木霉-芽孢杆菌固体共培养条件优化及对豇豆枯萎病防治效果研究

doi:10.16409/j.cnki.2095-039x.2025.02.064

摘要/Abstract

摘要： 由尖孢镰刀菌嗜导管专化型Fusarium oxysporum f.sp.vasinfectum引起的豇豆枯萎病严重影响我国豇豆的安全生产。为了有效防治该病，本研究通过对峙培养和亲和性筛选，获得了两株对豇豆枯萎病菌拮抗效果较好且互相亲和的拮抗菌，经鉴定分别为拟康宁木霉Trichoderma koningiopsis HN-006和贝莱斯芽孢杆菌Bacillus velezensis B-HN-1。在此基础上开展了两株拮抗菌固体共培养条件研究，经单因素和正交试验优化后获得最佳培养条件：共培养基质质量比为大米：稻壳：麦麸（1:1:1），含水量60%，木霉-芽孢杆菌接种质量比1:1.25（木霉2%，芽孢杆菌2.5%），芽孢杆菌延迟接种36h，共培养温度和时间分别为27.5℃和7 d，木霉孢子浓度达8.71×10⁹ CFU/g，芽孢杆菌活菌数达9.07×10¹⁰ CFU/g。通过对豇豆促生和盆栽防效试验发现，共培养物处理组促进了豇豆生长，且对枯萎病防效达（78.95±4.59）%。该研究提供了一种高效的木霉和芽孢杆菌共培养方案，为豇豆枯萎病防治提供了复合微生物制剂。

关键词: 拟康宁木霉, 贝莱斯芽孢杆菌, 共培养, 正交试验

Abstract: Fusarium wilt of cowpea, caused by Fusarium oxysporum, represents a significant constraint to the sustainable production of cowpea in China. To develop effective management strategies, two mutually compatible antagonistic strains exhibiting strong inhibitory activity against the pathogen were isolated using dual-culture assays and affinity-based screening. Molecular identification confirmed these isolates as Trichoderma koningiopsis HN-006 and Bacillus velezensis B-HN-1. Building upon these results, this study further optimized the solid-state co-culture conditions for the two antagonistic strains. Using single-factor experiments and orthogonal array testing, the optimal co-cultivation parameters were determined as follows: a substrate composed of rice, rice husk, and wheat bran in a 1:1:1 ratio, with a moisture content of 60%; an inoculation ratio of Trichoderma to Bacillus of 1:1.25 (corresponding to 2% Trichoderma and 2.5% Bacillus); delayed inoculation of Bacillus 36 hours after Trichoderma; and incubation at 27.5 ℃ for 7 days. Under these conditions, the spore concentration of Trichoderma reached 8.71 ×10⁹ CFU/g, and the viable cell count of Bacillus reached 9.07 ×10¹⁰ CFU/g. Subsequent plant growth promotion and pot experiment assays demonstrated that the co-culture product significantly enhanced cowpea growth and provided a control efficacy against wilt disease of (78.95 ± 4.59)%. This study establishes an efficient co-cultivation protocol for Trichoderma and Bacillus, and proposes a promising composite microbial inoculant for the management of cowpea Fusarium wilt.

Key words: Trichoderma koningiopsis, Bacillus velezensis, co-cultivation, orthogonal test

中图分类号:

S476

庄沈亮, 李坤, 侯巨梅, 刘铜. 木霉-芽孢杆菌固体共培养条件优化及对豇豆枯萎病防治效果研究[J]. 中国生物防治学报, 2025, 41(5): 1141-1153.

ZHUANG Shenliang, LI Kun, HOU Jumei, LIU Tong. Optimization of Solid-State Co-Culture Conditions of Trichoderma-Bacillus and Its Control Efficacy on Cowpea Fusarium Wilt[J]. Chinese Journal of Biological Control, 2025, 41(5): 1141-1153.

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