摘要
以马铃薯品种早大白和克新18号微型薯为外植体,利用根癌农杆菌介导法,将来源于Bt菌株Bt185的P2300-pa-8e抗虫基因转入马铃薯,并对其遗传转化的影响因素进行研究。获得抗性植株早大白19株、克新18号21株;经PCR检测,5株早大白、9株克新18号呈阳性;经Southern-blot杂交分析,4株早大白、6株克新18号呈阳性,证明P2300-pa-8e基因已整合到马铃薯基因组中。在网室内对转基因植株进行抗虫试验,转基因材料对鞘翅目害虫暗黑鳃金龟和华北大黑鳃金龟幼虫表现出抗性。
P2300-pa-8e gene was originated from Bt185,which exists in insect resistance transcription factor.In this study,P2300-pa-8e gene was transferred into the potato(Solanum tuberosum L.)cultivars,'Zaodabai'and'Kexin No.18'by agro-bacterium-mediated transformation,and influence factors of genetic transformation were optimized.Microtuber discs were used as explants.19 resistant plants of'Zaodabai'and 21 resistant plants of'Kexin No.18'were obtained.Resistant plants were blot analyzed by PCR detection,of which 5 and 9 plants were positive.And these resistand plants were analyzed by Southern-blot,of which 4 and 6 plants had a positive amplification.Southern blot showed that single copies of the P2300-pa-8e gene were inserted into the genome of the transgenic plants.By the result of insect-resistant experiment,the transgenic materials showed resistance to coleopteran pests Holotrichia parallela and the larvae of Holotrichia oblita(Fald.).
出处
《中国蔬菜》
北大核心
2011年第18期36-40,共5页
China Vegetables
基金
国家高技术研究发展计划项目(2009AA10Z103)
