Agrobacterium rhizogenes-mediated T-DNA transfer to chilli Capsicuum annuum cells

Ismanizan Ismail, . and Zamri Zainal, . and Nik Marzuki Sidik, . and Pung Yuh Fen, . (2000) Agrobacterium rhizogenes-mediated T-DNA transfer to chilli Capsicuum annuum cells. [Proceedings Paper]

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 g/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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Abstract

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 g/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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Item Type:	Proceedings Paper
Additional Information:	Available at Perpustakaan Sultan Abdul Samad Universiti Putra Malaysia 43400 UPM Serdang Selangor Malaysia. QK710.2 M239 2000 Call Number
AGROVOC Term:	Capsicum annuum
AGROVOC Term:	Chillies
AGROVOC Term:	Gene transfer
AGROVOC Term:	Gene expression
AGROVOC Term:	Agrobacterium rhizogenes
AGROVOC Term:	Explants
AGROVOC Term:	Genetic transformation
AGROVOC Term:	Roots
AGROVOC Term:	Root hairs
Geographical Term:	MALAYSIA
Depositing User:	Ms. Suzila Mohamad Kasim
Last Modified:	24 Apr 2025 05:15
URI:	http://webagris.upm.edu.my/id/eprint/12890

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