| 贝莱斯芽孢杆菌E69预防稻瘟病等多种真菌病害的潜力 |
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| 引用本文： | 沙月霞,隋书婷,曾庆超,沈瑞清.贝莱斯芽孢杆菌E69预防稻瘟病等多种真菌病害的潜力[J].中国农业科学,2019,52(11):1908-1917. |
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| 作者姓名： | 沙月霞 隋书婷 曾庆超 沈瑞清 |
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| 作者单位： | 1 宁夏农林科学院植物保护研究所,银川 7500112 中国农业大学植物保护学院,北京100193 |
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| 基金项目： | 宁夏农林科学院科技先导资金项目(NKYJ-16-26);国家重点研发计划(2017YFD0201606);宁夏回族自治区重点研发计划(2018BBF02018-06);宁夏农林科学院全产业链创新示范项目(QCYL-2018-09) |
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| 摘 要： | 目的 明确水稻内生菌贝莱斯芽孢杆菌（Bacillus velezensis）菌株E69对多种植物病原真菌的拮抗作用,尤其是对稻瘟病的生物防治效果,减少化学农药的使用。方法 贝莱斯芽孢杆菌菌株E69和枯草芽孢杆菌（B. subtilis,稻瘟病生物防治最常用微生物）菌株E66分离于水稻叶片内生细菌,采用对峙培养法测试菌株E69和E66及其发酵液、无菌上清液对稻瘟病菌（Magnaporthe oryzae）的拮抗作用,并测试两株内生芽孢杆菌对立枯丝核菌（Rhizoctonia solani）、镰孢菌（Fusarium spp.）、番茄灰霉病菌（Botrytis cinerea）、草莓炭疽病菌（Colletotrichum gloeospoioides）、烟草黑胫病菌（Phytophthora parasitica var. nicotianae）、链格孢菌（Alternaria alternate）、西瓜枯萎病菌（F. oxysporum）等11种植物病原菌的拮抗作用,温室条件下检测对水稻叶瘟的预防效果,田间试验检测对水稻叶瘟和穗颈瘟的预防效果,常规抑菌测试法研究对稻瘟病菌分生孢子萌发和附着胞形成的抑制作用。采用激光共聚焦显微镜观察绿色荧光蛋白标记后的工程菌株E69在水稻茎部的定殖情况。结果 菌株E69和E66对稻瘟病菌菌丝生长具有显著拮抗作用,温室条件下两个菌株对稻瘟病的预防效果分别为83.24%和76.57%,对叶瘟的田间预防效果分别为85.97%和79.76%,对穗颈瘟的田间预防效果分别为69.67%和68.82%,E69对叶瘟的预防效果显著高于75%的三环唑可湿性粉剂,对穗颈瘟的预防效果与三环唑无显著差异。菌株E69和E66对立枯丝核菌、镰孢菌、番茄灰霉病菌、草莓炭疽病菌、烟草黑胫病菌、叶枯病菌、西瓜枯萎病菌等11种植物病原菌有显著拮抗作用,E69的拮抗作用明显高于E66。菌株E69和E66能够强烈抑制稻瘟病菌分生孢子萌发和附着胞的形成,E69发酵液的抑菌效果分别为95.28%和94.16%,无菌上清液的抑菌效果分别为85.36%和84.31%;E66发酵液对分生孢子萌发和附着胞形成的抑菌效果分别为89.15%和87.38%,无菌上清液的抑菌效果分别为79.65%和72.45%。绿色荧光蛋白GFP78标记后的工程菌株E69在水稻茎部具有较好的定殖能力,可以稳定定殖在水稻茎部表皮、薄壁组织和维管束。结论 贝莱斯芽孢杆菌菌株E69是一种潜在的、预防效果明显的生防菌株,具有预防稻瘟病兼防纹枯病等多种真菌病害的应用潜力。
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| 关 键 词： | 贝莱斯芽孢杆菌 稻瘟病 植物病原真菌 生物防治 定殖能力 |
| 收稿时间： | 2019-01-29 |
Biocontrol Potential of Bacillus velezensis Strain E69 Against Rice Blast and Other Fungal Diseases |
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| SHA YueXia,SUI ShuTing,ZENG QingChao,SHEN RuiQing.Biocontrol Potential of Bacillus velezensis Strain E69 Against Rice Blast and Other Fungal Diseases[J].Scientia Agricultura Sinica,2019,52(11):1908-1917. |
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| Authors: | SHA YueXia SUI ShuTing ZENG QingChao SHEN RuiQing |
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| Institution: | 1 Institute of Plant Protection, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 7500112 College of Plant Protection, China Agricultural University, Beijing 100193 |
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| Abstract: | 【Objective】 The objective of this study is to clarify the antagonistic effect of Bacillus velezensis strain E69 isolated from rice endophytic bacteria against various fungal plant pathogens in vitro, especially the control efficacy of B. velezensis E69 on rice blast, and to reduce the use of chemical pesticides.【Method】 B. velezensis E69 and B. subtilis (most commonly used microorganisms in biological control of rice blast) E66 strains were isolated from endophytic bacteria of rice leaf tissue. The antagonistic effect of strains E69 and E66 and their fermented liquid, sterile supernatant against Magnaporthe oryzae was tested by confrontation culture method. Also, the antagonistic effect of strains E69 and E66 against other 11 pathogenic fungi, such as Rhizoctonia solani, Fusarium spp., Botrytis cinerea, Colletotrichum gloeospoioides, Phytophthora parasitica var. nicotianae, Alternaria alternate, F. oxysporum, et al., was determined. The preventive efficacy of E69 and E66 was tested for the control of rice leaf blast under greenhouse condition. The field experiments were conducted to evaluate the preventive efficacy of E69 and E66 against rice leaf blast and neck blast. The inhibitory effect of E69 and E66 against conidial germination and appressorial formation of M. oryzae was assessed in laboratory. The colonization of GFP-marked strain E69 in rice stem was observed by laser scanning confocal microscope.【Result】 Strains E69 and E66 significantly suppressed the mycelia growth rate of M. oryzae P131, the preventive efficacy against rice blast under greenhouse condition was 83.24% and 76.57%, respectively. The preventive efficacy of E69, E66 against rice leaf blast and neck blast in field was 85.97%, 79.76% and 69.67%, 68.82%, respectively. The preventive effect of E69 on leaf blast was significantly higher than that of 75% tricyclozole wettable powder, but there was no significant difference between the preventive efficacy of strain E69 and 75% tricyclazole powder against rice neck blast. E69 and E66 had significant antagonistic effects against 11 plant pathogens, such as R. solani, Fusarium spp., B. cinerea, C. gloeospoioides, P. parasitica var. nicotianae, A. alternate, F. oxysporum, et al. The antagonistic effect of E69 was higher than that of E66. E69 and E66 could strongly inhibit the conidial germination and appressorial formation of M. oryzae P131, the inhibitory effect of E69 fermented liquid was 95.28% and 94.16%, respectively, and the inhibitory effect of E69 sterile supernatant was 85.36% and 84.31%, respectively. The inhibitory effect of E66 fermented liquid was 89.15% and 87.38%, respectively, and the inhibitory effect of E66 sterile supernatant were 79.65% and 72.45%, respectively. The GFP-marked strain E69 showed good colonization ability in rice stem, and could be stably colonized in rice stem epidermis, parenchyma and vascular bundles. 【Conclusion】 B. velezensis strain E69 is a potential biocontrol strain with obvious preventive effects, which has the application potential of preventing rice blast, sheath blight and other fungal diseases. |
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| Keywords: | Bacillus velezensis rice blast plant pathogenic fungi biological control colonization ability |
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