热带作物学报2013，Vol.34Issue(2)：358-361,4.DOI:10.3969/j.issn.1000-2561.2013.02.030
巢式PCR检测琯溪蜜柚黑斑病病原菌
Detection of Pathogens Causing Citrus Black Spot of Guanxi Honey Pomelo by Nested PCR
摘要
Abstract
Citrus black spot is a fungal disease in most Guanxi honey pomelo. By using the specific primers, a unique 487 bp band was obtained by PCR amplification from genomic DNA of the pathogen causing citrus black spot of Guanxi honey pomelo and the method could accurately distinguish the honey pomelo black spot pathogen from the other disease pathogens of Sphaceloma fawcetti Jenk, Colletotrichum gleosporioides Penz and Mycosphaerella cirri White-Side in Guanxi honey pomelo. The sensitivity was determined by nested PCR. For the genomic DNA of honey pomelo black spot pathogen, the sensibility of nested PCR reached 100 fg/μL, 100 times higher than that of routine PCR. For the DNA of diseased tissues, the sensibility of nested PCR reached 100μg of DNA sample. Through nested PCR assay, the pathogen of honey pomelo black spot could be specifically detected form diseased and symptom latent infection honey pomelo tissues with the detection rate of 96.67% and 90%, respectively.关键词
琯溪蜜柚/黑斑病/内部转录间隔区/巢式PCR/检测Key words
Guanxi honey pomelo/ Citrus black spot/ ITS/ Nested-PCR/ Detection分类
农业科技引用本文复制引用
郑域茹,罗金水,张汉荣,卢松茂,林智明,王宏毅,李美桂..巢式PCR检测琯溪蜜柚黑斑病病原菌[J].热带作物学报,2013,34(2):358-361,4.基金项目
福建省公益类基本科研专项项目"琯溪蜜柚黑斑病巢式PCR检测技术的建立及应用"(No.2012GY01) （No.2012GY01）
福建省自然基金项目"琯溪蜜柚黑斑病病原菌RFLP检测鉴定技术"(No.2010J01113). （No.2010J01113）
