枝孢瓶霉属合成菌群对生姜青枯病的生物防治

Biocontrol effects of Cladophialophora spp. synthetic fungus resistant to ginger bacterial wilt

  • 摘要: 【目的】 评价4株枝孢瓶霉属(Cladophialophora spp.)深色有隔内生真菌(DSE)菌株的亲缘关系及其合成菌群对生姜青枯病的防治效果,为生姜青枯病生防菌剂的研制与应用提供科学依据。【方法】 以4株枝孢瓶霉属菌株LC3、LJ1、MS2和HX2组成的合成菌群(T4)为材料,通过ITS序列构建的系统发育进化树明确4株供试菌株间的亲缘关系;通过盆栽试验检测T4对生姜青枯病的防治效果;通过大田试验,对比化学药剂单独使用(CK)、生防菌与化学药剂配合使用(处理1)以及T4与化学药剂配合使用(处理2)3种处理对生姜抗病指标、生长指标和产量的影响;通过对应试剂盒的检测方法,测定T4对生姜防御酶活性的影响。【结果】 4株枝孢瓶霉属菌株在基于ITS序列构建的系统发育进化树上形成3个相互独立的进化分枝,其中菌株LC3与已知种Cladophialophora immunda的模式菌株聚在一起;菌株HX2单独形成一个进化枝,为新发表的新种Cladophialophora guangxiense;菌株LJ1和MS2聚在一起形成另一个独立的进化枝,鉴定为疑似新种Cladophialophora sp. nov. 1。盆栽试验结果显示,T4菌液处理的生姜苗平均株高较灭菌水浸根对照(CK)处理极显著提高58.5%(P<0.01);接种病原菌100 d时对生姜青枯病的防效为51.5%,青枯病发病率较CK降低22.2%。大田试验结果显示,处理2的青枯病发病率和病情指数分别较CK降低25.0%和68.6%,分蘖数、株幅和根状块茎长度分别增加50.4%、12.8%和36.4%,生物学产量和经济学产量分别增加50.0%和55.6%。在病原胁迫下,与CK相比,T4处理的生姜地上茎的过氧化物酶(POD)、超氧化物歧化酶(SOD)、多酚氧化酶(PPO)、过氧化氢酶(CAT)、几丁质酶和β-1,3-葡聚糖酶活性分别提高31.2%、25.1%、10.9%、7.1%、12.2%和32.1%。【结论】 枝孢瓶霉合成菌群T4对生姜青枯病具有良好的防病和治疗修复作用。枝孢瓶霉合成菌群T4可与杀菌剂配合使用,在生姜青枯病的生物防治方面具有良好的开发利用前景。

     

    Abstract: 【Objective】 To evaluate the molecular phylogeny of 4 Cladophialophora spp. dark septate endophyte( DSE)strains and the effects of their synthetic microbial community( T4) on preventing ginger bacterial wilt, which could provide theoretical basis for the exploitation and utilization of ginger bacterial wilt biocontrol agents.【Method】The synthetic microbial community(T4) composed of 4 strains of Cladophialophora spp.were used as the test material,and the rela‐tionship of the 4 tested strains was determined by phylogenetic tree based on ITS sequence.The effect of T4 on the control of ginger bacterial wilt was tested by pot experiment.Field tests were conducted to compare the effects of chemical agent alone(CK),biocontrol agent combined with chemical agent(treatment 1) and T4 combined with chemical agent(treat‐ment 2) on resistance indexes,growth indexes and yield of ginger.Finally,using the corresponding assay kit method,the effects of T4 on defense enzyme activity in ginger were determined. 【Result】 The 4 strains of Cladophialophora spp.formed 3 independent evolutionary clades on phylogenetic tree based on ITS sequences.Strain LC3 and the type strain of the known species Cladophialophora immunda were clustered together,strain HX2 formed a single clade,which was a newly published new species Cladophialophora guangxiense;strains LJ and MS2 were clustered together to form another independent clade,which were identified as a suspected new species Cladophialophora sp.nov.1.In potted experiment,the average plant height of ginger seedlings treated with T4 was extremely significantly increased by 58.5%compared with the Sterile water root dipping control(CK)(P<0.01,the same below).The control effect on ginger bacterial wilt was 51.5%after 100 d inoculated with pathogens,and the incidence rate decreased by 22.2%compared with CK.In the field test,compared with CK,the incidence rate and disease index of ginger bacterial wilt treated with treatment 2 were reduced by 25.0%and 68.6%respectively;the number of tillers,plant width and root tuber length increased by 50.4%,12.8%and 36.4%respectively;and the biological yield and economic yield increased by 50.0%and 55.6%respectively.Under pathogen stress,compared with the CK,peroxidase(POD),superoxide dismutase(SOD),polyphenol oxidase(PPO),catalase (CAT),chitinase and β-1,3-glucanase activities in ginger stalk increased by 31.2%,25.1%,10.9%,7.1%,12.2%and 32.1%respectively. 【Conclusion】 The synthetic microbial community T4 of Cladophialophora spp.has good effect on prevention and treatment of ginger bacterial wilt.The synthetic microbial community T4 of Cladophia‐lophora spp.can be used in combination with fungicides,which has a good development and utilization prospect in the biological control of ginger bacterial wilt.

     

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