浙江农业学报 ›› 2017, Vol. 29 ›› Issue (1): 81-88.DOI: 10.3969/j.issn.1004-1524.2017.01.12

• 植物保护 • 上一篇    下一篇

枯草芽孢杆菌STO-12抑菌活性及其抑菌物质分析

黄华毅1, 王佳琳2, 马荣2, 梁英梅3, 田呈明1, *   

  1. 1.北京林业大学 省部共建森林培育与保护教育部重点实验室, 北京 100083;
    2.新疆农业大学 林学与园艺学院, 新疆 乌鲁木齐 830052;
    3.北京林业大学 博物馆, 北京 100083
  • 收稿日期:2016-05-11 出版日期:2017-01-15 发布日期:2017-02-23
  • 通讯作者: 田呈明,E-mail:chengmt@bjfu.edu.cn
  • 作者简介:黄华毅(1989—),男,江西九江人,博士研究生,主要从事微生物与植物病害生物防治方面的研究。E-mail:hhy19890408@163.com
  • 基金资助:
    国家“十二五”科技支撑课题(2012BAD19B08); 国家大学生创新训练计划项目(201510758041)

Antifungal activities of Bacillus subtilis STO-12 and analysis on its antifungal substances

HUANG Huayi1, WANG Jialin2, MA Rong2, LIANG Yingmei3, TIAN Chengming1, *   

  1. 1. Key Laboratory for Silviculture and Conservation of Ministry of Education College of Forestry, Beijing Forestry University, Beijing 100083, China;
    2. College of Forestry and Horticulture, Xinjiang Agricultural University, Urumqi 830052, China;
    3. Museum of Beijing Forestry University, Beijing Forestry University, Beijing 100083, China
  • Received:2016-05-11 Online:2017-01-15 Published:2017-02-23

摘要: 枯草芽孢杆菌(Bacillus subtilis)STO-12是从枣树根际土壤中分离获得的一株拮抗细菌。通过平板对峙法和菌丝生长速率法分别探究了STO-12及其无菌培养滤液的抑菌活性,并分别采用滤纸片法和二分皿法初步分析了其抑菌活性物质,为STO-12的开发利用提供技术指导。结果表明:STO-12及其无菌培养滤液均能有效抑制枣缩果病菌(Alternaria tenuissima)、杨树炭疽病菌(Colletotrichum gloeosporioides)和杨树腐烂病菌(Cytospora chrysosperma)的菌丝生长,其中,无菌培养滤液对这3种病原真菌的抑制率分别达到84.87%(7 d)、38.19%(7 d)和100%(3 d);STO-12分泌的蛋白类和脂肽类物质均表现出了较好的抑菌活性,且都能使生长的菌丝产生膨大畸形,其中,蛋白类物质对枣缩果病菌、杨树炭疽病菌和杨树腐烂病菌的抑菌圈直径分别为12.33、9.50和16.50 mm,脂肽类物质抑制作用形成的抑菌圈直径分别为17.50、17.06和21.50 mm;细胞壁裂解酶特性检测发现,STO-12能产生纤维素酶和蛋白酶;此外,STO-12的挥发性气体也具有一定的抑菌作用,对这3种病原菌的抑制率分别达到38.96%(3 d)、9.35%(3 d)和82.14%(2 d)。以上结果表明,枯草芽孢杆菌STO-12是一株能拮抗多种病原真菌且分泌多种抑菌物质的高效拮抗菌,具有进一步开发应用的前景。

关键词: 枯草芽孢杆菌, 抑菌活性, 培养滤液, 抑菌蛋白, 脂肽

Abstract: Bacillus subtilis STO-12 was isolated from the rhizosphere soil of jujube plants as a biocontrol agent. In order to provide technical guidance for further exploitation and utilization of STO-12, the antifungal activities of STO-12 and its sterile culture filtrates were investigated by dual culture assay and colony diameter assay, respectively; and the antifungal substances of STO-12 were analyzed by paper disc method and two-compartment plate method, respectively. The results revealed that STO-12 and its sterile culture filtrates had strong antifungal activities, and the rates of sterile culture filtrates inhibited Alternaria tenuissima, Colletotrichum gloeosporioides and Cytospora chrysosperma were 84.87% (7 d), 38.19% (7 d) and 100% (3 d), respectively. Crude proteins and lipopeptides extracted from STO-12 exhibited high antifungal activities against Alternaria tenuissima, Colletotrichum gloeosporioides and Cytospora chrysosperma and the hyphae at the edge of inhibition zones developed swelling. For crude proteins, the average diameters of the inhibition zones for Alternaria tenuissima, Colletotrichum gloeosporioides and Cytospora chrysosperma were 12.33, 9.50 and 16.50 mm, respectively. For lipopeptides, the average diameters of the inhibition zones were 17.50, 17.06 and 21.50 mm, respectively. STO-12 could produce cellulase and protease. Moreover, volatiles produced by STO-12 also could inhibit the hyphal growth of Alternaria tenuissima, Colletotrichum gloeosporioides and Cytospora chrysosperma, and the inhibition rates were 38.96% (3 d), 9.35% (3 d) and 82.14% (2 d), respectively. Our results suggested that Bacillus subtilis STO-12 had strong antifungal activity, varied antifungal substances and wide antifungal spectrum, had the potential prospect of industrial development and field application.

Key words: Bacillus subtilis, antifungal activity, culture filtrate, antifungal proteins, lipopeptides

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