PROTOCOL DEVELOPMENT FOR GENETIC TRANSFORMATION IN Corchorus olitorius THROUTH Agrobacterium VECTORS

Abstract

T\~'O cts of the experiment were conducted at the Genetic Engineering Laboratory ienctic Resource c.111d 'ccd Division, Bangladesh Jute Research 111 titutc (BJRI) Dhaka, during the period of J Lily 2006 to J unc 2007, to fulf ll the objc ti \'CS of the study, 111 the first set of· the experiments, t \'O varieties of" C. olitorius were used to investigate their i11 i,i1140 regeneration potentiality, and in the second set of thee .periments .. two varieties of· C. ol itorius were used to observe their transformation abilit . Cotyledons (with arta hed petiole) were used as explant . 'I he percentage of' germinated seed (90.00o/o) on cotton-supported medium found to be 111t1cl1 higher tl1;.111 germinated seeds 68.JJo/t,) 011 <1g,1r-st1111J<)rtccl 111c<lit1111. l .. he seedlings gr0\\'11 011 cotton-supported 111ediu111 found to be 111t1cl1 more healthier than seedling grown 011 agar-supported 111cdiu111. Maxi111t1n1 110. of' plant regeneration IJ rccnt " as obtainedfrom the cotyl donary petioles of the varietic of C. olitorius 011 M agar olidifi d 111ediu111 supplemented ' ith 0.5 111g/l IAA and 3.0 111g/l BAP. Plant regeneration \i\'US also ob rved ·at di fferen t concentrarions of' i c 04 and i l wa f ound that, 56 111g/l gave th highe t result. r h efficiency of' plant reg ncration from the cotton- upportcd cedling wa: better than the agar- upported seedling . The plantlets produ ed roots 011 hormone free M 111ecJit1111 rapidly .... I'h effects of non-ionic surfactant (Pluronic 1::-68) 011 shoot regeneration for jut (C. olitoriuss cory edons wa as ed. St11JI)ll:111c11t,,tio11 of ulturc media with 0.01-0.08°/o (\7V/\1) P lure 11i I·-68 increased the111 'a11 percentage of' cotyledon producing shoot . 111 the second set of the experiments, a11 efficient <111d reproducible protocol 101· the production of transgenic jute plant was developed by inoculating .otyledonary petioles with 11. tuinefaciens ~ train LBA4404 carrying a binary vector J)l31121 which contains sclec: able marker gene 111;t/l conferring resistance to ka11a111yci11 and the GU reporter gene. After· co-cultivation and selection .. histochemical GUS assay wa performed i11 two varieties (0-9897 and 0-72). In the tr<111 sformcd explants, GUS reporter gene was exprc scd showing blue colour i11 the cxplant tissues, Non-transformed explants diet 11ot show any colour, Bct\~'\:CJ1 the varieties, 0-9897 sJ10\\'Cd the highest response to GUl assay (60.00%).