As. Pac. J. Mol. Biol. & Biotech., Dec 2001 Vol. 9(1) : 31-37

Genetic Transformation of Capsicum annuum by Agrobacterium rhizogenes

Ismanizan Ismail*, Zamri Zainal, Nik Marzuki Sidik and Pung Yuh Fen

School of Biosciences and Biotechnology, Faculty of Sciences and Technology, Universiti Kebangsaan
Malaysia, 43600, Bangi, Selangor Malaysia

(Received 23 September 2000 / Accepted 5 April 2001)

Abstract.

This study is the first report of protocol for transfer and expression of a foreign gene into Capsicum annuum L. by Agrobacterium rhizogenes mediated transformation. Agrobacterium rhizogenes strain A4 harbouring the plasmid pCAMB 1A 1301 containing 35SINT as a binary vector was used to infect the cotyledons and hypocotyls. The inoculated explants were transferred onto NIS media with 250 mg/l cefotaxime after 2 days of cocultivation. Putatively stable transformed roots and hairy roots were successfully obtained. GUS histochemical analysis has showed that the highest frequency of transformation were 8.9% from hypocotyls without preculture treatment and 9.3% from cotyledons with 2 days of precultures treatment. Transformed roots and hairy roots exhibited strong GUS activities predominantly along the vascular tissues as indicated by an intense blue color. Stable integration of the GUS gene was confirmed by partial amplification of GUS fragment (789 bp). Both GUS and PCR analysis have suggested the possible effect of co transfor mation and gene silencing in transformed roots and hairy roots.

Keywords: Agrobacterium rhizogenes, hairy roots, GUS expression, Capsicum annuum.