ganA在路德维希肠杆菌杀松材线虫中的功能解析

doi:10.16409/j.cnki.2095-039x.2022.11.020

中国生物防治学报 ›› 2023, Vol. 39 ›› Issue (5): 1094-1103.DOI: 10.16409/j.cnki.2095-039x.2022.11.020

ganA在路德维希肠杆菌杀松材线虫中的功能解析

王浩宇^2,3, 张弘弢^2,3, 张莹¹, 赵宇^2,3,7, 王传珍⁴, 张丽霞⁵, 王琦⁶, 吴迪^2,3, 于翠芳⁸, 李海勇⁴, 王爽¹, 牛犇^2,3

1. 北京市颐和园管理处, 北京 100091;
2. 东北林业大学林木遗传育种国家重点实验室, 哈尔滨 150040;
3. 东北林业大学生命科学学院, 哈尔滨 150040;
4. 烟台市森林资源监测保护服务中心, 烟台 264000;
5. 中农绿康(北京)生物技术有限公司, 北京 102101;
6. 中国农业大学植物病理学系, 北京 100193;
7. 牡丹江师范学院, 牡丹江 157000;
8. 青州市云驼风景区运行服务中心, 青州 262500

收稿日期:2022-06-22 出版日期:2023-10-08 发布日期:2023-10-25
通讯作者: 王爽,博士,正高级工程师,E-mail:kingnery05@163.com;牛犇,博士,教授,E-mail:ben_niu@nefu.edu.cn。
作者简介:王浩宇,硕士研究生,E-mail:1003810424@qq.com;张弘弢,硕士研究生,E-mail:915618646@qq.com;张莹,工程师,E-mail:820498539@qq.com。
基金资助:
国家重点研发计划（2022YFD1401900）；北京市公园管理中心科技课题（zx2021002）；黑龙江省自然科学基金（YQ2019C002）；国家自然科学基金（32071741）

Functional Analysis of ganA in the Nematocidal of Enterobacter ludwigii against Bursaphelenchus xylophilus

WANG Haoyu^2,3, ZHANG Hongtao^2,3, ZHANG Ying¹, ZHAO Yu^2,3,7, WANG Chuanzhen⁴, ZHANG Lixia⁵, WANG Qi⁶, WU Di^2,3, YU Cuifang⁸, LI Haiyong⁴, WANG Shuang¹, NIU Ben^2,3

1. Administrative Office of the Summer Palace, Beijing 100091, China;
2. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, China;
3. College of Life Science, Northeast Forestry University, Harbin 150040, China;
4. Yantai Service Center of Forest Resources Monitoring and Protection, Yantai 264000, China;
5. Sino Green Agri-Biotech Co., Ltd., Beijing 102101, China;
6. Department of Plant Pathology, China Agricultural University, Beijing 100193, China;
7. Mudanjiang Normal University, Mudanjiang 157000, China;
8. Occurrence Status and Control Strategy of Forestry Pests inYunto Scenic Spot of Qingzhou, Qingzhou 262500, China

Received:2022-06-22 Online:2023-10-08 Published:2023-10-25

摘要/Abstract

摘要： 生防微生物已被用于防控松材线虫病，但其杀线防病的分子机制尚不明确。本研究发现，经路德维希肠杆菌AA4处理后，松材线虫的校正死亡率达85%以上，菌株AA4能够显著减轻松树幼苗上松材线虫病的发生。为了探究AA4抑杀松材线虫的分子机制，本研究采用DNA同源重组技术，构建了菌株AA4 β-半乳糖苷酶基因ganA的敲除突变体，并对ganA在AA4抑杀线虫中的功能进行了解析。研究结果表明，敲除ganA后，突变体菌株抑杀松材线虫与防治松材线虫病的效果均显著降低，菌株的运动性与其在线虫上的定殖能力亦显著减弱。此外，与野生型菌株相比，ganA敲除突变体菌株限制线虫运动的能力显著减弱。因此，ganA可能通过调控AA4定殖松材线虫与限制线虫运动的能力，在AA4抑杀松材线虫中发挥重要功能。

关键词: 基因ganA, 路德维希肠杆菌, 松材线虫, 杀线活性, 定殖

Abstract: Biocontrol microorganisms have been used to control Bursaphelenchus xylophilus disease, but the molecular mechanism of nematicidal is not clear. This study found that the corrected mortality of B. xylophilus treated with E. ludwigii AA4 was more than 85%, and strain AA4 could significantly reduce B. xylophilus infection of pine seedlings. In order to explore the molecular mechanism of AA4 against B. xylophilus, the DNA homologous recombination technique was used to construct the β-galactosidase gene ganA, and the function of ganA in AA4 nematode inhibition was analyzed. The results showed that the B. xylophilus killing activity and B. xylophilus disease prevention in a ganA knockout mutant strain decreased significantly, and the mobility and ability to colonize nematodes of the mutant strain decreased significantly. In addition, compared with the wild-type strain, the ability of the mutant strain to restrict the movement of nematodes was significantly reduced. Therefore, ganA may participate in the process of AA4 killing B. xylophilus by regulating AA4’s ability to colonize B. xylophilus and restrict nematode movement.

Key words: ganA, Enterobacter ludwigii, Bursaphelenchus xylophilus, nematocidal activity, colonization

中图分类号:

S476

王浩宇, 张弘弢, 张莹, 赵宇, 王传珍, 张丽霞, 王琦, 吴迪, 于翠芳, 李海勇, 王爽, 牛犇. ganA在路德维希肠杆菌杀松材线虫中的功能解析[J]. 中国生物防治学报, 2023, 39(5): 1094-1103.

WANG Haoyu, ZHANG Hongtao, ZHANG Ying, ZHAO Yu, WANG Chuanzhen, ZHANG Lixia, WANG Qi, WU Di, YU Cuifang, LI Haiyong, WANG Shuang, NIU Ben. Functional Analysis of ganA in the Nematocidal of Enterobacter ludwigii against Bursaphelenchus xylophilus[J]. Chinese Journal of Biological Control, 2023, 39(5): 1094-1103.

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ganA在路德维希肠杆菌杀松材线虫中的功能解析

Functional Analysis of ganA in the Nematocidal of Enterobacter ludwigii against Bursaphelenchus xylophilus

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