中华猕猴桃华优遗传转化系统建立研究

为了更好的利用遗传转化技术进行猕猴桃品种改良研究，以中华猕猴桃华优组培苗幼嫩叶片为材料， 研究了外植体分化不定芽再生途径所需的培养条件及转化效率提高方法。结果表明，在叶片不定芽诱导培养基中加 入浓度为0.75~1.0 mg/L 的TDZ 时，叶片愈伤发生率达85.7%~90.1%，不定芽发生率达41.8%~44.6%；在TDZ 1.0 mg/L 基础上，补充0.1~0.4 mg/L IAA 可使叶片愈伤发生率提高到96.7%~100%，不定芽发生率提高到67.4%~71.9 %； 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，可使GUS 瞬时表达率及卡那抗性芽发生率分别 提高至27.9%、14.5%；外植体被农杆菌侵染前，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。GUS 染色与PCR 扩增检测结果显示，Shiva A基因已转化至中华猕猴桃华优转基因植株。

Study on establishment of transformation system mediated by Agrobacteriume in Actinidia chinensis Planch.cv Huayou.

Young leaves of plantlets were used as explants to investigate the factors influencing plants regeneration & transformation efficiency of Actinidia chinensis Planch.cv Huayou. The results showed that in medium plus 0.75~1.0 mg/L thidiazuron (TDZ), 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. when leaf explants were grown in medium containing 1.0 mg/L TDZ and 0.1-0.4 mg/L IAA. GUS Transient expression rate of explants & anti -Kan adventitious buds was significantly enhanced while 75 -100 mol/L AS was supplied in the Agrobacterium infecting cultures & co-cultivation medium. It was useful to increase GUS Transient expression rate of explants & anti-Kan adventitious buds that explants were precultured for 2-3 days before infected by Agobacterium. All regenerated plantlets were deemed putative transgenic by histochemical GUS assay and PCR detection.