首页 > 论文 > 激光生物学报 > 19卷 > 5期(pp:695-699)

苹果炭疽菌的分子鉴定与检测

Molecular Identification and Detection of Apple Anthracnose Pathogen

  • 摘要
  • 论文信息
  • 参考文献
  • 被引情况
  • PDF全文
分享:

摘要

测定苹果炭疽菌rDNA全序列, 比对苹果炭疽菌和其它炭疽菌ITS序列以及构建系统关系树, 发现苹果炭疽菌与胶孢炭疽菌的ITS序列相似性高达99.8 %, 并与胶孢炭疽菌聚在一起, 可以明确苹果炭疽菌应属于胶孢炭疽菌。进一步的序列比对发现, 苹果炭疽菌的18S rDNA 3’端比其它胶孢炭疽菌多出一段379 bp的序列, 根据这一特有片段设计引物CgF1与通用引物ITS4配对, 结果仅能从苹果炭疽菌中扩增出1 232 bp的特异性条带。用苹果炭疽菌接种离体苹果, 以接种发病的病组织总DNA为模板, 利用引物CgF1/ITS4进行PCR扩增, 同样可以扩增出1 232 bp的特异性条带, 而健康苹果组织DNA中未能扩增出任何条带, 表明该方法可用于苹果炭疽菌的鉴定和快速检测。

Abstract

The rDNA sequence of apple anthracnose pathogen was determined. Sequence comparing of ITS of Colletotrichum was conducted and a phylogenetic tree based on alignment of their ITS nucleotide sequences was constructed. The results showed that Cg-1 and Cg-2 shared the highest sequence similarity (99.8 %) with C.gloeosporioides and they could be grouped into one branch with C.gloeosporioides. It was confirmed that apple anthracnose pathogen should belong to C.gloeosporioides. It was found that there is a 379 bp sequence residue in 3’ ends of 18S rDNA of apple anthracnose pathogen comparing with the other C.gloeosporioides. Specific primer CgF1 was designed according to the 379 bp sequence, and only a 1 232 bp specific band could be amplified using primer pair CgF1 and ITS4. Apple fruits were inoculated with apple anthracnose pathogen. Total DNA was extracted from the diseased tissue, and an identical 1 232 bp band was amplified. It was suggested that the method could be applied for the detection and rapid identification of apple anthracnose pathogen.

投稿润色
补充资料

中图分类号:S43

DOI:10.3969/j.issn.1007-7146.2010.05.024

所属栏目:技术研究

基金项目:安徽省教育厅自然科学研究项目(KJ2007A095)

收稿日期:2009-12-28

修改稿日期:2010-07-29

网络出版日期:--

作者单位    点击查看

江 晶:安徽农业大学植物保护学院, 安徽 合肥 230036
江 彤:安徽农业大学植物保护学院, 安徽 合肥 230036
李 瑞:安徽农业大学植物保护学院, 安徽 合肥 230036
檀根甲:安徽农业大学植物保护学院, 安徽 合肥 230036

联系人作者:江晶(jiangjing917@163.com)

备注:江晶(1983-), 女, 硕士研究生,主要从事植物病原真菌研究。

【1】吴文平, 张志铭. 炭疽菌属(Colletotrichum Cda) 分类学研究 Ⅱ. 种的划分[J]. 河北农业大学学报, 1994, 17(2): 31-37.
WU Wen-ping, ZHANG Zhi-ming. Study on Taxonomy of Genus Colletotrichum Cda. Ⅱ. Division of Species [J]. Chinese Journal of Hebei Agricultural University, 1994, 17(2): 31-37.

【2】VON ARX J A. Die Arten der Gattung Colletotrichum Cda[J]. Phytopathol Z, 1957, 29: 413-468.

【3】VON ARX J A. A Revision of the Fungi Classified as Gloeosporium [J]. Biblioth Mycol, 1970, 24: 1-203.

【4】SUTTON B C.The Coelomycetes: Fungi Imperfecti with Pycnidia, Acervuli and Stromata [M]. England: Commonwealth Mycological Institute, 1980: 445.

【5】张志华, 洪葵. 核酸序列直接分析在真菌鉴定方面的应用[J].华南热带农业大学学报, 2006, 12(2): 39-43.
ZHANG Zhi-hua, HONG kui. Application of Nucleic ACID Sequence Analysis in Fungi Taxonmy[J]. Journal of South China University of Tropical Agricultural, 2006, 12 (2): 39-43.

【6】BROWN A E, SREENIVASAPRASAD S, TIMMER L W. Molecular Characterization of Slow-growing Orange and Key Lime Anthracnose Strains of Colletotrichum from Citrus as C.acutatum [J]. Phytopathology, 1996, 86(5): 523-527.

【7】MORIWAKI J, TSUKIBOSHI T, SATO T. Grouping of Colletotrichum Species in Japan Based on DNA Sequences [J]. J Gen Plant Pathol, 2002, 68(4): 307-320.

【8】MARTINEZ-CULEBRAS P V, QUEROL A, SUREZ-FERNANDEZ M B, et al. Phylogenetic Relationships among Colletotrichum Pathogens of Strawberry and Design of PCR Primers for Their Identification [J]. J Phytopathol, 2003, 151(3): 135-143.

【9】吴芳芳, 檀根甲. 苹果感染炭疽病菌后6种酶活性的变化[J]. 安徽农业大学学报, 2004, 31(1): 46-50.
WU Fang-fang, TAN Gen-jia. Changes in Six Kinds of Enzyme Activities in Apple Fruit Inoculated with C.gloeosprioides [J]. Chinese Journal of Anhui Agricultural University, 2004, 31 (1): 46-50.

【10】DAUCH A L, WATSON A K, JABAJI-HARE S H. Detection of the Biocontrol Agent Colletotrichum coccodes(183088) from the Target Weed Velvetleaf and from Soil by Stain-specific PCR Markers[J]. J Microbiol Methods, 2003, 55 (1): 51-64.

【11】HSIANG T, GOODWIN P H. Ribosomal DNA Sequence Comparisons of Colletotrichum graminicola from Turfgrasses and Other Hosts [J]. Eur J of Plant Pathol, 2001, 107(6): 593-599.

引用该论文

JIANG Jing,JIANG Tong,LI Rui,TAN Gen-jia. Molecular Identification and Detection of Apple Anthracnose Pathogen[J]. Acta Laser Biology Sinica, 2010, 19(5): 695-699

江 晶,江 彤,李 瑞,檀根甲. 苹果炭疽菌的分子鉴定与检测[J]. 激光生物学报, 2010, 19(5): 695-699

您的浏览器不支持PDF插件,请使用最新的(Chrome/Fire Fox等)浏览器.或者您还可以点击此处下载该论文PDF