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Evaluation of BOX-PCR and REP-PCR as Molecular Typing Tools for Antarctic Streptomyces
Current Microbiology ( IF 2.3 ) Pub Date : 2020-09-16 , DOI: 10.1007/s00284-020-02199-6
Marcela Proença Borba 1 , Ana Elisa Ballarini 1 , Joao Paulo Duarte Witusk 1 , Paris Lavin 2, 3 , Sueli Van Der Sand 1
Affiliation  

Molecular studies led to the resurgence of natural products research from genus Streptomyces, already known for their long history and importance for the pharmaceutical industry. However, species belonging to this genus are difficult to identify and the most commonly used techniques, which are based on 16S rRNA sequencing, do not discriminate between related species. In this work, amplification profiles generated from BOX-PCR and REP-PCR of 49 Antarctic soil streptomycetes were compared to evaluate the diversity present in the group and to characterize the bacterial isolates, along with some 16S rRNA amplifications. The BOX-A1R primer exhibit clearer amplification fragments, different from the amplification patterns obtained using the REP 1R and 2R primers. A higher diversity was observed with REP-PCR amplifications, even though a larger number of fragments was obtained with BOX-A1R primer amplifications. There are at least four isolates that showed great similarity (about 90%) in both techniques. In other hand, there are two others that are 90% similar in BOX-PCR, but distant in REP-PCR, showing only 40% of similarity. Results of the combination of BOX-PCR and REP-PCR represent a simple and low-cost method to discriminate between Streptomyces strains. There is no species identification with only the 16S rRNA, most isolates seem to be related to S. globisporus. Further studies added to the obtained results may provide better data to help the characterization of these microorganisms.

中文翻译:

BOX-PCR 和 REP-PCR 作为南极链霉菌分子分型工具的评估

分子研究使链霉菌属的天然产物研究重新兴起,链霉菌属以其悠久的历史和对制药行业的重要性而闻名。然而,属于该属的物种难以识别,并且基于 16S rRNA 测序的最常用技术不区分相关物种。在这项工作中,比较了 49 个南极土壤链霉菌的 BOX-PCR 和 REP-PCR 产生的扩增图谱,以评估该组中存在的多样性并表征细菌分离株,以及一些 16S rRNA 扩增。BOX-A1R 引物显示出更清晰的扩增片段,不同于使用 REP 1R 和 2R 引物获得的扩增模式。通过 REP-PCR 扩增观察到更高的多样性,尽管使用 BOX-A1R 引物扩增获得了大量片段。至少有四种分离株在两种技术中表现出极大的相似性(约 90%)。另一方面,还有另外两个在 BOX-PCR 中具有 90% 的相似性,但在 REP-PCR 中相距较远,仅显示 40% 的相似性。BOX-PCR 和 REP-PCR 组合的结果代表了一种简单且低成本的方法来区分链霉菌属菌株。只有 16S rRNA 没有物种鉴定,大多数分离株似乎与 S. globisporus 相关。对获得的结果进行进一步研究可能会提供更好的数据,以帮助表征这些微生物。另外两个在 BOX-PCR 中具有 90% 的相似性,但在 REP-PCR 中相距较远,仅显示 40% 的相似性。BOX-PCR 和 REP-PCR 组合的结果代表了一种简单且低成本的方法来区分链霉菌属菌株。只有 16S rRNA 没有物种鉴定,大多数分离株似乎与 S. globisporus 相关。对获得的结果进行进一步研究可能会提供更好的数据,以帮助表征这些微生物。另外两个在 BOX-PCR 中具有 90% 的相似性,但在 REP-PCR 中相距较远,仅显示 40% 的相似性。BOX-PCR 和 REP-PCR 组合的结果代表了一种简单且低成本的方法来区分链霉菌属菌株。只有 16S rRNA 没有物种鉴定,大多数分离株似乎与 S. globisporus 相关。对获得的结果进行进一步研究可能会提供更好的数据,以帮助表征这些微生物。
更新日期:2020-09-16
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