罗汉果遗传转化体系的建立  

Establishment of Genetic Trasformation System for Siraitia grosvenorii

作　　者：李文兰[1,2] 高永华[1] 李华英[3] 

机构地区：[1]桂林理工大学化学与生物工程学院,桂林541004 [2]武汉大学生命科学学院,武汉430072 [3]广西壮族自治区植保总站,南宁530022

出　　处：《分子植物育种》2010年第5期997-1002,共6页Molecular Plant Breeding

基　　金：广西自然科学基金(桂科青0728096)资助

摘　　要：本研究以罗汉果(Siraitia grosvenorii(Swingle)C.Jeffery)子叶为外植体,通过农杆菌介导法,探讨影响罗汉果转化的几个重要因素,建立罗汉果稳定的遗传转化再生体系。结果表明:农杆菌转化前对子叶进行预培养可以促进细胞分裂,预培养的最佳时间为1d,侵染时间以为20～30min为宜,共培养最佳时间为4d,附加100μmol/L的乙酰丁香酮(AS)可促进转化。通过潮霉素抗性不定芽分化和抗性不定根分化的两轮筛选及PCR检测,获得56株PCR阳性植株,阳性转基因植株频率为6.86%,初步证明目的基因已经整合到罗汉果基因组中。Using cotyledon of Siraitia grosvenorii as explants,an efficient and stable genetic transformation system mediated by Agrobacterium was established through investigating some factors influencing the transformation efficiency.The results showed that cotyledon pre-culture could promote cell division before Agrobacterium-mediated transformation.The highest transformation frequency was obtained,with the optimized pre-cultivate one day,the suitable infection duration 25 min,appropriate co-culture for4～6 days and 100 μmol/L of AS.Several putative transformants were obtained after two round of selection procedure.PCR analysis indicated that target gene had been integrated into the genome of Siraitia grosvenorii and its transgenic frequency was 6.86%.

关 键 词：罗汉果 农杆菌介导 遗传转化 

分 类 号：S567.239[农业科学—中草药栽培]