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A Cointegrate Ti Plasmid Vector for Agrobacterium tumefaciens - mediated Transformation of indica Rice cv Pusa Basmati 1

  • Subha S. Jacob
  • K. Veluthambi
Article

Abstract

An intermediate vector pSSJ1 was constructed by cloning a hph gene and a gus gene with catalase intron in pGV1500. pSSJ1 was cointegrated into a disarmed receptor Ti plasmid pGV2260 harboured in Agrobacterium tumefaciens strain C58C1RifR. The resulting A. tumefaciens strain C58C1RifR (pGV2260::pSSJ1) stably transformed Oryza sativa L. cv Pusa Basmati 1 scutellum-derived calli at 26% frequency. Introduction of the plasmid pSSJ3 (3′virB, virG and virC of pTiB0542) into A. tumefaciens C58C1RifR (pGV2260::pSSJ1) resulted in the elevation of acetosyringone-induced T -strand accumulation. Rice transformation efficiency of the cointegrate plasmid pGV2260::pSSJ1 increased from 26% to 33% in the presence of pSSJ3 and from 26% to 35% in the presence of pToK47 (complete virB, virG and virC). T-DNA integration in To plants was confirmed by Southern hybridization analysis. Inheritance analysis of the T0 plants with single-copy T-DNA insertions revealed segregation of hygromycin resistance in 3:1 ratio. The feasibility of rice transformation with a cointegrate Ti plasmid vector is clearly established.

Keywords

Agrobacterium cointegrate vector pTiBo542 T-strands transgenic rice 

Abbreviations

AS

acetosyringone

CaMV

cauliflower mosaic virus

CIM

callus-induction medium

2,4-0

2,4-dichlorophenoxyacetic acid

GUS

β-glucuronidase

Hyg

hygromycin

LB

left T-DNA border

MS

Murashige and Skoog

NAA

α-naphthaleneacetic acid

RB

right T-DNA border

RIM

root-induction medium

SIM

shoot-induction medium

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Copyright information

© Springer 2003

Authors and Affiliations

  1. 1.Department of Plant Biotechnology, School of BiotechnologyMadurai Kamaraj UniversityMaduraiIndia

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