农杆菌介导HaBADH基因转化马铃薯的研究  

Studies on Agrobactiu tumefacienciens-mediated Transformation of Patato with Betaine aldehyde Dehydrogenase Gene

作　　者：张丽[1] 程永芳[2] 巩檑[1] 甘晓燕[1] 聂峰杰[1] 陈虞超[1] 石磊[1] 张国辉[3] 李苗[1] 颉瑞霞[3] 宋玉霞[1] 

机构地区：[1]宁夏农林科学院农业生物技术研究中心,银川750002 [2]宁夏大学生命科学学院,银川750003 [3]宁夏农林科学院固原农科所,固原756000

出　　处：《分子植物育种》2016年第6期1462-1469,共8页Molecular Plant Breeding

基　　金：宁夏回族自治区育种专项(2014NYY201);宁夏回族自治区自然科学基金(NZ14210;NZ14269);宁夏农林科学院先导资金项目(NKYJ-14-05;NKYJ-15-32;NKYG-15-06;NKYJ-15-36;NKYQ-14-07)共同资助

摘　　要：本研究以马铃薯(Solanum tuberosum L.)试管薯作为转化受体,利用农杆菌介导法将从梭梭中获得的甜菜碱醛脱氢酶基因(HaBADH)转化马铃薯,采用正交试验设计优化了影响马铃薯转化的4个主要因子,建立了农杆菌介导的马铃薯高效转化体系,获得了转基因植株。研究结果表明,转化过程中,浸染时间15 min,菌液浓度OD600为1.0,筛选培养时卡那霉素浓度30 mg/L、品种为大西洋为最佳转化方案,抗性芽分化率最高达到了35.9%;4个因子中基因型对转化效率影响最大,菌液浓度的影响次之,卡那霉素浓度和浸染时间的影响最小。经PCR、RT-PCR及Southern blot检测,证明了HaBADH基因整合到马铃薯中,阳性转化率为45.3%。In this research we used the vitro tuber of potato as transformed receptor, and BADH(Betaine aldehyde dehydrogenase) gene derived from Haloxylon was transformed into the potato mediated by Agrobacterium tumefaciens. We established the efficient transformation system of potato mediated by Agrobacterium tumefaciens and accepted the transgenic plants, and the 4 main factors which affecting transformation efficiency was optimized through orthogonal experimental design. The results showed that the best treatment combination was Atlantic as genotype, infection time for 10 min, concentration of bacterium suspension for OD6001.0, and the concentration of kanamycin in selective culture 30 mg/L. The ratio of resistant buds differentiation was 35.9%. In the 4 factors,genotype was the greatest effect factor on the ratio of transformation, followed by the concentration of bacterium suspension, and the concentration of kanamycin and infection time with minimal impact. By PCR, RT-PCR and Southern blot identification proved that the HaBADH gene had been integrated into genome of potato and the positive transformation frequency was 45.3%。

关 键 词：马铃薯 转化 农杆菌 HaBADH基因 转基因植株 

分 类 号：S532[农业科学—作物学]