A 2-kb Mycovirus Converts a Pathogenic Fungus into a Beneficial Endophyte for Brassica Protection and Yield Enhancement

Mycoviruses are viruses that infect fungi,and hypovirulence-associated mycoviruses have the potential to control fungal diseases.However,it is unclear how mycovirus-mediated hypovirulent strains live and survive in the field,and no mycovirus has been applied for field crop protection.In this study,we found that a previously identified small DNA mycovirus(SsHADV-1)can convert its host,Sclerotinia sclerotiorum,from a typical necrotrophic pathogen to a beneficial endophytic fungus.SsHADV-1 downregulates the expression of key pathogenicity factor genes in S.sclerotiorum during infection.When growing in rapeseed,the SsHADV-1-infected strain DT-8 significantly regulates the expression of rapeseed genes involved in defense,hormone signaling,and circadian rhythm pathways.As a result,plant growth is promoted and disease resistance is enhanced.Field experiments showed that spraying DT-8 at the early flowering stage can reduce the disease severity of rapeseed stem rot by 67.6%and improve yield by 14.9%.Moreover,we discovered that SsHADV-1 could also infect other S.sclerotiorum strains on DT-8-inoculated plants and that DT-8 could be recovered from dead plants.These findings suggest that the mycoviruses may have the ability to shape the origin of endophytism.Our discoveries suggest that mycoviruses may influence the origin of endophytism and may also offer a novel strategy for disease control in which mycovirus-infected strains are used to improve crop health and release mycoviruses into the field.

Active DNA demethylation regulates MAMP-triggered immune priming in Arabidopsis

Plants recognize microbe-associated molecular patterns(MAMPs)to activate immune responses and defense priming to defend against pathogen infections.Transcriptional regulation of gene expression is crucial for plant immunity and is mediated by multiple factors,including DNA methylation.However,it remains unknown whether and how DNA demethylation contributes to immune responses in MAMPtriggered immunity.Here,we report that active DNA demethylation is required for MAMP-triggered immunity to bacterial pathogens.The rdd-2 triple mutant carrying mutations in ROS1,DML2,and DML3 that encode DNA glycosylases,which are key DNA demethylation enzymes,exhibits compromised immune responses triggered by the MAMPs fig22 and elf18.Genome-wide methylome analysis reveals that fig22 induces rapid and specific DNA demethylation in an RDD-dependent manner.The expression levels of salicylic acid signaling-related and phytoalexin biosynthesis-related genes are tightly associated with the fig22-induced promoter demethylation.The compromised accumulation of priming compounds and antimicrobial metabolites ultimately leads to a defense priming defect in the rdd-2 mutant.Our results reveal the critical role of active DNA demethylation in the MAMP-triggered immune response and provide unique insight into the molecular mechanism of fig22-modulated DNA demethylation.

Fusarivirus accessory helicases present an evolutionary link for viruses infecting plants and fungi

A significant number of mycoviruses have been identified that are related to plant viruses,but their evolutionary relationships are largely unexplored.A fusarivirus,Rhizoctonia solani fusarivirus 4(RsFV4),was identified in phytopathogenic fungus Rhizoctonia solani(R.solani)strain XY74 co-infected by an alphaendornavirus.RsFV4 had a genome of 10,833 nt(excluding the poly-A tail),and consisted of four non-overlapping open reading frames(ORFs).ORF1 encodes an 825 aa protein containing a conserved helicase domain(Hel1).ORF3 encodes 1550 aa protein with two conserved domains,namely an RNA-dependent RNA polymerase(RdRp)and another helicase(Hel2).The ORF2 and ORF4 likely encode two hypothetical proteins(520 and 542 aa)with unknown functions.The phylogenetic analysis based on Hel2 and RdRp suggest that RsFV4 was positioned within the fusarivirus group,but formed an independent branch with three previously reported fusariviruses of R.solani.Notably,the Hel1 and its relatives were phylogenetically closer to helicases of potyviruses and hypoviruses than fusariviruses,suggesting fusarivirus Hel1 formed an evolutionary link between these three virus groups.This finding provides evidence of the occurrence of a horizontal gene transfer or recombination event between mycoviruses and plant viruses or between mycoviruses.Our findings are likely to enhance the understanding of virus evolution and diversity.

Enrichment of bacteria involved in the nitrogen cycle and plant growth promotion in soil by sclerotia of rice sheath blight fungus

Rice sheath blight pathogen,Rhizoctonia solani,produces numerous sclerotia to overwinter.As a rich source of nutrients in the soil,sclerotia may lead to the change of soil microbiota.For this purpose,we amended the sclerotia of R.solani in soil and analyzed the changes in bacterial microbiota within the soil at different time points.At the phyla level,Proteobacteria,Acidobacteria,Bacteroidetes,Actinobacteria,Chloroflexi and Firmicutes showed varied abundance in the amended soil samples compared to those in the control.An increased abundance of ammonia-oxidizing bacterium(AOB)Nitrosospira and Nitrite oxidizing bacteria(NOB)i.e.,Nitrospira was observed,where the latter is reportedly involved in the nitrifier denitrification.Moreover,Thiobacillus,Gemmatimonas,Anaeromyxobacter and Geobacter,the vital players in denitrification,N2O reduction and reductive nitrogen transformation,respectively,depicted enhanced abundance in R.solani sclerotia-amended samples.Furthermore,asymbiotic nitrogen-fixing bacteria,notably,Azotobacter as well as Microvirga and Phenylobacterium with nitrogen-fixing potential also enriched in the amended samples compared to the control.Plant growth promoting bacteria,such as Kribbella,Chitinophaga and Flavisolibacter also enriched in the sclerotia-amended soil.As per our knowledge,this study is of its kind where pathogenic fungal sclerotia activated microbes with a potential role in N transformation and provided clues about the ecological functions of R.solani sclerotia on the stimulation of bacterial genera involved in different processes of N-cycle within the soil in the absence of host plants.