The Genome And Proteome Of Apis Mellifera Filamentous Virus

Honeybees are important pollinators in natural agroecosystem,playing a vital role in diversity of terrestrial plant and agricultural production.In recent years,the decline of honeybee colonies attracts the attention of scholars.Viruses are major threats for honeybee health,leading to deformed wing,paralysis,deformity and even death.This hinders the development of colony population.Apis mellifera filamentous virus is a large DNA virus?498 kb?with high epidemicity in bee colonies.Acute infectious of this virus causes milk white of hemolymph,depolymerization of fat body,climbing and even death.AmFV is prevalence in Apis mellifera in America,England,France,Switzerland as well as other countries.However,screening on honeybee of Chinese,it was found that AmFV has high prevalence rate?10%-80%?in Apis mellifera,while in the native species,Apis cerana showed lower infection rate?31.58%-85.48%?.This suggested AmFV might have potential harm to apicultural development.However,the genome and virion of this virus has not been fully understood,and there is undiscovered whether AmFV can infect other insects.This restricts the development of theoretical research and preventive treatment.This study investigated existence of AmFV in some pollinating insects and greater wax moth,purified virions by using optimization method,sequenced the genome of AmFV and identified viral proteins by mass spectrum.The experimental results are as follows:1.This paper made a survey on different pollination insects of hymenopteran insects and one honeybee parasite?Galleria mellonella?,found that AmFV was presence in Bombus terrestris,Apis andreniformis,Apis andreniformis in China,and Apis mellifera in Korea,and Galleria mellonella.This indicates AmFV had widely spread in different pollinator species.2.A purification protocol of AmFV was built using sucrose density gradient method,and AmFV microscope pictures were obtained in various conditions.The size of AmFV virion was measured at about 200×450 nm,and the nucleocapsid was about 40×3100 nm,triple folding into a “8” figure.3.This study sequenced the Chinese strain of AmFV,identified key protein using proteomics technique.The results show that Chinese strain of AmFV had 493265 nucleotides with 50.96% in “GC” content,encoding 253 Open reading frame?ORFs?.Bioinformatics analysis showed that AmFV may belonged to new nucleo-cytoplasmic large DNA viruses and was closely related to poxviruses.Moreover,we identified 83 proteins of AmFV,containing 31 proteins from cyst membrane,50 of nucleocapsid.According to the proteomics results,we made prokaryotic expression of seven virus protein?AmFV₀154,AmFV₀101,AmFV₀059,AmFV₀159,AmFV₀024,AmFV₀060,AmFV₀186?,and these laid foundations for antibody preparation and immunohistochemical study.The current study will help to analysis the component of genome and proteomics of AmFV,and to better understand the interreaction between AmFV and host,resulting in revealing virus pathogenic mechanism and protecting honeybees from pathogens.