Trichoderma harziensis Mixed with Different Microbial Agents for Cercospora Leaf Spot: Determination of Combined Virulence and Field Control Efficacy

doi:10.11924/j.issn.1000-6850.casb2025-0378

Abstract

Abstract:

In response to the severe damage caused by Cercospora leaf spot (Cercospora beticola) on sugar beets in Inner Mongolia, and the issues of chemical control leading to resistance and a lack of highly effective compound microbial agents for biological control, this study aimed to develop green control technology for this disease. Using Cercospora beticola as the target pathogen, the individual toxicity of five microbial agents was determined via the mycelial growth rate method. Promising agents were selected and combined with Trichoderma harzianum in different ratios. The optimal combination ratio was screened through joint toxicity tests (Co-toxicity coefficient method, CTC), followed by field efficacy trials. The results showed that: (1) the order of individual toxicity was Trichoderma harzianum (EC₅₀=0.10 μg/mL)> Paenibacillus polymyxa (EC₅₀=0.90 μg/mL) > Bacillus mucilaginosus > Bacillus amyloliquefaciens > Bacillus subtilis. (2) Combinations of T. harzianum with the other four agents all showed synergistic effects. Among them, the mixture of 5×10¹⁰ cfu/g P. polymyxa and 1×10¹⁰ cfu/g T. harzianum at a 6:4 ratio exhibited significant synergy, with an EC₅₀ value of 0.16 μg/mL and a co-toxicity coefficient of 139.35. (3) The field efficacy of this optimal combination reached 60.94%, significantly higher than that of individual agents (38.62%-50.64%). In conclusion, the compound agent of 5×10¹⁰ cfu/g P. polymyxa and 1×10¹⁰ cfu/g T. harzianum at a 6:4 ratio demonstrated excellent control efficacy against Cercospora leaf spot and can be recommended as a preferred option for its biological control. Future research should focus on elucidating the synergistic mechanisms of compound agents, optimizing formulation and application techniques to provide more robust technical support for the green control of Cercospora leaf spot in sugar beets.

Key words: Cercospora beticola, Trichoderma harzianum, microbial agent, virulence measurement, field control efficacy

GAO Bo, HAO Yongli, LI Zelin, DIAO Jiamei, LIU Qingpeng, ZHANG Wei. Trichoderma harziensis Mixed with Different Microbial Agents for Cercospora Leaf Spot: Determination of Combined Virulence and Field Control Efficacy[J]. Chinese Agricultural Science Bulletin, 2026, 42(3): 171-176.

Figures/Tables 6

References 30

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微生物菌剂名称	培养基系列浓度/(cfu/mL)
微生物菌剂名称	Ⅰ	Ⅱ	Ⅲ	Ⅳ	Ⅴ
5×10¹⁰cfu/g胶冻样芽孢杆菌	1.17×10⁵	5.86×10⁴	2.93×10⁴	1.46×10⁴	7.32×10³
1 × 10¹²cfu/g解淀粉芽孢杆菌	2.50×10⁷	6.25×10⁶	1.56×10⁶	3.91×10⁵	9.77×10⁴
5×10¹⁰cfu/g多粘类芽孢杆菌	5.36×10⁵	1.34×10⁵	3.35×10⁴	8.37×10³	2.09×10³
1×10¹⁰cfu/g哈茨木霉	6.25×10⁵	1.56×10⁵	3.91×10⁴	9.77×10³	2.44×10³
1 × 10¹²cfu/g枯草芽孢杆菌	4.69×10⁷	1.17×10⁷	2.93×10⁶	7.32×10⁵	1.83×10⁵

微生物菌剂名称	培养基系列浓度/(cfu/mL)
微生物菌剂名称	Ⅰ	Ⅱ	Ⅲ	Ⅳ	Ⅴ
5×10¹⁰cfu/g胶冻样芽孢杆菌	1.17×10⁵	5.86×10⁴	2.93×10⁴	1.46×10⁴	7.32×10³
1 × 10¹²cfu/g解淀粉芽孢杆菌	2.50×10⁷	6.25×10⁶	1.56×10⁶	3.91×10⁵	9.77×10⁴
5×10¹⁰cfu/g多粘类芽孢杆菌	5.36×10⁵	1.34×10⁵	3.35×10⁴	8.37×10³	2.09×10³
1×10¹⁰cfu/g哈茨木霉	6.25×10⁵	1.56×10⁵	3.91×10⁴	9.77×10³	2.44×10³
1 × 10¹²cfu/g枯草芽孢杆菌	4.69×10⁷	1.17×10⁷	2.93×10⁶	7.32×10⁵	1.83×10⁵

微生物菌剂	混合比例	培养基系列浓度
微生物菌剂	混合比例	Ⅰ	Ⅱ	Ⅲ	Ⅳ	Ⅴ
1×10¹²cfu/g解淀粉芽孢杆菌：1×10¹⁰cfu/g哈茨木霉	9:1	2.98×10⁷	1.49×10⁷	7.45×10⁶	3.72×10⁶	1.86×10⁶
5×10¹⁰cfu/g多粘类芽孢杆菌：1×10¹⁰cfu/g哈茨木霉	6:4	3.20×10⁵	1.60×10⁵	7.99×10⁴	4.00×10⁴	2.00×10⁴
1×10¹²cfu/g枯草芽孢杆菌：1×10¹⁰cfu/g哈茨木霉	8:2	2.80×10⁷	1.40×10⁷	7.00×10⁶	3.50×10⁶	1.75×10⁶

微生物菌剂	混合比例	培养基系列浓度
微生物菌剂	混合比例	Ⅰ	Ⅱ	Ⅲ	Ⅳ	Ⅴ
1×10¹²cfu/g解淀粉芽孢杆菌：1×10¹⁰cfu/g哈茨木霉	9:1	2.98×10⁷	1.49×10⁷	7.45×10⁶	3.72×10⁶	1.86×10⁶
5×10¹⁰cfu/g多粘类芽孢杆菌：1×10¹⁰cfu/g哈茨木霉	6:4	3.20×10⁵	1.60×10⁵	7.99×10⁴	4.00×10⁴	2.00×10⁴
1×10¹²cfu/g枯草芽孢杆菌：1×10¹⁰cfu/g哈茨木霉	8:2	2.80×10⁷	1.40×10⁷	7.00×10⁶	3.50×10⁶	1.75×10⁶

微生物菌剂名称	回归方程	EC₅₀/(μg/mL)	置信区间	相关系数(r)
5×10¹⁰cfu/g胶冻样芽孢杆菌	y=4.8467+1.0812x	1.39	1.05~1.83	0.990
1×10¹²cfu/g解淀粉芽孢杆菌	y=4.4249+1.1344x	3.21	2.00~5.17	0.987
5×10¹⁰cfu/g多粘类芽孢杆菌	y=5.0234+0.5270x	0.90	0.54~1.50	0.988
1×10¹⁰cfu/g哈茨木霉	y=5.4950+0.5033x	0.10	0.03~0.35	0.986
1×10¹²cfu/g枯草芽孢杆菌	y=4.4638+0.9037x	3.92	2.83~5.44	0.995