水葫芦论文：水葫芦生防菌Cercosporasp.FJ24的分离、鉴定与生物学特性_水葫芦毕业论文
水葫芦论文：水葫芦生防菌Cercosporasp.FJ24的分离、鉴定与生物学特性由刀豆文库小编整理，希望给你工作、学习、生活带来方便，猜你可能喜欢“水葫芦毕业论文”。
水葫芦论文：水葫芦生防菌Cercosporasp.FJ24的分离、鉴定与生物学特性
【中文摘要】水葫芦(Eichhornia craipes)的蔓延在几十个国家和地区造成了严重的环境问题,如何有效治理水葫芦受到全球范围内的广泛关注。其中,利用水葫芦致病真菌进行治理,是一种备受重视的途径。本文从福建省福州市郊区采集到被致病真菌强烈感染的水葫芦植株,从叶片上分离得到了具有强致病性的真菌菌株FJ24。在对其致病性能进行试验的基础上,对其生物学特性进行了研究。通过形态学鉴定,初步认为FJ24属于尾孢属。进一步地在分子生物学水平上,对其ITS区、EF-1区、β-Tub区、His-H3区序列进行PCR扩增,通过测序,对比Genebank中已经登录的尾孢属相应序列,构建系统发育树,分析其分类地位。结果如下:证实其在7天内能够使试验水葫芦植株达到100%的发病率,3天、7天、14天、21天、28天病情指数分别达到了23.33%、44.17%、78.33%、92.50%、100%。FJ24菌株在PDA培养基上菌落呈灰白色、边缘呈淡红色,簇状分布。菌丝大部分侵入培养基,边缘清晰。分生孢子梗单生或2~12根簇生,不分枝,浅褐色,宽度较规则、直立或略有弯曲,63.4~247.6×2.5~5.5μm。分生孢子单生,无色至浅褐色,有多个横隔,略有弯曲或无弯曲,线形、鞭型或蠕虫型,56.0~312.2×2.0~7.8μm。初步鉴定其为尾孢属。在PDA培养基上,FJ24菌株在全黑暗和光暗交替条件下生长速度较快,该菌在30°C下表现出最快生长速度,而且能长期保持,最适宜的PH条件为
中性偏碱性,在不同培养基上菌落形态存在一定差别,最适宜的培养基为MMA、MSDA、PDAY,该菌最适合利用的碳源和氮源分别为淀粉和乙酸铵。根据ITS区、EF-1区、β-Tub区、His-H3区序列的测序结果,构建系统发育树,结果表明Cercospora sp.FJ24菌株与Cercospora piaropi的多个水葫芦致病菌株存在着极高的同源性,序列相似度均达到了99%以上。生物学特性实验和分子生物学分析均表明,基本可以认为Cercospora sp.FJ24是属于Cercospora piaropi的。综上所述,本文分离得到的Cercospora sp.FJ24具有开发成为水葫芦生防制剂的潜力,同时由于该菌分离自我国本土,具有较高的安全性,因此对于水葫芦的生物防治具有一定的参考价值。
【英文摘要】Spread of Water Hyacinth(Eichhornia craipes)caused serious environmental problems in lots of countries and regions.The importance of controlling it was recognized throughout the world.Currently, controlling Water Hyacinth with pathogens gained more and more attention.A strain FJ24, with high pathogenicity against Water Hyacinth was isolated from heavily diseased Water Hyacinth collected from Fujian province.On the basis of determining the pathogenicity of FJ24 against Water Hyacinth, biological characteristics of the strain were studied.Using morphological identification , the strain was preliminarily considered as Cercospora.The strain was further analysed at the molecular level.Four
representative sequences of Cercospora sp.FJ24 including the ITS region,EF-1 region,β-Tub region and His-H3 region were sequenced and blasted with other signed Cercospora species on Genbank.Phylogenetic dendrogram were constructed to analyse the taxonomic position of Cercospora sp.FJ24.The results were as follows: The diseaed ratings of Cercospora sp.FJ24 reached 100% in 7 days;the diseased indexes were 23.33%,44.17%,78.33%,92.50% and 100% after 3 days ,7 days, 14 days, 21 days and 28 days respectively.Colony of Cercospora sp.FJ24 on PDA culture was hoar and its edge was light red, growing in cluster.Hypha almost invaded into the culture with clear edge.Conidiophores of Cercospora sp.FJ24 borne singly or in fascicles of 2 to 12, were light brown, septate, width regular, straight or lightly curved and no branched , measuring 63.4 to 247.6μm long and 2.5 to 5.5μm wide.Conidia developed singly , colourle or light brown with many transverse, measured 56.0 to 312.2μm long and 2.0 to 7.8μm wide.Preliminarily identify it as Cercospora.The colony of Cercospora sp.FJ24 grew best on PDA under the dark treatment of alternation illumination and darkne while the colony growed better on PDA under all darkne.The optimum temperature for colony longtime growth was found to be 30℃.The best colony growth was obtained at the condition of neutral pH or alkalescence.The fitting carbon and nitrogen source were amylum and ammonium acetate respectively.Cercospora sp.FJ24 presented varying morphological characteristics on different media tested, and the colonies grew best on MMA, PDAY and MSDA.Four phylogenetic trees were generated from the aligned ITS region,EF-1 region,β-Tub region and His-H3 region sequences.The treeing analyses showed that Cercospora sp.FJ24 presented highly homologous with many kinds of pathogenic strain in Cercospora piaropi.And the sequence similarities were above 99%.Biological characteristics of Cercospora sp.FJ24 and phylogenetic tree analyses indicated that Cercospora sp.FJ24 could be basically identified as Cercospora piaropi.The results listed above indicated that Cercospora sp.FJ24, isolated from this study, poeed the great potential to be developed into the fungal herbicide against Water Hyacinth.This research had positive significances for the biological control of exotic plants.【关键词】水葫芦 生防菌 Cercospora piaropi FJ24 鉴定 【英文关键词】Water Hyacinth Pathogenic strain Cercospora piaropi FJ24 Identification 【目录】水葫芦生防菌Cercosporasp.FJ24的分离、鉴定与生物
学特性12-2312-15摘要5-7ABSTRACT7-9第一章 前言1.1 水葫芦的生态学特性、分布和危害1.1.1 水葫芦的生态学特性12-1
31.1.3 我国水葫芦的危害情况
1.2.1 水葫芦的物理
1.3 1.1.2 水葫芦的分布13-1414-151.2 水葫芦的防治15-21化学防治15-161.2.2 水葫芦的生物防治16-21水葫芦的综合防治2121-23材料23准备2323-2
41.4 本研究的选题依据、目的和意义
2.1 实验第二章 实验仪器、材料与方法23-342.1.1 水葫芦病样采集232.1.3 菌株来源2
32.1.2 供试植株的2.2 主要仪器设备
2.3.1 基
2.3.3 2.3 固体培养基及其制备方法24-27
2.3.2 不同pH 值固体培养基24-25础培养基24不同C、N 源固体培养基2525-27272.4 方法27-342.4.2 接种试验27
2.3.4 其它种类固体培养基2.4.1 病原真菌的分离2.4.3 柯赫氏法则（Koch’s
2.4.5 Rule）验证27-28致病性测定2828-2929
2.4.4 植物病害统计方法282.4.6 影响菌落生长的因素2.4.7 DNA 提取292.4.8 ITS 区序列扩增
2.4.10 系统发育3.1 水葫芦病原
3.1.2 2.4.9 参考基因序列扩增29-30
第三章 结果与分析34-57分析30-34真菌FJ24 的分离34-36致病症状34-35
3.1.1 病原菌的分离34
3.1.3 柯赫氏法则验证353.1.4 致病
性测定35-3636-37征36-3737-47
3.2 水葫芦病原真菌FJ24 的形态学特征
3.2.2 分生孢子形态特3.2.1 菌落形态特征363.3 影响FJ24 菌落生长的因素研究3.3.1 光照对FJ24 菌落生长的影响37
3.3.2 不同光照条件下FJ24 生长曲线的测定37-38FJ24 菌落生长的影响38-39线的测定39-4040-4141-42
3.3.3 温度对
3.3.4 不同温度下FJ24 生长曲
3.3.5 PH 对FJ24 菌落生长的影响3.3.6 不同初始PH 下FJ24 生长曲线的测定3.3.7 碳源对FJ24 菌落生长的影响42
3.3.8 不同碳源下FJ24 生长曲线的测定42-43菌落生长的影响43-44测定4444-4545-4747-5750-52列54-57
3.3.9 氮源对FJ24
3.3.10 不同氮源下FJ24 生长曲线的3.3.11 不同培养基对FJ24 菌落生长的影响3.3.12 不同培养基中FJ24 生长曲线的测定3.4 Cercospora sp.FJ24 的分子生物学鉴定3.4.1 ITS 序列47-503.4.3 β-Tub 序列52-54第四章 讨论57-62
3.4.2 EF-1 序列3.4.4 His-H3 区序4.1 关于Cercospora sp.FJ24 的鉴定574.2 关于Cercospora sp.FJ24 的生物学
4.3 Cercospora
4.4 关于特性及其对实际应用的指导意义57-59sp.FJ24 菌株作为真菌除草剂的开发前景59-60Cercospora sp.FJ24 菌株的安全性60-61草剂的开发前景61-62
4.5 水葫芦真菌除
参考文献
第五章 结论62-63
63-68附录68-70致谢70-71攻读硕士学位期间发表的论文71
