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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。
Bursaphelenchus xylophilus is an important alien invasive species that has caused tremendous damage to pine forests in China. At present, there is not a very effective control method. Bacillus thuringiensis (Bt) crystal protein has a certain lethal effect on C. elegans and root knot nematodes. In order to verify whether Bt crystal protein has lethal effect on pine wood nematode, Agrobacterium tumefaciens-mediated transformation of filamentous fungi pHD-OH was constructed. Bacillus thuringiensis crystal protein (Bt) cry6A gene and green fluorescent protein sGFP gene were transformed into In Botrytis cinerea, the pine wood nematode was fed with B. cinerea transformants, and the effect of Bt crystal protein on the growth of the pine wood nematode population was analyzed. The results showed that sGFP gene and cry6A gene were transferred into Botrytis cinerea, and were stably expressed. Three cry6A gene transformants were single copy. The feeding of pine wood nematode with the cry6A gene transformants produced a lethal effect on the growth of pine wood nematode and inhibited the population growth of nematodes, while the sGFP transformants and wild type Botrytis cinerea had no significant effect on the population growth and mortality of pine wood nematode . This indicates that the cry6A gene has the potential of biocontrol against Bursaphelenchus xylophilus.