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以“津研四号”、“新津春四号”和“玉蛟龙”3种黄瓜为试材,通过比较影响发状根诱导率的条件,利用隶属函数综合评价农杆菌(A_4)对3种黄瓜发状根诱导能力,为建立一套高效的黄瓜发状根诱导体系提供依据。结果表明:3种黄瓜均成功诱导出发状根,转接培养后仍能正常生长。诱导条件分析表明,选择带叶茎段为外植体,菌液侵染10 min、菌液浓度为OD_(600)=0.3和乙酰丁香酮(acetosyringone)浓度为10 mg/L时诱导效果最好。隶属函数分析表明,A_4对3种黄瓜发状根诱导能力依次为“玉蛟龙”>“津研四号”>“新津春四号”。PCR分子鉴定表明,A_4菌Ri质粒的T-DNA基因已成功整合到3种黄瓜核基因组中。
The cucumber cultivars with three kinds of cucumber (Jinyan No.4, Xinjinchun No.4 and Yujiao Long) were used to evaluate the effects of Agrobacterium tumefaciens by using the membership function (A_4) on cucumber hairy root induction, to establish a set of efficient cucumber hairy root induction system provide the basis. The results showed that all the three kinds of cucumbers successfully induced the root-like roots, which could still grow normally after the transfer culture. Induction conditions showed that the best explants were the explants with the stem segments of leaf explant inoculated for 10 min, with OD 600 (600) = 0.3 and acetosyringone 10 mg / L . The membership function analysis showed that the inducement ability of A_4 to three kinds of cucumber hairy roots was “Jade Dragon”> “Jinyan No.4”> “Xinjinchun No.4”. PCR molecular identification showed that the T-DNA gene of the A_4 bacterium Ri plasmid has been successfully integrated into the three cucumber nuclear genomes.