Download (26Mb) - OAR@ICRISAT

AM3 explant: This was a by-product during the preparation of AM2 explant as
indicated above. Removal of AM2 explants resulted in shoot tip with eplcotyl region. Further
removal of shoot tip provided [lie AM3 explont that contalned au~llnry nieristeni on either side
of the eplcotyl and was cultured on SIM.
AM4 explant: The de-coated seeds were gerlti~n;itcd OII SIM at ;I dclisity of 10 to 15
per plate. They were grown for about a week ~tlit~l :inlllaiy bud was prollilnent. Then the
axiilary bud \\as rcltioved up to tlic base two cuts \\,ere give11 as 111 the casc oTprt.pol.;~tioll of
AM2 explont. The resultant axillary nieristeiti explant, were sub cillturcd on tlic same
niediuni for another 6-7 days. Tile b,lsc ofaxillary bud enlarged and sotiic ~ii~~ltiplc shoot buds
cnierged. Tlie emerglng shoot buds were reniovcd apolil by canli~lly acr,lpping the shoot buds
irith ;I sharp blade Tlir res~ilt;inr cxplnlit corit:i~li~tig cotyledoli uitli ;I bulge on \he
cotyiedu~iaty ~iode rcg~oli \\CIS cultttrcd 011 IIU~IIIUII~ lice MS.
The ~iirdia tbr itiductiott ul'~iiult~ple ~Iiou!) liutil tlie givc~i cx~luttt~ ciili be d~v~ded in
rive m;~jur cl;isscs. I. TDZ-ba~cd ;tiiil 2. DAl'-bt~scd. Thc\c conslats oScltllcl- 'TDZ or UAI' as
pril~cipnl niultlplc slioot iliti~~ctilig cytuk~n~ii ;III~ a111c acc~s\ory C ~ ~ O ~ I I ;ilid ~ I I ~UXIIIS ~ S werc
i~icludcd ~n tile tnedi;] ;is per tlis rcquircn~ciit. Tl~c orhcr cyt~kilillls illid d~~xillj ilddcd were 2-
iP (5 and I0 ~IM ill TDZ-b;ised mctli;~), k~llelili (2 ;iild 5 liZ1 I 'rU%-b:~scd:ti?d 2 2nd 4 pM in
BAP-bnscd illsd~;~ conib~n.itio~is). Optinliutlon of'tllc i~~tiuct~oli nicdi~~m was dolie initially
w~tlt tnaturt. elnbtyu 3x1, and I;iter w~tli ,~xillnr) meristem explant, Ah12. Unless otherwise
mentioncd. the explants were cultiiled 011 tlic 111duc1iori iiiedium Sor abuut IX to 20 days alicr
wliicli they werc tri~nsverred to the clongat~on ~nediulti. An average of 6 to X expl;l~lts werc
c~llt~lrcd per plate and the re,puliies werc atudlcd.