Identification And Function Analysis Of DsRNA From Botryosphaeria Dothidea

Botryosphaeria dothidea is an important pathogen that damages the disease of forest fruit and has serious damage.As an important biocontrol factor,fungal virus is expected to be found and applied in this kind of disease pathogen.In this study,19 B.dothidea strains were used as starting strains.Through the extraction,isolation,identification,and function analysis of 13 strains of fungal viruses,the taxonomic status and function of B.dothidea fungal viruses were identified,and the B.dothidea fungal viruses were identified.The system evolution and new ideas for biological control provided the basis.The main findings are as follows:Screening for efficient isolation of dsRNA viruses Using sdau11-122 and sdau11-64 as tested strains,water-saturated phenol was selected as extraction agent and fungal RNA extraction method was used for comparison.The convenience of operation and quality of dsRNA extraction were used as criteria to screen out The fungal RNA extraction method is an efficient method for extracting B.dothidea dsRNA virus.The extraction time of this method is cut in half and the extraction quality is good.Seventeen other strains such as sdau11-65 and sdau11-71 were used for dsRNA extraction according to this method,which validated the high efficiency of the method and found that the virus has diversity.Identify the taxonomic status of dsRNA viruses in some strains of B.dothidea Through the whole-genome sequencing and BLAST comparison,it was found that the RdRp size of the sdau11-122 strain was 1408 bp,and the CP gene size was 1562 bp.By homology comparison,the homology with Botryosphaeria dothidea partitivirus 1 reached 99%.Paritiviridae(Partitivirus),named BdPV2;sdau11-64 has a viral sequence fragment of 5161 bp,which was found to be homologous to Sphaeropsis sapinea RNA virus 1 after comparison.99%,belonging to the genus Victorivirus of the family Totiviridae,was named Botryosphaeria dothidea Victorivirus 2(BdVV2),the fungal virus that was first discovered in Botryosphaeria dothidea.Defined the function of 13 fungal viruses The 13 wild strains were prepared with protoplasts and combined with virazole to obtain detoxified strains.Virus particles were extracted and transfected to obtain regenerated strains.Wild strains,detoxified strains and regenerated strains were tested for colony color,colony growth characteristics,and artificial thorns.Injury vaccination method,the results showed that: the color difference of sdau11-119 and sdau11-65 between the virus-free strains and wild strains showed little difference;other strains had some differences in color;sdau11-122,sdau11-71,sdau12-60 and other strains were wild,There was no difference in the growth rate of detoxified and regenerated mycelium.The pathogenicity measured wild and regenerated strains with large lesions,while the detoxified strains had smaller lesions,indicating that the virus had a promoting effect on pathogenic bacteria;sdau11-64,The growth rate of sdau11-119,sdau11-86,sdau11-80,sdau12-40,etc.were higher in the detoxified strains than in the wild and regenerated mycelium,and the pathogenicity was determined to be smaller in the wild strains and regenerated strains,while the strains were detoxified after detoxification.Larger spots,weaker virulence,weaker pathogenicity,and important biological control value.