TY - JOUR

T1 - Novel molecular resources for single-specimen barcoding of enigmatic crustacean y-larvae

AU - Dreyer, Niklas

AU - Olesen, Jørgen

AU - Grygier, Mark J.

AU - Eibye-Jacobsen, Danny

AU - Savchenko, Alexandra S.

AU - Fujita, Yoshihisa

AU - Kolbasov, Gregory A.

AU - Machida, Ryuji J.

AU - Chan, Benny K.K.

AU - Palero, Ferran

N1 - Publisher Copyright: © 2024 The Author(s) (or their employer(s)) Published by CSIRO Publishing.

PY - 2024

Y1 - 2024

N2 - Despite discovery more than 100 years ago and documented global occurrence from shallow waters to the deep sea, the life cycle of the enigmatic crustacean y-larvae isincompletely understood and adult forms remain unknown. To date, only 2 of the 17 formally described species, all based on larval stages, have been investigated using an integrative taxonomic approach. This approach provided descriptions of the morphology of the naupliar and cyprid stages, and made use of exuvial voucher material and DNA barcodes. To improve our knowledge about the evolutionary history and ecological importance of y-larvae, we developed a novel protocol that maximises the amount of morpho-ecological and molecular data that can be harvested from single larval specimens. This includes single-specimen DNA barcoding and daily imaging of y-nauplii reared in culture dishes, mounting of the last naupliar exuviae on a slide as a reference voucher, live imaging of the y-cyprid instar that follows, and fixation, DNA extraction, amplification and sequencing of the y-cyprid specimen. Through development and testing of a suite of new primers for both nuclear and mitochondrial protein-coding and ribosomal genes, we showcase how new sequence data can be used to estimate the phylogeny of Facetotecta. We expect that our novel procedure will help to unravel the complex systematics of y-larvae and show how these fascinating larval forms have evolved. Moreover, we posit that our protocols should work on larval specimens from a diverse array of moulting marine invertebrate taxa.

AB - Despite discovery more than 100 years ago and documented global occurrence from shallow waters to the deep sea, the life cycle of the enigmatic crustacean y-larvae isincompletely understood and adult forms remain unknown. To date, only 2 of the 17 formally described species, all based on larval stages, have been investigated using an integrative taxonomic approach. This approach provided descriptions of the morphology of the naupliar and cyprid stages, and made use of exuvial voucher material and DNA barcodes. To improve our knowledge about the evolutionary history and ecological importance of y-larvae, we developed a novel protocol that maximises the amount of morpho-ecological and molecular data that can be harvested from single larval specimens. This includes single-specimen DNA barcoding and daily imaging of y-nauplii reared in culture dishes, mounting of the last naupliar exuviae on a slide as a reference voucher, live imaging of the y-cyprid instar that follows, and fixation, DNA extraction, amplification and sequencing of the y-cyprid specimen. Through development and testing of a suite of new primers for both nuclear and mitochondrial protein-coding and ribosomal genes, we showcase how new sequence data can be used to estimate the phylogeny of Facetotecta. We expect that our novel procedure will help to unravel the complex systematics of y-larvae and show how these fascinating larval forms have evolved. Moreover, we posit that our protocols should work on larval specimens from a diverse array of moulting marine invertebrate taxa.

KW - barcoding

KW - biodiversity

KW - Facetotecta

KW - larval barcoding

KW - molecular resources

KW - new primers

KW - systematics

KW - y-larvae

U2 - 10.1071/IS23018

DO - 10.1071/IS23018

M3 - Journal article

AN - SCOPUS:85187644623

VL - 38

JO - Invertebrate Systematics

JF - Invertebrate Systematics

SN - 1445-5226

IS - 3

M1 - IS23018

ER -