| Nasonia is the second genus of Hymenoptera to have been subjected to whole-genome sequencing,assembly,and annotation,after Apis mellifera.Nasonia vitripennis(Hymenoptera: Pteromalidae),is becoming a model organism for studying the developmental and evolutionary genetics,as well as for studying the interactions between insect hosts and symbionts owing to its overall ease of laboratory use,short generation time,tolerance for inbreeding,and straightforward rearing.Wolbachia,an intracellular Gram-negative bacterium,is the most common endosymbiont in insects and can affect the host's reproductive modifications,fitness,etc.The reproductive effect of Wolbachia on N.vitripennis is just CI(cytoplasmic incompatibility).Insects rely on the innate immune system to mount defense responses against pathogenic invasions.Transinfection of bacteria or fungi could substantially alter the expression of host immune system components.However,few studies have focused on the effects of native Wolbachia infection.We studied the proteomics of N.vitripennis from three native Wolbachia–infected and three antibiotic-treated Wolbachia-free samples using isobaric tags for relative and absolute quantification-liquid chromatography tandem mass spectrometry,accompanying with two-sex life table.Major research result as follows:1.The impact of native infection with Wolbachia on the life characteristics of N.vitripennis was collected through the two-sex life table.The results showed that native infection of Wolbachia can significantly affect the developmental duration,longevity,and fecundity,while no significant effect on the intrinsic rate of growth,limit growth rate,and net reproductive rate.2.iTRAQ proteomics sequencing technology was used to sequence the proteomics of the N.vitripennis.A total of 3109 proteins were identified from 22946 unique peptides based on the N.vitripennis database with peptide FDR?0.01,and 3096 proteins were quantified in all six libraries,including three Wolbachia-infected and three Wolbachia-free N.vitripennis libraries.All 3096 proteins were annotated using Blast2 GO.The proteins were identified associated with a wide range of biological process(BP),molecular function(MF)and cellular component(CC).2578 proteins were annotated by 9297 GO terms.Notably,there were 13 proteins participated in immune system process and 15 proteins involved in reproductive process.None of them were significantly differently expressed.3.There were only 23 proteins that were significantly differentially expressed between Wolbachia-infected and Wolbachia-free N.vitripennis based on the proteomic analysis,including 15 upregulated and 8 downregulated proteins.Function annotation of these proteins revealed that all proteins involved in catalytic activity and binding in the MF category,and classification based on BP showed that the majority of proteins were associated with metabolic process.The results of qPCR showed that except NV11606 and SP6,other genes encoding significant differentially expressed proteins(NV11923,NV11878,NV18383,NV16308 and NV21224-PA)and genes for immune-related or reproductive-related proteins(NV16743,NV10430,NV15633,NV13603,NV12583 and NV10830)showed consistent changes in the proteome.The result of interaction relationship among 23 differently expressed proteins,13 proteins participated in immune system process and 15 proteins involved in reproductive process displayed that differentially expressed proteins can't indirectly influence immune-related proteins or reproductive proteins by interaction.But there are complex interactions between several immune-related proteins or reproductive proteins.4.To explore the pathways that the quantified proteins involved,all 3096 proteins were analyzed by using KEGG pathway database.The result showed that 1975 proteins were assigned to 1962 KEGG orthology(KO)and involved in 337 maps.Fatty acid metabolism involves a total of 16 pathways.The proteins involved in the glycerophospholipid metabolism pathway,glycerolipid metabolism,and fatty acid degradation were relatively highly expressed.There were no differences in protein expression between carnivorous N.vitripennis and D.melanogaster,Aedes aegypti,Solenopsis invicta,and Apis mellifera.In this experiment,the effects of native infection of Wolbachia on N.vitripennis were analyzed comprehensively from the characteristics of life and proteome,which provided a theoretical basis for the mutual adaptability of host and symbiotic organisms during native symbiosis between insect host and endosymbiotic bacteria. |
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