ITS_OTU

Six oak bark pieces (~1g) from four oak trees (three pieces from oak 1 and one piece from each of the other trees) in Northern Germany were collected. In addition an infusion (20g oak bark in a sterile tea bag in 300 mL water for 24 hours) for each tree was prepared. DNA was extracted from all samples using the Soil DNA Kit from Omega Bio-Tek. The resulting DNA was sent to LGC Genomics (GmbH, Berlin, Germany) for amplification of the fungal ITS (ITS1, 5.8S and ITS2) sequences using ITS1f (CTTGGTCATTTAGAGGAAGTAA) and ITS4 (TCCTCCGCTTATTGATATGC) primers using the 454 GS FLX+ Titanium Sequencer (Roche). We used the “pairwise.seqs” command in mothur for the OTU-based analysis with ignoring the penalization of the sequence ends. The shared file shows the distribution of OTUs in the ten groups at 97% and 95% identity level.

本研究从德国北部的4株橡树中共采集6份树皮样本，每份约1g，其中1号橡树提供3份，其余3株橡树各提供1份。此外，为每株橡树制备树皮浸提液：将20g橡树树皮装入无菌茶包，加入300mL蒸馏水浸泡24小时。所有样本均采用Omega Bio-Tek公司的土壤DNA提取试剂盒（Soil DNA Kit）完成DNA提取。提取获得的DNA被送往位于德国柏林的LGC Genomics（GmbH）公司，使用引物ITS1f（序列：CTTGGTCATTTAGAGGAAGTAA）与ITS4（序列：TCCTCCGCTTATTGATATGC）对真菌内转录间隔区（ITS，包含ITS1、5.8S及ITS2区段）进行扩增，并通过罗氏（Roche）454 GS FLX+ Titanium测序仪完成测序。本研究使用mothur软件中的"pairwise.seqs"命令开展基于操作分类单元（OTU）的分析，分析过程中忽略序列末端的罚分处理。共享文件展示了10个样本组在97%与95%序列相似度阈值下的OTU分布情况。