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为探讨二化螟Chilo suppressalis(Walker)肠道微生物多样性与抗药性的关系,本研究采用基于16S rDNA的变性梯度凝胶电泳(DGGE)和16S rDNA文库序列分析方法,检测和分析了二化螟4个抗药性水平不同的种群幼虫中肠细菌群落多样性。生测结果表明,以二化螟黑龙江种群(HIJ种群)作为相对敏感品系,连云港种群(LYG种群)对杀虫单、毒死蜱、三唑磷的抗性为低抗至中抗水平,瑞安种群(RA种群)和诸暨种群(ZJ种群)的抗性为中抗至极高抗水平,这3个种群对阿维菌素均为敏感水平。16S rDNA文库序列分析表明,PCR扩增得到的16S rDNA基因代表了二化螟幼虫中肠内21种细菌系统发育型,其中大多数属于链球菌属Streptococcus。在不同抗药性水平二化螟种群中,幼虫中肠微生物群落除ZJ种群的Lactococcus garvieae,L.lactissubsplactis和Ochrobactrum anthropic等3种菌较丰富外,其余均以肠球菌属Enterococcus为主。DGGE图谱显示,HIJ种群条带较为单一,LYG种群条带最为丰富,ZJ种群与RA种群条带丰富度相似。4个种群均出现Enterococcus faecium,E.hirae和Arthrobacter sp.等细菌,且以肠球菌属Enterococcus为主。结果显示了不同抗药性水平的二化螟种群中肠细菌群落的丰富度存在差异,推测可能与二化螟不同抗药性差异有关。
In order to explore the relationship between intestinal microflora and resistance in Chilo suppressalis (Walker), the 16S rDNA-based denaturing gradient gel electrophoresis (DGGE) and 16S rDNA sequence analysis were used to detect and analyze two Bacterial community diversity in midgut of larvae of four populations with different resistance levels. The results of biopsy showed that resistance to insecticide single, chlorpyrifos and triazophos in Liyonggang population (LYG population) was low to moderately resistant to the populations of Heilongjiang population (HIJ population) RA population) and Zhuji population (ZJ population) were resistant to extremely high resistance levels, all of which were sensitive to avermectin. 16S rDNA sequence analysis showed that the 16S rDNA gene amplified by PCR represented 21 bacterial species in the midgut of the larvae of the Chilo suppressalis larvae, most of which belonged to Streptococcus. In different resistance levels of rice stem borer population, the larval midgut microbial community in addition to the ZJ population of Lactococcus garvieae, L.lactissubsplactis and Ochrobactrum anthropic three kinds of bacteria are more abundant, the rest are Enterococcus Enterococcus. The DGGE maps showed that the bands of HIJ were single, the bands of LYG were the most abundant, and the bands of ZJ and RA were similar. Enterococcus faecium, E.hirae and Arthrobacter sp. Were found in all four populations, and Enterococcus faecalis was the dominant enterococcus. The results showed that there was a difference in the abundance of intestinal bacterial communities in the rice stem borer (RSB) populations with different drug resistance levels, which may be related to the different resistance of the rice stem borer, Chilo suppressalis.