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

Abstract

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

CLC Number:

S895.3

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.

References 33

[1]	KLEIN A M, VAISSIōRE B E, CANE J H, et al.Importance of pollinators in changing landscapes for world crops[J].Proc Roy Soc B Biol Sci, 2007, 274(1608):303-313.
[2]	MATTHIJS S, DE WAELE V, VANDENBERGE V, et al.Nationwide screening for bee viruses and parasites in Belgian honey bees[J].Viruses, 2020, 12(8):890.
[3]	HRISTOV P, SHUMKOVA R, PALOVA N, et al.Honey bee colony losses:why are honey bees disappearing?[J].Sociobiology, 2021, 68(1):e-5851.
[4]	CHAUZAT M P, JACQUES A, LAURENT M, et al.Risk indicators affecting honeybee colony survival in Europe:One year of surveillance[J].Apidologie, 2016, 47(3):348-378.
[5]	LOURENÇO A P, GUIDUGLI-LAZZARINI K R, DE FREITAS N H A, et al.Immunity and physiological changes in adult honey bees (Apis mellifera) infected with Nosema ceranae:The natural colony environment[J].J Insect Physiol, 2021, 131:104237.
[6]	TOKAREV Y S, HUANG W F, SOLTER L F, et al.A formal redefinition of the genera Nosema and Vairimorpha (Microsporidia:Nosematidae) and reassignment of species based on molecular phylogenetics[J].J Invertebr Pathol, 2020, 169:107279.
[7]	HUANG W F, JIANG J H, CHEN Y W, et al.A Nosema ceranae isolate from the honeybee Apis mellifera[J].Apidologie, 2007, 38(1):30-37.
[8]	GOBLIRSCH M.Nosema ceranae disease of the honey bee (Apis mellifera)[J].Apidologie, 2018, 49(1):131-150.
[9]	FRIES I, FENG F, DA SILVA A, et al.Nosema ceranae n.sp.(Microspora, Nosematidae), morphological and molecular characterization of a microsporidian parasite of the Asian honey bee Apis cerana (Hymenoptera, Apidae)[J].Eur J Protistol, 1996, 32(3):356-365.
[10]	HIGES M, MEANA A, BARTOLOMÉ C, et al.Nosema ceranae (Microsporidia), a controversial 21 st century honey bee pathogen[J].Environ Microbiol Rep, 2013, 5(1):17-29.
[11]	WU Y Q, ZHENG Y F, CHEN Y N, et al.Apis cerana gut microbiota contribute to host health though stimulating host immune system and strengthening host resistance to Nosema ceranae[J].Roy Soc Open Sci, 2020, 7(5):192100.
[12]	PANEK J, PARIS L, RORIZ D, et al.Impact of the microsporidian Nosema ceranae on the gut epithelium renewal of the honeybee, Apis mellifera[J].J Invertebr Pathol, 2018, 159:121-128.
[13]	HUANG S K, YE K T, HUANG W F, et al.Influence of feeding type and Nosema ceranae infection on the gut microbiota of Apis cerana workers[J].mSystems, 2018, 3(6):e00177-18.
[14]	HUANG Q, EVANS J D.Targeting the honey bee gut parasite Nosema ceranae with siRNA positively affects gut bacteria[J].BMC Microbiol, 2020, 20(1):258.
[15]	ZHANG Y K, LU X X, HUANG S K, et al.Nosema ceranae infection enhances Bifidobacterium spp.abundances in the honey bee hindgut[J].Apidologie, 2019, 50(3):353-362.
[16]	PTASZYŃSKA A A, PALEOLOG J, BORSUK G.Nosema ceranae infection promotes proliferation of yeasts in honey bee intestines[J].PLoS One, 2016, 11(10):e0164477.
[17]	XING W H, ZHOU D D, LONG Q, et al.Immune response of eastern honeybee worker to Nosema ceranae infection revealed by transcriptomic investigation[J].Insects, 2021, 12(8):728.
[18]	CHAIMANEE V, CHANTAWANNAKUL P, CHEN Y P, et al.Differential expression of immune genes of adult honey bee (Apis mellifera) after inoculated by Nosema ceranae[J].J Insect Physiol, 2012, 58(8):1090-1095.
[19]	DUSSAUBAT C, BRUNET J L, HIGES M, et al.Gut pathology and responses to the microsporidium Nosema ceranae in the honey bee Apis mellifera[J].PLoS One, 2012, 7(5):e37017.
[20]	HUANG W F, SOLTER L F, YAU P M, et al.Nosema ceranae escapes fumagillin control in honey bees[J].PLoS Pathog, 2013, 9(3):e1003185.
[21]	BADAOUI B, FOUGEROUX A, PETIT F, et al.RNA-sequence analysis of gene expression from honeybees (Apis mellifera) infected with Nosema ceranae[J].PLoS One, 2017, 12(3):e0173438.
[22]	VIDAU C, PANEK J, TEXIER C, et al.Differential proteomic analysis of midguts from Nosema ceranae-infected honeybees reveals manipulation of key host functions[J].J Invertebr Pathol, 2014, 121:89-96.
[23]	RAWLINGS N D, BARRETT A J, THOMAS P D, et al.The MEROPS database of proteolytic enzymes, their substrates and inhibitors in 2017 and a comparison with peptidases in the PANTHER database[J].Nucleic Acids Res, 2018, 46(D1):D624-D632.
[24]	HOUDELET C, SINPOO C, CHANTAPHANWATTANA T, et al.Proteomics of anatomical sections of the gut of Nosema-Infected western honeybee (Apis mellifera) reveals different early responses to Nosema spp. Isolates[J].J Proteome Res, 2021, 20(1):804-817.
[25]	ALAUX C, BRUNET J L, DUSSAUBAT C, et al.Interactions between Nosema microspores and a neonicotinoid weaken honeybees (Apis mellifera)[J].Environ Microbiol, 2010, 12(3):774-782.
[26]	LIU T P.Ultrastructure of the midgut of the worker honey bee Apis mellifera heavily infected with Nosema apis[J].J Invertebr Pathol, 1984, 44(3):282-291.
[27]	NAUG D, GIBBS A.Behavioral changes mediated by hunger in honeybees infected with Nosema ceranae[J].Apidologie, 2009, 40(6):595-599.
[28]	MAYACK C, NAUG D.Energetic stress in the honeybee Apis mellifera from Nosema ceranae infection[J].J Invertebr Pathol, 2009, 100(3):185-188.
[29]	MAYACK C, NAUG D.Parasitic infection leads to decline in hemolymph sugar levels in honeybee foragers[J].J Insect Physiol, 2010, 56(11):1572-1575.
[30]	HUGHES A L.Evolution of the βGRP/GNBP/β-1, 3-glucanase family of insects[J].Immunogenetics, 2012, 64(7):549-558.
[31]	EVANS J D, ARONSTEIN K, CHEN Y P, et al.Immune pathways and defence mechanisms in honey bees Apis mellifera[J].Insect Mol Biol, 2006, 15(5):645-656.
[32]	HRISTOV P, SHUMKOVA R, PALOVA N, et al.Factors associated with honey bee colony losses:a mini-review[J].Vet Sci, 2020, 7(4):166.
[33]	CASTELLI L, BRANCHICCELA B, GARRIDO M, et al.Impact of nutritional stress on honeybee gut microbiota, immunity, and Nosema ceranae infection[J].Microb Ecol, 2020, 80(4):908-919.

Proteomic Analysis of Midgut of Apis mellifera Worker Infected with Vairimorpha ceranae (Nosema ceranae)

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