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作 者:毛娟[1] 陆璐[2] 陈佰鸿[1] 禇明宇 赵长增[1]
机构地区:[1]甘肃农业大学农学院,兰州730070 [2]西北师范大学生命科学学院,兰州730070
出 处:《园艺学报》2013年第6期1101-1109,共9页Acta Horticulturae Sinica
基 金:甘肃省农牧厅生物技术专项(GNSW-2004-04)
摘 要:为进一步明确甜瓜果实软化的分子机理,构建了 ACC 氧化酶Ⅰ基因(CmACOⅠ)启动子与GUS 基因融合的植物表达载体,采用根癌农杆菌介导法转化甜瓜‘甘甜一号’。通过卡那霉素抗性和 PCR检测筛选呈阳性的转化植株,取不同组织进行 X-Gluc 染色。结果表明,转化植株的根、茎、叶、花、果实等器官组织经 X-Gluc 染色后,只在花药组织和成熟果果皮中出现蓝色斑点,其余组织均未检出,表明甜瓜 CmACOⅠ启动子能够驱动 GUS 基因在转基因甜瓜花药和成熟果果皮中特异表达。To provide an important reference for further clarifying the molecular mechanism of fruit softening, GUS histochemical staining method was employed to detect tissue specific regulation of the CmACOI promoter in melon (Cucumis melo L.) . The plant expression vector with a fusion gene, Cm.4COIpromoter and GUS gene was established and melon 'GT-I' was transformed by using Agrobacterium-mediated transformation system. Then the roots, stems, leaves, flowers, fruits and other organs in transformed plants during different growth stages were dyed with X-Glue. The transformed plants with Kanamysin resistance and positive detection by PCR were obtained. It showed that some blue spots in the anther tissue and peel organ of the after-ripening fruit in the plants with a fusion gene ( CmACO] promoter and GUS gene) were detected by using X-Glue dye, while the other organs did not appear blue spots. It showed that GUS gene was driven by melon CrnACOI promoter and expressed soecifieallv in anther tissue and heel organ of the. nfter-rinaning fruit.
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