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
Recommended Citation
Kaur, Jatinder; Laude, Rita P.; Mendioro, Merlyn S.; Castillo, Eugene M.; Hernandez, Jose E.; and Datta, Swapan K., "Production and molecular characterization of herbicide-resistant transgenic rice" (2005). Journal Article. 4219.
https://www.ukdr.uplb.edu.ph/journal-articles/4219