Transformation of indica rice with synthetic cryia(C) gene. | [Biologia • 1998]

Author(s):

1. Tayyab Husnain: National Centre of Excellence Molecular Biology, University of Punjab, Lahore, Pakistan

2. Farzana Khanum: National Centre of Excellence Molecular Biology, University of Punjab, Lahore, Pakistan

3. Tahira Fatima: National Centre of Excellence Molecular Biology, University of Punjab, Lahore, Pakistan

4. Esther Khan: National Centre of Excellence Molecular Biology, University of Punjab, Lahore, Pakistan

5. Sheikh Riaziddin: National Centre of Excellence Molecular Biology, University of Punjab, Lahore, Pakistan

6. Illimar Altosaar: BioChemistry Department, University of Ottawa, Ottawa, Canada

Abstract:

Meristem tissue explants of Oryza sativa variety Basmati 370 were bombarded with tundgsten particles coated with a 3:1 mixture of two plasmid DNAs, one containing hygromycin marker as a 1031 bp fragment in pROB5 vector, the other containing synthetic crylA(c) gene as a 1845 bp fragment in pUC vector. The bombarded explants were grown for six weeks on selection medium containing 30mg/1 of hygromycin and finally screened for inherent insect resistance against 2nd instar larvae of rice leaf folder (Cnaphalocrocis medinalis). Out of 10,000 bombarded tissues, 39 plants were selected on the hygromycin containing medium and 62% of the selected plants exhibited the mortality of the insects. To study the inheritance pattern of the transgene, T1, T2 and T3 generations of plants were further analysed for the presence of the crylA(c) gene, estimation of the copy number and protein expression using PCR amplification, Southern blot analysis, Western blot analysis, and insect bioassay. On analysis of eight T1 plants, three showed a 655bp internal fragment of crylY(c) while the full length fragment (1845bp) was found integrated as 2-5 copies in two plants. The same plants exhibited resistance in insect bioassays where both mortality and supression in weight and length of the insects was observed. Further, a positive Western blots confirmed the expression of a 68Kd CrylA(c) protein.

Page(s): 180-192

DOI: DOI not available

Published: Journal: Biologia, Volume: 44, Issue: 1--2, Year: 1998

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