期刊
JOURNAL OF HORTICULTURAL SCIENCE & BIOTECHNOLOGY
卷 84, 期 2, 页码 199-203出版社
HEADLEY BROTHERS LTD
DOI: 10.1080/14620316.2009.11512504
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资金
- Natural Science Foundation for the University of Jiangsu Province, China [04KJA210158]
- Natural Science Foundation of Jiangsu Province, China [BK2002403]
An efficient Agrobacterium-mediated transformation system was developed for cucumber (Cucumis sativus L. 'Jinyan') using stem nodes as explants. Stem nodes were isolated from cucumber seedlings 10 d after germination, wounded four-to-five-times with a knife, infected for 30 min with Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pCAMBIA 1303, then co-cultured for 5 d. Adding a washing step after co-culturing, and using 25 mg l(-1) meropenem to suppress bacterial growth in the regeneration medium, eliminated any overgrowth of Agrobacterium and significantly improved the efficiency of transformation when 15 mg 1(-1) hygromycin was applied as a selection agent. Finally, a transformation efficiency of 7.1% was achieved with a high frequency (50%) of single-copy T-DNA integration 2 months after co-culture. On average, two-to-three independent transformation events occurred from each original explant.
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