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exceed 5 cni Icngtl~. Loiigcr shoots posed tlic sbo\c-ii~r~it~o~ied I>~UOICIII I I I C ~ ~ ~ I lcilgtl~ ~ C ~ in the culture tubes. Shoots III;II d~d IIUI rout aliould 1101 st.i) ior lollger riil~c 111111s aanie culture rnediiiill and be tKlnsli.rrcd tu 111c licrl~ IIIC~IIII~~ ,ilicr II~C tirht 0;11cl1 II;I~ IULIC~. Care should be riikrii iiut lo ;illon tlic aliouta 10 II~L~~III~ IIYIIL'I./I!~~II~II! bi'c;iii~c SLICII shoots do no1 rout \\ell a~lci Ii.i\c tu be c,iriicd IU IC ~)II~IZS ! ul'r~ot~~~g. tli~rdcn~iig iiiiil trC~~is~>li~~it~it~o~i (11' cliick~~c:~ li:i\ I)cc11 ,I ,L~~IL!LIS IJILIIIICIII 1 ~C IVI 10 ~~~cl'licic~it 111 \ 11ro $ri~(iiiig by utilizing tlic roo1 sysle111 I'rolii tile prc-gcr~i~in;~~e~l sccd1111gs (Ki~~liii;i~~~~~rll~y CI ill. ~iiiiiiber ~t'Iic~~~~le~ic~l I>~;IIII\. 'l'lic co~ii~ircIie~~\~\ c I)IOIO~OIi)i l~;i~.clc~i~~ig ,IIII,IIIIS ivctc l;ick or gro\+~h of roo ti^^^ ~~,ICIII t~r~cl 1i~1111icli1y ;I~~~III~,III/;III~~II (~IIIE 10 111c cllic~c~~cy ~~f\I;itic hydlopo~n~cs s)ste~ii III,II CII~LI~C\ IC ~IO~III 111' roo^ \y\Ic111 C;III lhc coi~\~dvrv~I ;IS llic best nicthod for hardcn~np. I'lii. ollicr 111c1Iiud ui II;I~L~SIIIII~ ,111d II;III\~)~~I~~I;I~IOII III~OLI~II tlircc different stages \baa al,u cific~~~it ~ n rc11;lblc. d Ihu\rci'cr, \ ii~~pl~c~lj, ~i~ti. ;III~ III 1ot;iI tllc efficiency of Ilydropoilic\ a)slenii\ si~pcr~(~t In ;I~~I~IUII. Ilic 3-bI;igc n~ctliod cil>plu~s hunlldily eblabll\]inlcll[ bj uu\crlily ,ICI III~I~ gr;~dii:~l O/>CI~III~ 111 lllc I~IIC'T \I;I~cs, which I\ vey case sensiri\e E~cn iiliiiur \;~r~:il~u~i\ ul' Il~~iii~dily III:I~;I~~I~CIII re\~lltcd 111 clllicr
wilting or drylng of tl~e pliints. This prublci~i \L;I\ 11eg;itct1 III tile I~>drul>oi~~cs syslcn~. However, a carckil cua~irinatioi~ US gro\\tli u1'1Iic \tc111,illti loor ~!~I~III. Ic;iS ~l~ospl~olugy should be dul~r ~hllc hordei~ing IIIC sooleil ~~I.IIII\ III at.itic II!~I.UPUIIIC~ I'uttii~g IIIC~IIIIII was one ot' tlic Inlportellll l;ictt11.\ 111;il ,~l'Scct ?IIIc~\~~c;I II;I~~~CIIIII~ ,II:J ~\I.~~~I\~IIIICIII. Chlckpca ge~lurdll) prckrs bl,ick soil liu\tc\cr. gio\\lli 01' S~ILII r!\Iclll \\.I, g~c,i~ly 111hib1ttd by tll~s pou~~lg IIIC~ILIIII 111 :111 ilic tli~cc SI;I~CS tIc\crlbcd 111 IIIC I~l>u\~ WCIIOII\ Sand \\as used :it Ihiglic~ ps~~~~itiui~s IU I)SU\ iilc ~CIICI JCI;I~IOII ii~~d ~~I;III\cI) lcs. llllitlllll~ of bi;~ck soil \\as ;~ddctl lu I~~,IIIII;III~ ~IIC \\.I~cI 11ultli1ig C,II);ICI~~ 01I1cr I;ICIOS~ l~kc temperature, liglit 111te11bity. pIioto/)1'11t1tl, IIII~;I~IUII :illil \\CIC cilw \t,~i~tI~i~tli/~d OI)~IIII;I/ green huusc or LIY ~tl~er li;iraI~ ~IIYI~UIIIII~III C,III OC ~CIIIII~CIII;I~ tu IIIC ILI~C~CIIIII~ 01' chlcl\lx". OII~ li~iliti~t~un ~1'111~ CIIICI\I)C;I L~\I;I~~I\~III~CIII btucIl~\ 111 I/I;II IIIC IC~IIIICIIICIII (11' walk-ln type gro\vtli cIi;~i~~bsr, ~VIIICII 111;1l,c\ II,I~~~CIIIII~ III~ICC\ \OIIIC~~.~I,I~ I.CSIIICIC(/ 10 Seiv lnstitiiteb t11:lt c;111 iit'ii~~~l tile iiiciIil> Sliit~c II~~~OI)OIIIC\ \)\IC.III 15 LIII cfIi~,ie111 \y\t~111 that can \harden 111 LIIIO prod~i~td ~IIIc~~)c;I ~~I,IIII\ 111 illc inori~~,iI CIIIILI~C 1.00111 CUI~~I~I~II\ without conil)sol~~i/iiig tile 111tqr11y ,iil~l v~l,~li~j. 01' IIIC pl:iill. 111 ~ilro ~ICIILC~ cl~ickl~ei~ pla~~ts sl>o!\ed :ill cii~ly tlt~\~crii~g ;iiltl II~:IIIISII~ 111 \OI~IC III\~;IIIC,C\ IIIC ~I;IIII> Ilo~vcrcd 111 culture tubes while they \rere rootliig. 1111s siiggc\l\ III,II il~iltl cu~~tl~tioii~ of lcnlpclalurc, light intensity iind l~~iil~icl~ty ~>r~vidctl 11) tlic CIIIILII~ roo111 111ig11t ci~tii.it~ ~C~II~CLIVL: ge11c11clemc~~ts Sur Ilo\+cr~~lg ;ind III,I~LI~II) 'I III\ r,ltc ul'i~~~~t~irit~ \\a\ I~IUII~ 10 be bluib'cr In case of tjeld-so\\il sci'~ll111g~. 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
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~
Page 32 and 33:
them. About 10% of the Earth's land
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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
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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
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 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 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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