东方蜜蜂微孢子虫感染西方蜜蜂工蜂中肠的蛋白质组学分析

doi:10.11843/j.issn.0366-6964.2023.04.033

摘要/Abstract

摘要： 通过分析西方蜜蜂工蜂感染东方蜜蜂微孢子虫Vairimorpha ceranae（Nosema ceranae）后，中肠蛋白质组的差异，探讨病原与中肠细胞的互作机制。给西方蜜蜂5日龄工蜂自由取食含10⁴个·μL^-1的孢子蔗糖液48 h后，在30℃恒温饲养18 d （23日龄）。结果表明，感染组工蜂存活率仅为34.81%，显著低于对照组的71.85%（P<0.05），感染后工蜂中肠蛋白浓度显著下降（P<0.05），但两组的平均单只日均糖液消耗量没有显著差异（P>0.05）。通过串联质谱标签（TMT）定量蛋白分析系统比较了感染组与对照组工蜂的中肠蛋白质组差异，发现差异显著的蛋白共565个，其中感染中肠显著上调的蛋白有301个，显著下调的蛋白有264个。差异蛋白GO富集分析显示，感染对中肠细胞的细胞进程、能量代谢、细胞形态、蛋白质复合物、分子结合和催化活性等有显著影响；KEGG通路注释发现富集到氨酰-tRNA生物合成、内质网中的蛋白质加工、不同类型的N-聚糖生物合成显著上调，部分抗氧化相关的蛋白显著上调；而显著下调蛋白富集到氨基酸代谢和蔗糖代谢等通路。东方蜜蜂微孢子虫感染后期显著减弱西方蜜蜂工蜂的消化和免疫相关蛋白表达，这些差异显著蛋白为后续开展东方蜜蜂微孢子虫与西方蜜蜂的互作蛋白筛选与验证提供了重要支撑。

关键词: 东方蜜蜂微孢子虫, 蛋白质组, 串联质谱标签, 西方蜜蜂, 中肠

Abstract: This study aims to reveal the interaction mechanism between the pathogen and the midgut cells by analyzing the differential expressed proteome of the midgut after Apis mellifera worker infected with/without Vairimorpha ceranae(Nosema ceranae). The 5-day old workers were fed ad lib with sucrose solution containing 10⁴spore·μL^-1 for 48 h, then raised in the incubator under 30℃ for 18 days(23 days old). The results showed that the survival rate of workers in infected group was only 34.81%, which was significantly lower than that of the control, 71.85% (P<0.05). The midgut protein concentration was significantly decreased either (P<0.05), however, there was no significant difference in the daily consumption of sucrose solution per bee (P>0.05). The comparison of the midgut proteomes of the infected bee and the control by using tandem mass tags (TMT) showed that, 565 proteins were significantly differentially expressed, in which 301 proteins were up-regulated and 264 proteins were down-regulated in the infected midgut. The enrichment of different protein GO functional annotations showed that the cell process, energy metabolism, cell morphology, protein complexes, molecular binding, and catalytic activity in the infected midgut cells were significantly affected. Through the enrichment of the KEGG pathway, it was found that aminoacyl-tRNA biosynthesis, protein processing in the endoplasmic reticulum, and various types of N-glycan biosynthesis were significantly up-regulated, as well as some antioxidant proteins. The proteins enriched in amino acid metabolism and sucrose metabolic pathways were significantly down-regulated. It was revealed that the infection of V.ceranae at late stage significantly compromised the protein expression relatived to the digestion and immunity. These significant differential proteins provides a strong support for the subsequent screening and verification of interaction proteins between V. ceranae and A. mellifera.

Key words: Vairimorpha ceranae(Nosema ceranae), proteome, tandem mass tags, Apis mellifera, midgut

中图分类号:

S895.3

陈礼, 黄伟峰, 黄少康. 东方蜜蜂微孢子虫感染西方蜜蜂工蜂中肠的蛋白质组学分析[J]. 畜牧兽医学报, 2023, 54(4): 1703-1712.

CHEN Li, HUANG Weifone, HUANG Shaokang. Proteomic Analysis of Midgut of Apis mellifera Worker Infected with Vairimorpha ceranae (Nosema ceranae)[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1703-1712.

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