1种苎麻根腐病线虫的鉴定

Identification of a nematode isolate from rot root of ramie

【目的】对分离的一种苎麻根腐病线虫进行形态和分子生物学鉴定,为病原线虫分子生物学检测技术的研发及苎麻根腐病的防治奠定基础。【方法】从感染苎麻根腐病植株根中分离出一种植物病原线虫,利用显微技术观察线虫的形态特征,对其进行初步鉴定;之后采用分子生物学技术,提取单条线虫的DNA,利用通用保守性引物扩增了线虫核糖体DNA内部转录间隔区(rDNA-ITS)序列,并用软件DNAstar和DNAMAN进行序列分析,对该植物病原线虫进行进一步鉴定。【结果】根据形态特征,将获得的苎麻根腐病线虫初步鉴定为咖啡短体线虫。序列分析结果表明,该线虫rDNA-ITS序列与咖啡短体线虫湖北种群、越南种群、台湾种群、日本种群及尼日利亚种群的同源性依次为99.8%,99.0%,98.0%,97.8%和94.6%。该线虫rDNA-ITS序列在GenBank注册号为HQ403649。【结论】形态及分子生物学鉴定结果表明,分离的苎麻根腐病线虫为咖啡短体线虫(Pratylenchus coffeae)。

【Objective】 The study conducted morphological and molecular identification of a plant parasitic nematode isolated from rot root of ramie,which may lay a foundation for developing a molecular detection method of pathogen and integrated control.【Method】 A plant pathogenic nematode was isolated from rot root of ramie in Yuanjiang.The nematode was investigated under the microscope for initial identification.rDNA-ITS region was amplified using DNA extracted from single nematode and sequenced.The sequence was analysed by DNAstar and DNAMAN software for further identification.【Result】 The nematode was preliminarily identified as Pratylenchus coffeae by morphological characters.Furthermore,the identity of rDNA-ITS sequence is 99.8%,99.0%,98.0%,97.8% and 94.6% with populations from Hubei（China）,Vet Nam,Taiwan,Japan and Nigeria,respectively.The GenBank accession number of this rDNA-ITS sequence is HQ403649.【Conclusion】 The morphological and molecular character indicated that the nematode was P.coffeae.