中华猕猴桃华优遗传转化系统建立研究-广东农业科学.pdfVIP

38 东农业科学 2014 年第 苑 期 中华猕猴桃华优遗传转化系统建立研究 荣生 杜晓莉 霍秀娟 韩美丽 马跃峰 覃建林 郭成林 西农科院植物保护研究所/ 西作物病虫害生物学重点实验室袁 西 南宁 530007 冤 摘要 为了更好的利用遗传转化技术进行猕猴桃品种改良研究袁以中华猕猴桃华优组培苗幼嫩叶片为材料袁 研究了外植体分化不 芽再生途径所需的培养条件及转化效率提高方法遥 结果表明袁在叶片不 芽诱导培养基中加 入浓度为 0.75~1.0 mg/L 的T DZ 时袁 叶片愈伤发生率达 85.7% ~90.1% 袁 不 芽发生率达 41.8% ~44.6% 曰 在 T DZ 1.0 mg/L 基础上袁补充 0.1~0.4 mg/L IAA 可使叶片愈伤发生率提高到 96.7% ~100% 袁不 芽发生率提高到 67.4% ~71.9 % 曰 在农杆菌侵染液与共培养基中同时加入 75~100 滋mol/L 浓度的AS 袁可使 GUS 时表达率及卡那抗性芽发生率分别 提高至 27.9% 尧14.5% 曰外植体被农杆菌侵染前袁的预培养 2~3 d 有利于提高 GUS 表达率与卡那抗性芽发生率遥 GUS 染色与 PCR 扩增检测结果显示袁Shiva A 基因已转化至中华猕猴桃华优转基因植株遥 关键词 猕猴桃曰 不 芽曰 再生曰 转化 中图分类号 Q789 文献标识码 A 文章编号 1004-874X 2014冤07原0038-05 Study on establishment of transformation system mediated by Agrobacteriume in Actinidia chinensis Planch.cv Huayou. LU Rong-sheng袁DU Xiao-li袁HUO Xiu-j uan 袁HAN Mei-li, MA Yue-feng, QIN Jian-lin, GUO Cheng-lin (Institute of Plant Protec tion, GXA A S/ Guangxi Key Laboratory of Biology for Crop Disease s and Inse c t Pe s ts , Nanning 530007, China) A bstract: Young leaves of plantlets were used as explants to investigate the factors influencing plants regeneration transformation efficiency of A c tinidia c hine ns is Planch.cv Huayou. T he results showed that in medium plus 0.75 ~1.0 mg/L thidiazuron (T DZ), callus formation rate reached 85.7% -90.1% and differentiation of buds rate reached 41.8% to 44.6%. Relatively high callus formation rate ( 96.7% -100 %) differentiation of buds rate ( 67.4% -71.9 %) were achived. w hen leaf explants were gro