Production and molecular characterization of herbicide-resistant transgenic rice

Issue Date

12-2005

Abstract

The delivery and integration of herbicide resistant gene (bar) into Indica and Japonica cultivars of Oryza sativa was done using biolistic transformation method. One Indica (PR106 and one Japonica (Taipei 309) cultivar was used for particle bombardment. Both cultivars responded well on MS medium with 2,4-D (2 mg/L), casein hydrolysate (500 mg/L and maltose (30 mg/L) used for callus induction from immature embryos and mature seeds. Different explants used in a series of bombardment experiments. The explants were calli derived from cultured anthers, mature seeds and immature embryos. The calli from mature seeds were found to be the excellent explant for particle bombardment with plasmid pUBA. Taipei 309 showed better regeneration efficiency than PR106. Phosphinothricin (PPT) was used as a selectable marker for bar to select herbicide-resistant calli on the media. PPT @2.5 mg/L was found sufficient to select putative embryogenic herbicide resistant calli in six cycles of selection. Five out of 26 plants of Taipei309 were found to be herbicide resistant. Southern hybridization revealed the integration of bar gene into these lines. Three plants of PR106 were also found PCR positive for bar gene. Phosphinothricin acetyl transferase (PAT) assay confirmed the protein activity in Taipei309 and PR106 lines. Using leaf pasting assay, leaf tips of all the putative transformants of Taipei 309 and PR 106 were used for rapid screening for resistance to the herbicide Basta. This method was found more rapid for screening putative transformants than molecular and biochemical analyses.

Source or Periodical Title

Philippine Journal of Crop Science

ISSN

0115-463x

Volume

30

Issue

3

Page

23-30

Document Type

Article

Frequency

tri-quarterly

Physical Description

tables, pictures

Language

English

This document is currently not available here.

Share

COinS