An inbred strain of cucumber (Cucumis sativus L. cv. S516)
was propagated by artificial self-pollination, and used for
transformation. pCAMBIA1300-RD29A-LOS5 (from Professor
Gong, College of Biological Science of China Agricultural
University) plasmid was integrated into
Agrobacterium rhizogenes EHA3101. Seeds were surface
sterilized and sown on MS medium containing 30 g L-1
sucrose and 2.5 g L-1 gelrite. Cotyledons from the germinated
seeds were separated and cut into 1 cm2 piece to be
used as explants. The explants were dipped into the
pCAMBIA1300-RD29A-LOS5 Agrobacterium suspension
for 15 min, and placed on the shoot induction media containing
2 mg L-1 6-benzylaminopurine (BA), 1 mg L-1 ABA,
and cultured for 3 d at 25°C in darkness. Then the explants
were transferred to germination media containing 2 mg L-1
BA, 1 mg L-1 ABA, 5 mg L-1 hygromycin, 100 mg L-1 claforan,
and cultured for 14 d. Regenerated shoots were placed on
selection media containing 2 mg L-1 BA, 1 mg L-1 ABA, 25
mg L-1 hygromycin, 100 mg L-1 claforan, for another 14 d.
The shoots showing resistance to hygromycin were cultured
till full growth in MS medium containing 50 mg L-1
claforan (Tabei et al. 1998).
An inbred strain of cucumber (Cucumis sativus L. cv. S516)was propagated by artificial self-pollination, and used fortransformation. pCAMBIA1300-RD29A-LOS5 (from ProfessorGong, College of Biological Science of China AgriculturalUniversity) plasmid was integrated intoAgrobacterium rhizogenes EHA3101. Seeds were surfacesterilized and sown on MS medium containing 30 g L-1sucrose and 2.5 g L-1 gelrite. Cotyledons from the germinatedseeds were separated and cut into 1 cm2 piece to beused as explants. The explants were dipped into thepCAMBIA1300-RD29A-LOS5 Agrobacterium suspensionfor 15 min, and placed on the shoot induction media containing2 mg L-1 6-benzylaminopurine (BA), 1 mg L-1 ABA,and cultured for 3 d at 25°C in darkness. Then the explantswere transferred to germination media containing 2 mg L-1BA, 1 mg L-1 ABA, 5 mg L-1 hygromycin, 100 mg L-1 claforan,and cultured for 14 d. Regenerated shoots were placed onselection media containing 2 mg L-1 BA, 1 mg L-1 ABA, 25mg L-1 hygromycin, 100 mg L-1 claforan, for another 14 d.The shoots showing resistance to hygromycin were culturedtill full growth in MS medium containing 50 mg L-1claforan (Tabei et al. 1998).
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