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Forskningsartikel2019Vetenskapligt granskadÖppen tillgång

Establishing arthropod community composition using metabarcoding: Surprising inconsistencies between soil samples and preservative ethanol and homogenate from Malaise trap catches

Marquina, Daniel; Esparza-Salas, Rodrigo; Roslin, Tomas; Ronquist, Fredrik

Sammanfattning

DNA metabarcoding allows the analysis of insect communities faster and more efficiently than ever before. However, metabarcoding can be conducted through several approaches, and the consistency of results across methods has rarely been studied. We compare the results obtained by DNA metabarcoding of the same communities using two different markers - COI and 16S - and three different sampling methods: (a) homogenized Malaise trap samples (homogenate), (b) preservative ethanol from the same samples, and (c) soil samples. Our results indicate that COI and 16S offer partly complementary information on Malaise trap samples, with each marker detecting a significant number of species not detected by the other. Different sampling methods offer highly divergent estimates of community composition. The community recovered from preservative ethanol of Malaise trap samples is significantly different from that recovered from homogenate. Small and weakly sclerotized insects tend to be overrepresented in ethanol while strong and large taxa are overrepresented in homogenate. For soil samples, highly degenerate COI primers pick up large amounts of nontarget DNA and only 16S provides adequate analyses of insect diversity. However, even with 16S, very little overlap in molecular operational taxonomic unit (MOTU) content was found between the trap and the soil samples. Our results demonstrate that none of the tested sampling approaches is satisfactory on its own. For instance, DNA extraction from preservative ethanol is not a valid replacement for destructive bulk extraction but a complement. In future metabarcoding studies, both should ideally be used together to achieve comprehensive representation of the target community.

Nyckelord

environmental DNA; insects; Malaise traps; metabarcoding; nondestructive extraction; preservative ethanol

Publicerad i

Molecular Ecology Resources
2019, Volym: 19, nummer: 6, sidor: 1516-1530
Utgivare: WILEY

    UKÄ forskningsämne

    Ekologi
    Bioinformatik och systembiologi

    Publikationens identifierare

    DOI: https://doi.org/10.1111/1755-0998.13071

    Permanent länk till denna sida (URI)

    https://res.slu.se/id/publ/102048