期刊
ARCHIVES OF BIOLOGICAL SCIENCES
卷 59, 期 4, 页码 311-317出版社
INST BIOLOSKA ISTRAZIVANJA SINISA STANKOVIC
DOI: 10.2298/ABS0704311N
关键词
Agrobacterium tumefaciens; genetic transformation; beta-glucuronidase; hygromycin B; Lotus corniculatus
类别
Cotyledons from 6-day-old Lotus corniculatus cv. Bokor seedlings, transversally cut into two halves, were capable of regenerating buds without intervening callus formation. The explants were co-cultivated with the Agrobacterium tunlefaciens LBA4404/pTOK233 superbinary vector carrying the uidA-intron gene and the genes hpt and nptII. They were cultured for 14 days on a regeneration medium, then subjected to a stepwise hygromycin B selection procedure consisting of gradually increasing antibiotic concentrations (5-15 mg L-1) over 21 weeks. Transformed shoots were obtained within 5 months after co-cultivation. Out of 124 initially co-cultivated explants, 52 (42%) plants survived hygromycin B selection. The presence of transgenes in regenerated plants was verified by beta-glucuronidase histochemical assays and PCR analysis for the presence of uidA gene sequences. Hygromycin B-resistant and PCR-positive T0 plants were cultured in the greenhouse to produce flowers and seeds. The obtained data demonstrate that the reported transformation protocol could be useful for introducing agriculturally important genes into the new L. corniculatus cultivar Bokor.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据