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首页> 外文期刊>European Journal of Plant Pathology >Application of PCR-RFLP technique to species identification and phylogenetic analysis of Streptomyces associated with potato scab in Brazil based on partial atpD gene sequences
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Application of PCR-RFLP technique to species identification and phylogenetic analysis of Streptomyces associated with potato scab in Brazil based on partial atpD gene sequences

机译：PCR-RFLP技术在基于局部ATPD基因序列的基于部分ATPD基因序列的菌马铃薯结痂鉴定与系统发育和系统发育分析

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摘要

In the present study the atpD gene was evaluated as an alternative molecular marker for differentiation of Streptomyces species causing potato scab and to establish the phylogenetic relationships within this group. PCR-RFLP experiments using HaeIII and CfoI restriction enzymes allowed differentiation of most Streptomyces species, except the genomospecies S. reticuliscabiei and S. turgidiscabies which showed identical profiles between them. Phylogenetic relationships within Streptomyces spp. were also examined by comparing partial atpD gene sequences (858 nt) and the strains were separated in three main groups: Group I, supported by bootstrap values of 79%, comprised of S. scabiei, S. europaeiscabiei, S. stelliscabiei, S. reticuliscabiei, S. turgidiscabies and S. acidiscabies; Group II, supported by bootstrap values of 100%, comprised of S. sampsonii; and Group III supported by bootstrap values of 100%, comprised of S. caviscabies, S. setonii, S. luridiscabiei and S. puniciscabiei. The atpD gene sequence similarity observed among the species in this study ranged from 89.2% to 100%. Twenty-nine Streptomyces isolates obtained from potato growing areas in Brazil were analyzed by PCR-RFLP of atpD gene technique and identified as S. scabiei (15 strains), S. caviscabies/ S. setonii (12) and S. sampsonii (2). Using this tool S. europaeiscabiei strains were also detected from import material from the Netherlands. The presence of genes associated with the S. turgidiscabies pathogenicity island (PAI) was also determined and potato tuber slice assays were performed to confirm the pathogenicity of the strains. The data obtained herein indicated that the PCR-RFLP of atpD gene technique can be used for diagnosis and identification of phytopathogenic Streptomyces strains, representing a rapid and inexpensive tool. Moreover, this gene can be useful for phylogenetic analysis of Streptomyces spp. associated with potato scab.
机译：在本研究中，ATPD基因被评价为替代分子标记，用于分化导致马铃薯结痂的细胞瘤性物种并建立该组内的系统发育关系。使用HAEIII和CFOI限制酶的PCR-RFLP实验允许分化大多数链霉菌物种，但基因组织S. Reticuliscabiei和S. Turgidiscabies，在它们之间显示出相同的轮廓。 STELTOMYCES SPP中的系统发育关系。还通过比较部分ATPD基因序列（858nt）来检查（858nt），并且在三个主要组中分离菌株：I基团，由Bootstrap值支持79％，由S.Scabiei，S. Europaeiscabiei，S. Stelliscabiei，S。 Reticuliscabiei，S. Turgidiscabies和S. Cateriscabies; II集团由191％的引导价值支持100％，由S. Sampsonii组成;和第三组由举止的引导价值100％支持，由S. Caviscabies，S. Setonii，S. luridiscabiei和S.Puniciscabiei提供。本研究中物种中观察到的ATPD基因序列相似度范围为89.2％至100％。通过ATPD基因技术的PCR-RFLP分析了从巴西的马铃薯生长区域获得的二十九种链霉菌分离物，并确定为S.Scabiei（15株），S. Caviscabies / S. Setonii（12）和S. Sampsonii（2） 。使用该工具S. Ouropaeiscabiei菌株也被从荷兰的进口材料中检测到。还确定了与S. Turgidiscabies致病性岛（PAI）相关的基因的存在，并进行马铃薯块茎切片测定以确认菌株的致病性。本文获得的数据表明ATPD基因技术的PCR-RFLP可用于诊断和鉴定植物疗法链霉菌菌株，代表一种快速和廉价的工具。此外，该基因可用于链霉菌SPP的系统发育分析。与马铃薯结痂有关。

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