利用PCR-RFLP技术诊断马铃薯腐病线虫Diylenchus destructor

Niloufar Mahmoudi, E. Pakina, Liudmila A. Limantceva, A. Ivanov
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引用次数: 2

摘要

2019年,在俄罗斯莫斯科地区对线虫进行的一次调查中,从马铃薯植株的块茎中发现了一种已知的线虫。Destructor属是植物寄生线虫中最有问题的属之一。该属的众多物种已被各种来源引用。由于许多物种的形态相似,且缺乏分离特征,破坏D.destructor的鉴定很困难。利用分子分类学和系统发育学对其进行鉴定。在目前的研究中,为扩增ITS rRNA基因的PCR-RFLP说明性模型提供了两种可以识别马铃薯块茎中的D.destructor的酶。对收集的群体进行了跨越ITS1-ITS2两个区域的rDNA序列的分析。用TaqI酶消化ITS1-5.8S-ITS2区的PCR产物产生三个片段;100、190、550,并且使用Tru1I,产生两个片段;300和480bp。将获得的DNA序列与在其他国家分离的人群的GenBank中保存的DNA序列进行比较。结果表明,从不同寄主植物物种分离的种群之间没有区别,包括在俄罗斯联邦发现的种群。ITS rRNA的新序列以登录号MN122076、MN658597、MN658599、MN657637和MN658638存放在GenBank中。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Diagnosis of potato rot nematode Ditylenchus destructor using PCR-RFLP
During an investigation of nematodes in the Moscow region of Russia in 2019, a known species Ditylenchus destructor was recovered from tubers of potato plants. The genus Destructor is one of the most problematic genera of plant-parasitic nematodes. The numerous species reported for this genus have been cited from various sources. Due to the morphological similarity of many species and the lack of separation characteristics, the identification of D. destructor is difficult. Molecular taxonomy and phylogeny were used to confirm the identification. In the current study, PCR-RFLP illustrative models for the amplification of the ITS-rRNA gene were provided with two enzymes that could recognize D. destructor in potato tubers. Analysis of the rDNA sequences spanning both ITS1-ITS2 regions was carried out on the collected populations. The digestion of the PCR product of the ITS1-5.8S-ITS2 region with the enzyme TaqI produced three fragments; 100, 190, 550, and with Tru1I, two fragments were produced; 300 and 480 bp. The obtained DNA sequences were compared with those DNA sequences deposited in GenBank of populations isolated in other countries. The results showed no distinction between populations isolated from different host plant species, including populations found in the Russian Federation. New sequences from ITS-rRNA were deposited in the GenBank under accession number MN122076, MN658597, MN658599, MN658637, MN658638.
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