Rossmann Simeon, Lysøe Erik, Skogen Monica, Talgø Venche, Brurberg May Bente
Division of Biotechnology and Plant Health, Norwegian Institute of Bioeconomy Research (NIBIO), Ås, Norway.
Department of Plant Sciences, Norwegian University of Life Sciences (NMBU), Ås, Norway.
Front Microbiol. 2021 Mar 25;12:637068. doi: 10.3389/fmicb.2021.637068. eCollection 2021.
Plants with roots and soil clumps transported over long distances in plant trading can harbor plant pathogenic oomycetes, facilitating disease outbreaks that threaten ecosystems, biodiversity, and food security. Tools to detect the presence of such oomycetes with a sufficiently high throughput and broad scope are currently not part of international phytosanitary testing regimes. In this work, DNA metabarcoding targeting the internal transcribed spacer (ITS) region was employed to broadly detect and identify oomycetes present in soil from internationally shipped plants. This method was compared to traditional isolation-based detection and identification after an enrichment step. DNA metabarcoding showed widespread presence of potentially plant pathogenic and species in internationally transported rhizospheric soil with being the overall most abundant genus observed. Baiting, a commonly employed enrichment method for species, led to an increase of golden-brown algae in the soil samples, but did not increase the relative or absolute abundance of potentially plant pathogenic oomycetes. Metabarcoding of rhizospheric soil yielded DNA sequences corresponding to oomycete isolates obtained after enrichment and identified them correctly but did not always detect the isolated oomycetes in the same samples. This work provides a proof of concept and outlines necessary improvements for the use of environmental DNA (eDNA) and metabarcoding as a standalone phytosanitary assessment tool for broad detection and identification of plant pathogenic oomycetes.
在植物贸易中远距离运输的带有根系和土块的植物可能携带有植物病原卵菌,从而引发威胁生态系统、生物多样性和粮食安全的疾病爆发。目前,能够以足够高的通量和广泛的范围检测此类卵菌存在的工具并非国际植物检疫检测体系的一部分。在这项研究中,靶向内部转录间隔区(ITS)区域的DNA宏条形码技术被用于广泛检测和鉴定国际运输植物土壤中存在的卵菌。在富集步骤之后,该方法与传统的基于分离的检测和鉴定方法进行了比较。DNA宏条形码技术显示,在国际运输的根际土壤中广泛存在潜在的植物病原卵菌物种,其中腐霉属是观察到的总体上最丰富的属。诱饵法是一种常用于检测腐霉属物种的富集方法,它导致土壤样品中的金藻数量增加,但并未增加潜在植物病原卵菌的相对丰度或绝对丰度。根际土壤的宏条形码分析产生了与富集后获得的卵菌分离株相对应的DNA序列,并正确地鉴定了它们,但并不总是能在相同样品中检测到分离出的卵菌。这项工作提供了一个概念验证,并概述了将环境DNA(eDNA)和宏条形码技术用作独立的植物检疫评估工具以广泛检测和鉴定植物病原卵菌所需的改进措施。