球囊菌侵染对意大利蜜蜂幼虫肠道免疫与解毒相关基因表达及蛋白质、脂质和糖类含量的影响

doi:10.16380/j.kcxb.2021.05.004

摘要/Abstract

摘要：

【目的】球囊菌Ascosphaera apis是一种真菌性病原体，可以侵染意大利蜜蜂Apis mellifera ligustica幼虫，导致蜂群患白垩病。本研究通过球囊菌侵染意大利蜜蜂幼虫，探究对意大利蜜蜂幼虫肠道免疫解毒相关基因表达及蛋白质、脂质和糖类含量的影响，分析球囊菌侵染胁迫下意大利蜜蜂免疫应答反应与营养物质的关系。【方法】实验室饲养意大利蜜蜂幼虫，处理组每头3日龄幼虫接种2×10⁶孢子/mL球囊菌，对照组饲喂正常食物;收集6日龄幼虫肠道样品。通过qRT-PCR测定收集的肠道样品中免疫、解毒、发育相关基因和病原基因的表达，并利用生化方法测定肠道样品中蛋白质、脂质、葡萄糖和糖原含量。【结果】球囊菌侵染意大利蜜蜂后，幼虫肠道免疫相关基因LOC406144, Apid1, Def1, Def2, LOC406142, PGRP-SA, Pgrp-s2, PGRP-S3, PGRP-lc, GNBP-1, GNBP3, PPOact, LOC552247, domeless, KAT2A和bsk显著上调表达，cactus显著下调表达；解毒相关基因Cat, GSTS3, Cyp4g11, LOC725294, Ampka-r1, LOC411223, LOC409791, AmNOS和Sod2显著上调表达，CYP6AS14, CPR14, CYP306A1和LOC100576555显著下调表达；发育相关基因usp显著上调表达，VGMC, HEX70b和Vg显著下调表达；病原基因BQCV capsid protein和Ascosphaera apis 28S rRNA表达量显著上升；蛋白质与脂质含量显著下降，葡萄糖与糖原含量显著上升。【结论】球囊菌侵染意大利蜜蜂幼虫后影响其幼虫肠道中免疫、解毒、发育相关基因和病原基因的表达，影响意大利蜜蜂的免疫应答及能量应激反应，加速蛋白质、脂质的流失及葡萄糖和糖原能量储备，影响机体正常的生理活动。

关键词: 意大利蜜蜂, 球囊菌, 免疫相关基因, 解毒相关基因, 免疫应答, 营养成分

Abstract: 【Aim】 Ascosphaera apis is a fungal pathogen that infects Apis mellifera ligustica larvae and causes chalkbrood in honeybee colonies. This study aims to investigate the effects of A. apis infection on the expression of genes related to intestinal immunity and detoxification and the contents of proteins, lipids and carbohydrates in the gut of A. m. ligustica larvae, and to analyze the relationship between the immune response and nutrients of A. m. ligustica under A. apis infection. 【Methods】 The A. m. ligustica larvae were raised in the laboratory. Each 3-day-old larva was inoculated with 2×10⁶ spores/mL of A. apis in the treatment group, and that in the control group was fed with the normal food. Then, the gut samples of 6-day-old larvae were collected. The expression levels of genes related to immune, detoxification and development and genes of pathogens in the gut samples were detected by qRT-PCR, and the contents of proteins, lipids, glucose, and glycogen in the gut samples were determined by biochemical methods. 【Results】 After A. apis infection, the expression levels of immune-related genes LOC406144, Apid1, Def1, Def2, LOC406142, PGRP-SA, Pgrp-s2, PGRP-S3, PGRP-lc, GNBP-1, GNBP3, PPOact, LOC552247, domeless, KAT2A and bsk in the larval gut of A. m. ligustica were significantly up-regulated and that of cactus was significantly down-regulated. The expression levels of detoxification-related genes Cat, GSTS3, Cyp4g11, LOC725294, Ampka-r1, LOC411223, LOC409791, AmNOS and Sod2 in the larval gut of A. m. ligustica infected by A. apis were significantly up-regulated and those of CYP6AS14, CPR14, CYP306A1 and LOC100576555 were significantly down-regulated. The expression level of development-related gene usp in the larval gut of A. m. ligustica infected by A. apis was significantly up-regulated and those of VGMC, HEX70b and Vg were significantly down-regulated. The expression levels of pathogen genes BQCV capsid protein and Ascosphaera apis 28S rRNA in the larval gut of A. m. ligustica infected by A. apis were significantly up-regulated. The contents of proteins and lipids in the larval gut of A. m. ligustica infected by A. apis were significantly down-regulated, and those of glucose and glycogen were significantly up-regulated. 【Conclusion】 The A. apis infection influences the expression of genes related to immune, detoxification and development and genes of pathogens in the gut of A. m. ligustica larvae, affecting the immune response and energy stress response and accelerating the loss of proteins and lipids and the energy reservation of glucose and glycogen, and these may result in abnormal physiological activities of A. m. ligustica larvae.

Key words: Apis mellifera ligustica; Ascosphaera apis, immune-related gene, detoxification-related gene, immune response, nutrients

候梦赏, 赵必安, 邱园妹, 万昆林, 梁立强, 李志国, 苏松坤. 球囊菌侵染对意大利蜜蜂幼虫肠道免疫与解毒相关基因表达及蛋白质、脂质和糖类含量的影响[J]. 昆虫学报, 2021, 64(5): 574-584.

HOU Meng-Shang, ZHAO Bi-An, QIU Yuan-Mei, WAN Kun-Lin, LIANG Li-Qiang, LI Zhi-Guo, SU Song-Kun. Effects of Ascosphaera apis infection on the expression of immune- and detoxification-related genes and contents of proteins, lipids and glucoses in the gut of Apis mellifera ligustica larvae[J]. Acta Entomologica Sinica, 2021, 64(5): 574-584.

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球囊菌侵染对意大利蜜蜂幼虫肠道免疫与解毒相关基因表达及蛋白质、脂质和糖类含量的影响

Effects of Ascosphaera apis infection on the expression of immune- and detoxification-related genes and contents of proteins, lipids and glucoses in the gut of Apis mellifera ligustica larvae

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