PROTOCOL DEVELOPMENT FOR GENETIC TRANSFORMATION IN Corciwnis olitori its THROUGH Agrobacteriwn VECTORS

Abstract

Two sets of' (lie experi nents were conducted at the Genetic Engineering Laboratory, Genetic Resources and Seed Division, Bangladesh Jute Research institute (13J1U). Dhaka, during the period of' July 2006 to June 2007, to fulfi I the objectives of the study. In the first set of the experiments, two varieties of C. o/itorius were used to investigate their in vitro regeneration potentiality, and in the second set of the experiments, two varieties of C. olilorius were used to observe their transformation ability. Cotyledons (with attached petiole) were used as explants. The percentage of germinated seeds (90.00%) on cotton-supported medium found to be much higher than germinated seals (68.33%) on agar-supported medium. The seedlings grown on cotton-supported medium found to be much more healthier than seedlings grown on agar-supported medium. Maximum no. of plant regeneration percent was obtained From the eutyledonary petioles of the varieties of C. olitorws on MS agar solidified medium supplemented with 0.5 ing/l IAA and 3.0 mg/I BAP. Plant regeneration was also observed at different concentrations oflTeSO4 and it was found that. 56 mg/I gave the highest result. The efficiency of plant regeneration froni the cotton-supported seedlings was better than the agar-supported seedlings. The plantlets produced roots on hormone free MS medium rapidly. The effects of non—ionic surfaetants (Pluronie F-OS) on shoot regeneration for jute (C. olitorius) coty edons was assessed. Supplementation of culture me(' in with 0.01 -0.08% (w'v) Pluri. nic F-OS increased the mean percentage of cotyledon producing shoots, in the second set of the eXperil)lentS, an efficient and reproducible protocol for the production of transgenic jute plant was developed by inoculating cotyledonary petioles with A. tu,aefciciens strain LBA4404 carrying a binary vector pBl 121, which contains selccable marker gene nptll conferring resistance to kanamycin and the GUS reporter gene. After co-cultivation and selection. histochemical GUS assay was performed in two varieties (0-9897 and 0-72). In the transformed explants, GUS reporter gene was expressed showing blue colour in the explant tissues. Non-transformed explants did not show any colour. I3etwcen the varieties, 0-9897 showed the highest response to GUS assay (60.00%).