META-BARCODING BASED ASSESSMENT OF WATER ENVIRONEMTAL DNA TO REVEAL ABUNDANCCE OF FRESHWATER INSECTS USING NEXT-GENELATION SEQUENCING
-
- YEAGASHI Sakiko
- 山梨大学 学院総合研究部工学域土木環境工学系
-
- HOSOKAWA Hiroki
- 愛媛大学 工学部環境建設工学科
-
- WATANABE Kozo
- 愛媛大学 工学部環境建設工学科
Bibliographic Information
- Other Title
-
- 河川水中の環境DNAの次世代シーケンス解析を利用した水生昆虫の群集構造および生息個体数推定の可能性：従来型定量評価手法と比較して
- カセン スイチュウ ノ カンキョウ DNA ノ ジセダイ シーケンス カイセキ オ リヨウ シタ スイセイ コンチュウ ノ グンシュウ コウゾウ オヨビ セイソク コタイスウ スイテイ ノ カノウセイ : ジュウライ カタ テイリョウ ヒョウカ シュホウ ト ヒカク シテ
Search this article
Abstract
In order to examine that eDNA abundance could predict abundance of source freshwater insects, we performed meta-barcording analysis of freshwater insects using water environmental DNA (eDNA) in Shigenobu River, Ehime, Japan by Next generation sequencing (NGS) analysis. The eDNA, which is released from various organisms, is one of the most stractiong way to reveal and monitor species diverstity of aquatic animals. Although NGS analysis was employed meta-barcoding analysis for eDNA, a few studies reported the relationship between abundance of freshwater insects and regerated reads generated from NGS. At first, we collected 2L of river water from 12 sites and freshwater insects from 11 sites, 3 times. The water samples were filterd by 0.2 um membrane filter. DNA extraction was performed by Phenol-Chloroform method and ethanol presipitation. Extracted DNAs were purified by DNeasy Blood and Tissue kit (Qiagen) again. Chytochrome Oxidase I reagion was amplified using universal primer for Insects. NGS analysis was performed by MiSeq using v3 reagents (illumina). After quality control and reads screaning using primer sequence, operational taxonomic units (OTU) were clustered. The source animals of OTUs were searched in NCBI data base by BLAST. In result, we detected almost al families, which collected from field, from eDNA. The abudance of freshwater insects was compared with reads abundance. In result, we detected a lot of non-target species (e.g. microorganisms, crastaceans, inland insects). When high number of insect reds, PCR procedure should be improved. The number of freshwater insects showed positive relationship to read abundance from eDNA. It supported that eDNA analysis using NGS predict the abundance of freshwater insects.
Journal
-
- Journal of Japan Society of Civil Engineers, Ser. G (Environmental Research)
-
Journal of Japan Society of Civil Engineers, Ser. G (Environmental Research) 73 (7), III_139-III_147, 2017
Japan Society of Civil Engineers