山新杨转Bt+蜘蛛杀虫肽基因的分析  

Transformation of Populus davidiana×P. bollena with Bt+spider Toxin Gene

作　　者：左丽丽[1] 王志英[1] 梁臣[2] 谢淑萍[3] 

机构地区：[1]东北林业大学,哈尔滨150040 [2]黑龙江省鸡西市林业局 [3]黑龙江省海林市林业局

出　　处：《东北林业大学学报》2009年第7期112-114,共3页Journal of Northeast Forestry University

基　　金：黑龙江省科技攻关项目(GB06B303-4);黑龙江省重点基金项目(ZJN04-0101)资助

摘　　要：以山新杨(Populus davidiana Dode×P.bollena Lauche)为受体材料,在筛选遗传转化条件的基础上,利用农杆菌介导叶盘转化法进行了Bt+蜘蛛杀虫肽基因转化研究。结果表明:在叶片转化时,卡那霉素的临界质量浓度为10mg/L;生根筛选培养时卡那霉素的临界质量浓度为15mg/L;利用质量浓度为600mg/L的头孢噻肟钠脱菌,对叶片生长和不定芽形成影响不大。山新杨遗传转化的工程菌菌液浓度以0.1～0.2(OD600)为最佳,其叶片分化率为73%～77%、抗性芽率为20%～23%;浸染时间以2～5min为宜,其叶片分化率和抗性芽率最高。在共培养基中加入200μmol/L的乙酰丁香酮,其叶片分化率和抗性芽率分别比对照提高1.43倍和2.61倍。对转化植株的PCR检测结果呈阳性,初步证明外源基因Bt+蜘蛛杀虫肽导入并整合进了山新杨基因组。The gene of Bt ＋ spider toxin was transformed using Populus davidiana Dode × P. bollena Lauche as transgenie acceptors by Agrobacterium tumefaciens mediated method. The leaf discs were immersed into 0.1 - 0.20D600 of bacteria solution for 2 - 5 min, which was the optimal condition for infection. The threshold concentration of kanamycin was 10 mg/L for differentiation and 15 mg/L for rooting. 600 mg/L of cefotaxime could inhibit the growth of bacteria without inhibition to the differentiation of leaf discs. As results, the disc differentiation rate was 73 to 77 percent, 20 to 23 percent of which generated buds resistant to kanamycin. When 200 μmol/L of acetosyringone was supplemented into the medium, the disc differentiation rate increased by 1.43 times of the control, and the rate of buds resistant to kanamycin also increased by 2.61 times of the control. PCR result shows that the exogenous gene has integrated into the genome of the poplar.

关 键 词：山新杨 Bt+蜘蛛杀虫肽基因 遗传转化 

分 类 号：S763.306.5[农业科学—森林保护学] S792.119[农业科学—林学]