Transformation of basmati rice (Oryza sativa Linn) with bacterial genes by particle bombardment. | [Pakistan Journal of Plant Sciences • 1995]

Author(s):

1. Tayyab Hussain: Centre for Advanced Molecular Biology, University of Punjab, Lahore, Pakistan

2. Farzana Khanum: Centre for Advanced Molecular Biology, University of Punjab, Lahore, Pakistan

3. Sheikh Riazuddin: Centre for Advanced Molecular Biology, University of Punjab, Lahore, Pakistan

4. Milton P. Gordon: BioChemistry Department, University of Washington, Seattle, Washington, USA

Abstract:

In these studies, various parameters including distance between filter unit and tissues, ratio of DNA to tungsten particles, age of the tissues, intervention of accelerated particles with nylon mesh and drying of the tissues before bombardment were optimized for transient and stable expression of marker genes in rice using a home-made particle acceleration gun. The Beta-glucuronidase gene was introduced into leaf bases for transient expression while the hph which confers hygromycin resistance was introduced to mature embryos for stable transformation. Under the established conditions, up to 70% of leaf bases showed transient expression as analyzed by histochemical assay. In addition, mature embryos were also bombarded under optimum conditions with DNA-coated tungsten particles. After 5 days, the excised leaf bases from embryo derived plants were selected on MS medium containing hygromycin to obtain fast growing green plants. Southern hybridization showed stable integration of the marker gene in the genome of these plants.

Page(s): 219-228

DOI: DOI not available

Published: Journal: Pakistan Journal of Plant Sciences, Volume: 1, Issue: 2, Year: 1995

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