Efficient plant regeneration and Agrobacterium-mediated transformation via somatic embryogenesis in melon (Cucumis melo L.) 哈密瓜的有效植株再生和体胚诱导农杆菌转型 We established an efficient plant regeneration and transgenic system using somatic embryogenesis in two melon cultivars: Vedrantais (Cucumis melo L., var. cantalupensis) and Earl’s Favourite Fuyu A (C. melo L., var. reticulatus). These model cultivars are used for genetic and development studies of melon. As explants, 1- to 4-mm2 sections of mature seeds were used, including the cotyledon and hypocotyl. To induce embryo formation, the explants were cultured in embryo-induction (EI) medium, which consisted of liquid MS medium containing 2 mg/l 2,4-d and 0.1 mg/l BA. After 4 weeks of culture, an average of 116.7 and 148.6 embryos per seed were produced for Vedrantais and Earl’s Favourite Fuyu A, respectively. The frequency of germination 2 weeks after transfer into MS liquid medium was 80% in Vedrantais and 35% in Earl’s Favourite Fuyu A. More than 60% of the regenerated plants were diploid, and exhibited normal phenotypes when grown in the field. Using this protocol, Agrobacterium-mediated transformation in Vedrantais was developed. Calli and embryos were obtained in EI selection medium containing 25 mg/l kanamycin and 375 mg/l Augmentin 9 weeks after bacterial inoculation, and over 80% of these were transgenic. Putative transgenic plants were regenerated from these embryos in 1.0% agar-solidified MS medium containing 50 mg/l kanamycin, and were grown in a greenhouse. Transformation was confirmed with a histochemical GUS assay, PCR analysis, and Southern blot analysis of the transgenic plants. We obtained transgenic plants successfully and the transgenic rate exceeded 2.3%, which is sufficient for practical use. 我们利用体细胞胚胎在Vedrantais (Cucumis melo L., var. cantalupensis)和Earl’s Favourite Fuyu A (C. melo L., var. reticulatus)，建立有效植株再生和基因转殖系统，并且使用在甜瓜的基因发展研究上，培植体，是使用１～４平方公厘的成熟种子切片，并且包含子叶和胚轴。接着诱导体胚形成。培植体在EI培养基中培养，此为MS液态培养基再加入2 mg/l 2,4-d和0.1 mg/l BA.。经过四个礼拜的培养之後，Vedrantais和Earl’s Favourite Fuyu，每个种子的平均胚胎数分别为116.7和148.6，换到液态培养基两个礼拜後，Vedrantais的发芽率是８０％而Earl’s Favourite Fuyu的发芽率是30%。此成长过程中，超过60%的再生殖株是二倍体跟正常表现型。在实验显示，农杆菌转殖在Vedrantais的生长情形较好。最後经过九个礼拜的细菌培养，产生的癒合组织和胚胎基因改变超过80%，推测这些转基因植物从胚胎再生，是在1.0% ＭＳ固态培养基加入50 mg/l kanamycin并且在温世长大，使用ＧＵＳ组织化学染色法、ＰＣＲ分析法和南方墨点法分析做基因分析。我们成功的得到了转基因植物，可以作为实用的更超出了2.3% 有的地方怪怪的,请大大帮我看一下 --

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