Agrobacterium rhizogenes-mediated T-DNA transfer to chilli (Capsicuum annuum) cells
2000
Ismanizan Ismail | Zamri Zainal | Nik Marzuki Sidik | Pung, Yuh Fen
This study is the first report of protocol for transfer and expression of foreign gene into Capsicuum annuum L. by Agrobacterium rhizogenes. Agrobacterium rhizogenes strain A4 harbouring the plasmid pCAMBIA containing 35SINT as binary vector was used to infect the cotyledons and hypocotyls which served as organ explants. The inoculated explants were transferred onto MSO media with 250 µg/ml cefotaxime after 2 days of cocultivation, Putative stably transformed roots were successfully obtained as early as one week after inoculation. GUS histochemical analysis has showed that the highest frequency of transformation were 9.8% from hypocotyls without preculture treatment and 12.5% from cotyledons with 2 days of precultures treatment. The selected roots exhibited strong GUS activities predominantly along the vascular tissues as indicated by a deep blue color. Stably integration of the GUS gene was confirmed by amplification of 789 bp GUS fragment. Both GUS and PCR analysis have suggested the effect of co-transformation and gene silencing phenomenon in the transformed roots and hairy roots.
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