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Figure 4.1 A - D inducl~on of sonlatic enlbryos from mature embryo axis and leallei explants ;itler 6 weeks of cullure on the 111edi1111l conla~ni~lg 2,4,j-T as pri~~c~pal growth regulator. A, B a ~ C. ~ d induction of enlbryos from Inlalure einbryo axis expla~il, D. Induction of soinatic embryos froin leaflet esplant (arrows iiidicotl: globular embryos). Figure 4.2 A, B lnductioil of sonlatic cmbryos SI-oin inlaiurc c111bryu asis CXPIOIII by uring 2.4-D as principal growth regi~lnlor. .A. Top \,icw of tlie einbryo axis sllowiiig inultiple globular embryos, B. Letefiil view of enibryo axis showing multiple globular embryos formed from plumiile region (arrows indicate globular embryos). Figure 4.3 A - E Effect of concentration of B.4P on ~nullrple shoot regeneration fro111 inalurc cinbryo axis explant after 15 d:iys of culture on slioot ~nduclion medi~lm. A. Multiplc slioot regeneration at 10 j~kl HAF concei~tralio~l, U. M~illipli: slloot rcgcncr~~ion will1 30 pM BAP, C. Miiltiple sliuot reguneration \$'it1140 pM BAP, D. Multiple sl~ool regeneralion with 50 pM BAP, E. Mult~plc slluot rcgenelntion w11Il I00 1tM BAP. Figure 4.4 A - F Multiple shoot regenel-ation fro111 dil'fcren~ cxplnnts al tile third week of culture 011 sliooi iiiduclio~i ~iicdioni .A. Shoot lip sllowi~~g rn11111plc slioots cincrgitig fro111 ~neristeii~at~c region. B. Axillary bud showing nlultiple shools or~ginaling fro111 lip and basal regions, C. Mature enlbryo axis sliowing mulliple shoots cinerging rrom sllout tip as well as axillary bud regiu~is, D. AM1 explant showing ~liultiplc shoots from axillary region, E. AM2 explant showing swollen brisal region and multiple shoots emerging synchronously as cluster, F. AM4 explant show~ng synchroi~ous multiple shoots from different places oraxiIlai.y niel.istem region.
Figure 4.5 A - I; Stages of ~nultiple stloot regelleration from AM4 explant alter culture on the sl~oot induction medium A. Explant sl~owing the area of axillary bud re~noval (arrow sliows the sire of surgery), B. Muluple sl~oot buds oriyin;ltiiig fro111 the area loft by axillary bud removal (arrow sl~ows the s110ots buds e~ilcrging fru111 axillary ~nlcriste~l~ area). C. Esplii~~l showing swollen area after removal ofregcnc~at~~~g shoot buds (this stage of explant sas na~nsd 3s AM4 cxpla~it) akcr 7 days of culture OII shoot i~iduct~o~l rncdlulll, D. Regc~ierat~o~~ of ~il~iltiplc sl~oot biltls li.0111 tl~ cut ~po~tiul~s uftlic rcgclleratlng iiastiv after 6 days ofci~lture on MS, E. Exubcra~lt nlultiplc sl~oot yrowtli Sron~ tile rcgcncrating tlssue after 10 to I? days of culture on MS, 1;. Mult~plc sl~oots originaiiny fro^^^ different p;~rts of the regellcrating area aCter 12 to 14 days of culture on MS. Figure 4.6 A - C Various stages of tlongatio~i of regenerating sl~oot buds on shoot elo~lgation 111edium; A. Elongut~on of young shoot 011 shoot clo~igatio~~ medic1111 1 (SEMI) ancr 1 week uf culture, B. Elol~gatio~i of sliool on shoot elollgation 111cdiu111 2 (SEM2) nlicr 2 wccks of culture, C. Elongallon of sl~oots 011 SEM2 after 3 wecks. Figure 4.7 A , B Rooting of eloiipaled slioots on root induction ~niedill~~~ co~~tailli~lg 5 pM IBA, A. Rooting 011 filter paper bridye i~lllrlcrsed ill liquid root ind~iclio~~ nicdium, 8. Rooting OII solid root induction ~nlcdii~n~ co~~iaininy 5 pM IBA. Figure 4.8 Stage 1 Ilardening of tissue ci~lture gro\an cli~ckpea after transferring to 8 cnl pot containi~lg 2-4 mm sand.
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 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
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 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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