La Spada Federico, Cock Peter J A, Randall Eva, Pane Antonella, Cooke David E L, Cacciola Santa Olga
Department of Agriculture, Food and Environment, University of Catania, 95123 Catania, Italy.
The James Hutton Institute, Invergowrie, Dundee DD2 5DA, UK.
J Fungi (Basel). 2022 Mar 22;8(4):330. doi: 10.3390/jof8040330.
Isolation techniques supplemented by sequencing of DNA from axenic cultures have provided a robust methodology for the study of communities in agricultural and natural ecosystems. Recently, metabarcoding approaches have emerged as new paradigms for the detection of species in environmental samples. In this study, Illumina DNA metabarcoding and a conventional leaf baiting isolation technique were compared to unravel the variability of communities in different environments. Overall, 39 rhizosphere soil samples from a natural, a semi-natural and a horticultural small-scale ecosystem, respectively, were processed by both baiting and metabarcoding. Using both detection techniques, 28 out of 39 samples tested positive for . Overall, 1,406,613 internal transcribed spacer 1 (ITS1) sequences and 155 isolates were obtained, which grouped into 21 taxa, five retrieved exclusively by baiting ( and ), 12 exclusively by metabarcoding (//-like; unknown sp. 1; unknown sp. 2; unknown sp. 3; unknown sp. 4; unknown sp. 5) and four with both techniques (, , and ). Both techniques complemented each other in describing the variability of communities from natural and managed ecosystems and revealing the presence of rare or undescribed taxa.
通过对纯培养物的DNA进行测序来补充的分离技术,为研究农业和自然生态系统中的群落提供了一种强大的方法。最近,元条形码方法已成为检测环境样本中物种的新范例。在本研究中,对Illumina DNA元条形码技术和传统的叶诱捕分离技术进行了比较,以揭示不同环境中群落的变异性。总体而言,分别从自然、半自然和园艺小规模生态系统中采集了39个根际土壤样本,并用诱捕法和元条形码技术进行了处理。使用这两种检测技术,39个样本中有28个检测出呈阳性。总体而言,获得了1,406,613条内部转录间隔区1(ITS1)序列和155个分离株,它们分为21个分类单元,其中5个仅通过诱捕法获得(和),12个仅通过元条形码技术获得(//样;未知种1;未知种2;未知种3;未知种4;未知种5),4个通过两种技术都获得(、、和)。这两种技术在描述自然和人工管理生态系统中群落的变异性以及揭示稀有或未描述的分类单元的存在方面相互补充。