利用从斯里兰卡分离株克隆的重复DNA序列开发用于立枯丝核菌的DNA探针和基于PCR的诊断检测方法。
Development of a DNA probe and a PCR based diagnostic assay for Rhizoctonia solani using a repetitive DNA sequence cloned from a Sri Lankan isolate.
作者信息
Weerasena O V D S Jagathpriya, Chandrasekharan N Vishvanath, Wijesundera Ravi L C, Karunanayake Eric H
机构信息
Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Colombo, Colombo 8, Sri Lanka.
出版信息
Mycol Res. 2004 Jun;108(Pt 6):649-53. doi: 10.1017/s095375620400989x.
Rhizoctonia solani is a destructive fungal pathogen of many economically important plants all over the world and the causative organism of sheath blight of rice in many tropical countries including Sri Lanka. A repetitive sequence from the genome of R. solani was cloned and characterized with a view to develop a DNA probe and a PCR diagnostic assay for detection of the fungus. The cloned sequence was 1550 bp long and appeared to be interspersed throughout the genome. The cloned sequence hybridized only to R. solani DNA and was sensitive enough to detect 100 pg of R. solani genomic DNA. PCR primers were designed from the cloned sequence and it was possible to develop a PCR assay for the specific detection of the fungal DNA with 10 pg sensitivity.
立枯丝核菌是一种对全球许多具有重要经济价值的植物具有毁灭性的真菌病原体，也是包括斯里兰卡在内的许多热带国家水稻纹枯病的致病生物。为了开发用于检测该真菌的DNA探针和PCR诊断方法，从立枯丝核菌的基因组中克隆并鉴定了一个重复序列。克隆的序列长1550 bp，似乎散布在整个基因组中。该克隆序列仅与立枯丝核菌DNA杂交，灵敏度足以检测100 pg立枯丝核菌基因组DNA。根据克隆的序列设计了PCR引物，并开发出了一种PCR检测方法，可特异性检测真菌DNA，灵敏度为10 pg。
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