Soil prokaryotic and eukaryotic diversity in translocated meadows, surrounding area and control meadows

To combine the conservation of valuable wet meadows with the reclamation of a post-industrial area, 1.3 hectares of turf were translocated from a build-up area to an abandoned quarry. Three of the Molinion meadows were compared four years following their translocation to habitats surrounding translocated meadows and control meadows (remained near the donor site). The composition of the main groups of organisms in the soil: bacteria, archaea, fungi, microfauna and unicellular algae, using a barcoded amplification of the soil DNA followed by Illumina sequencing was performed. Genomic DNA was extracted with the Soil DNA Purification Kit (EURx) using 0.25 g of mixed soil. For the amplification of the prokaryotes, the universal primers 515F and 926R targeting the hypervariable V4-V5 region of the 16S rRNA gene were used with a forward primer tagged. For the detection of the prominent soil eukaryotes, including Fungi, Chlorophyta, Alveolata, Rhizaria, Stramenopila and groups of mesofauna (such as Nematoda, Annelida and Collembola), the PCR targeting ITS2 marker gene was carried out using a mixture of 19 modified ITS3ngs forward primers and a tagged degenerate ITS4ngs reverse primer in line with Tedersoo et al., 2016.

为实现珍贵湿草甸的保护与后工业区土地修复的协同目标，研究团队将1.3公顷的草皮从建筑堆积区域移栽至一处废弃采石场。移栽四年后，研究人员对3处莫利尼欧草甸（Molinion meadows）开展对比研究，对照样本涵盖移栽草甸周边生境中的草甸，以及留存于供体场地附近的对照草甸。本研究采用土壤DNA条形码扩增结合Illumina测序（Illumina sequencing）技术，对土壤中主要生物类群——细菌、古菌、真菌、微型动物及单细胞藻类的群落组成进行表征。实验称取0.25g混合土壤样品，使用EURx品牌土壤DNA纯化试剂盒（Soil DNA Purification Kit）提取基因组DNA。针对原核生物的扩增，研究采用靶向16S rRNA基因高变区V4-V5的通用引物515F与926R，并对正向引物进行标签修饰。为检测土壤中主要真核生物类群，包括真菌界（Fungi）、绿藻门（Chlorophyta）、囊泡虫类（Alveolata）、根足类（Rhizaria）、不等鞭毛类（Stramenopila）及中型土壤动物类群（如线虫动物门（Nematoda）、环节动物门（Annelida）、弹尾目（Collembola）），研究参照Tedersoo等（2016）的实验方案，采用19条修饰后的ITS3ngs正向引物与带标签的简并ITS4ngs反向引物组合，针对ITS2标记基因开展PCR扩增。