摘要杭白菊（white Chrysanthemum morifolium）作为我国传统的栽培药用植物，是浙江省八大名药材“浙八味”之一。近年来由于病害问题日益突出，化学农药不合理使用加剧，造成农药污染和药品残留超标，并引发病原真菌严重的耐药性问题，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。本文以杭白菊重要病害叶枯病的病原真菌尖孢镰刀菌（Fusarium oxysporum）为研究对象，对其进行4种常见杀菌剂的药性评估。并进一步测定尖孢镰刀菌对常用苯并咪唑类药剂多菌灵和三唑类药剂戊唑醇的敏感性，发现对于多年使用的化学农药多菌灵存在明显抗性。其中两个高抗菌株Fo 027和Fo 028其β-微管蛋白氨基酸序列198位由正常谷氨酸（E，Glu）突变为谷氨酰胺（Q，Gln）；一个中抗菌株Fo 014其氨基酸序列167位由正常苯丙氨酸（F，Phe）突变为酪氨酸（Y，Tyr），引起蛋白构象上的改变。本研究认为极有可能是病原真菌β-微管蛋白上的点突变造成了多菌灵抗性，同时未发现戊唑醇抗性菌株，认为该药物可以替代多菌灵作为桐乡地区杭白菊病害防治的主要药剂。72380

毕业论文关键词：苯并咪唑类药物； 真菌病害； 尖孢镰刀菌； 抗性机制；点突变

Abstract White Chrysanthemum morifolium was the traditional cultivated medicine in China, and was one of the famous medicinal materials “Zhe ba wei” in Zhejiang Province。 In recent years due to disease problems have become increasingly prominent, the irrational use of chemistry pesticides increased, resulting in pesticide pollution and drug residues, and lead to serious fungal pathogen resistance, there are significant risks of White Chrysanthemum morifolium production and food safety。  It is major hidden danger to the health of white chrysanthemum production and food safety。 Fusarium oxysporum, an important pathogen of leaf blight, was studied in this paper, and the drug resistance of 4 common fungicides was evaluated。 And it was found that the obvious resistance to the chemical pesticides used for many years, through the further determination of sensitivity of Fusarium oxysporum on commonly used benzimidazole fungicides carbendazim and triazole agents tebuconazole。 There are two resistant strains Fo 027 and Fo 028, and the beta tubulin amino acid sequence 198 by normal glutamate (Glu) e mutation glutamine (Q, Gln); a medium antibacterial plant Fo 014, the amino acid sequence of 167 by normal phenylalanine (F, Phe) mutation of tyrosine (Tyr) y, caused a change in the conformation of the protein。 This study suggests that most likely pathogenic fungi beta tubulin point mutations caused the carbendazim resistance。 While tebuconazole resistant strains was not found, and it is considered that the medicine can be used as the main agent to prevent and control the disease of White Chrysanthemum morifolium in Tongxiang area。

Keyword: Benzimidazole； Fungal diseases； Fusarium oxysporum； Resistance mechanism；Point mutation

目    录

中文摘要 2

英文摘要 3

1  引言 6

2  杭白菊病原真菌药物敏感性测定 7

2。1材料与方法 7

2。1。1供试材料 7

2。1。2四种杀菌剂的初步测定 7

2。1。1尖孢镰刀菌对戊唑醇和多菌灵的EC50值测定 8

3 抗药分子机制研究 8

3。1材料与方法 8

3。1。1供试材料