摘要杭白菊(Chrysanthemum morifolium)系菊科(Asteraceae)多年生草本植物，是我国传统中药，是浙江省八大名药材“浙八”之一。浙江省为杭白菊的主产区，其生产的杭白菊因品质好、观赏性强而享誉国内外。近年来，随着栽种面积逐年增加，杭白菊的病害问题日益突出，加之化学农药的不合理使用，造成农药污染和药品残留超标，并引发病菌的耐药等问题，对杭白菊的健康生产及食用者的安全健康都存在重大隐患。因此，开展杭白菊上重要致病真菌的分离、鉴定，对合理开展病害防治具有十分重要的意义。43888

本研究利用传统形态学的方法，并结合rDNA - ITS序列的分析，分离并鉴定了寄宿于杭白菊的重要病原真菌。结果表明，引起杭白菊真菌病害的病原菌主要有尖孢镰刀菌(Fusarium oxysporum)、球毛壳菌(Chaetomium globosum)、木贼镰刀菌(Fusarium equiseti)、拟康宁木霉(Trichoderma koningiopsis)。其中，鉴定的尖孢镰刀菌的数量最多，达37.5%。首次发现球毛壳菌能引起杭白菊的叶枯病，并为提出较为有效的防治措施奠定基础。

毕业论文关键词：杭白菊；真菌病害；病原菌分离鉴定

Abstract Chrysthemum morifolium is a herb of Asteraceae and has being used as a traditional Chinese medicine, and was one of the famous medicinal materials ‘Zebawei’ in Zhejiang Province. Chrysthemum morifolium produced in Zhejiang Province has large amount of output at home and abroad, because of its good quality and highly ornamental. In recent years, with the planted area increased, the disease is becoming increasingly prominent. Coupled with the irrational use of chemical pesticides, causing medicine residues exceeding. And causing the problems such as drug resistance of the fungal. It is major hidden danger to the health of Chrysthemum morifolium production and food safety. So it is necessary to isolate and identify the major pathogens of diseases control.

The pathogens of diseases were identified with traditional morphology and molecular biology for rDNA - ITS sequence. Result showed that the major disease on Chrysthemum morifolium is caused by Fusarium oxysporum, Chaetomium globosum, Fusarium equiseti and Trichoderma koningiopsis. Among them, the number of Fusarium oxysporum reached 37.5%. It is the largest number in this research. Besides, This is the first report of leaf blight disease of Chrysthemum morifolium caused by Chaetomium globosum in China, which would be important significance for the control of plant diseases on Chrysthemum morifolium.

Keyword: Chrysthemum morifolium; Fungal disease; Pathogen isolation and identification

目    录

摘   要 I

Abstract II

1  引言 1

2  材料与方法 3

2.1 供试材料 3

2.2 病原菌的分离与纯化 3

2.3 病原菌的形态鉴定 3

2.4 病原菌的分子鉴定 4

2.4.1 基因组DNA 的提取和检测 4

2.4.2 PCR扩增及测序分析 5

2.5 致病性测定 5

2.5.1 叶片准备 5

2.5.2 真菌孢子制备 5

2.5.3 致病菌侵染实验 5

3