~iate~ili~~ila ,ilicr the i.s~iiuv:~l ol',~\~llo~~y bud. 'I'hc dey \+Iicii ;ixill,~ry but1 \+';is rc~iiuvcd \\;IS eu~isiilercd as d.ly-I ;\lid tile slud~cs were coiiduclcd 1111 7 diiyb. Fig 4. I3 A tu H ~ll~\\'b 11c\cIu~i111~1it ol' I~~C~I~ICIIIUI~~ ;II dlff~~~lll ~ 1 ~ ~~;l.\l~~;l~) ~ 1 ~ ~ 5 lll~~l\l~lll ;lrC;l \\ 11~1C 15 10 ?(I lll~Vl\lClll~llt~~ ~~L'\c~u~JL'~~ ~1~1\~~!~11 4 to 6 days oi'culrurc on tlic slioot IIIC~UCIIOI? I~IC(/IIII~~ 1.1s 4.1) I5 sIio\v\ ;I \11iiilI ~p~rlio~i 01 axillary bud rc~iiaii~iiig cvcci alicr ~lic cvcislu~l of tile gruw~lig ,~\~llary but1 and dcvelopmeii~ of rnerlstelllo~d\ ;I[ the base of 11. .fliis rcsiilt alluiv\ l~islolt~g~c;~l c\ldcncc lor llie cnicrgc!ise of I~ILIIIIIIIC ;1tl\c1it111011\ 4 1 1 ~ 1 bt~d~ I'ro111 llic b;is;~l 11;1itll' IIIC ,~\~ll:iry bud :is sho\+~i 111 lllc 1'1g 4 -I U MCI.I\ICIII;I~IC :1et1\11y 01 IIIC ;ICII\CI! IIIVI~IIII~ ;III~ grt)iviiip cell:, could be ubscr\,cil 1i.ol11 tllc e\pl;~~~ts obt,~iiicti I~.OIII d;i)-5 tl~ii\,ii.db (1.1~ 4.13 I), 1'. F lllld I{). Tliese tIi\isio~ib rcullcd 111 [lie Ibr~ii,~r~o~i ol' II~C~I~ICII~~IIII. /OIICI IIl;it ~C\LIIIC~ ill llic f'urniatlon of 11lul1il)ie ili(10t bud> oh >liu\vll 111 ilic l:ig 4 13 (;, lb~g 4 1.1 A to I sliuws nicrlstcniatic ;icti\wl! ol'd~i i~l~~ig ecI1 iIi;~t C\CIIIII;III~ ~c~cIuI~c~I ill10 ~ ~iiilt~lll~ sliouts. Ccnetic tl~;insfuriii;it~on iraa carr~cd uiir by LI,III~ b ~ul~\ti~ ;IS IVCI 3s Afirohoi!i,~.ili~~~- lncdiilted lllelhoda In boll) llie proccd\ire> llic p~~l,~l~\c rr:iii~l'orniaiira \vcrc obtalllcd by selccti~lg the trallhl'urlllallt, liilllg il/illI ii\ lll~ s~~c~l~lble lll~.l.k~~. gCI1S iilld k:~naillycill 2s llle antjblouc br selcc[lon Conlrul cxplanls \r.crc ubcd lu tesl thc Ictl1,il tluse (LD 50) by culturing AM4 explants oil MS \rltI~ ~~III~IIII~CII~ (5, 10, 15, 20. 25 mid 30 11iuL) with
varying concenlr;~lions ol'TD% (0. 2. 4. 10 jlhl). 6 lo 8 cspl.~~it\ \rcrc ci~lluled per pl;~tc (Table 4.15). TDZ \\;is eniployed so .IS lo Itst tile clli.c~ ol' TI)% ti11 c~lli;~~~ci~ig survibal of thecultured explants 011 k;~~ianiyc~ii COI~I:IIIIIII~ III~LIIIIII~ hi~r\l\:~I \\.I, IC IIIC;I\II~L' tlf ;III~ collcel~~lble gro\\lll 01' tI1c ~\l~I~llll ~1lcI~lLllllg llllllll~llc \lltlcrhl>lellcc of chlorophyll plgliicnt In llic eul~l,1111. Tl~oi~yl~ llic c\l~lLl~~r p~o\\~l~ \\;I.; 111li11111cd c\cn ;it 20 11i9'1 ka~ia~n)ci~~ COI~C~IIII~II~~II. cliloro~~l~yll 1pig111c11t 11crh1\1etI (or re\\ ti~~irc cI;iy, ,~ttcr wli~cli expl;l~~ta blcuclictl. F.\~>I~IIII sro~111 ;111d aI1tlo1 b11tI II~~~~ICI~OII \\;I\ COII~PICICI~ lnliibitcd :11it1 CX~I~IIII ~OI~II~ICICI) blisc~cl~cil 111c C~JI~~CIIII.;IIIIJII LII' 30 111s I. I\;III;IIII)ICIII .A gradu;il dccrc;~sc In si~rvi\;~l I.;I~C US tlie ~YIII;IIII\ \\,I\ li1i111\1 \\II/I illcre;~hc ill L;III,IIIIYCIII co~ice~~tr;~t~o~i. Grti\vtl~ lctli,~l tloac \\$la fi\c
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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
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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-
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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
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Figure 4.1 A - D inducl~on of sonla
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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
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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
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introduction into protoplasts using
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evident by 1983 that the Agrobacter
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The above promoters were found to b
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gained an illstant popi~larity owi~
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legume crop species. Cotyledon expl
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1994). Clonal propagation of FI int
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were induced to for111 callus when
Page 56 and 57:
' the ernblyos into plantlets on MS
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and nprII for selection. PCR and So
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Agrobacleriuiii and ill otlier expe
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ioteclinologicsl i~iiprovemel~t of
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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
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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 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 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
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Table 4.3 111ducti011 of en~bryos O
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'Table 4.6 Dificrent combinations o
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Table 4.8 Effect oI'TDZ, 2-iP :~nd
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Table 4.10 Effect of i~lclusio~~ or
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Table 4.12 Induction of multiple sh
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Table 4.14 Effect of media constitu
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Figure 3.1 Dii~gralll~l~i~tic rcpre
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Figure 3.3 A - C Diagrn~nn~atic rep
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I . Nhu 1 51 ?C,%- .pal5125 &pa1512
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I
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Induction of somatic embryos from m
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I.iy111. J.5 SI:i;i.\ iil ~ ~ l ~ ~
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IIistologic:~l stutlic\ of dcveiopr
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Rc\[ricfiot~ ;111;1l)si\ ur ill? ~
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I'C'R :~llipLiiir:iriuil 01'700 bl)
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Fixi1r.c 17 \r:15 -l)K,.\ ;!lid J"
Page 272:
1:igurr 4.22 Sui~lllcrn :III:II!s~$
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