杨梅枯萎病拮抗内生菌的分离及其抑菌机制研究

doi:10.15933/j.cnki.1004-3268.2021.04.013

河南农业科学 ›› 2021, Vol. 50 ›› Issue (4): 96-105.DOI: 10.15933/j.cnki.1004-3268.2021.04.013

所属专题：拮抗菌的筛选专题

杨梅枯萎病拮抗内生菌的分离及其抑菌机制研究

曹鹏飞¹，刘青娥²

（1.丽水市农林科学研究院，浙江丽水 323000； 2.丽水学院，浙江丽水 323000）

收稿日期:2020-07-31 出版日期:2021-04-15 发布日期:2021-04-15
通讯作者: 刘青娥(1976-)，女，浙江龙泉人，副教授，主要从事生物资源开发利用研究工作。E-mail:qinger91@126.com
作者简介:曹鹏飞(1978-)，男，陕西旬邑人，高级农艺师，主要从事植物病理学研究工作。E-mail:297473247@qq.com
基金资助:
丽水市科技计划重大研发项目（2020ZDYF01）；丽水市公益性项目（2016GYX15）

Isolation of Antagonistic Endophytes against Myrica rubra Wilt and Their Antagonistic Mechanism

CAO Pengfei¹,LIU Qing’e²

(1.Lishui Institute of Agricultural Science,Lishui 323000,China; 2.Lishui University,Lishui 323000,China)

Received:2020-07-31 Published:2021-04-15 Online:2021-04-15

摘要/Abstract

摘要： 为分离筛选拮抗内生菌用于杨梅枯萎病的防治，以健康的杨梅、鱼腥草、凤尾草和金银花为材料，从中筛选出拮抗优势内生菌株，并通过测定经优势菌株作用后菌体细胞膜、细胞壁、呼吸代谢、RNA含量和拓扑异构酶Ⅰ活性的变化，研究其抑菌机制。结果表明，从4种植物中共分离筛选出杨梅枯萎病拮抗内生菌14株，其中FWB1J 和YMB1J菌株的抑菌效果最好，经鉴定FWB1J为木霉属（Trichoderma）、YMB1J为假丝酵母属（Candida），2株菌株的MIC均为0.400 0 g/mL，EC₅₀分别为0.034 7、0.029 0 g/mL，EC₉₀分别为0.315 5、0.254 6 g/mL。经2株优势内生菌株胞外代谢产物处理后，病原菌的N-乙酰葡萄糖胺和几丁质酶活性均极显著高于对照，在处理36 h时达到最大，其中YMB1J菌株 EC₉₀组的N-乙酰葡萄糖胺含量比对照高176.700 μg/g；胞外可溶性糖、可溶性蛋白渗漏量逐渐增加，在处理10 h时渗漏量达最大值；麦角甾醇含量极显著低于对照组，FWB1J菌株EC₉₀组比对照降低了82.460 μg/g；病原菌RNA含量始终低于对照，处理至60 h时FWB1J菌株EC₉₀组RNA含量最低，较对照低16.450 μg/g；琥珀酸脱氢酶（SDH）和苹果酸脱氢酶（MDH）活性极显著低于对照，处理至72 h时，FWB1J菌株质量浓度EC₉₀处理组2种酶活性较对照分别降低了2.750、318.330 U/mg。综上，FWB1J和YMB1J菌株可破坏杨梅枯萎病病原菌细胞壁完整性，增加细胞膜通透性，抑制核酸合成和呼吸代谢，进而抑制病原菌生长，甚至促进其死亡。

关键词: 杨梅, 枯萎病, 拮抗内生菌, 抑菌活性, 抑菌机制

Abstract: In order to isolate and screen antagonistic endophytes for the control of M.rubra wilt disease,healthy M.rubra, Houttuynia cordata, phoenix and honeysuckle were used as materials to screen the antagonistic endophytic strains.The changes of cell membrane,cell wall,respiratory metabolism,RNA content and topoisomerase Ⅰ activity were determined to study the antagonistic mechanism.The results showed that 14 strains of antagonistic endophytes against M.rubra wilt were isolated from four plants,of which the FWB1J strain and YMB1J strain had the best antibacterial effect.And these two strains were
identified as Trichoderma sp.and Candida sp.respectively. Their MICs were 0.400 0 g/mL,their EC₅₀ were 0.034 7 g/mL and 0.029 0 g/mL respectively,their EC₉₀ were 0.315 5 g/mL and 0.254 6 g/mL,respectively.After being treated with the extracellular metabolites of two dominant endophytic strains,the content of N-acetylglucosamine and chitinase activity of pathogen were significantly higher than those of the control,and reached the maximum at 36 h,among them,the content of N-acetylglucosamine in the EC₉₀ group of YMB1J strain was 176. 700 μg/ g higher than that of the control.The leakage of extracellular soluble sugar and soluble protein increased gradually,and reached the maximum at 10 h.The content of ergosterol was significantly lower than that of the control group,and the EC₉₀ group of FWB1J strain was 82.460 μg/g lower than the control.The content of RNA in pathogen was always lower than that of the control group,and the RNA content of the EC90 group of FWB1J strain was the lowest,which was 16.450 μg/g lower than that of the control group at 60 h.The activities of succinate dehydrogenase and malate dehydrogenase were significantly lower than that of the control group,and the activities of the two enzymes in the EC₉₀group of FWB1J strain were decreased by 2.750 U/mg and 318.330 U/mg at 72 h,respectively.Theses indicated that FWB1J and YMB1J strains could destroy the integrity of cell wall, increase the permeability of cell membrane, inhibit nucleic acid synthesis and respiratory metabolism,and then inhibit the growth of the pathogen and even promote their death.

Key words: Myrica rubra, Wilt disease, Antagonistic endophyte, Antagonistic activiy, Antagonistic mechanism

中图分类号:

S436.67

曹鹏飞, 刘青娥. 杨梅枯萎病拮抗内生菌的分离及其抑菌机制研究[J]. 河南农业科学, 2021, 50(4): 96-105.

CAO Pengfei, LIU Qing’e. Isolation of Antagonistic Endophytes against Myrica rubra Wilt and Their Antagonistic Mechanism[J]. Journal of Henan Agricultural Sciences, 2021, 50(4): 96-105.

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杨梅枯萎病拮抗内生菌的分离及其抑菌机制研究

Isolation of Antagonistic Endophytes against Myrica rubra Wilt and Their Antagonistic Mechanism

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