丛枝菌根真菌提高感染青枯菌...和稳定性及有益菌属相对丰度_张嘉慧.pdf

丛枝菌根真菌提高感染青枯菌番茄根际土壤细菌群落丛枝菌根真菌提高感染青枯菌番茄根际土壤细菌群落多样性和稳定性及有益菌属相对丰度多样性和稳定性及有益菌属相对丰度张嘉慧1，邢佳佳1，彭丽媛1，邬奇峰2，陈俊辉1，徐秋芳1，秦华1*（1浙江农林大学环境与资源学院，浙江杭州311300；2杭州市临安区农林技术推广中心，浙江杭州 311300）摘要:【目的目的】青枯病是由茄科雷尔氏菌(Ralstonia solanacearum,亦称青枯菌)诱导产生的一种高温高湿型土传病害，土壤温度高、湿度大时易于青枯菌的繁殖进而引发青枯病。丛枝菌根真菌(arbuscularmycorrhiza,AM)可能通过调控根际微生物区系对病原体产生影响，我们研究了 AM 真菌对青枯菌入侵条件下土壤细菌群落的影响。【方法方法】以番茄(Solanum lycopersicum)为试材进行盆栽试验，供试 AM 真菌为摩西管柄囊霉(Funneliformismosseae)M47V，供试病原菌为茄科雷尔氏菌 QL-RS1115(GenBank：GU390462)。催芽 5 日的番茄种子，接种AM 菌剂的为菌根苗，未接种 AM 真菌的为非菌根苗。在番茄幼苗生长 30 天时，一半菌根苗和非菌根苗接种青枯菌，另一半不接种青枯菌，共 4 个处理。在接种青枯菌后 1 天和 14 天，采集番茄样品，采用抖土方法采集根际土壤，利用实时荧光 PCR 分析番茄根际青枯菌数量，采用 16SrRNA 高通量测序探究土壤细菌群落多样性和结构稳定性。【结果结果】在接种青枯菌初期(1 天)，非菌根苗接种青枯菌(TRAMF)和菌根苗接种青枯菌(TR+AMF)两组处理的根际土壤细菌群落结构发生明显改变，Chao1 指数、Shannon 指数和 Simpson 指数显著降低(P0.05)，共现网络的节点数和连接数明显减少，模块化程度降低，共现网络简化表明细菌群落结构的稳定性降低。接种青枯菌 14 天后，不动杆菌属(Acinetobacter)、鞘氨醇单胞菌属(Sphingomonas)、溶杆菌属(Lysobacter)、假单胞菌属(Pseudomonas)等有益细菌属在感染青枯菌的番茄根际富集，细菌共现网络的节点数和连接数增加，模块化程度提高，表明细菌群落稳定性得到恢复。与非菌根苗相比，菌根苗接种青枯菌(TR+AMF)和菌根苗未接种青枯菌(TN+AMF)两个处理番茄根际土壤中青枯菌丰度显著降低(P0.05)。AM 真菌显著提高 Chao1 指数和 Shannon 指数(P0.05)，提高了感染青枯菌番茄根际土壤中黄杆菌属(Flavobacterium)、黄色土源菌属(Flavisolibacter)、噬胞菌属(Cytophaga)和苔藓杆菌属(Bryobacter)的相对丰度，同时增加了共现网络的节点数和连接数，并促进番茄根际细菌物种之间的良性互作，提高细菌网络的复杂程度。【结论结论】感染青枯菌的番茄根际会富集不动杆菌属(Acinetobacter)、鞘氨醇单胞菌属(Sphingomonas)、溶杆菌属(Lysobacter)、假单胞菌属(Pseudomonas)等有益菌属以提高其抗病性，恢复细菌多样性和群落稳定性。接种 AM 真菌可显著降低番茄根际土壤中青枯菌的丰度，特别是侵染青枯菌后提高番茄根际的黄杆菌属(Flavobacterium)、黄色土源菌属(Flavisolibacter)、噬胞菌属(Cytophaga)和苔藓杆菌属(Bryobacter)的相对丰度，进而抑制土壤中青枯菌的生长，并通过提高细菌的多样性和丰富度，促进番茄根际细菌物种之间的稳定共生和良性互作，从而提高细菌群落对青枯菌的抵抗能力。关键词:根际土壤；青枯菌；丛枝菌根真菌；黄杆菌属；细菌群落多样性；细菌群落稳定性Arbuscular mycorrhizal fungi improves diversity and stability of bacterialcommunity and abundance of beneficial bacteria genus in the rhizosphereof tomato infected with Ralstonia solanacearumZHANGJia-hui1,XINGJia-jia1,PENGLi-yuan1,WUQi-feng2,CHENJun-hui1,XUQiu-fang1,QINHua1*(1 College of Environment and Resources,Zhejiang A&F University,Hangzhou,Zhejiang 311300,China;2 AgroforestryTechnology Extension Centre of Linan District,Hangzhou,Zhejiang 311300,China)植物营养与肥料学报2023,29(1):120131doi:10.11674/zwyf.2022260JournalofPlantNutritionandFertilizershttp:/www.plantnutrifert.org收稿日期：20220511接受日期：20221020基金项目：浙江省重点研发计划项目（2022C02046）。联系方式：张嘉慧E-mail:；*通信作者秦华E-mail:Abstract:【Objectives】Bacterialwiltisasoil-bornediseasewithhightemperatureandhumidity,whichiscausedbyRalstonia solanacearum.Arbuscularmycorrhiza(AM)fungihavethepotentialtoinhibitspecificpathogensinsoilbyregulatingthemicrobialcommunityofthehostrhizosphere.However,theeffectsofAMfungionsoilbacterialcommunityinfectedbyRalstonia solanacearumisstillunclear.【Methods】Potculturemethodwasusedintheresearch,thetestplantwastomato(Solanum lycopersicum),theAMfunguswasF.mosseaeM47V,andthepathogenwasRalstonia solanacearumQL-RS1115(GenBank:GU390462).MycorrhizalseedlingswerepreparedbyinoculatingtomatoseedswithF.mosseaeM47Vafter5daysofgermination,andnon-mycorrhizalseedlingswithoutAMfungusinoculation.Halfofthemycorrhizalandnon-mycorrhizaltomatoseedlingsof30-days-oldwereinoculatedwithpathogen,whiletheotherhalfconsistedofonlyAMtreatments.At1and14daysafterinoculation,tomatoplantsandrhizospheresoilweresampled.Real-timePCRwasusedtoanalyzethenumberofbacterialspeciesintomatorhizosphere,and16SrRNAhigh-throughputsequencingwasusedtoexplorethediversityandstructuralstabilityofsoilbacterialcommunity.【Results】Onthe1stdayofpathogeninoculation,theChao1index,ShannonindexandSimpsonindexofbacterialcommunitystructureinrhizospheresoilsunderthetwoQL-RS1115inoculationtreatmentsreducedsignificantly(P0.05),thenumberofnodesandconnectionsofco-occurrencenetworksignificantlydecreased,andthedegreeofmodularitydecreased.Thesimplifiedco-occurrencenetworkindicatedthatthestabilityofbacterialcommunitystructuredecreased.Onthe14thdayofpathogeninoculation,thebeneficialbacteriasuchasAcinetobacter,Sphingomonas,Lysobacter,andPseudomonaswereenrichedinrhizospheresoil,thenumberofnodesandconnectionsofbacterialco-occurrencenetworkincreased,andthedegreeofmodularityincreased,indicatingthatthestabilityofbacterialcommunitywasrestored.Comparedwithnon-mycorrhizalseedlingstreatments,boththemycorrhizalseedlingswithandwithoutpathogenshowedsignificantlylowerabundanceofpathogeninrhizospheresoil(P0.05).AMfungisignificantlyincreasedtheChao1indexandShannonindex(P0.05),andincreasedtherelativeabundanceofFlavobacterium,Flavisolibacter,Cytophaga,andBryobacterintherhizospheresoiloftomatoinfectedwithpathogen.Atthesametime,thenumberofnodesandconnectionsoftheco-occurrencenetworkwereincreased,andthebenigninteractionbetweenbacterialspeciesintomatorhizospherewaspromoted,andthecomplexityofthebacterialnetworkwasimproved.【Conclusions】TomatoinfectedwithRalstonia solanacearuminducestheenrichmentofbeneficialbacteria,includingAcinetobacter,Sphingomonas,Lysobacter,andPseudomonasintherhizospheretorestorethemicrobialdiversityandstability.InoculationofAMfungisignificantlydecreasestherelativeabundanceofRalstonia solanacearumbutincreasesthatofFlavobacterium,andfurtherimprovesthediversityandstabilityofbacterialco