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作 者:史京花[1] 吕慧能[1] Daina Simmonds 盖钧镒[1] 喻德跃[1]
机构地区:[1]南京农业大学农学院,南京210095 [2]Agriculture and Agri-Food Canada,Genomics & Biotechnology,Bldg.21,960 Carling Ave.,Ottawa,Ontario K1A 0C6,Canada
出 处:《分子细胞生物学报》2006年第1期71-76,共6页Journal of Molecular Cell Biology
基 金:本研究得到国家留学基金项目(编号:(2001)3048)"948"项目(编号201013(A))的资助。~~
摘 要:植物离体培养是植物基因操作中的重要一环.也是植物个体发育研究中基因表达研究的有益参考体系。在继代培养过程中发生的遗传变异有时会使再生植株丧失优良的性状而需加以控制和避免。为此,首先需要了解培养过程中的遗传变异情况。We cultured soybean immature cotyledons to induce somatic embryos and set up soybean somatic embryo lines by culturing the induced somatic embryos in liquid medium on shaker. Regenerated plants of normal fertility were easily obtained with the cultures of various ages by culturing the somatic embryos on differentiation media. DNAs were isolated from the embryogenic cultures after 5, 9, 15 or 17 months' suspension and from 42 plants regenerated from somatic embryos of various culturing ages. 102 SSR markers covering soybean genome almost evenly were chosen to check variation of microsatellite DNA during suspension culture and differentiation. Among the eight DNA samples of soybean somatic embryos of various ages and 42 DNA samples of regenerated plants, there was no any variation of the major bands of the 102 SSR markers during one year' s subcuhuring and differentiation. There were only six Weaker subsidiary bands of five SSR markers among the 102 SSR markers added in four of the fifty DNA samples. Two of them happened to the same regenerated plant differentiated from the 9-month embryogenic cultures. Three happened to the two DNA samples from somatic embryos irregular in morphology of the 5-month embryogenic cultures. The last subsidiary band variation happened to a DNA sample of the 17-month embryogenic cultures. The results show that stable microsatellites were maintained during the suspension culture and differentiation while we made the cultures highly embryogenic potential and easy to regenerate.
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