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首页> 外文期刊>Nematology >Development of an efficient PCR-based diagnosis protocol for the identification of the pinewood nematode, Bursaphelenchus xylophilus (Nematoda : Aphelenchoididae)
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Development of an efficient PCR-based diagnosis protocol for the identification of the pinewood nematode, Bursaphelenchus xylophilus (Nematoda : Aphelenchoididae)

机译：开发一种基于PCR的高效诊断方案，用于鉴定松材线虫，松材线虫（线虫：Aphelenchoididae）

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摘要

Pine wood wilt disease caused by the pine wood nematode, Bursaphelenchus xylophilus, has been a serious problem in the southern regions of Korea. Efficient diagnosis of B. xylophilus from infected pine wood specimens is critical for the management of this pest. Traditional microscopic examination often results in an erroneous identification because a closely related non-pathogenic species, B. mucronatus, has a great degree of morphological similarity to B. xyophilus. In an attempt to search for reliable molecular markers for the discrimination of these species, we have cloned the 5S rRNA genomic DNA fragments containing both coding and intergenic spacer (IGS) regions from B. xylophilus and B. mucronatus through a homology-probing PCR strategy. Sequence analyses revealed that coding sequences of the 5S rRNA gene from the two species are almost identical (98.3% homology) but that the IGS sequences differ substantially between the species. Based on the IGS sequence differences (69.7% homology), we designed species-specific primer sets and developed a PCR-based diagnosis protocol for the identification and discrimination of the two nematode species on a molecular basis.
机译：由松木线虫Bursaphelenchus xylophilus引起的松木枯萎病在韩国南部地区已成为严重的问题。从受感染的松木标本中有效诊断木糖双歧杆菌对于控制该害虫至关重要。传统的显微镜检查通常会导致错误的识别，因为密切相关的非致病性物种粘液芽孢杆菌与木糖双歧杆菌在形态学上有很大的相似性。为了寻找可靠的分子标记来区分这些物种，我们通过同源性探测PCR策略克隆了木糖双歧杆菌和粘液双歧杆菌的5S rRNA基因组DNA片段，其中既包含编码区又包含基因间隔区（IGS）。 。序列分析显示，来自两个物种的5S rRNA基因的编码序列几乎相同（98.3％同源性），但是两个物种之间的IGS序列有很大不同。基于IGS序列差异（69.7％的同源性），我们设计了物种特异性引物组，并开发了基于PCR的诊断方案，用于在分子基础上鉴定和区分两种线虫。

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