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三七叶际微生物的分离鉴定及生防功能研究OA北大核心CSTPCD

Isolation and identification of phyllosphere microbiome of Panax notoginseng(Burk.)F.H.Chen and its biocontrol functions

中文摘要英文摘要

[目的]从三七叶际分离筛选对三七叶部黑斑病病原菌具有较强抑制效果的生防菌株,为开发安全高效的生防菌剂提供菌种资源,也为中药材病害的生态防控提供新思路.[方法]采用连续稀释法从1年生健康三七叶片中分离可培养叶际微生物,通过平板对峙法测定分离菌株对三七叶部黑斑病菌人参链格孢菌(Alternaria panax)菌株SL17的拮抗效果,筛选对人参链格孢菌具有生防作用的叶际微生物菌株,结合ITS和16S rDNA测序对活性菌株进行分类鉴定,挑选抑菌效果最佳的菌株进一步进行广谱抑菌能力测定.[结果]从健康三七叶际共分离纯化得到125株分离物,通过平板对峙法从中筛选获得50株对人参链格孢菌具有明显抑制效果的拮抗菌株,其中26株为真菌,24株为细菌.26株拮抗真菌中,2株三七叶内生真菌菌株NZ-14和NZ-23的抑菌效果最佳,其中康氏木霉菌(Trichoderma konin-gii)菌株NZ-14可完全抑制人参链格孢菌菌丝生长,抑制率达100.00%;24株拮抗细菌中,2株三七叶内生细菌菌株NX-20和NX-23的抑菌效果最佳,其中枯草芽孢杆菌(Bacillus subtilis)菌株NX-20对人参链格孢菌的抑制率达71.13%.叶际拮抗菌对三七主要病原菌抑制活性测定结果显示,菌株NZ-14、NZ-23、NX-20和NX-23对槭菌刺孢菌(Mycocentrospora acerina)菌株DMS5、茄腐镰刀菌(Fusarium solani)菌株F3、锈腐菌(Cylindrocarpon destructans)菌株RS006均具有良好的抑制能力,抑制率为39.26%~78.97%.[结论]三七叶际存在大量具有潜在抑菌能力的微生物,其中2株木霉菌菌株NZ-14、NZ-23和2株芽孢杆菌菌株NX-20、NX-23的抑制效果最好,对三七叶部和根部主要病害病原菌均有较强的抑制作用,可作为三七病害生物防控的微生物资源.

[Objective]The purpose of the study was to isolate and screen biocontrol strains from Panax notoginseng(Burk.)F.H.Chen phyllosphere with strong inhibition effect against black spot disease of P.notoginseng leaves,in order to provide microbial resources for the development of safe and efficient biocontrol fungicides,as well as to provide reference for the ecological management of diseases affecting Chinese medicinal materials.[Method]Cultivable phyllosphere micro-biomes were isolated from healthy one-year-old P.notoginseng leaves using serial dilution method.The antagonistic effects of the isolated strains against Alternaria panax strain SL17,a pathogen causing black spot disease on P.notogin-seng leaves,were evaluated using plate confrontation method.Phyllosphere microbiome strains with biocontrol effects against A.panax were screened.The active strains were classified and identified by ITS and 16S rDNA sequencing,and the strains with the best inhibition effects were further selected for the determination of broad-spectrum inhibition ability.[Result]A total of 125 isolates were isolated and purified from healthy P.notoginseng phyllosphere,from which 50 an-tagonistic strains with obvious inhibition effects against A.panax were obtained using plate confrontation method,inclu-ding 26 fungal strains and 24 bacterial strains.Among the 26 antagonistic fungi,two P.notoginseng leaf endophytic fungal strains NZ-14 and NZ-23 showed the best inhibitory effects,of which Trichoderma koningii NZ-14 could completely in-hibit the mycelial growth of A.panax with an inhibition rate of 100.00% .Among the 24 antagonistic bacterial isolates,two strains of P.notoginseng leaf endophytic bacteria NX-20 and NX-23 had the best inhibition effect,with Bacillus subti-lis NX-20 having an inhibition rate of 71.13% against A.panax.The results of the inhibitory activity assays of the phyllo-sphere antagonists against the main fungal pathogens of P.notoginseng showed that the strains NZ-14,NZ-23,NX-20 and NX-23 had good inhibition activity against Mycocentrospora acerina strain DMS5,Fusarium solani strain F3,and Cylindrocarpon destructans strain RS006,with inhibition rates ranging from 39.26% to 78.97% .[Conclusion]There are many microbiomes in the phyllosphere of P.notoginseng with potential inhibition activity,among which the two Tricho-derma strains NZ-14 and NZ-23 and the two Bacillus strains NX-20 and NX-23 have the best inhibition effects and strong inhibitory activity against the main fungal pathogens of P.notoginseng leaves and roots,and can be used as microbial re-sources for biological control of P.notoginseng diseases.

禄蓉;张秀;田光祥;陈朝苍;赵云思;朱书生;杨敏

云南农业大学植物保护学院/农业生物多样性与病虫害控制教育部重点实验室/云南生物资源保护与利用国家重点实验室,云南昆明 650201

植物保护学

三七叶际微生物黑斑病拮抗菌木霉菌芽孢杆菌生物功能

Panax notoginseng(Burk.)F.H.Chenphyllosphere microbiomeblack spotantagonistic bacteriaTrichodermaBacillusbiological function

《南方农业学报》 2024 (004)

1035-1045 / 11

国家自然科学基金项目(32060719);云南省农林联合重点项目(202101BD070001-003);云南省中青年后备人才项目(202005AC160045) National Natural Science Foundation of China(32060719);Yunnan Agriculture and Forestry Joint Key Project(202101BD070001-003);Yunnan Young and Middle-aged Reserve Talent Project(202005AC160045)

10.3969/j.issn.2095-1191.2024.04.012

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