A study of macroinvertebrate communities in Bolshiye Koty Bay of Lake Baikal using DNA metabarcoding

L. S. Kravtsova, T. E. Peretolchina, T. I. Triboy, I. A. Nebesnykh, A. E. Tupikin, M. R. Kabilov
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Abstract

The diversity of macroinvertebrates, the structure of their communities in Bolshiye Koty Bay (Lake Baikal) was studied by a DNA metabarcoding approach using an Illumina MiSeq system. Internal primer mlCOIintF in combination with jgHCO2198 of the Folmer fragment of the COI gene were used for macroinvertebrate metabarcoding. A total of 118009 reads of the COI gene fragment (at least 313 bp in length) were obtained. The correlation of the Spearman coefficient (S = 0.6, p < 0.05) with the abundance of macroinvertebrates in the samples before DNA extraction showed that the number of reads can serve as an indirect characteristic of the abundance of a species (operational taxonomic unit, OTU). 115 OTUs belonging to the higher taxa of macroinvertebrates were identified: Porifera, 1; Platyhelminthes, 3; Annelida, 38; Arthropoda, 55; Mollusca, 18. At a high level of resolution (with homology with GenBank reference sequences ≥ 95 %, coverage ≥ 90 %), 46 taxa of macroinvertebrates comprising three communities were registered: one dominated by molluscs (Choanomphalus conf. maacki) and two dominated by chironomids (Orthocladius grega rius Linev., Sergentia baicalensis Tshern.). Communities are characterized by low species diversity according to Shannon (from 0.7 to 1.2 bits), high concentration of dominance according to Simpson (from 0.5 to 0.7) and low evenness according to Pielou (from 0.3 to 0.4). Dominants and subdominants in the communities account for 91 to 96 % of COI gene fragment reads. The spatial distribution of the dominant species identified in the communities is influenced by the geomorphological features of the bottom and the composition of sediments in the area studied. The approach proposed for studying the structure of macroinvertebrate communities based on DNA metabarcoding and next generation sequencing can be recommended for express assessment of the state of aquatic ecosystems in the monitoring.
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基于DNA元条形码的贝加尔湖Bolshiye Koty湾大型无脊椎动物群落研究
利用Illumina MiSeq系统,采用DNA元条形码方法研究了贝加尔湖Bolshiye Koty湾大型无脊椎动物的多样性及其群落结构。利用内部引物mlCOIintF与COI基因Folmer片段的jgHCO2198结合进行大型无脊椎动物元条形码编码。共获得COI基因片段118009个reads(长度至少为313 bp)。Spearman系数的相关性(S = 0.6, p <0.05),与DNA提取前样品中大型无脊椎动物的丰度比较表明,reads数可以作为物种丰度(operational taxonomic unit, OTU)的间接特征。共鉴定出大型无脊椎动物高等分类群115个otu: Porifera, 1;扁形动物门,3;环节动物门,38岁;节肢动物门,55岁;软体动物类,18岁。在高分辨率(与GenBank参考序列同源性≥95%,覆盖率≥90%)下,共登记到3个群落的46个大型无脊椎动物类群:一个以软体动物(Choanomphalus confs . maacki)为主,两个以手性动物(Orthocladius grega rius Linev)为主。黄芩(Sergentia baicalensis Tshern);Shannon的物种多样性指数为0.7 ~ 1.2 bits, Simpson的优势度指数为0.5 ~ 0.7 bits, Pielou的优势度指数为0.3 ~ 0.4 bits,群落的均匀度指数为低。群落中的显性和亚显性占COI基因片段reads的91% ~ 96%。群落中优势种的空间分布受研究区域底部地貌特征和沉积物组成的影响。提出的基于DNA元条形码和下一代测序的大型无脊椎动物群落结构研究方法,可用于监测中水生生态系统状态的快速评价。
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