摘要
通过生物测定得到了对多种重要农林害虫高活性的苏云金杆菌菌株Bt CYZ-4,对其基因型鉴定表明:该菌株基因型较丰富。该菌株对粘虫(Mythimna separata)初孵幼虫48h LC50为6.6×106个/mL(芽孢密度);对美国白蛾(Hyphantria cunea)、甜菜夜蛾(Spodoptera exigua)和棉铃虫(Helicoverpa armigera)初孵幼虫72h LC50分别为2.9×106、2.25×105和4.7×106个/mL;对林业害虫杨扇舟蛾(Clostera anachoreta)二龄幼虫72h LC50为1.7×106个/mL,同时该菌株对卫生害虫淡色库蚊(Culex pipiens paliens)也表现出较好的活性,24h LC50为2.01×105个/mL。利用cry1、cry2、cry3、cry7、cry8、cry9和vip3A基因引物对该菌株进行基因型鉴定,并对其扩增片段进行限制性酶切分析,结果表明,该菌株含有cry1Ac、cry1Ab、cry2Ab和vip3A基因,不存在cry3、cry7、cry8、cry9基因,其它基因型仍有待鉴定。SDS-PAGE分析表明,晶体蛋白质的分子质量主要为130、70、57、27ku,营养期为88ku。该菌株是一株具有应用潜力的Bt野生株。
A Bacillus thuringiensis （Bt） isolate CYZ-4, which was high lethal to some pests of Lepidoptera and Diptera, was screened by bioassay and DPS analysis. And its gene types of cry1, cry2, cry3, cry7, cryS, cry9 , vip3A and the protein types were identified by PCR-RFLP and SDS-PAGE. The bioassay result indicated that the isolate was more lethal to these pests compared with Bt subsp, kurstaki HD1. The LC50 of Bt CYZ-4 isolate against Mythimna separate was 6.6 × 10^6 cells/ mL in 48 hours. The LC50 values of the isolate against 1st larvae of Hyphantria cunea （Drury） , Spodoptera exigua, and Helicoverpa armigera were 2.9 × 10^6, 2.25 × 10^5, and 4.7× 10^6 cell/mL, and that against 2nd larvae of Clostera anachoreta was 1.7 × 10^6 cells/mL in 72 hours; while it also had insecticidal activity against Culex pipiens pallens, and the LC50 was 2.01 × 10^5 cells/mL in 24 hours. The genotypes of the Bt isolate were identified by the methods of PCR-RFLP and SDS-PAGE. vip3A, crylAc, cry1Ab and cry2Ab were identified in Bt CYZ-4, and cry3, cry7, cryS, cry9 were not found, while other genes still need to be identified. SDS-PAGE analysis also showed that the strain produced crystal proteins of 130, 70, 57 and 27 ku, and 88 ku at vegetative stage. Results prove that Bt CYZ-4 is a potential wild strain against some pests in oractice.
出处
《东北林业大学学报》
CAS
CSCD
北大核心
2009年第9期104-105,共2页
Journal of Northeast Forestry University
基金
河北省太行山区有机苹果栽培技术研究与示范项目(09230602D)
河北农业大学博士基金资助
