草莓胶孢炭疽菌拮抗细菌贝莱斯芽孢杆菌JK3的鉴定及其抗菌活性

doi:10.3969/j.issn.1004-1524.2020.05.11

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (5): 831-839.DOI: 10.3969/j.issn.1004-1524.2020.05.11

草莓胶孢炭疽菌拮抗细菌贝莱斯芽孢杆菌JK3的鉴定及其抗菌活性

冯江鹏^1,2, 邱莉萍³, 梁秀燕^1,2, 陈碧秀^1,2, 夏海洋¹, 彭春龙², 钟永军^1,2,*

1.台州学院生物制药研究所,浙江台州 318000;
2.台州学院生命科学学院,浙江台州 318000;
3.台州市农业科学研究院生物技术研究所,浙江临海 317000

收稿日期:2019-12-10 出版日期:2020-05-25 发布日期:2020-05-29
通讯作者: *,钟永军,E-mail:yjzhong@tzc.edu.cn
作者简介:冯江鹏(1996—),男,甘肃酒泉人,本科生,专业为生物科学。E-mail:320834878@qq.com
基金资助:
浙江省自然科学基金青年基金 (LQ18D050001); 台州市科技计划(1701ny06); 浙江省大学生科技创新活动计划暨新苗人才计划(2019R436011); 2018年台州市211人才工程(第三层次择优); 台州学院校培育基金(2017PY027)

Identification of antagonistic bacteria Bacillus velezensis JK3 against anthracnose of strawberry and its antipathogenic activity

FENG Jiangpeng^1,2, QIU Liping³, LIANG Xiuyan^1,2, CHEN Bixiu^1,2, XIA Haiyang¹, PENG Chunlong², ZHONG Yongjun^1,2,*

1. Institute of Biopharmaceuticals, Taizhou University, Taizhou 318000, China;
2. School of Life Sciences, Taizhou University, Taizhou 318000, China;
3. Biotech Research Institute, Taizhou Academy of Agriculture Sciences, Linhai 317000, China

Received:2019-12-10 Online:2020-05-25 Published:2020-05-29

摘要/Abstract

摘要： 利用平板对峙法从堆肥中分离筛选到1株对草莓胶孢炭疽菌具有较强拮抗作用的细菌株系JK3。通过生理生化特征、16S rDNA和rpoB序列分析,将JK3菌株鉴定为贝莱斯芽孢杆菌。利用离体草莓叶片测定了JK3发酵上清液对草莓炭疽病的防治效果。结果显示:JK3发酵上清液对草莓胶孢炭疽菌引起的草莓炭疽病有明显预防效果,但治疗作用较差;提前24 h或接种胶孢炭疽菌孢子的同时喷施JK3发酵上清液,防治效果超过95%;接种胶孢炭疽菌孢子后24 h喷施发酵上清液,防治效果为11.5%。对JK3发酵上清液进行不同的温度、pH和蛋白酶处理,结果显示,发酵上清中的抑菌物质对pH和蛋白酶处理不敏感,但高温可降低其抑菌活性,推测主要抗菌活性成分不是蛋白质。由于芽孢杆菌多产生脂肽类抗菌物质,利用相关合成基因检测引物进行PCR扩增并测序,结果显示,JK3菌株含有脂肽类抗菌物质生物合成相关基因srfAA、bymB、fenD、bacA和ituC。以上结果表明,贝莱斯芽孢杆菌JK3有开发为防治胶孢炭疽菌引起的草莓炭疽病的生防制剂潜力。

关键词: 草莓炭疽病, 胶孢炭疽菌, 贝莱斯芽孢杆菌, 生物防治, 拮抗活性

Abstract: A bacterial isolate JK3, which was isolated from compost, showed stronger antagonistic activity against Colletotrichum gloeosporioides causing strawberry anthracnose in dual culture assay. The isolate JK3 was identified as Bacillus velezensis by physiological and biochemical characterization, 16S rDNA and rpoB gene sequence analysis. The inhibitory effects of JK3 fermented liquid on C. gloeosporioides were evaluated with the detached strawberry leaves. JK3 fermented liquid showed significant protection on strawberry anthracnose but poor therapeutic effect. More than 95% control efficiency was achieved by spraying JK3 fermented liquid 24 h before inoculation with the spore suspension of C. gloeosporioides or simultaneous inoculation. Only 11.5% control efficiency was resulted by spraying JK3 fermented liquid 24 h after inoculation with the pathogen. The antagonistic activity of the fermented liquid was stable to different pH values and proteases, but sensitive to high temperature. Thus implied that active component was not related to proteins. It has been reported that most isolates of Bacillus spp. produce active lipopeptides. The existences of lipopeptide biosynthetic genes in JK3 were examined by PCR amplification with proper primer pairs and DNA sequencing. Results showed that the JK3 contained srfAA, bymB, fenD, bacA and ituC genes. The above data indicated that B. velezensis JK3 had a potential to be developed as biocontrol agent for controlling strawberry anthracnose caused by C. gloeosporioides.

Key words: strawberry anthracnose, Colletotrichum gloeosporioides, Bacillus velezensis, biocontrol, antagonistic activity

中图分类号:

S476⁺.1

冯江鹏, 邱莉萍, 梁秀燕, 陈碧秀, 夏海洋, 彭春龙, 钟永军. 草莓胶孢炭疽菌拮抗细菌贝莱斯芽孢杆菌JK3的鉴定及其抗菌活性[J]. 浙江农业学报, 2020, 32(5): 831-839.

FENG Jiangpeng, QIU Liping, LIANG Xiuyan, CHEN Bixiu, XIA Haiyang, PENG Chunlong, ZHONG Yongjun. Identification of antagonistic bacteria Bacillus velezensis JK3 against anthracnose of strawberry and its antipathogenic activity[J]. , 2020, 32(5): 831-839.

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草莓胶孢炭疽菌拮抗细菌贝莱斯芽孢杆菌JK3的鉴定及其抗菌活性

Identification of antagonistic bacteria Bacillus velezensis JK3 against anthracnose of strawberry and its antipathogenic activity

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