Transformation of Thai rice varieties with genes relating to disease resistance
1996
Porntip Tinjuangjun
Four Thai rice varieties namely Supanburi 60 (SP60), Supanburi90 (SP90), RD 21 and Khao Dawk Mali 105 (KDML105) were transformed with genes relating to disease resistance. Mature embryos were cultured on MS and modified MS media for callus induction. Modified MS consisting of 2 mg/l 2,4-D and 8 mM L-proline yielded 44.40 percent embryogenic callus induction, the highest yield for KDML105. High percentage (33.33 percent) of plantlet regeneration for KDML105 was obtained from the R4 medium, the MS medium supplemented with 1 mg/l IAA and 4 mg/l BA.Protoplasts were isolated from cell suspension culture, mature leaf and young leaf of KDML105. Results showed that cell suspension culture served as good source for protoplast isolation. Isolated protoplasts can be cultured up to 2 weeks before the development ceased and the protoplasts eventually died. Calli and scutella of KDML105 were bombarded by DNA coated tungsten and gold particles using Bio Rad gun and particle inflow gun. Plasmid DNAs used for transformation were pAct1-D with gus,pIG121Hm with gus and hph, and pUBA with bar. Transient expression of GUS was more frequently observed in bombarded scutella than in calli. Selection of transformed tissues were made on selective media containing 0-50 mg/l bialaphos or 25 and 50 mg/l hygromycin. Non-transformed calli died within 2 weeks while transformed calli remained viable. Regeneration of transformed calli however, was not yet successful.
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