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stem and lravca Secol~d;lrq ~'IIICII~~ \\err .~lso ~pru~~~otcd b! 1111s IIC;IIIIICIII 1'111s sort 01' pron~otion of veget~~t~vc gru\vtl ~:IIIIS ~I~IIII~C~III~C \\IIIIc I~~~IIIII;I~IIIII~ :IIILI 1proccss111g of l~ivaluable transgenic pli~~lts. 5.1.3 Histological slutlica ol'i~~ultiplc slioul tlr\rlol)i~iel~t I'I.UIII .\A14 c\pl:~l~t: L)c\cluj>~i~c~~t of ~n~~lt~plc ~Iluurh i1t1111 :\\I4 C\~I~,IIII \\:I, iti111~1 I,I I~c ;IC~\CIIIIIILI~I\ [~Ilcllulyl~~;~ll! I-~o\\c\cs. IIICII. :I~\CIIII~IU~I> 11~11111.~ !\.I\ LOII~~III~C~ .II'ICI IIIC I1i~tolt1~1c;il obscr\;~t~u~li c;~rr~ctI utit .II d1ll2~11t sliigc\ 01' gr~\\tIi uI' 111c C\~~~.IIII 11r~i.1tllv IC ,\hl-l ehplal~t LIC\CIUJ)III~III oc~,~irj 111 t\\u ~I;I~L', I. 111) 10 .I\III;II! but1 ICIIILIV,~~ ,111~12. 1111 ti) 111c Stiipe ufrt1110\~1l 111'111~1lll~1lc JIIUOI buli UII~III~I~III~ fio111 ~C~CIICI.;I~III~ .I~C:I I'~C\CIICC 01' ax11l;lry bud c\crlh I)IC\\LI~C 01' ,II)IC;I~ ~UII~III~IIICC OII IC II~\LIC 01' CIII)ICLI~III;I~~ IIO~C regloll. Ile~icc. ;IS c\l~cctcd I~CIII~IV;I~ 01';1\1ll;iry hut1 IIC~.IIC\ t11c ;I~)IC;I~ ~~IIIIIII,IIICC :IC/ d~i'l'trc~~~ regiulib III rlrc LIIII~I 111 IIOII-III~I.I\ICIII;~IIC C ~ ~ IC\LII~\ ~ i 11 C~III l)c suggested that re~lloval ot'rnt~lt~plc >Iiuur bud, 011 t1.1)-5 UI d,~y-c~ ;III~ C~-LLIIIII;I~I~III lilr 72 ~OLIS~ COLII~ giic bcttcr 1icqt1c11cy ~~'~CIICIIL, ~I;III~~~IIII;I~I~II~ 5.2 Genetic tra~~sl'urnlatioa: TransSirni~ng ~IC ALll c\l)lallt\ 1t11 111c b111;iry co~i\lri~ct\ contalnlllg Ili('~:ylAb and SBTI genes, p~uduced cll~ckpca Irdll>gvlllcs. 111 ;lccurdancc IVIIII lllc convci~l~onal
molecul~ir analysis of tlic I'III~~~I~IC ~I,!III~, 111c lpIa111s \vcrc 111111;illy scrcc~icd for GUS histochemical assay and tiboul 6U"u oi tlie ~pl~i~ilr LIIO\ICJ PU~III\C ~e;iitii)ii tlii/i\ gelit act~vity has clearly demo~~sti.atetl it! v;iactil~r tishlic, l'llc 1cr1111n;11 !o!111g lc;~~lcts hiit,\vcd a clear blue color ;lctl\ ity oi'tlic i~icurpol.;i~ed III,/:\ gcilc Ilu\\c\cr. tile 1.11cr >lager ol'llie n~olccul~ir ~II~:II)SIS hlio\!cd ~OI~IC \a!r~,~blc ~cholt\ III ICYIII\ co~itir~ii;il~on iiiid Soulliein ;~il,ilyaih. ,)I'IIIII~~I)L~V OI'~~),III\~~ 111 Itc'l< 111col.pol.nied gcllc 111\;ir1;1bly cu~ilir~llb tli~ I~YCICIICC 01'ilie IC\I)C~II\C gelic III IC ge11v111e ui llie lraiisgeilic plalil. Ho\vc\s~, tluc lo suliic \,irl;niulia 1111l\iio\\11 tllc l)i'l< rc\~~Its showed home variable results 111 llic J~III~IIILL' I~,III\I~III~~,IIII~ ' 1 1 1 ~ j~l,i~it\ \VIIII Ui('rl,ll/h gene sllowcd n 50% ~l.;in~lbr~~i;iriw ilctlucncy \\1111 ~c\l,cct lo llic ~ipill kclie In lllc I'CI< conlir~ii:il~o~i ;ili;ilyals I:lic )11;1111\ 0111 01' 10 >IIO\VC~ ;IIII~I~I~~C;III~III UI'IIIC 700 bl) Ii~;ig11ic111 tlinl \vu\ y~cc~lic to illc ~i/i/lI gclic 511111;11.1) tiic ~I~II til'0 tc\lc~I lpl;~iili \\ 1111 \/)I/ \IIII\VUII amplllicaiiu~l ot'llle Ical)ccII\c li,ig~iic~it, \/l~lIi IC\I)CCI lo I.! L11 II;I~I~ICII~ :~~i~pl~lic;~l~~ii IY01ii tile 111i1.k S~II~, 111erc \\.IS \l~gliIly IIICYC;I~~(I lrct~~~c~~cy X 0111 01 I0 /~il'r~'lA/~ lpl:1111s slio\ved a~npl~lii.;ii~v~~ iind 5 11111 ul' 0 .S/j77 I~I~IIII\ >ll~i+ctl ~IIC ~CII)~III\U lil~~l~l~li~d Srag~~icnt It c;i11 be 5~1ggcslctI lic~c lI1,il ioiiic 111111111 IIICI II~CLII~I~CII~ 111 II~c rc,!cIIoII liilxlurc ni~ght be inlerfcr~ii~ \VI~II 11ic ii~i~~)l~Iic;~t~tl~~, llc~)cc, tieqt~ci~c! ul co~ilirr~ied transfor~nat~o~~ trltli respccl IJ ilic 11ii11 kcr ge~lcb e;111 Ibc fixed III 1111. ra~lgc 01'50 10 70% 01' the piitatlve tra~isfor~n;~n~.r. ~~tuc11 lo~er lictluclicy ui tr;~~isiirrii~;~l~u~i was ol)servud w~tli respect lo llle sgronumicall!, II~I~IO~~~III~ ;III~ i113ec1 ~~>I~I;II~CC gc11l.i U/( rj,lilb and SBTI. Only four oul of 10 tcs~ed 5;1111pich iroln llic pla~its 1~111ilbl.111cd w~rl~ Iir('i1IA0 showed
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
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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
Page 22 and 23:
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
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
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larvae of certaln Insects were unab
Page 82 and 83:
3.1 Pl:~nt 111nteri:il sllrl cultur
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0.5. 2.0 and 5.0 pM were added. 111
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Lec!/i?is: Young anci juve~iile lea
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AM3 explant: This was a by-product
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PM) 2nd GAJ (2 5 PM), eltller singl
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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 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 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
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'Table 4.6 Dificrent combinations o
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Table 4.8 Effect oI'TDZ, 2-iP :~nd
Page 212 and 213:
Table 4.10 Effect of i~lclusio~~ or
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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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