黄瓜‘中国龙’毛状根诱导体系的建立

Establishment of hairy root induction system of cucumber‘Chinese Long'

黄瓜是全球范围内重要的经济作物。建立黄瓜基因组测序品种‘中国龙’的毛状根诱导体系，可为今后分析黄瓜基因功能、挖掘黄瓜优良基因提供技术支撑。本文以‘中国龙’为材料，从种子表面灭菌方法、发根农杆菌的侵染浓度和共培养时间3方面，研究发根农杆菌菌株K599对‘中国龙’毛状根诱导的情况。结果表明，75％（φ）的乙醇先处理种子30s，再用5％的次氯酸钠灭菌5—10min的种子消毒方法，农杆菌菌液OD600值在1．2左右时进行侵染，共培养2天，是获得‘中国龙’毛状根的适宜组合方法。PCR扩增结果证实，经这样诱导出的黄瓜毛状根中，发根农杆菌Ri质粒的rolC基因已在黄瓜毛状根基因组中整合并得到表达。试验探索出了一套适合‘中国龙’毛状根诱导的方法。

Cucumber （Cucumis sativus L.） is an important worldwide economic crop. Establishing a reliable cucumber hairy root induction system of ＇ Chinese Long＇ which was used for denovo sequencing, is significant for cucumber functional genomics research. In this article, ＇Chinese Long＇ was used to study the induction of cucumber hairy roots by Agrobacterium rhizogenes （K599） through 3 different aspects, namely, seed surface sterilization method, bacterium concentration （shown as OD600 value）, and the co-culture time. The results showed that, seed surface sterilization with 75% alcohol for 30 s, 5% NaClO for 5-10 min, OD600 value of bacterium is about 1.2 for infection, and then co-culture for 2 d, are the suitable condition for cucumber hairy root induction. The PCR amplification of rolC gene showed that the Ri plasmid of A. rhizogenes was integrated into the genome of induced cucumber hairy roots and was expressed. An optimal method has been explored to induce hairy roots of ＇ Chinese Long＇.