Analysis of tissue dependent DNA yield for optimal sampling of micro-moths in large-scale biodiversity surveys

Higher taxa often show increasing species richness towards tropical low latitudes, a pattern known as the latitudinal biodiversity gradient (LBG). A rare reverse LBG (with greater richness towards temperate high latitudes) is exhibited by Gracillariidae leaf-mining moths, in which most described species occur in northern temperate areas. We carried out the first assessment of gracillariid species diversity in two Neotropical regions to test whether the relatively low tropical species diversity of this family is genuine or caused by insufficient sampling and a strong taxonomic impediment. Field surveys in six French Guianan and one Ecuadorian site produced 516 gracillariid specimens that were DNA barcoded to facilitate identification and to match larvae inside leaf mines with adults. Species delineation from sequence data was approximated using Automatic Barcode Gap Discovery and Refined Single Linkage Analysis through the Barcode Index Number system, and the proportion of described/undescribed species was estimated after comparison with types of 83% of described species. Locally, alpha-diversity far exceeds that of any known temperate fauna, with as many as 108 candidate species (59.3% as singletons) collected at one site, and with an estimated species richness lower bound of 240 species. Strikingly, at least 85% of the species collected as adults were found to be undescribed. Our sampling represents the most thorough survey of gracillariid species diversity in the Neotropics to date and the results from both our molecular and morphological analyses indicate that the current reverse LBG seen in this group is an artefact of insufficient sampling and a strong description deficit in the Neotropics.

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“…Adult moths were mounted on microlepidopteran setting boards immediately after collection (see Lopez‐Vaamonde et al . for full protocol).…”

Section: Methodsmentioning

confidence: 99%

DNA barcoding reveals a largely unknown fauna of Gracillariidae leaf‐mining moths in the Neotropics

Lees

Kawahara

Rougerie³

et al. 2013

Molecular Ecology Resources

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“…However, the coverage for the Madagascan lepidopteran fauna is very sparse. Nevertheless, the use of Barcode Index Numbers (BINs) (Ratnasingham and Hebert 2013) as proxies for species allows the assessment of hyperdiverse groups that are taxonomically poorly known, such as micromoths (Schmidt et al 2017;Aagaard et al 2017;Lees et al 2013;Lopez-Vaamonde et al 2012). As of 29 June 2018 (including the current study) there were 2852 DNA barcode BINs for Madagascar out of a total of 113 161 lepidopteran BINS, according to a search of the BIN database in the public portal of BOLD.…”

Section: Introductionmentioning

confidence: 97%

DNA barcodes reveal deeply neglected diversity and numerous invasions of micromoths in Madagascar

Lopez‐Vaamonde

Sire

Rasmussen

et al. 2019

Genome

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Madagascar is a prime evolutionary hotspot globally, but its unique biodiversity is under threat, essentially from anthropogenic disturbance. There is a race against time to describe and protect the Madagascan endangered biota. Here we present a first molecular characterization of the micromoth fauna of Madagascar. We collected 1572 micromoths mainly using light traps in both natural and anthropogenically disturbed habitats in 24 localities across eastern and northwest Madagascar. We also collected 1384 specimens using a Malaise trap in a primary rain forest at Andasibe, eastern Madagascar. In total, we DNA barcoded 2956 specimens belonging to 1537 Barcode Index Numbers (BINs), 88.4% of which are new to BOLD. Only 1.7% of new BINs were assigned to species. Of 47 different families found, Dryadaulidae, Bucculatricidae, Bedelliidae, Batrachedridae, and Blastobasidae are newly reported for Madagascar and the recently recognized Tonzidae is confirmed. For test faunas of Canada and Australia, 98.9%-99.4% of Macroheterocera BINs exhibited the molecular synapomorphy of a phenylalanine in the 177th complete DNA barcode codon. Non-macroheteroceran BINs could thus be sifted out efficiently in the Malaise sample. The Madagascar micromoth fauna shows highest affinity with the Afrotropics (146 BINs also occur in the African continent). We found 22 recognised pests or invasive species, mostly occurring in disturbed habitats. Malaise trap samples show high temporal turnover and alpha diversity with as many as 507 BINs collected; of these, astonishingly, 499 (98.4%) were novel to BOLD and 292 (57.6%) were singletons. Our results provide a baseline for future surveys across the island.

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“…Additionally, reliable storage methods were described and discussed by Knölke et al (2005) and Lopez-Vaamonde et al (2012) .…”

Section: Resultsmentioning

confidence: 99%

A streamlined collecting and preparation protocol for DNA barcoding of Lepidoptera as part of large-scale rapid biodiversity assessment projects, exemplified by the Indonesian Biodiversity Discovery and Information System (IndoBioSys)

Schmidt¹,

Hausmann²,

Araújo³

et al. 2017

BDJ

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Here we present a general collecting and preparation protocol for DNA barcoding of Lepidoptera as part of large-scale rapid biodiversity assessment projects, and a comparison with alternative preserving and vouchering methods. About 98% of the sequenced specimens processed using the present collecting and preparation protocol yielded sequences with more than 500 base pairs. The study is based on the first outcomes of the Indonesian Biodiversity Discovery and Information System (IndoBioSys). IndoBioSys is a German-Indonesian research project that is conducted by the Museum für Naturkunde in Berlin and the Zoologische Staatssammlung München, in close cooperation with the Research Center for Biology – Indonesian Institute of Sciences (RCB-LIPI, Bogor).

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Analysis of tissue dependent DNA yield for optimal sampling of micro-moths in large-scale biodiversity surveys

Cited by 12 publications

References 52 publications

DNA barcoding reveals a largely unknown fauna of Gracillariidae leaf‐mining moths in the Neotropics

DNA barcoding reveals a largely unknown fauna of Gracillariidae leaf‐mining moths in the Neotropics

DNA barcodes reveal deeply neglected diversity and numerous invasions of micromoths in Madagascar

A streamlined collecting and preparation protocol for DNA barcoding of Lepidoptera as part of large-scale rapid biodiversity assessment projects, exemplified by the Indonesian Biodiversity Discovery and Information System (IndoBioSys)

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