柱花草种子内生拮抗菌的筛选、鉴定及抗炭疽菌作用

doi:10.16409/j.cnki.2095-039x.2025.02.003

摘要/Abstract

摘要： 柱花草是世界热带地区广泛种植的优质豆科牧草，炭疽病是其主要病害。目前，主要采用化学农药防治柱花草炭疽病，尚无有效防治柱花草炭疽病的生物农药。为筛选防治柱花草炭疽病害的拮抗菌，利用热研2号柱花草太空诱变抗病株系种子与柱花草炭疽菌共培养靶向分离获得可培养内生菌株。平板对峙试验发现，10株内生菌株对柱花草炭疽病菌有较强抑制作用，其中ZW21菌株的拮抗作用最强，对柱花草炭疽菌HK04、DZ19和W2菌丝的抑制率分别为70.10%、73.41%和76.86%，对其他4种常见热带植物胶胞炭疽菌的抑菌率也可达70%以上。通过形态学、生理生化和分子生物学方法，鉴定ZW21菌株为蜡样芽胞杆菌Bacillus cereus。扫描电镜观察和胞外酶活试验表明，ZW21具有使病原菌菌丝膨大、扭曲和畸形等拮抗作用，且可通过分泌纤维素酶、淀粉酶和蛋白酶等物质破坏病原菌结构，从而起到抑制病原菌生长的作用。柱花草叶片离体接菌试验表明，ZW21发酵液对W2病菌的相对防效可达89.02%，有潜力用于创制防治柱花草炭疽病的生物农药。

关键词: 柱花草, 炭疽病, 内生细菌, 蜡样芽胞杆菌, 拮抗作用

Abstract: Stylosanthes (stylo) is a leguminous forage widely planted in tropical regions of the world. Anthracnose caused by Colletotrichum is the major disease that greatly limits the growth of stylo. Currently, chemical pesticides are the major approaches to control stylo anthracnose, and there is no effective biological pesticides. In order to screen antagonistic bacteria against Colletotrichum, endophytic strains were isolated by co-culturing the seed extract from stylo resistant line with C. gloeosporioides strain. The plate confrontation test showed that 10 strains could significantly inhibit the mycelial growth of C. gloeosporioides, and the ZW21 strain had the strongest antagonistic effect, with the inhibition rates of 70.10%, 73.41%, and 76.86% on the C. gloeosporioides strains HK04, DZ19, and W2, respectively. In addition, the ZW21 strain could also effectively inhibit the mycelial growth of four different C. gloeosporioides strains isolated from major tropical plants. The scanning electron microscope examination showed that the strain ZW21 caused the swelling, distortion and deformity of mycelia. The extracellular enzyme detection showed that the ZW21 strain could produce cellulase, protease, cellulase, xylanase and etc., destroying the structure of pathogen and limiting its growth. Based on physiological, biochemical and molecular biological analysis, strain ZW21 was identified as Bacillus cereus. The inoculation experiments on the detached stylo leaves showed that the fermentation broth of ZW21 strain had a biocontrol efficiency of 89.02%. These results indicate that ZW21 is expected to be used to create bio-pesticides for the prevention and control of stylo anthracnose.

Key words: Stylosanthes, anthracnose, endophytic bacteria, Bacillus cereus, antagonism

中图分类号:

S476

许云凤, 张吉昌, 罗丽娟, 蒋凌雁. 柱花草种子内生拮抗菌的筛选、鉴定及抗炭疽菌作用[J]. 中国生物防治学报, 2025, 41(1): 132-142.

XU Yunfeng, ZHANG Jichang, LUO Lijuan, JIANG Lingyan. Screening and Identification of Antagonistic Strains from Endophytic Bacteria Isolated from Seeds of Stylosanthes and Exploration of Inhibitory Effects against Colletotrichum gloeosporioides[J]. Chinese Journal of Biological Control, 2025, 41(1): 132-142.

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