宏基因组解析溴氰虫酰胺对小菜蛾肠道菌群结构的影响

Effect of cyantraniliprole on structure of intestinal microflora of Plutella xylostella(Linnaeus)revealed by metagenomic sequencing

【目的】探究溴氰虫酰胺胁迫对小菜蛾肠道酶活、微生物菌群、相关功能基因及抗性基因的影响,为双酰胺类杀虫剂处理小菜蛾肠道微生物的结构和功能研究提供参考。【方法】采用宏基因组测序技术研究浓度为0.15 mg/L溴氰虫酰胺浸叶饲喂小菜蛾后其肠道微生物群落结构及其功能的变化情况。【结果】溴氰虫酰胺浸叶饲喂后小菜蛾肠道中羧酸酯酶和谷胱甘肽-S-转移酶活性显著增高,而多功能氧化酶活性显著降低。溴氰虫酰胺未处理与处理小菜蛾肠道的微生物分别注释到34和35个门、66和64个纲、120和125个目、200和214个科、261和294个属、347和401个种,其中微孢子门(Microsporidia)、厚壁菌门(Firmicutes)为处理与对照组的共有优势菌门,小孢子虫属(Nosema)、肠球菌属(Enterococcus)为处理组与对照组的共有优势菌属,但溴氰虫酰胺处理后2种优势菌属的相对丰度均有降低。小菜蛾肠道微生物在eggNOG与KEGG数据库中分别注释到11 682和6437个功能基因,分布于水化合物转运与代谢、翻译后修饰、蛋白质翻转、遗传信息处理等功能类群;在CAZy和CARD数据库中分别注释到410个碳水化合物活性酶基因及18类311个抗生素耐药性本体。溴氰虫酰胺处理改变了肠道微生物菌群的代谢功能,其在eggNOG与KEGG数据库、CAZy数据库中及CARD数据库中可注释的功能基因均增加,分别为15 432和8591个功能基因、577个碳水化合物活性酶基因及22类419个抗生素耐药性本体。【结论】溴氰虫酰胺浸叶处理增加小菜蛾肠道羧酸酯酶和谷胱甘肽-S-转移酶活性、降低多功能氧化酶活性,降低肠道优势细菌的相对丰度,但增加碳水化合物活性酶基因、抗生素耐药性等功能基因的数量。

【Objective】The effect of cyantraniliprole on the gut enzyme,microbial composition,relative function genes and resistance genes of Plutella xylostella is explored,which provides reference for the study of microbial composition and function in gut of P.xylostella treated with diamide insecticides.【Method】Through metagenomic sequencing technology,the changes of intestinal microbial community structure and function of P.xylostella after feeding 0.15 mg/L cyantraniliprole-soaked leaves were studied.【Result】The activities of carboxylesterase and glutathione-S-transferase in the gut of P.xylostella fed with cyantraniliprole-soaked leaves increased significantly,while the activities of mixed-functional oxidase decreased significantly.A total of 35 phyla,64 classes,125 orders,214 families,294 genera and 401 species were identified in the gut of P.xylostella treated with cyantraniliprole,while 34 phyla,66 classes,120 orders,200 families,261 genera and 347 species were identified in the the gut of P.xylostella with non-treatment of cyantraniliprole(CK).Microsporidia and Firmicutes were the com-mon dominant phyla,and Nosema and Enterococcus were the dominate genera in the gut of P.xylostella in the treated group and CK,while the relative abundance of common dominant genera in the treated group were lower than that in CK.A total of 11682 and 6437 functional genes were annotated in the egg NOG and KEGG databases,respectively,which distributed in functional groups of carbohydrate transport and metabolism,posttranslational modification,protein turnover,genetic information processing and other groups;410 carbohydrate-active en-zyme genes and 18 categories of 311 antibiotic resistance ontologies were annotated in the CAZy and CARD databases,respective-ly.Cyantraniliprole treatment changed the metabolic function of the intestinal microflora of P.xylostella.The intestinal microflora of P.xylostella were annotated with 15432 and 8591 functional genes,577 carbohydrate-active enzyme genes,and 419 antibiotic resistance on-tologies in 22 categories in the egg NOG and KEGG database,CAZy database,and CARD database,respectively.【Conclusion】Cyantranilip-role treatment increases intestinal carboxylesterase and glutathione-S-transferase activities,decreases mixed-functional oxidase activity,and reduces the relative abundance of dominant intestinal microorganisms in gut,but increases the number of functional genes such as carbohy-drate-active enzyme genes and antibiotic resistance in P.xylostella.