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Phylotype and sequevar determination and AFLP fingerprinting of Ralstonia solanacearum strains causing bacterial wilt of potato in southeastern Iran
European Journal of Plant Pathology ( IF 1.7 ) Pub Date : 2020-05-28 , DOI: 10.1007/s10658-020-02018-5
Ali Mollae , Akbar Hosseinipour , Mahdi Azadvar , Hossain Massumi , Fatemeh Ebrahimi

Bacterial wilt of potato caused by Ralstonia solanacearum is a severe threat to potato cultivation in Kerman province, located in southeastern Iran and the third largest potato producer in the country. The present study intends to determine the precise phylogenetic position, sequevar, and genetic diversity of R. solanacearum strains associated with the disease. To this end, the major potato-growing regions of the province (Jiroft and Laleh Zaar) were surveyed to collect tuber samples from affected potato plants showing typical wilt symptoms. Twenty-three strains were collectively purified and subjected to phenotypic and genotypic characterization. Biochemical tests showed that all the strains belonged to R. solanacearum biovar 2A. Furthermore, phylotype-specific multiplex polymerase chain reaction results indicated that the strains belonged to phylotype II. For sequevar typing as well as the determination of the phylotype subcluster (IIA/IIB), polymerase chain reaction amplicons of the endoglucanase precursor gene of 11 representative strains were sequenced. In phylogenetic analysis, the strains were grouped with the reference strains of phylotype IIB-1 retrieved from the GenBank. Then, to assess genetic diversity, 19 selected strains were subjected to genomic fingerprinting, i.e., amplified fragment length polymorphism (AFLP) using seven primer pair combinations. The AFLP data showed heterogeneity among the strains of R. solanacearum and were grouped into four clusters. With our knowledge, this is the first report of the phylotype, and sequevar determination (i.e., IIB-1) in addition to AFLP-based genomic fingerprinting of R. solanaceraum strains causing potato wilt in southeastern Iran.



中文翻译:

伊朗东南部引起马铃薯枯萎病的青枯雷尔氏菌菌株的型型和后代测定及AFLP指纹图谱

Ralstonia solanacearum引起的马铃薯细菌性枯萎病严重威胁着位于伊朗东南部,该国第三大马铃薯生产国的克尔曼省的马铃薯种植。本研究旨在确定精确的系统发育位置,sequevar,和遗传多样性- [R 与青枯菌菌株有关的疾病。为此,对该省主要的马铃薯种植区(Jiroft和Laleh Zaar)进行了调查,以从受影响的马铃薯植株中采集块茎样品,这些样品显示出典型的枯萎症状。总共纯化了二十三个菌株,并进行了表型和基因型表征。生化试验表明,所有菌株均属于青枯biovar 2A。此外,系统型特异性多重聚合酶链反应结果表明,该菌株属于系统型II。为了进行后代分型和确定系统型亚簇(IIA / IIB),对11个代表性菌株的内切葡聚糖酶前体基因的聚合酶链反应扩增子进行了测序。在系统发育分析中,将菌株与从GenBank检索到的系统型IIB-1参考菌株进行分组。然后,为了评估遗传多样性,使用七个引物对组合对19个所选菌株进行了基因组指纹分析,即扩增的片段长度多态性(AFLP)。AFLP数据显示青枯菌菌株之间的异质性并分为四个集群。据我们所知,这是首次导致伊朗东南部马铃薯枯萎的青枯雷尔氏菌菌株的系统型和后代测定(即IIB-1)报告,此外还有基于AFLP的基因组指纹分析。

更新日期:2020-05-28
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