Study on the Genetic Transformation Conditions of Begonia wallichiana L.with Leaf Disc Method and Corresponding Identification Techniques

[Objectives]This study was conducted to improve the Agrobacterium-mediated genetic transformation system of Begonia wallichiana.[Methods]With sterilized tube seedling leaves as the recipient material and GFP as the reporter gene,optimization experiments were carried out in terms of infection time and method,co-cultivation time and method,and PCR detection technology.[Results]The transformation effect was better under the conditions of shaking Agrobacterium liquid,infection time of 1-2 h,and co-cultivation on sterilized filter paper for 2 d.After co-cultivation,the recipient material was first subjected to recovery culture,and then used for Hyg gradient screening,which was conducive to obtaining resistant transformants.The designed specific PCR detection technology could quickly identify false positives in resistant regenerated plants,and the proportion of transgenic plants was 16.7%.[Conclusions]The research results provide a new technical reference for the genetic transformation of ornamental plants.

茶树茎段不定芽高效发生体系的建立

茶树(Camellia sinensis)是重要的经济作物,杂合度高且变异度大,其高效离体再生体系鲜见报道。以舒茶早茎段为起始外植体,进行不定芽高效发生影响因子研究。结果表明,MS+2 mg·L^(-1)6-BA为定芽诱导的最适配方,定芽诱导率为84.44%,吸收底盘膨大率为80%,利于后续不定芽诱导;MS+2 mg·L^(-1)6-BA+0.2 mg·L^(-1)NAA+0.1 mg·L^(-1)KT+1 mg·L^(-1)脯氨酸为不定芽增殖诱导的适宜配方,不定芽诱导率为88.89%,平均芽数为7.8。不定根诱导的适宜配方为1/2MS+3 mg·L^(-1)IBA,生根率为85.56%。采用RAPD和ISSR技术对再生植株进行分子检测,在连续2代离体再生植株中未发现明显变异。