老鼠簕真菌病病原的分离鉴定及生物学特性

马林; 袁宗胜; 郑瑜婷; 郑雄; 陈维峰; 王思凡; 谢宝贵; 刘芳

doi:10.19303/j.issn.1008-0384.2023.11.009

老鼠簕真菌病病原的分离鉴定及生物学特性

doi: 10.19303/j.issn.1008-0384.2023.11.009

马林^1,,
袁宗胜²,
郑瑜婷³,
郑雄¹,
陈维峰¹,
王思凡⁴,
谢宝贵¹,
刘芳^1, ,

1.
福建农林大学生命科学学院，福建　福州，350002
2.
闽江学院地理与海洋学院，福建　福州，350108
3.
古田县食用菌研发中心，福建　宁德，352200
4.
福建农林大学未来技术学院，福建　福州，350002

基金项目: 国家自然科学基金项目（32002108）；闽江学院引进人才预研项目（33004328）

详细信息

作者简介:
马林（1997 — ），男，硕士研究生，主要从事食用菌和农业微生物研究，E-mail：worldmalinmm@gmail.com

通讯作者:
刘芳（1980 — ），女，讲师，主要从事食用菌和农业微生物研究，E-mail：fjliufang@163.com

中图分类号: S436
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- 文章访问数: 404
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- 被引次数: 0
出版历程
- 收稿日期: 2023-07-22
- 修回日期: 2023-08-16
- 网络出版日期: 2023-12-21
- 刊出日期: 2023-11-28

Isolation, Identification, and Biological Characteristics of a Fungal Pathogen on Acanthus ilicifolius

1.
College of Life Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
2.
College of Geography and Oceanography, Minjiang University, Fuzhou, Fujian　350108, China
3.
Research and Development Center of Edible Fungi of Gutian, Ningde, Fujian　352200, China
4.
College of Future Technology, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China

摘要

摘要: 目的明确老鼠簕（Acanthus ilicifolius）叶斑病的病原种类及其生物学特性。方法从福建省漳江口红树林国家级自然保护区采集病叶，采用离体组织分离法对病叶病原菌进行分离，采用形态学与分子生物学（ITS和EF1-α基因序列分析）的方法鉴定其病原菌种类，并对该病原菌的菌丝在不同温度、pH、碳源和氮源等条件下的生物学特性进行研究，以及对过氧化氢酶（CAT），超氧化物歧化酶（SOD）、过氧化物酶（POD）、漆酶、纤维素酶（CL）、β-葡萄糖苷酶（β-GC）进行活性检测。结果经鉴定该病原菌为真菌界子囊菌门镰刀菌科的轮枝镰刀菌（Fusarium verticillioides）。这是轮枝镰刀菌侵染老鼠簕引发叶斑病的首次报道。生物学特性研究显示该病原菌生长的最适温度25 ℃，最适pH为10，最佳碳源为淀粉，最佳氮源为酵母粉。3种氧化还原酶中，过氧化物酶（POD）活性最高；3种水解酶中，纤维素酶（CL）活性最高。结论分离鉴定了福建省漳江口老鼠簕叶斑病病原菌为轮枝镰刀菌（Fusarium verticillioides），明确了病原菌的生物学特性。
- 老鼠簕 /
- 叶斑病 /
- 鉴定 /
- 轮枝镰刀菌 /
- 生物学特性
Abstract: Objective Pathogen of the leaf spot disease on Acanthus ilicifolius was isolated and identified with its biological characteristics determined. Method Microbes were isolated from the diseased A. ilicifolius leaves collected from the Zhangjiangkou Estuary Mangrove National Nature Reserve in Fujian Province by the detached tissue method. Morphological and molecular biological techniques including ITS and EF1-α gene sequence analysis were employed to identify the pathogen. Optimal growth conditions of the identified fungus with respect to temperatures, pH, carbon sources, and nitrogen sources, in addition to its activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), laccase, cellulase (CL), andβ-glucosidase (β-GC) were determined. Result For the first time Fusarium verticillioides in the Ascomycota phylum was identified as the responsible pathogen of the disease. It thrived at 25°C and pH 10 on a medium using starch as carbon source and yeast powder as nitrogen source. POD was the oxidoreductase, and CL the hydrolase, that showed the highest activity in the fungus. Conclusion The pathogen that caused the leaf spot disease on bougainvillea in the Zhangjiang Estuary was identified as F. verticillioides. The biological characteristics of the strongly adaptable fungus were determined.
- Acanthus ilicifolius /
- leaf spot disease /
- identification /
- Fusarium verticillium /
- biological characteristics

HTML全文

图 1 老鼠簕叶斑病症状

A为自然发病叶片症状；B为室内离体接种后叶片正面症状；C为室内离体接种后叶片背面症状。

Figure 1. Symptoms on A. ilicifolius infected by leaf spot disease

A: Symptoms on diseased leaves in nature; B: symptoms shown on face of diseased leaf after in vitro inoculation; C: symptoms shown on back of diseased leaf after in vitro inoculation.

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图 2 老鼠簕叶斑病病原菌形态特征

A为菌落的正面形态；B为菌丝的圆形缠绕形态；C为孢子聚集结构；D为小分生孢子；E为大分生孢子；F为厚垣孢子。

Figure 2. Morphology of pathogen causing leaf spot disease on A. ilicifolius

A: Top view of fungal colony; B: circular winding mycelial growth; C: aggregated spores; D: microconidia; E: macroconidia; F: chlamydospore.

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图 3 基于EF-1α基因序列的系统发育树

Figure 3. Phylogenetic tree based on EF-1α gene sequences

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图 4 基于ITS基因序列的系统发育树

Figure 4. Phylogenetic tree based on ITS gene sequences

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图 5 病原菌在不同温度的生长情况

在显著性水平为0.05的情况下，不同小写字母表示显著差异。下同。

Figure 5. Growth of pathogen at different temperatures

Data with different lowercase letters indicate significant differences at level of 0.05. Same for below.

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图 6 病原菌在不同pH的生长情况

Figure 6. Growth of pathogen at different pH

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图 7 病原菌在不同碳源的生长情况

Figure 7. Growth of pathogen on different carbon sources

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图 8 病原菌在不同氮源的生长情况

Figure 8. Growth of pathogen on different nitrogen sources

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图 9 病原菌的氧化还原酶活性

Figure 9. Oxidoreductase activity of pathogen

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图 10 病原菌的水解酶活性

Figure 10. Hydrolase activity of pathogen

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