通过农杆菌介导将菜豆几丁质酶基因导入马铃薯

INTRODUCTION OF CHITINASE GENE FROM KIDNEY BEAN INTO POTATO BY AGROBACTERIUM MEDIATED TRANSFORMATION

选用两个生产上主栽马铃薯品种“鲁引 1号”和“鲁引 4号”的试管微型薯为材料 ,通过农杆菌介导成功地将菜豆几丁质酶基因导入到马铃薯中。试管微型薯薯片与农杆菌共培养 3d后 ,转到含卡那霉素 10 0mg/L的分化培养基上诱导不定芽分化。待抗性芽长到 1 0～ 1 5cm高时 ,转入含卡那霉素 10 0mg/L的液体培养基中进行生根筛选。经过分化和生根两轮筛选的转化植株的PCR检测结果均为阳性 ,而未经转化的对照植株为阴性 ,证明该筛选系统是可靠的。早熟品种鲁引 1号的转化频率明显高于晚熟品种克新 4号 ,其最高转化频率为 4 1 0 % ,平均每个外植体可得到 3 85株转化苗。

Microtubers of two leading potato cultivars Luyin 1 and Kexin 4 were used as exlants in the study.The chitinase gene from kidney bean was introduced into potato by Agrobacterium mediated transformation.After three day's coculture with Agrobacterium tumefaciens,the discs of in vitro grown microtubers were transferred onto selection medium containing 100mg/L kanamycin to induce buds.When the shoots reached 1 0～1 5cm high,they were transferred into liquid medium containing 100mg/L kanamycin to select by rooting.Through the two cycles of selection,the PCR reactions of all transformed plants were positive,which indicated that the selection system was reliable.The transformation frequency of the early cultivar Luyin 1 was obviously higher than that of the late cultivar Kexin 4,with the highest frequency of 41 0% and 3 85 plants per explants.