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Botanical microbiomes on the cheap: Inexpensive molecular fingerprinting methods to study plant-associated communities of bacteria and fungi.
Applications in Plant Sciences ( IF 3.6 ) Pub Date : 2020-04-15 , DOI: 10.1002/aps3.11334
David Johnston-Monje 1, 2 , Jessica Lopez Mejia 1
Affiliation  

High‐throughput sequencing technologies have revolutionized the study of plant‐associated microbial populations, but they are relatively expensive. Molecular fingerprinting techniques are more affordable, yet yield considerably less information about the microbial community. Does this mean they are no longer useful for plant microbiome research? In this paper, we review the past 10 years of studies on plant‐associated microbiomes using molecular fingerprinting methodologies, including single‐strand conformation polymorphism (SSCP), denaturing gradient gel electrophoresis (DGGE), amplicon length heterogeneity PCR (LH‐PCR), ribosomal intergenic spacer analysis (RISA) and automated ribosomal intergenic spacer analysis (ARISA), and terminal restriction fragment length polymorphism (TRFLP). We also present data juxtaposing results from TRFLP methods with those generated using Illumina sequencing in the comparison of rhizobacterial populations of Brazilian maize and fungal surveys in Canadian tomato roots. In both cases, the TRFLP approach yielded the desired results at a level of resolution comparable to that of the MiSeq method, but at a fraction of the cost. Community fingerprinting methods (especially TRFLP) remain relevant for the identification of dominant microbes in a population, the observation of shifts in plant microbiome community diversity, and for screening samples before their use in more sensitive and expensive approaches.

中文翻译:

廉价的植物微生物群落:便宜的分子指纹图谱方法,用于研究与植物相关的细菌和真菌群落。

高通量测序技术彻底颠覆了与植物相关的微生物种群的研究,但它们相对昂贵。分子指纹技术更实惠,但产生的微生物群落信息却少得多。这是否意味着它们不再对植物微生物组研究有用?在本文中,我们使用分子指纹技术回顾了过去十年来与植物相关的微生物组的研究,包括单链构象多态性(SSCP),变性梯度凝胶电泳(DGGE),扩增子长度异质性PCR(LH-PCR),核糖体基因间间隔物分析(RISA)和自动化核糖体基因间间隔物分析(ARISA),以及末端限制性片段长度多态性(TRFLP)。在比较巴西玉米的根际细菌种群和加拿大番茄根系的真菌调查中,我们还将提出TRFLP方法与使用Illumina测序生成的结果并列的数据。在这两种情况下,TRFLP方法都能以与MiSeq方法相当的分辨率水平获得所需结果,但成本却很少。社区指纹方法(尤其是TRFLP)对于确定种群中的优势微生物,观察植物微生物组群落多样性的变化以及在以更敏感,更昂贵的方式使用样品之前对其进行筛查具有重要意义。TRFLP方法以与MiSeq方法相当的分辨率达到了预期的结果,但成本却很小。社区指纹方法(尤其是TRFLP)对于确定种群中的优势微生物,观察植物微生物组群落多样性的变化以及在以更敏感和更昂贵的方式使用样品之前对其进行筛查具有重要意义。TRFLP方法可在与MiSeq方法相当的分辨率水平上产生所需的结果,但成本却很少。社区指纹方法(尤其是TRFLP)对于确定种群中的优势微生物,观察植物微生物组群落多样性的变化以及在以更敏感,更昂贵的方式使用样品之前对其进行筛查具有重要意义。
更新日期:2020-04-15
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