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1、 编 号： 审定成绩： 重庆邮电大学毕业设计（论文）设计（论文）题目：棉花黄萎病菌的分离及培养技术研究学 院 名 称 ： 生物信息学院学 生 姓 名 ： 专 业 ： 生 物 技 术班 级 ： 学 号 ： 指 导 教 师 ： 江怀仲答辩组 负责人 ：填表时间： 2015 年 5月重庆邮电大学教务处制重庆邮电大学本科毕业设计（论文） 摘 要棉花黄萎病（Verticillium Wilt of Cotton）是一种土传性的维管束系统病害，严重影响棉花产量和质量，有人称它为“癌症”。病害发生的关键因子，除了品种抗病性较低和适宜的气候条件外，土壤中棉花黄萎病菌的大量积累是棉花黄萎病发生流行的主要原因。据
2、国外报道，棉花黄萎病菌形成的休眠结构微菌核，可以在土壤中常年存活，并成为危害侵染循环中最重要的侵染源。在全球因黄萎病所造成的经济损失平均每年达10亿多美元，筛选利用抗病品种是防治黄萎病最经济有效的途径之一，而品种的筛选与鉴定及相关研究都需要分离和培养黄萎病菌种。本研究通过查阅现有文献和结合现有实验条件，采集四川简阳棉花地里耕作层土壤、棉花根际土壤作为病菌分离材料，然后用最简单有效的逐步稀释法稀释样本，并将稀释液涂布在PDA培养基上。根据生长出来的菌落形态和颜色等特征，挑取单菌落再用平板培养法分离出纯化的棉花黄萎病菌。然后将所得的棉花黄萎病菌用不同液体培养基对照培养，从而选择出一种最利于棉花黄萎
3、病菌生长的培养基，以利于将分离纯化出的棉花黄萎病菌培养成可供应用的黄萎菌液，供相关研应究用。将采集的四份土壤样本，稀释成10-1、10-2、10-3不同浓度，涂布平板和平板培养后得到了4株纯化的真菌，经过鉴定最终得到了1株纯化的棉花黄萎病菌。然后用不同液体培养基对棉花黄萎病菌对照培养，得出淘米水培养基培养的棉花黄萎病菌以微菌核形式为主；察氏液体培养基培养出来的棉花黄萎病菌含有较多菌丝和孢子；马丁氏培养基培养出来的棉花黄萎病菌以含孢子为主。【关键词】棉花黄萎病菌 分离和培养 液体培养基 ABSTRACTVerticillium Wilt of Cotton is one of the main
4、diseases which affecting the yield and quality of the cotton. The key factor of the disease, in addition to less resistant varieties and appropriate climate conditions, the accumulation of a large number of Verticillium dahliae in soil is the main reason which cotton Verticillium wilt is popular. Ac
5、cording to foreign reports, sleeping structure micro-sclerotia which Verticillium dahliae formed can survive in soil all the year round and become the most important infection source of against infection cycle. In the world, the economic loss caused by Verticillium wilt account for one billion U.S.
6、Dollars average every year, one of the most cost-effective ways which control Verticillium wilt is filtering and using resistant varieties, while screening and identification of the species and related research all need to isolate and culture Verticillium species. In this study, according to search
7、the existing literature with the existing experimental conditions, we collected farming soil and rhizosphere soil from cotton of Sichuan Jianyang as bacteria isolated material, and then gradually dilute the sample, and coating the dilution on the PDA medium. Based on the growth characteristics of co
8、lony, such as shape and color, we can pick a single colony and culture it on the medium. In this way, we can isolate and select the purified Verticillium dahliae. And then Verticillium dahliae were cultured with different liquid medium. So that selecting one of the most favorable medium for the grow
9、th of Verticillium dahliae, in order to culture the isolated and purified Verticillium dahliae into yellow wilt bacterium，for research applications.Diluting the four soil samples which collected from Jianyang, into different concentrations of 10-1,10-2,10-3.Plating culture after coating plate, we ge
10、t four sets of purified fungi. After identification, I get a set of purified Verticillium dahliae. Then using different liquid media to culture Verticillium dahliae, finally, from the reproduction of Verticillium dahliae, we can draw a conclusion, after using Taomi Shui medium to culture Verticilliu
11、m dahliae, obtained a lot of micro-sclerotia; using Czapek liquid medium to culture, obtained many mycelium and spores; using Mading Shi medium to culture, obtained many spores.【Key words】Verticillium dahliae Isolation and Culture liquid medium目 录前 言 1第一章 棉花黄萎病菌概述 2第一节 棉花黄萎病菌的形态特征 .2第二节 棉黄黄萎病发病症状 .2第三节 棉花黄萎病菌的传播及防治方法 .4第四节 研究目的及意义.5第二章 棉花黄萎病菌的分离.6 第一节 实验材料 .6 第二节 实验方法 .8第三节 实验结果与分析 .12第三章 棉花黄萎病菌的培养 16 第一节 实验材料 .16 第二节 实验方法 .17 第三节 培养结果 .18 第四节 本章小结 .21结 论 .23 致 谢24参考文献 25附 录 26 一、英文原文26二、英文翻译36- 44 -前 言
