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Establishment of an efficient regeneration system using heading leaves of Chinese cabbage (Brassica rapa L.) and its application in genetic transformation

大白菜抽穗叶片高效再生体系的建立及其在遗传转化中的应用

关键词:
来源:
Horticulture, Environment, and Biotechnology
全文链接1:
http://agri.ckcest.cn/file1/M00/02/9D/Csgk0FvzdzqATyw8ABcoTwEy-KE937.pdf
全文链接2:
类型:
学术文献
语种:
英文
原文发布日期:
2018-07-06
摘要:
To preserve and propagate valuable Chinese cabbage (Brassica rapa L.) using mature vegetative organs without seeds, we established a high efciency regeneration system from heading leaf explants and further explored its application in genetic transformation. Our results showed that maximum shoot regeneration of cultivars ‘Beijing New No. 3’ (88.46±1.65%) and ‘Chengyangqing’ (73.77±1.73%) were obtained in the presence of 17.76 μM 6-benzylaminopurine (BA) and 5.37 μM α-naphthylacetic acid (NAA). The addition of silver nitrate (AgNO3) at 11.77 μM was essential for shoot regeneration from heading leaf explants. Genotypic diferences in shoot regeneration frequencies (from 47.24±3.65 to 88.46±1.65%) were observed amongst eight Chinese cabbage cultivars, in addition to the number of shoots per explant (from 1.31±0.02 to 2.02±0.05) in Murashige and Skoog medium containing 17.76 μM BA, 5.37 μM NAA and 11.77 μM AgNO3. Low temperature (4 °C) had an efect on in vitro preservation of leafy heads with a delay in leaf wilting, and there were no signifcant diferences in shoot induction frequency within 24 h for cultivars ‘Beijing New No. 3’ and ‘Chengyangqing’. In the genetic transformation experiments using selection with kanamycin (17.17 μM), a transformation efciency of 0.6–1.2% was achieved, as assessed from PCR and Southern blot results. The above results suggested that the heading leaf explants can not only achieve efcient seed-independent propagation of Chinese cabbage, but also provide a feasible platform for genetic transformation.
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