Mycoparasitism of Nematode-Trapping Fungus Monacrosporium ellipsosporumand Its Biochemical Basis

Monacrosporium ellipsosporum, a nematode-trapping fungus, was isolated by baiting with sclerotiaof Sclerotinia sclerotiorum in soil from a tobacco field in Yuxi, Yunnan Province. Colonizationfrequency of the sclerotia by the fungus was 18% in natural soil. Reinoculation tests byplacing surface-sterilized sclerotia on fungal cultures for two weeks and then surface-sterilized again led to 32% sclerotia be infected. Dual culture tests in PDA plates did notgive rise to a suppression zone between the colonies of M. ellipsosporum and its counterpartfungi S. sclerotiorum and Rhizoctonia solani, suggesting there was little or no nutritionalcompetition and absent of antifungal compounds. However, M. ellipsosporum could grow overabsent of S. sclerotiorum and R. solani, and significantly inhibited their growth on agarplates. Scanning electron and light microscopic observations showed that hyphae of M. ellipsosporumgrew along and appressed on hypha of S. sclerotiorum and coiled around hyphae of R. solani.Assays of cell wall-degrading enzymes showed that M. ellipsosporum grew well in chitin agarmedia, with clear transparent hydrolysis zones. Activities of total chitinase, exo-chitinase,β-1, 3-glucanase and protease were 140.2±11.9, 82.9±4.1, 111.2±7.6 and 76.1±4.3 U respect-ively, after incubation for 4 days at 30 ℃ in liquid media containing ground sclerotia of S.sclerotiorum as sole nutrient source. These enzymes might be important in the mycoparasiticactivity of M. ellipsosporum.

Classification of dendrocola nematode-trapping fungi

Pine wilt disease,caused by Bursaphelenchus xylophilus and Bursaphelenchus mucnatus,is a serious quarantined disease.Arboreal nematode-trapping fungi of P inus spp.are effective predators on nematodes and have strong host adaptability.The development of these fungal resources may be an effective way to control pine wood nematodes.We collected 515 samples of pine wilt disease from the areas of Ninghai City(Zhejiang province),Shuangbai County(Yunnan province),and Daxing'anling(Heilongjiang province),China.Through isolation,culture and identification,6 species of nematode-trapping fungi(A rthrobotrys cladodesr,A.oligospora,A.musiformis,A.dendroides,Dactylellina ellipsospora,Monacrosporium thaumasium)were identified for predation against B.xylophilus,and 9 species(Arthrobotrys dactyloides,A.cladodes r A.oligospora A.dendroides,Dactylellina ellipsospora,Dactylella asthenopaga,D.leptospora,Arthrobotrys superba,Monacrosporium drechseri)were identified for predation against B.mucnatus.This study provides information in the classification of arboreal predator nematodes and provides an important basis for future biological control of pine wood nematodes.

The lifestyle transition of Arthrobotrys oligospora is mediated by microRNA-like RNAs

The lifestyle transition of fungi,defined as switching from taking organic material as nutrients to pathogens,is a fundamental phenomenon in nature.However,the mechanisms of such transition remain largely unknown.Here we show microRNA-like RNAs(milRNAs)play a key role in fungal lifestyle transition for the first time.We identified milRNAs by small RNA sequencing in Arthrobotrys oligospora,a known nematode-trapping fungus.Among them,7 highly expressed milRNAs were confirmed by northern-blot analysis.Knocking out two milRNAs significantly decreased A.oligospora’s ability to switch lifestyles.We further identified that two of these milRNAs were associated with argonaute protein QDE-2 by RNA-immunoprecipitation(RIP)analysis.Three of the predicted target genes of milRNAs were found in immunoprecipitation(IP)products of QDE-2.Disruption of argonaute gene qde-2 also led to serious defects in lifestyle transition.Interestingly,knocking out individual milRNAs or qde-2 lead to diverse responses under different conditions,and qde-2 itself may be targeted by the milRNAs.Collectively,it indicates the lifestyle transition of fungi is mediated by milRNAs through RNA interference(RNAi)machinery,revealing the wide existence of miRNAs in fungi kingdom and providing new insights into understanding the adaptation of fungi from scavengers to predators and the mechanisms underlying fungal infections.