设施番茄根围土样中木霉菌多样性及功能活性分析
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摘要
为探索设施栽培条件下蔬菜根围土壤中木霉种群多样性特征及其抗常见植物病原真菌及有机磷农药降解活性,从连续20多年种植番茄的设施大棚根围土样中以及番茄根系内部分离木霉菌,并以tef1-α序列和系统发育分析进行种类鉴定,估算各菌株的相对种群密度;对其生防和农残降解活性进行了测定。结果共分离不同木霉菌株27株,结合形态学特征和tef1-α序列分析,鉴定出分属6个不同的木霉种,即绿色木霉(Trichoderma virens)、深褐木霉(T.atrobrunneum)、西蒙斯木霉(T.simmonsii)、深绿木霉(T.atroviride)、厚木霉(T.crassum)和1个疑似木霉新种(T.sp.nov.);木霉各菌株产孢簇常呈鲜绿色到暗绿色,分布不均匀,常形成同心圆结构。对峙条件下这些木霉对土壤中常见土传病原菌立枯丝核菌(Rhizoctonia solani)、大丽轮枝菌(Verticillium dahliae)、尖孢镰孢菌(Fusarium oxysporum)均有普遍的不同程度的抑菌活性;除深褐木霉两个菌株对毒死蜱的降解效果不明显外,其它种类的木霉均有菌株能有效降解毒死蜱,木霉菌株在根围土壤中的相对种群密度仅为104 cfu/g,且菌株间差异较大。研究结果表明设施农业土壤中木霉菌具有丰富的种质资源多样性,在形态特征及功能活性方面表现明显的菌株差异;木霉对3种常见植物病原真菌有普遍的抑菌作用,少数菌株对常用有机磷农药降解作用明显,可以发挥生防和农化残留降解方面的协同效应。该结果对于研究设施农业土壤中木霉的多样性,指导功能菌株筛选及应用,更好挖掘利用木霉菌资源有着较好的借鉴意义。
This work aimed to explore the diversity of Trichoderma from the rhizosphere soil of vegetables under facility culture aswell as their aniti-phytopathogenic fungi and organophosphorus-degrading activity. Trichoderma were isolated from the rhizosphere soil andthe roots of the sampled tomato from facility culture fields for more than 20 years of cultivation,and were identified by tef1-α sequencingcombined with phylogenetic analysis. Besides,the relative population density of each strain was estimated,and the biocontrol activity and thedegradation activity were further studied. Results showed that 27 different strains were isolated,and then identified based on the morphologicalcharacteristics and tef1-α,they belonged to T. virens,T. atrobrunneum,T. simmonsii,T. atroviride,T. crassum and one suspected newspecies T. sp. nov. Conidiation of Trichoderma strains usually was distributed unevenly,more or less concentric,and bright green to dull green.All the strains showed varied levels of inhibiting activity to several plant pathogenic fungi such as Rhizoctoma solani,Fusarium oxysporum andVerticillium dahlia. In the degradation experiment of chlorpyrifos,except strains belonging to T. atrobrunneum,strains in other Trichodermaspecies effectively degraded chlorpyrifos. The relative population density of Trichoderma in the rhizosphere soil was 104 cfu/g,and there wassignificant difference among different strains. The results indicate that there is rich Trichoderma germplasm diversity in the soil under facilityculture,and they show obvious strain difference in morphological characteristics and functional activities. Trichoderma strains have generalinhibiting activity to three plant pathogenic fungi,and some of them can effectively degrade organophosphorus pesticide. Thus,they play asynergistic effect of biocontrol and biodegradation of pesticides and fertilizers. The results of this paper are of great significance for the study ofthe adaptability of Trichodema in the soil of facility agriculture and the exploitation and utilization of Trichoderma resources.
This work aimed to explore the diversity of Trichoderma from the rhizosphere soil of vegetables under facility culture aswell as their aniti-phytopathogenic fungi and organophosphorus-degrading activity. Trichoderma were isolated from the rhizosphere soil andthe roots of the sampled tomato from facility culture fields for more than 20 years of cultivation,and were identified by tef1-α sequencingcombined with phylogenetic analysis. Besides,the relative population density of each strain was estimated,and the biocontrol activity and thedegradation activity were further studied. Results showed that 27 different strains were isolated,and then identified based on the morphologicalcharacteristics and tef1-α,they belonged to T. virens,T. atrobrunneum,T. simmonsii,T. atroviride,T. crassum and one suspected newspecies T. sp. nov. Conidiation of Trichoderma strains usually was distributed unevenly,more or less concentric,and bright green to dull green.All the strains showed varied levels of inhibiting activity to several plant pathogenic fungi such as Rhizoctoma solani,Fusarium oxysporum andVerticillium dahlia. In the degradation experiment of chlorpyrifos,except strains belonging to T. atrobrunneum,strains in other Trichodermaspecies effectively degraded chlorpyrifos. The relative population density of Trichoderma in the rhizosphere soil was 104 cfu/g,and there wassignificant difference among different strains. The results indicate that there is rich Trichoderma germplasm diversity in the soil under facilityculture,and they show obvious strain difference in morphological characteristics and functional activities. Trichoderma strains have generalinhibiting activity to three plant pathogenic fungi,and some of them can effectively degrade organophosphorus pesticide. Thus,they play asynergistic effect of biocontrol and biodegradation of pesticides and fertilizers. The results of this paper are of great significance for the study ofthe adaptability of Trichodema in the soil of facility agriculture and the exploitation and utilization of Trichoderma resources.
引文
[1]杨淏然,马兆红,司智霞.山东寿光设施蔬菜土壤修复对策与实例[J].中国蔬菜,2016,6:1-5.
[2]Yang L,Zhao YH,Zhang BX,et al.Isoation and characterizationof a chlorpyrifos and 3,5,6-trichloro-2-pyridinol degradingbacterium[J].FEMS Microbiol Lett,2005,251:67-73.
[3]Tripathi P,Singh PC,Mishra A,et al.Trichoderma:a potentialbioremediator for environmental clean up[J].Clean TechnEnviron Policy,2013,15:541-550.
[4]张广志,杨合同,等.毒死蜱降解木霉菌对几种重要植物病原真菌的生防活性[J].菌物学报,2014,33(6):1292-1301.
[5]张广志,张新建,李红梅,等.土壤中木霉的分离及其对毒死蜱降解特性研究[J].生物技术通报,2016,32(6):205-210.
[6]陈捷.木霉菌生物学与应用研究—回顾与展望[J].菌物学报,2014,33(6):1129-1135.
[7]Martin JP.Use of acid,rose gengal and streptomycin in the platemethod for estimating soil fungi[J].Soil Science,1950,69:215-232.
[8]Jaklitsch WM,Samuels GJ,Dodd SL,et al.Hypocrea rufa/Trichoderma viride:a reassessment,and description of five closelyrelated species with and without warted conidia[J].Studies inMycology.2006,56:135-177.
[9]Bissett J.A revision of the genus Trichoderma.III.Infragenericclassification[J].Can J Bot,1991,69:2372-2417.
[10]Park MS,Oh SY,et al.Trichoderma songyi sp.nov.,a new speciesassociated with the pine mushroom(Tricholoma matsutake)[J].Antonie Van Leeuwenhoek.2014,106:593-603.
[11]Chaverri P,Gazis RO,Samuels GJ.Trichoderma amazonicum,anew endophytic species on Hevea brasiliensis and H.guianensisfrom the Amazon basin[J].Mycologia,2011,103:139-151.
[12]Kumar S,Stecher G,Tamura K.MEGA7:Molecular evolutionarygenetics analysis version 7.0 for bigger datasets[J].MolecularBiology and Evolution,2016,33:1870-1874.
[13]Lan WS,Cong J,Jiang H,et al.Expression and characterization ofcarboxylesterase E4 gene from peachpotato aphid(Myzus persicae)for degradation of carbaryl and malathion[J].Biotechnol Letters,2005,27:1141-1146.
[14]石利利,林玉锁,徐亦钢,等.高尔夫球场土壤和水中毒死蜱农药残留的测定[J].农业生态环境,2000,16(3):35-38.
[15]Bissett J,Gams W,Jaklitsch W,et al.Accepted Trichoderma namesin the year 2015[J].IMA fungus,2015,6(2):263-295.
[16]Qin WT,Zhuang WY.Four new species of Trichoderma withhyaline ascospores in the Brevicompactum and Longibrachiatumclades[J].Mycosystema,2016,35(11):1317-1336.
[17]Qin WT,Zhuang WY.Seven wood-inhabiting new species of thegenus Trichoderma(Fungi,Ascomycota)in Viride clade[J].Scientific Reports,2016,6:27074.doi:10.1038/srep27074.
[18]Qin WT,Zhuang WY.Seven new species of Trichoderma(Hypocreales)in the Harzianum and Strictipile Clades[J].Phytotaxa,2017,305(3):121-139.
[19]Chen K,Zhuang WY.Three new soil-inhabiting species ofTrichoderma in the stromaticum clade with test of their antagonismto pathogens[J].Curr Microbiol,2017,74:1049-1060.
[20]Jaklitsch WM,Voglmayr H.Biodiversity of Trichoderma(Hypocreaceae)in Southern Europe and Macaronesia[J].Studies in Mycology,2015,80:1-87.
[2]Yang L,Zhao YH,Zhang BX,et al.Isoation and characterizationof a chlorpyrifos and 3,5,6-trichloro-2-pyridinol degradingbacterium[J].FEMS Microbiol Lett,2005,251:67-73.
[3]Tripathi P,Singh PC,Mishra A,et al.Trichoderma:a potentialbioremediator for environmental clean up[J].Clean TechnEnviron Policy,2013,15:541-550.
[4]张广志,杨合同,等.毒死蜱降解木霉菌对几种重要植物病原真菌的生防活性[J].菌物学报,2014,33(6):1292-1301.
[5]张广志,张新建,李红梅,等.土壤中木霉的分离及其对毒死蜱降解特性研究[J].生物技术通报,2016,32(6):205-210.
[6]陈捷.木霉菌生物学与应用研究—回顾与展望[J].菌物学报,2014,33(6):1129-1135.
[7]Martin JP.Use of acid,rose gengal and streptomycin in the platemethod for estimating soil fungi[J].Soil Science,1950,69:215-232.
[8]Jaklitsch WM,Samuels GJ,Dodd SL,et al.Hypocrea rufa/Trichoderma viride:a reassessment,and description of five closelyrelated species with and without warted conidia[J].Studies inMycology.2006,56:135-177.
[9]Bissett J.A revision of the genus Trichoderma.III.Infragenericclassification[J].Can J Bot,1991,69:2372-2417.
[10]Park MS,Oh SY,et al.Trichoderma songyi sp.nov.,a new speciesassociated with the pine mushroom(Tricholoma matsutake)[J].Antonie Van Leeuwenhoek.2014,106:593-603.
[11]Chaverri P,Gazis RO,Samuels GJ.Trichoderma amazonicum,anew endophytic species on Hevea brasiliensis and H.guianensisfrom the Amazon basin[J].Mycologia,2011,103:139-151.
[12]Kumar S,Stecher G,Tamura K.MEGA7:Molecular evolutionarygenetics analysis version 7.0 for bigger datasets[J].MolecularBiology and Evolution,2016,33:1870-1874.
[13]Lan WS,Cong J,Jiang H,et al.Expression and characterization ofcarboxylesterase E4 gene from peachpotato aphid(Myzus persicae)for degradation of carbaryl and malathion[J].Biotechnol Letters,2005,27:1141-1146.
[14]石利利,林玉锁,徐亦钢,等.高尔夫球场土壤和水中毒死蜱农药残留的测定[J].农业生态环境,2000,16(3):35-38.
[15]Bissett J,Gams W,Jaklitsch W,et al.Accepted Trichoderma namesin the year 2015[J].IMA fungus,2015,6(2):263-295.
[16]Qin WT,Zhuang WY.Four new species of Trichoderma withhyaline ascospores in the Brevicompactum and Longibrachiatumclades[J].Mycosystema,2016,35(11):1317-1336.
[17]Qin WT,Zhuang WY.Seven wood-inhabiting new species of thegenus Trichoderma(Fungi,Ascomycota)in Viride clade[J].Scientific Reports,2016,6:27074.doi:10.1038/srep27074.
[18]Qin WT,Zhuang WY.Seven new species of Trichoderma(Hypocreales)in the Harzianum and Strictipile Clades[J].Phytotaxa,2017,305(3):121-139.
[19]Chen K,Zhuang WY.Three new soil-inhabiting species ofTrichoderma in the stromaticum clade with test of their antagonismto pathogens[J].Curr Microbiol,2017,74:1049-1060.
[20]Jaklitsch WM,Voglmayr H.Biodiversity of Trichoderma(Hypocreaceae)in Southern Europe and Macaronesia[J].Studies in Mycology,2015,80:1-87.