Introducing the CrylA(c) gene into basmati rice and transmitting transgenes to R3 progeny
2003
Gosal, S.S. | Gill, R. | Sindhu, A.S. | Dhaliwal, H.S. | Christou, P.I.
Synthetic crylA(c) driven by the maize ubiquitin I promoter has been successfully introduced into basmati rice. Scutellar tissues of Pusa Basmati 1, excised after 6-7 d from mature seeds and cultured on proliferation medium [MS + 2,4-D (2.5 mg L¯¹) + proline (560 mg L¯¹)], were bombarded using gold particles coated with crylA(c) DNA on osmoticum medium [MS + 2,4-D (2.5 mg L¯¹) + mannitol (0.4 M)]. Bombarded tissues, after 2 d of growth on proliferation medium, were subjected to hygromycin selection (30 mg L¯¹) for two cycles of 2 wk each. Plants regenerated on hygromycin (30 mg L¯¹) containing medium were screened using the histochemical GUS assay. PCR, Southern, and western analysis of R0 and R1 plants confirmed the stable integration and expression of crylA(c). Segregation analysis during R1, R2, and R3 generations confirmed the Mendelian inheritance for selectable marker, reporter, and crylA(c) genes. Insect bioassays during R1 and R2 generations have shown enhanced resistance to yellow stem borer.
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