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摘要浙麦冬是浙江省八大名药材“浙八”之一，用于治疗阴虚肺燥、咳嗽痰粘、肠燥便秘，也作为绿地观赏植物栽培。然而，近年来麦冬病害问题日益突出。化学农药的不合理使用造成的农药污染和药品残留超标，以及引发病菌的耐药性等问题，对药材的安全生产及食用者的健康都造成重大隐患。因此，如何正确识别麦冬植物病原菌，如何进行无害化治理，提高药材的安全卫生质量，以达到最佳药理品质，是当前麦冬病害防治研究工作的重点。本研究采集了9株表现枯萎，黑褐色斑点症状的麦冬植株，经病原菌分离纯化，结合形态学分析及ITS-PCR技术完成鉴定工作。分离获得病原真菌9株，分属5个种，包括链格孢菌（Alternaria sp.）、日本曲霉（Aspergillus japonicus）、束状炭疽菌（Colletotrichum dematium）、木贼镰刀菌（Fusarium equiseti）和尖孢镰刀菌（Fusarium oxysporum）。将分离到的真菌接种到健康的浙麦冬植株上，完成科赫氏法则验证。研究发现日本曲霉是引起浙麦冬褐斑病的病原菌，为麦冬病害的防治提供了理论依据。51944
Abstract Liriope platyphylla is one of the eight famous herbs called as "Zhe ba wei" in Zhejiang province, used in the treatment of lung dryness due to Yin deficiency, dry cough sputum viscosity, intestinal constipation, are cultivated as ornamental plants in Greenland. However, in recent years, the problems of plant diseases of Liriope platyphylla has become increasingly prominent. Unreasonable use of chemical fungicides and pesticides caused pollution, residue and resistance problems. This will be a major hidden danger for production of genuine medicinal in Zhejiang and even food health. Therefore, how to correctly identify the medicinal plant pathogens, how to carry out the harmless treatment, and improve the safety quality of Liriope platyphylla, to achieve the best quality of pharmacology, is the current focus of prevention and control of medicinal plant disease. This study collected 9 plants withered, dark brown spotting symptoms of Liriope platyphylla plant. Pathogenic fungi were isolated and identified by morphological analysis and ITS-PCR technology. 9 fungal strains were isolated. And 5 species of fungi were identified, including Alternaria sp., Aspergillus japonicas, Colletotrichum dematium, Fusarium equiseti and F. oxysporum. The isolated fungus was inoculated onto healthy Liriope platyphylla, completing Koch's postulates verification. We found that Aspergillus japonicus caused the brown spot of Liriope platyphylla. This work provide theoretical basis for prevention and treatment of the disease of Liriope platyphylla.
毕业论文关键词： 浙麦冬； 真菌病害； 褐斑病； 分离；鉴定
Keyword: Liriope platyphylla； fungal disease； brown spot； separation； identification
目 录
摘要 2
1 引言 5
2 浙麦冬致病真菌的分离与鉴定 6
2.1 材料与方法 6
2.1.1 供试材料 6
2.1.2 病原菌的分离鉴定 6
2.1.2.1 病原菌的分离与纯化 6
2.1.2.2 病原菌的形态鉴定 7
2.1.2.3 病原菌的分子鉴定 7
2.1.2.3.1 基因组DNA提取与检测 7
2.1.2.3.2 PCR扩增及测序分析 8
2.1.2.4 致病性测定 8
2.1.2.4.1 真菌孢子制备 8
2.1.2.4.2 致病菌侵染实验