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林业科学 ›› 2015, Vol. 51 ›› Issue (8): 67-73.doi: 10.11707/j.1001-7488.20150809

• 论文与研究报告 • 上一篇    下一篇

2种生防菌株对杨树溃疡病原葡萄座腔菌的抑制作用

杨蕾1,2, 周国英1, 梁军1,2,3   

  1. 1. 中南林业科技大学经济林培育与保护教育部重点实验室 长沙 410004;
    2. 中国林业科学研究院森林生态环境与保护研究所 国家林业局森林保护重点实验室 北京 100091;
    3. 南京林业大学南方现代林业协同创新中心 南京 210037
  • 收稿日期:2014-10-30 修回日期:2015-03-16 出版日期:2015-08-25 发布日期:2015-09-10
  • 通讯作者: 梁军
  • 基金资助:

    国家"十二五"科技支撑计划课题"商品林重大病虫害监测预警与防控技术研究示范"(2012BAD19B08);湖南省研究生科研创新项目"杨树溃疡病生防菌研究"(CX2014B328);林业公益性行业科研专项"重大森林病虫灾害防控技术的关键理论基础"(201204501)。

Inhibitory Effects of Two Biocontrol Fungous Strains on Poplar Canker Botryosphaeria dothdea

Yang Lei1,2, Zhou Guoying1, Liang Jun1,2,3   

  1. 1. Key Laboratory of Non-Wood Forest Cultivation and Conservation of Ministry of Education, Central South University of Forestry & Technology Changsha 410004;
    2. Key Laboratory of Forest Protection of State Forestry Administration Research Institute of Forest Ecology, Environment and Protection, CAF Beijing 100091;
    3. Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University Nanjing 210037
  • Received:2014-10-30 Revised:2015-03-16 Online:2015-08-25 Published:2015-09-10

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摘要：

[目的] 从生防菌株产生的拮抗作用酶及生防菌株对病原菌生长的影响2方面研究黄绿木霉(YGF9)和木贼镰刀菌(LX6F2)对杨树溃疡病病菌葡萄座腔菌的抑菌机制,为杨树溃疡病的生物防治提供参考。[方法] 通过生防菌株在几丁质培养基、纤维素培养基、β-l,3-葡聚糖培养基和酪蛋白培养基上的反应和生长情况,判定生防菌株能否产生几丁质酶、纤维素酶、β-1,3-葡聚糖酶和蛋白酶;通过显微镜观察2种生防菌株对病原菌菌丝生长和孢子萌发的影响。[结果] 菌株YGF9和LX6F2代谢过程中均能使β-1,3-葡聚糖培养基中的蓝色消失,说明2种菌株具有产β-1,3-葡聚糖酶的能力;2种菌株可以在酪蛋白培养基上形成水解圈,说明2种菌株具有产蛋白酶的能力;2种菌株在几丁质培养基和纤维素培养基上没有产生水解圈和透明圈,说明2种菌株可能没有产几丁质酶和纤维素酶的能力。菌株LX6F2的菌丝可以与病原菌菌丝平行生长、缠绕或穿插生长在病菌菌丝上,从而使得病原菌菌丝扭曲、变形、异常膨大;菌株YGF9的菌丝使得病原菌菌丝发生折叠、畸形、断裂,从而阻碍病原菌菌丝的生长;2种菌株的发酵产物均可导致杨树溃疡病菌葡萄座腔菌的孢子皱缩畸形、孢子内部结构发生变化,从而使病原菌孢子不能正常萌发;抑菌物质使病原菌芽管变短、扭曲畸形,从而芽管上长出的菌丝不能向前伸展,使孢子失去侵袭能力;生防菌株对孢子萌发的抑制率可达94%以上。[结论] 2种生防菌株可通过自身产生的拮抗作用酶和影响病原菌菌丝生长和孢子萌发来达到对葡萄座腔菌的抑制作用。本试验中未发现2种生防菌株产生几丁质酶和纤维素酶,在后续研究中需用不同的试验方法进行验证。此外,本文仅对生防菌株产生拮抗作用酶进行了定性研究,没有计算其产酶量,若今后需要分离提取这些拮抗作用酶,则需要进一步对生防菌株的产酶量进行定量分析。

关键词: 杨树溃疡病, 黄绿木霉, 木贼镰刀菌, 葡萄座腔菌, 生防菌株, 抑菌效果

Abstract:

[Objective] Poplar canker is one of the major biological disaster of poplar plantations in China. Techniques of biological control to restrain poplar canker are sustainable and effective. Trichoderma aureoviride strain YGF9 and Fusarium equiseti strain LX6F2 are two biocontrol strains which were isolated from poplar tissue and soil, respectively. They may have the potential to prevent from and control poplar canker. In order to explore the mechanism of these strains in controlling disease, biocontrol efficacy of the strain YGF9 and the strain LX6F2 on poplar canker caused by Botryosphaeria dothidea were investigated with its antagonism enzyme and inhibition of pathogen's growth. [Method] Production of chitinase, cellulose enzyme, β-1, 3-glucanase and protease by the two strains was determined through the reaction and growth status of the two biocontrol strains grown on chitin medium, cellulose medium, β-1, 3-glucan culture medium and casein medium. Effects of the two biocontrol strains on the pathogen hyphal growth and spore germination were observed through a microscope. [Result] The results showed that the strains YGF9 and LX6F2 could make β-1, 3 glucanase medium's blue color disappear in metabolic process, indicating that the two strains could produce β-1, 3-dextranase. The two strains had the ability to form hydrolyzed circle on casein medium, suggesting that the two strains had the ability to produce protease; The hydrolyzed circle and transparent circle were not generated by the two strains in chitin medium and cellulose hydrolysis medium, inferring that the two strains were not able to produce chitinase and cellulase. The strain LX6F2 hypha grew parallelly to pathogen hyphae, winded or interspersed with pathogen hyphae, so as to make the pathogen hyphae distortion, deformation and enlargement abnormal. The strain YGF9 hypha made pathogen hyphae folding, deformity, fracture, thereby preventing pathogen hyphal growth. The fermentation products of the two biocontrol strains could lead to spores of poplar canker pathogen B. dothidea deformity, and change in the spore internal structure, thus the pathogen spores couldn't germinated normally. The antifungous substances could make germ tubes shorter and twist deformity, so that germ tubes couldn't stretch forward. The inhibition rate of the biocontrol strains to pathogenic spore germination could reach to more than 94%. [Conclusion] It was found that the two biocontrol strains could produce antagonism enzymes and inhibit pathogen's growth, which can interpret why the strain YGF9 and strain LX6F2 could prevent from poplar canker. Chitinase and cellulase produced by the two biocontrol strains were not found in the present experiment, for which, the further study is required by using different experimental methods for validation. In addition, the present study only studied the ability of the two biocontrol strains producing antagonism enzyme in qualitative analysis, the amount of enzyme production, the separation and application of these antagonism enzymes, as well as quantitative analysis of the amount should be studied later.

Key words: poplar canker, Trichoderma aureoviride, Fusarium equiseti, Botryosphaeria dothidea, biocontrol strains, inhibitory effect

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