GUO Xuyu,KANG Yue,LUO Chunxu,et al.Isolation and Identification of Grape Pathogenic Fungi and Screening of Antagonistic Bacteria[J].HEILONGJIANG AGRICULTURAL SCIENCES,2025,(07):51-60.[doi:10.11942/j.issn1002-2767.2025.07.0051]
葡萄病原真菌分离鉴定及拮抗细菌筛选
- Title:
- Isolation and Identification of Grape Pathogenic Fungi and Screening of Antagonistic Bacteria
- 文章编号:
- 9
- 文献标志码:
- A
- 摘要:
- 2023年7月，中国黑龙江省齐齐哈尔市园艺研究所葡萄园中发现葡萄叶片出现边缘黄化、黑斑等病害症状。为明确发病原因，采用组织分离法从自然发病的叶片中分离并纯化病原真菌，结合形态学特征、ITS-rRNA序列及系统发育分析对病原真菌进行了鉴定，同时采用离体叶片接种法验证了菌株的致病性，并筛选了对葡萄病原真菌有拮抗效果的细菌。基于形态学和ITS序列分析，将病原真菌PT9-4鉴定为Alternaria alternata，PT10-2鉴定为Curvularia lunata，PT12-1鉴定为Diaporthe gulyae，PT12-5鉴定为Fusarium oxysporum。致病性测定结果表明，这4株真菌均对葡萄叶片具有致病性，并完成了科赫法则的验证。对峙试验表明选用的3株拮抗细菌对4株病原真菌的抑制效果最高达到78.48%。细菌发酵液平板试验表明，3株拮抗细菌发酵液显著抑制了病原真菌菌丝生长，抑制率达到60%以上。
- Abstract:
- In July 2023, symptoms such as leaf edge yellowing and the presence of black spots were observed on grape leaves in a vineyard located at the Horticultural Research Institute of Qiqihar City, Heilongjiang Province, China. In order to determine the causative agents of the observed disease, this study employed a tissue isolation method to isolate and purify pathogenic fungi from naturally infected leaves. The identification of these pathogenic fungi was conducted based on their morphological characteristics, ITS-rRNA sequences, and phylogenetic analysis. The pathogenicity of the isolated strains was confirmed through a detached leaf inoculation method, and bacteria exhibiting antagonistic effects against the grape pathogenic fungi were subsequently screened. Morphological and ITS sequence analyses identified the pathogenic fungi as follows, PT9-4 was identified as Alternaria alternata, PT10-2 as Curvularia lunata, PT12-1 as Diaporthe gulyae, and PT12-5 as Fusarium oxysporum. The pathogenicity assays revealed that all four fungal strains were pathogenic to grape leaves, thereby fulfilling Koch’s postulates. Antagonistic experiments indicated that the three selected antagonistic bacteria effectively inhibited the four pathogenic fungi, achieving a maximum inhibition rate of 78.48%. Furthermore, plate experiments utilizing bacterial fermentation broth demonstrated that the fermentation broths of the three antagonistic bacteria significantly inhibited the mycelial growth of the pathogenic fungi, with an inhibition rate exceeding 60%.
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备注/Memo
收稿日期：2025-04-21 基金项目：黑龙江省省属本科高校基本科研业务费（145309327）。 第一作者：郭絮语（1999-），女，硕士研究生，从事葡萄病害生物防治研究。E-mail：xxg_3257@163.com。 通信作者：王芳（1981-），女，博士，副教授，从事植物抗病性研究。E-mail：wangfangnd@hotmail.com。