转Bt cry6A基因的灰葡萄孢菌对松材线虫致死效应分析  

Lethal Effect Analysis of Bortytis cinerea Transformants with Bacillus thuringiensis cry6A Gene on Bursaphelenchus xylophilus

作　　者：王殿东[1,2] 周梦溪[3] 罗中钦[1] 谢丙炎[1] 杨宇红[1] 茆振川[1] 刘二明[3] 陈国华[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081 [2]长江师范学院生命科学与技术学院,重庆408100 [3]湖南农业大学生物安全科学技术学院,长沙410128

出　　处：《中国生物防治学报》2013年第3期443-449,共7页Chinese Journal of Biological Control

基　　金：国家“973”计划项目(2009CB119201);国家自然科学基金项目(30900926);转基因生物新品种培育重大专项重点项目(2009ZX08009-034B)

摘　　要：松材线虫是重要的外来入侵物种,给我国松林造成了巨大的破坏,目前尚没有十分有效的防治方法。苏云金芽胞杆菌(Bt)晶体蛋白对秀丽杆线虫及根结线虫具有一定的致死作用。为验证Bt晶体蛋白是否对松材线虫具有致死作用,本研究构建农杆菌介导的丝状真菌转化载体pHD-OH,将苏云金芽胞杆菌晶体蛋白(Bt)cry6A基因和绿色荧光蛋白sGFP基因转化至灰葡萄孢菌中,用灰葡萄孢菌转化子喂饲松材线虫,分析Bt晶体蛋白对松材线虫种群增长的影响。结果表明,sGFP基因和cry6A基因均转入灰葡萄孢菌中,且能够稳定表达,3个cry6A基因转化子为单拷贝。cry6A基因转化子喂饲松材线虫后,对松材线虫产生致死效应,抑制线虫的种群增长,而sGFP转化子和野生型灰葡萄孢菌对松材线虫的种群增长量和死亡率没有显著影响。这表明cry6A基因对松材线虫具有生防潜力。The pine wood nematode was an alien invasive species and causes severe damage to pine forestry in China. However, no effective control method could apply to pine wood nematode. Crystal proteins of Bt had lethal effect on model nematode and root knot nematode. In order to reveal the lethal effect of crystal protein of Bt on pine wood nematode, the vector pHD-OH for Agrobacterium tumefaciens-mediated transformation was constructed and used to transform sGFP gene and cry6A gene into Bortytis cinerea, and then the fugal transformant was feed by Bursaphelenchus xylophilus, to reveal the effect of cry6A gene transformant on dynamic of population of Bursaphelenchus xylophilus. The results showed that sGFP gene and cry6A gene was expressed steadily in the transformants and cry6A gene in transformant was single copy. The three transformants with cry6A gene could kill Bursaphelenchus xylophilus, so inhibit the population increase of the nematode, but the wild type and sGFP gene transformant had no impaction on population of Bursaphelenchus xylophilus. The conclusion revealed that cry6A gene has potential biocontrol to Bursaphelenchus xylophilus.

关 键 词：苏云金芽胞杆菌 灰葡萄孢菌 松材线虫 致死效应 

分 类 号：Q784[生物学—分子生物学] S763.49[农业科学—森林保护学]