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红掌胶胞炭疽菌的分子检测
Molecular detection of Colletotrichum gloeosporioides in Anthurium andraeanum
【摘要】 胶胞炭疽菌是引起红掌炭疽病的病原菌。根据GenBank中炭疽属不同种的ITS序列差异,设计了胶胞炭疽菌的特异性引物E1/E2,由此建立的PCR检测体系可以从38个胶胞炭疽菌菌株中扩增得到329 bp的特异性条带,而扩增其它近似或相关菌株时没有相应的特异性条带。该检测体系对胶胞炭疽菌基因组DNA的扩增灵敏度达到10 pg。将引物E1/E2与ITS区通用引物进行套式PCR扩增后,检测灵敏度至少提高10 000倍。当土中胶胞炭疽菌分生孢子达到200个/g土时可检测出。进一步利用此检测体系对携带病原菌的灌溉水、发病组织进行检测,均能快速稳定地检测出病原菌。
【Abstract】 Colletotrichum gloeosporioides caused anthracnose in Anthurium andraeanum.Based on diffe-rences in internal transcribed space(ITS) sequences of Colletotrichum genus,a pair of species-specific pri-mers,E1 and E2,was synthesized.The primers amplified a single PCR band of 329 bp with DNA extracted from C.gloeosporioides isolated from A.andraeanum,while other relative strains had no corresponding band.The detection sensitivity was 10 pg of genomic DNA.Using ITS1/ITS4 as the first round primers and E1/E2 as the second round primers,the detection sensitivity increased 10 000-fold to 10 fg.The detection sensitivity for the soil pathogens was 200 conidia/g soil.The PCR-based method developed here could stably and quickly detect the pathogen from water samples and diseased plant.
【Key words】 Colletotrichum gloeosporioides; Anthurium andraeanum; molecular detection; nest PCR;
- 【文献出处】 植物病理学报 ,Acta Phytopathologica Sinica , 编辑部邮箱 ,2008年02期
- 【分类号】S436.8
- 【被引频次】36
- 【下载频次】470