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humidity uf 50% 111 the dayrinie a11d uier 80Uh 111 the 111gh1 'rllc pl:1111s ivrre inltlally exposed to humidity as lilgli as over 90% b) ~I~ILIC 01. cuver~~ig 1lic111 w~tll PI'ISIIC b;igs. Tliry were sloicly i~cclltilnti~cd to tllc i~~iib~clil IIIIII~I~II~ b) grililu;illy c~l~cliilig [lie cover. Prolonged coierlng. 1.c.. oicr c\liohi~rv 111 li~gl~ I~III~II~II) c:ii~~cd \\IIIIII~ ;111d dccrc;~sed exposure caused preinillllrc dl.).lllg, b011i ~.~il~ll~iig III dciilli of Illc ~llii~il. IICIIC~ llic iili~11~ hu~nidlty accliniat~?at~ol~ procesh rcqi~~retl ;1bou1:I ~iiunlli ;III~ i11111c1\1 C;IK bc1i)rc c~itcril~g illto the stage 3. lhc pl:ilil. Iicl;ce, tllc e1illl.e II;I~~CI;III~ I)IOCC~\ \v;I\ duli~ ;II ,I II~III IIIICII>II~ OS 10.000 lo 12,000 lux ill ll~e C~II\ II.~II'" gron Ill cli;~~iibcr. I)lrccr CS/)U~LI~U 10 llic SIIIIII~III 111 llie grcc11 house caused dry111g 01' 111 ~I:IIII. lY1111 ~C~~ICCI IU IIIC I~IIOIO~)CI IU~. \I;I~U I W:I> lii;~~~itillll~d at I0 Iioilr l~glit ;IIILI4 Ihot~r dark, \t~igc 2 ,II 12 liu~ir l~gl;~ ,ilc cultllrc growl; cl;~
addition to tile tubes collt:lllllllg llcli~itl 11ltdii1111. 111 th~s 111st11od IC 1011 1101.11011 01'11111 plilllt was exposed to tllc ULII side COI~~I~IUI~~ ,111d hlo\\ly c ~ e e l ~ \\IIIIS ~ l gro\\illg ~ ~ ~ ~ I:igure ~ ~ 4.10 ~ d H, C sliows onulilcl mc~hud ol'sl;~l~c l~)drol~o~!~i.a s)sIcl!i ill i\lilcl~ 1I1c rouiillg ayslclil \+as completely 111111i~'racd [lie 4 a11e11g111 A~'IIOII'\ ~O~~IIIOII. 111 1111, 111crI10~\. IC I~~,IIII \\I18 direclly cxpoaed 10 1I1e ;III!~ICIII ~UII~IIIUII$ IYVIII ~IIC lir>r cI,I\ ll~c\er, (1ry111g\\;I> pl.evented bccauac ol' Ihc Il(l111d 11;11i1rc 01' 111c IIULISII~IIII~ IIIC~IIIIII. 'Illla ~i~e~li~d CZIII supple~ne~ll rile atagc 1 ;111d 2 ~Isacr~be~l ,~bi)\c;111d ll!~ plil~il\ CJII dlrc CIIIPII))C~I ~~I~CCIIIII~ \\ I~CII IIICIC \\:la 110 g~u!\lli ol'lli~ 10ti1 \)\~cI~I. \\IIICII dirsctlj i11111blti11g lllc g10\\111 of I11e ~I;IIII. ' ~III~ IIICI~IU~ \\;la IIII~~UI~,III\ III ~II~IJIICIII~ tllc survi\;il ticq~~u~lcy by irouble>llout~ng In 1Iic IIJI~CIIII!~ I)IUCCI\ IIICII~IOIIC~ 111 llie III;I~C~~~I~S :~nd methods section. C;ro\vlh of llic i'uor jysli.~!!, wliicl! 1s .I Ley I:lctor Ibr Ills II;I~~CIIIII~ appeared III nbou~ 15 ti;lys alicr IC I~.III~~)~:IIII,I~I~~II i1!10 ?O CIII 1po1.;. lllc Ilu\~ei':, kcre renloved aa sooll tiley ;ippcorcil ior ;lboi~t 3 \vi.cL\ >u ;I$ 10 II~CI~.I\~ llic icgelanvc grow111 and lliiis ilici'ease the ~ii~mbci' ol' accda per ~I,IIII I~CIIIU\,II ul' Il~i\\'cr\ :II\u rcs~111cd III IIIU 111creabe of br:111cl1111g ;III~ IC ~I;IIII~ ;i5$11111e~l
Page 1:
GENETIC TRANSFORMATION OF Cicer ari
Page 4 and 5:
Certificate Ger/$ed /La/ /Ae eldire
Page 6 and 7:
I deem il as a great pleastire to e
Page 8 and 9:
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
Page 28 and 29:
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
Page 34 and 35:
observations GkI Shull's experiment
Page 36 and 37:
(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
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 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 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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