云南地区铁线莲叶枯病病原菌的鉴定和主要生物学特性

doi:10.3969/j.issn.1004-1524.2022.07.12

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (7): 1449-1456.DOI: 10.3969/j.issn.1004-1524.2022.07.12

云南地区铁线莲叶枯病病原菌的鉴定和主要生物学特性

杨玲¹^,²(), 沙楠景², 潘鹏举², 吴伯志¹^,^*()

1.云南农业大学农学与生物技术学院,云南昆明 650201
2.云南省高校林木生物技术重点实验室(西南林业大学),云南昆明 650224

收稿日期:2021-02-23 出版日期:2022-07-25 发布日期:2022-07-26
通讯作者: 吴伯志
作者简介:* 吴伯志,E-mail: bozhiwu@outlook.com
杨玲(1974—),女,重庆人,博士研究生,主要从事作物品种生态研究。E-mail: 394057384@qq.com
基金资助:
云南省高校林木生物技术重点实验室项目基金(51700201);云南省高校林下生物资源保护与利用科技创新团队(51400605)

Identification and main biological characteristics of pathogen of Clematis leaf blight in Yunnan

YANG Ling¹^,²(), SHA Nanjing², PAN Pengju², WU Bozhi¹^,^*()

1. College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201,China
2. Key Laboratory for Forest Biotechnology in Universities of Yunnan Province, Southwest Forestry University, Kunming 650224,China

Received:2021-02-23 Online:2022-07-25 Published:2022-07-26
Contact: WU Bozhi

摘要/Abstract

摘要：

为明确引起云南省铁线莲叶枯病病原菌的种类,从患叶枯病的植株上采集病叶,采用常规组织分离法进行病原菌的分离纯化,通过观察病原菌的形态学特征,结合核糖体内转录间隔区(ITS)和大亚基(LSU)双基因序列分析对病原菌的种类进行鉴定,采用科赫法则进行验证。并用不同培养基、温度、光照、pH、碳源和氮源对病原菌的主要生物学特性进行了测定。结果表明:铁线莲叶枯病的致病菌为Spencermartinsia sp.,按照科赫法则接种植株的叶片均发病。马铃薯葡萄糖琼脂培养基(PDA)和马铃薯蔗糖琼脂培养基(PSA)为其适宜培养基,生长最适温度为25 ℃,不同光照处理下生长不受影响,生长最适pH为6.5~7.5,最佳碳源为葡萄糖和蔗糖,最佳氮源为硝酸钠。

关键词: 铁线莲叶枯病, 病原鉴定, 生物学特性

Abstract:

In order to identify the pathogen of Clematis leaf blight in Yunnan Province, the typical diseased leaves were collected from the diseased plants. The pathogen was isolated and purified by tissue separation method, and identified by morphological characteristics combined with sequence analysis of ribosomal internal transcribed spacer (ITS) and large subunit (LSU), and verified by Koch postulates. The main biological characteristics of the pathogen were determined with different media, temperature, light, pH, carbon and nitrogen sources. The results showed that the pathogen of Clematis leaf blight was Spencermartinia sp., according to Koch postulates, all the leaves inoculated with the pathogen were infected. Potato dextrose agar (PDA) and potato saccharose agar(PSA) were the suitable media, the optimum temperature for growth was 25 ℃, the growth was not affected under different light treatments, the optimum pH for growth was 6.5-7.5, the best carbon source was glucose and sucrose, and the best nitrogen source was sodium nitrate.

Key words: Clematis leaf blight, pathogen identification, biological characteristics

中图分类号:

S435.67

杨玲, 沙楠景, 潘鹏举, 吴伯志. 云南地区铁线莲叶枯病病原菌的鉴定和主要生物学特性[J]. 浙江农业学报, 2022, 34(7): 1449-1456.

YANG Ling, SHA Nanjing, PAN Pengju, WU Bozhi. Identification and main biological characteristics of pathogen of Clematis leaf blight in Yunnan[J]. Acta Agriculturae Zhejiangensis, 2022, 34(7): 1449-1456.

图/表 7

图1 铁线莲叶枯病症状与回接验证 A,自然发病植株的症状;B,回接植株发病症状;C,对照。

Fig.1 Comparison of diseased and inoculated plants of Clematis A, Symptoms of naturally diseased plant; B, Symtoms of the inoculated plant; C, Control.

图2 病原菌的形态学特征 A,第5天菌落的正面;B,第5天菌落的背面;C,松散聚集的分生孢子器;D,分生孢子器;E,分生孢子;F,产孢细胞及幼嫩的孢子。

Fig.2 Morphological characteristics of the pathogen A, The front of colony on the 5th day; B, The back of colony on the 5th day; C, Loose and aggregated conidiomata; D,Pycnidia; E, Conidia; F, Conidiogenous cells and young conidia.

图3 基于ITS和LSU联合构建的系统发育树加粗表示模式菌株;★表示试验所得数据;图中斜杠前数值表示支持率,斜杠后数值表示自举值。

Fig.3 Phylogram generated from maximum likeihood analysis based on combined ITS and LSU Type species were given in bold typeface;★ represented data from the study; In the figure, the value before the slash represented the support value, and the value after the slash represented the ultrafast bootstrap value.

图4 不同培养基对病原菌生长的影响不同小写字母表示各处理间差异显著(P<0.05)。下同。

Fig.4 Effect of different media on the pathogen growth Different lowercase letters represent significant difference at 0.05 level. The same as below.

图5 不同温度对病原菌生长的影响

Fig.5 Effect of different temperatures on the pathogen growth

图6 不同pH值对病原菌生长的影响

Fig.6 Effect of different pH values on the pathogen growth

图7 碳源和氮源对病原菌生长的影响 A,碳源对病原菌生长的影响;B,氮源对病原菌生长的影响。

Fig.7 Effect of carbon source and nitrogen source on the pathogen growth A, Effect of carbon source on the pathogen growth; B, Effect of nitrogen source on the pathogen growth.

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云南地区铁线莲叶枯病病原菌的鉴定和主要生物学特性

Identification and main biological characteristics of pathogen of Clematis leaf blight in Yunnan

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