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从思茅松毛虫Dendrolimu kikuchii Matsumura4龄幼虫肠道环境中分离、纯化、培养,获得11株好氧细菌菌株。以细菌基因组DNA为模板,用细菌通用引物(27f、1492r)对模板进行PCR扩增,并用4种限制性内切酶HaeⅢ、HindⅢ、HinfⅠ、TaqⅠ对PCR产物进行ARDRA(amplified rDNA restriction analysis)多态性分析,在84%的相似水平上可分成6大类群,多样性丰富。对6大类群的代表菌株进行16SrDNA序列测定,分析表明:分离到的11株好氧细菌中4N05、4N08和4N11归属克雷伯氏杆菌属(Klebsiellasp.),4N02归属Lysinibacillus属;4N03和4N09确定到种,分别是γ-变形杆菌(Gamma proteobacterium)和枯草芽孢杆菌(Bacillus subtilis strain);4N04、4N06、4N07和4N01、4N10也可以确定到种,其中前3株是短短芽孢杆菌(Brevibacillus brevisstrain),后2株是沼泽短芽孢杆菌(Brevibacillus limnophilusstrain),并且这5株菌都归属短芽孢杆菌属(Brevibacillus sp.)。通过研究,既可以为松毛虫的生物防治提供微生态理论依据,又丰富了微生物资源库。
Isolation, purification and culture of the 4-instar larvae of Dendrolimus kikuchii Matsumura larvae of Dendrolimus kikuchii were performed to obtain 11 strains of aerobic bacteria. Bacterial genomic DNA was used as a template to amplify the template by universal bacterial primers (27f, 1492r). The PCR products were amplified with four restriction endonucleases (HaeⅢ, HindⅢ, HinfⅠ, TaqⅠ) The morphological analysis can be divided into 6 groups at a similar level of 84%, with rich diversity. The 16S rDNA sequence analysis of representative strains of 6 groups showed that among the 11 aerobic bacteria isolated, 4N05, 4N08 and 4N11 belonged to Klebsiella sp., 4N02 belonged to Lysinibacillus; 4N03 and 4N09 were identified To the species, respectively, Gamma proteobacterium and Bacillus subtilis (Bacillus subtilis strain); 4N04,4N06,4N07 and 4N01,4N10 can also be identified species, of which the first three strains are Brevibacillus brevisstrain ), The latter two strains are Brevibacillus limnophilusstrain, and all five strains belong to Brevibacillus sp. Through the study, it can not only provide theoretical basis for the biological control of pine caterpillars, but also enrich the microbial resource base.