Research Article |
Corresponding author: Bernard Landry ( bernard.landry@ville-ge.ch ) Academic editor: Richard Mally
© 2023 Bernard Landry, Julia Bilat, James Hayden, M. Alma Solis, David C. Lees, Nadir Alvarez, Théo Léger, Jérémy Gauthier.
This is an open access article distributed under the terms of the CC0 Public Domain Dedication.
Citation:
Landry B, Bilat J, Hayden J, Solis MA, Lees DC, Alvarez N, Léger T, Gauthier J (2023) The identity of Argyria lacteella (Fabricius, 1794) (Lepidoptera, Pyraloidea, Crambinae), synonyms, and related species revealed by morphology and DNA capture in type specimens. ZooKeys 1146: 1-42. https://doi.org/10.3897/zookeys.1146.96099
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In this study the aim was to resolve the taxonomy of several species of Argyria Hübner (Pyraloidea, Crambinae) with previously unrecognised morphological variation. By analysing the DNA barcode (COI-5P) in numerous specimens, the aim was to reconstruct phylogenetic relationships between species, to provide better evidence for synonymies, and to circumscribe their geographical distribution. Using an innovative DNA hybridisation capture protocol, the DNA barcode of the lectotype of Argyria lacteella (Fabricius, 1794) was partially recovered for comparison with the 229 DNA barcode sequences of Argyria specimens available in the Barcode of Life Datasystems, and this firmly establishes the identity of the species. The same protocol was used for the following type specimens: the Argyria abronalis (Walker, 1859) holotype, thus confirming the synonymy of this name with A. lacteella, the holotype of A. lusella (Zeller, 1863), syn. rev., the holotype of A. multifacta Dyar, 1914, syn. nov. newly synonymised with A. lacteella, and a specimen of Argyria diplomochalis Dyar, 1913, collected in 1992. In addition, nine specimens of A. lacteella, A. diplomochalis, A. centrifugens Dyar, 1914 and A. gonogramma Dyar, 1915, from North to South America were sampled using classical COI amplification and Sanger sequencing. Argyria gonogramma Dyar, described from Bermuda, is the name to be applied to the more widespread North American species formerly identified as A. lacteella. Following morphological study of its holotype, Argyria vestalis Butler, 1878, syn. nov. is also synonymised with A. lacteella. The name A. pusillalis Hübner, 1818, is considered a nomen dubium associated with A. gonogramma. The adult morphology is diagnosed and illustrated, and distributions are plotted for A. lacteella, A. diplomochalis, A. centrifugens, and A. gonogramma based on slightly more than 800 specimens. For the first time, DNA barcode sequences are provided for the Antillean A. diplomochalis. This work provides a modified, improved protocol for the efficient hybrid capture enrichment of DNA barcodes from 18th and 19th century type specimens in order to solve taxonomic issues in Lepidoptera.
Argyria centrifugens Dyar, Argyria diplomochalis Dyar, Argyria gonogramma Dyar, COI barcodes, Crambidae, historical DNA, hybrid enrichment, species delimitation
The name Tinea lacteella Fabricius, 1794, and its synonyms, have been applied to small white moths of the genus Argyria Hübner collected in the New World since
Thus, because morphological and DNA barcode variation was observed in Argyria specimens that otherwise share a similar (ca. 11 mm) wingspan and previously unrecognised external diagnostic characters, we found it necessary to try to fix the identity of A. lacteella and the species similar to it, and to better understand their synonymy and geographical distribution. We aimed to do that by integrating both the COI barcode data available in BOLD and the type specimens of the species as well as those pertaining to synonymised names.
Until recently it has been impossible to recover genetic information from old museum specimens because the DNA they contain is degraded and occurs in very low quantities compared to contaminant DNA from other organisms (
In this study, we adapted hybrid enrichment methods to target the COI barcode in old museum specimens. We designed probes along the entire nucleotide sequence of the COI barcode and synthesised our own RNA probes. We extracted historical DNA from four type specimens dating back to the 18th, 19th, and early 20th centuries and an additional specimen collected in 1992. To successfully recover the COI barcode from this degraded, fragmented and contaminant-rich DNA, we combined hybrid enrichment capture and next-generation sequencing. We performed this sophisticated approach for these precious specimens because classic PCR amplification attempts were unsuccessful. In parallel, we amplified the DNA barcode for nine additional samples and integrated them with all available Argyria sequences in the BOLD database. Combining phylogenetic inferences, species delimitation approaches based on sequence data and morphology, we propose a new classification of several Argyria species. This study shows that innovative methods of museomics can solve complex taxonomic questions still debated. More generally, it reconciles the modernity of innovative molecular approaches with the biological heritage that museums have been preserving for centuries.
The original description of A. lacteella and subsequent citation of the name by
MGCL McGuire Center for Lepidoptera and Biodiversity, Gainesville, Florida, USA;
VOB V. O. Becker collection, Camacan, Bahia, Brazil;
Specimens from which DNA was not extracted were dissected following
Photographs were taken with a variety of devices in five institutions (
To ascertain the identity of Argyria lacteella, the DNA of its unique (as far as known) type specimen housed in the
In the
DNA extraction on historical samples were performed using PCR & DNA Cleanup Kit (Monarch). The protocol was adapted from
Data pertaining to specimens sampled with hyRAD and Sanger protocols at
ID | Sample | Year of collect | Extraction method | Tissue | DNA conc. (ng/ul) | #_reads | #_fragments | Mean fragment size (bp) | % COI barcode |
---|---|---|---|---|---|---|---|---|---|
COI capture | |||||||||
CRA01 |
|
before 1859 | non-destructive | abdomen | 53.60 | 3.100,910 | 1.559,505 | 83.25 | 99.8 |
CRA02 | NHMUK013696753 |
before 1863 | non-destructive | leg | 0.40 | 8.889 | 1.583 | 81.88 | 96.3 |
CRA03 |
|
1784–1789 | destructive | leg | 0.50 | 3.101 | 60 | 77.00 | 51.4 |
CRA04 |
|
1992 | destructive | leg | 2.94 | 862.323 | 183.175 | 103.51 | 100 |
CRA07 | USNM Argyria multifacta (Dyar, 1914) | 1911 | destructive | 2 legs | 0.17 | 14.538 | 10.686 | 49.40 | 93.1 |
PCR amplification | |||||||||
CRA05 |
|
2021 | destructive | leg | 0.81 | 100 | |||
CRA06 |
|
2021 | destructive | leg | 0.79 | 100 | |||
BLDNA137 |
|
2018 | destructive | leg | 100 | ||||
BLDNA138 |
|
2018 | destructive | leg | 100 | ||||
BLDNA141 |
|
2018 | destructive | leg | 100 | ||||
BLDNA065 |
|
2004 | destructive | leg | 100 | ||||
JEH20210604A |
|
2021 | destructive | leg | 100 | ||||
JEH20210604C |
|
2020 | destructive | leg | 100 | ||||
JEH20210604D |
|
2015 | destructive | leg | 100 |
For additional samples, the DNA barcode was amplified by PCR. For CRA05 and CRA06 (Table
Raw reads were cleaned using Cutadapt (
To investigate the phylogeny of Argyria species, the sequences from all the samples including the keyword “Argyria” were retrieved from the Barcode of Life Data System (BOLD) (Suppl. material
Three different methods were used to investigate species delimitation: Automatic Barcode Gap Discovery (ABGD) (
The sequence dataset is available on BOLD (DS-ARGYRIA). Raw reads are available on the NCBI SRA BioProject PRJNA914237.
Historical DNA extraction showed different yields mainly related to the age of the specimens but also to the type of tissue and the extraction method, i.e., destructive or non-destructive (Table
Phylogenetic inference has been performed on the whole COI barcode alignment including BOLD sequences, barcodes amplified for this study and sequences recovered using our historical DNA capture approach. The samples corresponding to the two species A. rufisignella and A. nummulalis have been used as outgroups, and their common node is well supported (Fig.
Phylogenetic inferences including 174 Argyria COI barcode sequences, i.e., 160 barcodes from BOLD, 9 COI sequences amplified by PCR (in bold), and 5 COI sequences obtained using capture from historical specimens (in red). Nodal support expressed in SH-aLRT and ultrafast bootstrap (UFBoot) is given as indicated in the caption except for nodes when SH-aLRT < 80 and/or UFBoot < 95. Species delimitation results including morphology, ABGD, GMYC, and PTP are indicated by different colours to represent the species proposed.
Tinea lacteella
Fabricius, 1794: 313. Type locality: “Americae insulis” (USA Virgin Island of Saint Croix; see Remarks).
=albana (Fabricius, 1798 (Pyralis). Unnecessary replacement name.
=abronalis Walker, 1859: 969 (Zebronia??). Type locality: Brazil, Rio de Janeiro.
=lusella (Zeller, 1863: 51) (Catharylla). Type locality: St. Thomas Island [USA Virgin Islands]. Syn. rev.
=vestalis Butler, 1878: 494, 495. Type locality: Jamaica. Syn. nov.
=multifacta Dyar, 1914: 317. Type locality: Panama, Porto Bello. Syn. nov.
Lectotype
of Tinea lacteella (Fig.
Holotype
of Zebronia? abronalis (Figs
Holotype
of Catharylla lusella (Fig.
Holotype
of Argyria vestalis (Fig.
Holotype
of Argyria pusillalis variety multifacta (Fig.
238 specimens (see Suppl. material
This is a small satiny white moth of 9.5–14 mm in wingspan. The forewing brown markings are median triangles on the costa and dorsal margin usually linked by a thin straight line sometimes slightly thicker on the discal cell as a spot, but sometimes inconspicuous, another triangle subapically on costa, usually separated by a thin white line from a short oblique dash anteriorly, and a wavy terminal line (Figs
Phylogenetic inference based on COI barcode alignment reveals a large clade grouping the A. lacteella samples. This clade is relatively homogeneous since the percentage of divergence within this species remains low with an average of 1.25% (Fig.
Widespread in the Western Hemisphere from the US State of Florida north to Alachua County in the north, across Central America and the Antilles, in South America to Argentina in the south, as well as on the Galápagos Islands (Fig.
The first label associated with the lectotype of A. lacteella (Fig.
The locality of origin is an additional complication associated with A. lacteella. The locality of Fabricius’ Pyralis albana (1798) is mentioned as “Americae insulis” [American islands] whereas that of A. lacteella (1794) is “Americae meridionalis arboretis” [South American arboretum]. Given that “Dr. Pflug” is mentioned in the original description of A. lacteella, it is reasonable to conclude that “Americae insulis” was a correction for “Americae meridionalis arboretis”. This is because Paul Gottfrid Pflug (1741–1789), a medical doctor, lived in the Caribbean island of Saint Croix (United States Virgin Islands) during the last five years of his life, where he collected insects that he sent to Denmark. He is mentioned often by Fabricius as a specimen collector (O. Karsholt, pers. comm. to BL, 3 June 2021). Therefore, A. lacteella/albana is from an American island (Americae insulis) that is probably Saint Croix.
As confirmed by Copenhagen Museum former curator Ole Karsholt and present curator Thomas Pape, only one type specimen presently exists for lacteella/albana (Fig.
The type specimen of A. lacteella (Fig.
The original description of Catharylla lusella
The original description of Argyria vestalis does not mention more than one specimen and the
Argyria multifacta was described as a variety of A. pusillalis for which “All the specimens have the median band continuous across the wing” (
This species evidently became established in Florida, USA in the 1970s and consequently, earlier records from Florida (
The vesica of a male specimen from Florida, USA (not illustrated here) was successfully everted by J. Baixeras, who wrote the following to BL on 17 October 2022: “After a lot of manipulation I was able to evert what seems like a rather tubular vesica bearing a single row of non-deciduous cornuti tightly arranged like in a “gun charger” mode. The vesica seems to be somewhat convoluted at the base (I do not think it an artefact), then straight. The cornuti are extended all over the length of the vesica except in the terminal part, close to the genital opening. The basal convolution is interesting and, if my surmise is correct, should be correlated with some structure in the female, either a pocket, broadening sclerotisation or, in some cases, some corrugated area allowing expansion during insertion.” The basal convoluted bend at the base of the vesica reflects the shape of the basal section of the female ductus bursae, which is indeed corrugated (Figs
Argyria gonogramma
Dyar, 1915: 87–88. Type locality: Bermuda.
=pusillalis Hübner, 1818: 30, [36], [38], figs 167, 168. Type locality: [USA, Maryland] Baltimore. Nomen dubium.
= pussillalis [sic] Hübner, 1818: 28; original misspelling.
Argyria lacteella
(Fabricius, 1794):
Holotype
♂ (Figs
411 specimens (see Suppl. material
In this small satiny-white moth measuring between 10.5 and 13.5 mm in wingspan, the median markings of the forewing (Figs
Phylogenetic inference reveals that Argyria gonogramma constitutes a homogeneous clade. The monophyletic clade is identified in both GMYC and PTP species delimitation approaches, but it is not found in the ABGD approach and is grouped with A. lacteella (Fig.
Bermuda, Bahamas, widespread in the Eastern USA, from North Carolina in the North to the south of Florida, west to eastern Texas (Fig.
The specimen of A. gonogramma labelled ‘Type’ in the
Argyria pusillalis Hübner is associated here with A. gonogramma and not with A. lacteella as in
Argyria pusillalis was originally named “pussillalis” (Hübner 1818: 28), then mentioned as “pusillalis” on page 30 and on two indices (pages [36] and [38]), and finally as “pussillalis” again on the plate with the illustrations. Given that “pusillus” is Latin for small, it seems reasonable to believe that the original spelling “pussillalis” was in error.
Argyria gonogramma is a North American native species that was previously misidentified as A. lacteella and that has been collected in the United States since the late 1800’s. The earliest specimens in the
That this species is native to the Southeastern U.S., or at least was established long before A. lacteella, is shown by earlier collecting dates for specimens in the
The earliest record of A. lacteella in 1979 in the USA (Florida) supports the conclusion that
Based on collected series of specimens both Argyria gonogramma and A. lacteella now occur in sympatry and fly on the same dates in Florida, for example at Archbold Biological Station in Highlands County or in Pinellas County.
The single moth at the basis of the Vermont record has been dissected and is correctly determined, but it is far outside the range since we know of no other record of A. gonogramma north of North Carolina. It was collected in sandplain habitat (M. Sabourin, pers. comm. to JH, 29 August 2022), which is consistent with the species’ habitat preference in Florida.
Argyria diplomochalis
Dyar, 1913: 113. Type locality: [USA] Culebra Island, Puerto Rico.
Argyria diplamachalis [sic]: Schaus, 1940: 400.
Lectotype
♂ (Fig.
41 (see Suppl. material
Measuring 10–13 mm in wingspan this species (Figs
The phylogenetic clade corresponding to the species A. diplomochalis comprises only three samples sequenced for this study. It appears that no sequence available in the BOLD database corresponds to this species. All three species delimitation approaches identify this clade (Fig.
Antilles, from Cuba in the West to Dominica in the Lesser Antilles in the east (Fig.
Described from 12 cotypes from Culebra Island and Bayamon, Puerto Rico, a lectotype is designated here to ensure that the name continues to refer to this species exclusively.
Examination of specimens of “A. diplomochalis” cited by
Argyria centrifugens
Dyar, 1914: 318. Type locality: Panama, Canal Zone, Paraiso.
Holotype
♂ (Figs
87 specimens (see Suppl. material
Argyria centrifugens (Fig.
Female genitalia of Argyria species 24 Argyria lacteella (holotype of A. abronalis, with spermatophore inside corpus bursae;
Phylogenetic inference reveals that the species A. gonogramma constitutes a distinct lineage separate from the species A. insons. The three species delimitation methods identified this species but also identified a subcluster separating the sample BLDNA141. This specimen originates from Colombia while all the other specimens come from Costa Rica. The observed genetic divergence is certainly related to a geographical divergence. A genetic study including samples from more distant localities such as Brazil would better characterise the genetic variability of this species.
Central and South America, from Honduras to Colombia and Brazil. Records from the central west coast of Florida are possibly recent introductions (Fig.
The species was described from a specimen labelled “Type” of an unspecified sex and two other specimens, “Also two others, Cabima, May, 1911 (Busck).” (
One female specimen identifiable as A. centrifugens was collected in Florida (Largo, Pinellas County), 1 Feb. 1995, by J.-G. Filiatrault, deposited in the
We were able to resolve complex taxonomic questions for Argyria using an innovative DNA hybridisation capture protocol to recover high percentages of the DNA barcode of 18th–20th century type specimens. Thus, we were able to solve taxonomic problems regarding synonymies of multiple names applied to the same species. Furthermore, we compiled distribution maps based on refined identities and specimens from multiple museums, leading to other questions regarding responses to environmental change through time. For example, we provided evidence to refine the type locality of Argyria lacteella as St Croix Island, whereas the three recent (2021) specimens we examined from that island belong to A. diplomochalis (see Suppl. material
James Hogan (
Identity of Argyria lacteella (Fabricius) revised
Data type: word document
Species names, code numbers and collecting data
Data type: specimens names and collecting data
Explanation note: This table contains the species names, code numbers, and collecting data of all of the specimens examined in this study and that were used to generate the distribution maps.