Research Article |
Corresponding author: Katja Hogendoorn ( katja.hogendoorn@adelaide.edu.au ) Academic editor: Michael Ohl
© 2015 Katja Hogendoorn, Mark Stevens, Remko Leijs.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Hogendoorn K, Stevens M, Leijs R (2015) DNA barcoding of euryglossine bees and the description of new species of Euhesma Michener (Hymenoptera, Colletidae, Euryglossinae). ZooKeys 520: 41-59. https://doi.org/10.3897/zookeys.520.6185
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This paper launches an open access DNA barcoding project “AUSBS” under the Barcoding of Life Datasystems (BOLD). The aims of the project are to help scientists who lack the necessary morphological knowledge to identify known species using molecular markers, to aid native bee specialists with the recognition of species groups that morphologically are difficult to define, and, eventually, to assist with the recognition of new species among known species. Using integrative taxonomy, i.e. morphological comparison to type specimens in Australian museum collections combined with phylogenetic analysis of a fragment of the mitochondrial DNA cytochrome
barcoding, species discovery, euryglossine bees, Bush Blitz survey, conservation
Australia is facing a dramatic and unprecedented loss of biodiversity (
Australian native bees serve as a case in point. Despite their environmental and economic importance as pollinators of native plants (e.g.
To make native bees more accessible to the scientific community, an open access project “AUSBS” has been initiated in the Barcoding of Life Datasystems (BOLD,
Naturally, this initiative will only be successful if there is a wide coverage of species in the barcoding database. It is intended to regularly update the database with entries of additional species. So far, 271 sequences have been added to the project, covering 120 species in four of the five Australian bee families (Megachilidae, Apidae, Colletidae and Halictidae) that were collected during Bush Blitz surveys – intensive short-term surveys of remote, protected areas funded by the Australian Federal Government (
In this publication the results of DNA barcoding are focused on species within the Euryglossinae (Colletidae). This subfamily has been relatively well studied through descriptions of new species by
The bee specimens studied in this paper were collected during six intensive short term (1–2 week) Bush Blitz surveys (
For DNA analyses a single middle leg was removed from up to five specimens per morpho-species. These legs were stored in 100% ethanol to allow preservation of the DNA, and submitted to BOLD for DNA barcoding using the cytochrome c oxidase subunit 1 gene. Specimen details, including collecting dates and locality information can be accessed in BOLD under the project Australian Bee Survey (accession numbers AUSBS001-12 to 190-12 and AUSBS191-13 to 380-13, e.g. http://www.boldsystems.org/index.php/Public_RecordView?processid=AUSBS131-12, and http://www.boldsystems.org/index.php/Public_RecordView?processid=AUSBS205-13). Using only BOLD BIN compliant sequence data (
We attempted to key all collected euryglossine specimens to species. The specimens were compared to all type specimens and all other relevant reliably named material at the Queensland Museum, the Western Australian Museum, the Australian National Insect Collection, and the South Australian Museum. Based on these morphological comparisons, several species, including four species in the genus Euhesma, were identified as new. These four species are described here.
The descriptions of the species and the morphological terminology follow the format used by
BOLD Barcoding of Life Database
SAMA South Australian Museum, Adelaide
WAM Western Australian Museum
BOLD barcoding of Euryglossinae resulted in DNA barcode data for 87 specimens comprising 40 species. Of these, morphological examination resulted in the identification of 17 species, 6 species were recognized as new and 17 species were identified to genus or species group level (Suppl. material
Phylogenetic relationships among Euhesma species collected on Eremophila flowers, based on BOLD sequence data, analysed using MrBayes (GTR-inv+gamma, partitioned by codon, 8M generations). Posterior probabilities for nodes are shown when > 0.7. The species described in this publication are underlined. The three digits preceding taxon names refer to Barcoding of Life Database: AUSBS###-12/13 specimens (
A number of specimens were identified to belong to Euhesma species groups that previously were revised by
One of the four new species described here belongs to the Euhesma walkeriana-species group (
Three other new species belong to a group of Euhesma species that are associated with flowers of Eremophila (
While the molecular results group a fifth species of Euhesma, represented by a single barcoded male (RL1788A-AUSBS065) and some additional male and female specimens, with Euhesma species caught on Eremophila, it has a number of characters that are not consistent with this group, i.e. head and mouth parts not elongated and not caught on Eremophila. Thus, this species did not fit into any of the other known Euhesma groups and therefore the description was deferred until additional data allow improved justification for the position of this species relative to other Euhesma species.
Within the subfamily Euryglossinae, the genus Euhesma Michener contains a large number (65) of highly diverse species (Michener 2007). The genus was erected by Michener (2000) as a “dumping ground” (
The fifteen known species in the walkeriana species group (
As only a single species is added to this group, we do not produce a modified key, but suggest modifying the key produced by
4 (3) | Labrum yellow | 5 |
– | Labrum brown/black | 5A |
5A (4) | Mandibles transparent white with dark tips | E. albamala sp. n. |
– | Mandibles yellow, brown or black | 6 |
Holotype: ♀, RL1807C, Cane River Conservation Park, Western Australia, 22.0936°S, 115.3507°E, 26 June 2011, R. Leijs, on flowers of a red flowering Grevillea (WAM). BOLD: AUSBS082-12. Paratypes: ♂, RL1811, same date and locality as holotype (WAM). BOLD: AUSBS009-12; 4♀, all same date and locality as holotype (SAMA 32-033287, -288, -289, -290).
Most like E. spinola; however, the combination of the white mandibles with dark tips and the simple claws distinguishes this from all other species. The absence of both pothook and clubbed setae is a character shared with E. lobata, E. spinola and E. walkeriana.
Female. Length approximately 5.0 mm; wing length about 3.2 mm. Head width 1.5 mm. Relative head measurements: width 50, length 41; clypeal length 11; lower interocular distance 30; upper interocular distance 28; interantennal distance 8; antennocular distance 7; interocellar distance 10; ocellocular distance 8.
Supraclypeal area, frons and mesosoma with metallic bronze-green sheen; mandibles translucent white with dark tips; labrum brown; legs amber with coxae and tibiae dark brown, and hind basitarsi pale yellow; face with long white hairs; forelegs with coxal lobes weakly developed with long hyaline unbranched setae; clubbed and pothook setae absent; hind basitibial plate with carina; hind-tibia with a row of tubercles beyond basitibial area; claws simple; metasoma brown with posterior margins of terga translucent, creating a banded effect; pygidial plate broadly spathulate.
Male. Length approximately 3.8 mm; wing length approximately 3 mm. Head width 1.2 mm. Relative head measurements: width 50, length 45; clypeal length 10; lower interocular distance 30; upper interocular distance 30; interantennal distance 8; antennocular distance 7; interocellar distance 10; ocellocular distance 8. Coloration and pubescence as in female, but pronotum with metallic purple sheen; metasoma dark brown with posterior margins of terga translucent. The apical five flagellar segments of the male look crumpled, however this could be an artifact. Terminalia as in Fig.
The specific name refers to the white mandibles.
The bees in the three known species groups associated with Eremophila, the atra (eight species), the alicia (eight species) and the coppinensis (four species) species groups, have modified heads and/or labial palps that suggest foraging on narrow, tubular flowers (
To allow easy identification of the Euhesma species associated with Eremophila,
1 | Labial palps enormously extended, nearly as long as or longer than head | 2 |
– | Labial palps much shorter than head | atra-group 14 |
2(1) | Labial palps with segment 2 much shorter than segment 4 | alicia-group 6 |
– | Labial palps with segment 2 subequal to or longer than segment 4 | coppinensis-group 3 |
3(2) | Supraclypeal area and clypeus medianly concave; body length about 5 mm | E. walkeri |
– | Neither supraclypeal area nor clypeus medianly concave; body length about 6 mm | 4 |
4(3) | Labial palps with segments 2 and 4 subequal in length | 5 |
– | Labial palps with segment 2 clearly longer than segment 4 | E.coppinensis |
5(4) | Labial palps longer than head | E. macrayae |
– | Labial palps as long as head | E. newmanensis |
6(2) | Labial palp segment 3 clearly longer than segment 4 | 7 |
– | Labial palp segment 3 not clearly longer than segment 4 | 8 |
7(6) | Supraclypeal area, clypeus medianly and frons medianly concave | E. pantoni |
– | Supraclypeal area, clypeus and frons not concave | E. alicia |
8(6) | Labial palp segments 3 and 4 together much shorter than head | 9 |
– | Labial palp segments 3 and 4 together about as long as head | 11 |
9(8) | Upper margin of clypeus indistinct | E. aulaca sp. n. |
– | Upper margin of clypeus distinct | 10 |
10(9) | Body length about 5 mm | E. yellowdinensis |
– | Body length about 6 mm | E. wiluna |
11(8) | Tibiae of all legs golden | E. cuneifolia |
– | Tibiae of all legs mostly dark brown | 12 |
12(11) | Fronto-clypeal suture clearly evident | E. meeka |
– | Fronto-clypeal suture not clearly evident | 13 |
13(12) | Clypeus with a median longitudinal furrow | E. sulcata |
– | Clypeus without a median longitudinal furrow | E. granitica |
14(1) | Only known from Northern Territory and Queensland | E. sturtiensis |
– | Only known from Western Australia and South Australia | 15 |
15(14) | Tibiae, tarsi and terminal gastral terga golden brown | 16 |
– | Tibiae, tarsi and terminal gastral terga predominantly dark brown | 17 |
16(15) | First recurrent vein distal to first submarginal crossvein | E. aurata |
– | First recurrent vein interstitial with first submarginal crossvein | E. lyngouriae sp. n. |
17(15) | Labial palp longer than antennal flagellum | 18 |
– | Labial palp not longer than antennal flagellum | 19 |
18(17) | Supraclypeal area almost glabrous and highly polished | E. symmetra |
– | Supraclypeal area neither glabrous nor polished | E. nalbarra |
19(17) | Dorsal surface of clypeus concave medianly (indented) | E. atra |
– | Dorsal surface of clypeus with a slight median longitudinal furrow | E. scoparia |
– | Dorsal surface of clypeus in no way concave | 20 |
20(19) | Labial palps clearly longer than maxillary palps | E. leonora |
– | Labial palps and maxillary palps about equal in length | 21 |
21(20) | Fronto-clypeal suture distinct | E. balladonia |
– | Fronto-clypeal suture absent on clypeus | E. micans sp. n. |
Holotype: ♀, SAMA 32-03385, Bon Bon Station, South Australia, 30.5250°S, 135.5917°E, 27 October 2010, R. Leijs, on flowers of Acacia victoriae (SAMA). BOLD: AUSBS135-12. Paratype: ♂, SAMA 32-03386, same date and locality as holotype (SAMA). BOLD: AUSBS136-12.
Most like E. leonora; however this is the only species in which the frons above the antennae is shining, and the facial fovae are narrow and not curved towards the ocelli.
Female. Length approximately 4.5 mm; wing length approximately 3.1 mm. Head width 0.9 mm. Relative head measurements: width 50; length 64, clypeal length 17; lower interocular distance 32; upper interocular distance 33; interantennal distance 13; antennocular distance 5; interocellar distance 16; ocellocular distance 10. Anterior margin of clypeus truncate, upper margin slightly concave; other areas of clypeus and frons convex with depressions centrally, around the anterior ocelli and antennal implants; lower part of facial fovae broadened, upper part narrow, not bent towards ocelli; antennal scapes anteriorily flattened; malar space short; labial palp segments increasing in length in the order 2, 1, 3, 4. Clypeus and frons above antennae shiny with punctures wide apart; facial fovae, interocellar area and scutum with dense reticulation and dull.
Head black; antennae brown with flagella yellowish ventrally; labial palp segments 1,2 dark brown, segments 3, 4 light brown; legs yellowish with femora dark brown. First recurrent vein of forewing interstitial with first submarginal crossvein. Scattered long white hairs on frons, clypeus, antennal scapes, vertex, mandibles, posterior genae, sides of thorax, venter and gastral tergum 5.
Male. Length approximately 3.6 mm; wing length approximately 2.6 mm. Head width 0.8 mm. Relative head measurements: width 50; length 57, clypeal length 24; lower interocular distance 29; upper interocular distance 34; interantennal distance 10; antennocular distance 9; interocellar distance 14; ocellocular distance 10. Forewings and labial palps as in female; final three flagellar segments with indentations ventrally; inner hind tibial spur finely pectinate; frons above antennae shining with punctures wide apart. All labial segments dark; legs dark brown with fore tibia and all tarsae yellowish. Terminalia as in Fig.
The specific name refers to the shiny frons.
Holotype: ♀, SAMA 32-033284, Bon Bon Station, South Australia, 30.2389°S, 135.5098°E, 26 October 2010, R. Leijs, on flowers of Swainsona stipularis (SAMA). BOLD: AUSBS137-12.
♂, unknown.
Most like E. aurata, however, forewing first recurrent vein interstitial with first submarginal crossvein, margin between clypeus and supraclypeal area lacking furrow, all abdominal segments amber, lacking brown bands.
Female. Length approximately 6.0 mm; wing length approximately 3.5 mm. Head width 1.6 mm. Relative head measurements: width 50; length 50; clypeal length 14; lower interocular distance 28; upper interocular distance 32; interantennal distance 12; antennocular distance 6; interocellar distance 12; ocellocular distance 8. Anterior margin of clypeus truncate, upper margin slightly concave, lacking median furrow; frons with median area elevated; facial fovae broad, upper part slightly bent towards ocelli; antennal scapes anteriorily flattened; malar space short; labial palp segments increasing in length in the order 1 = 3, 2. Clypeus and frons above antennae; facial fovae interocellar area and scutum with dense reticulation and dull. Head black; antennae brown with flagella yellowish ventrally; labial palp segments 1 dark brown, 2 and 3 yellowish; legs and gaster amber, tergal fovae and fore femora posteriorly dark brown. First recurrent vein of forewing interstitial with first submarginal crossvein. Scattered long white hairs on frons, clypeus, antennal scapes, vertex, mandibles, posterior genae, sides of thorax, venter and gastral tergum 5.
The type lacks labial palp segment 4.
The specific name refers to the amber coloured legs and gaster of the female.
Holotype: ♀, SAMA 32-033281, Bon Bon Station, South Australia, 30.8440°S, 135.5389°E, 27 October 2010, R Leijs (SAMA). BOLD: AUSBS133-12.
Paratype(male). ♂, SAMA 32-033282, same date and locality as holotype. Paratypes: 3♀, Bon Bon Station, South Australia, 30.7416°S, 135.3665°E, on Eremophila scoparia; 1♀, Bon Bon Station, South Australia, 30.5250°E, 135.5917°E; 7♀+1♂ in ethanol RL1636 all same date as holotype (SAMA). BOLD: AUSBS131,2,4-12.
The species is most like E. yellowdinensis, but the clypeus has a distinctive deep median furrow and the total length of the labial palps is only slightly shorter than the head.
Female. Length approximately 5.5 mm; wing length approximately 3.5 mm. Head width 1.6 mm. Relative head measurements: width 50; length 50; clypeal length 14; lower interocular distance 28; upper interocular distance 32; interantennal distance 12; antennocular distance 6; interocellar distance 12; ocellocular distance 8. Clypeus with deep median furrow, anterior margin truncate, upper margin indistinct; facial fovae broad, upper part slightly bent towards ocelli; malar space short; labial palp segments increasing in length in the order 1,2,3=4. Clypeus and frons above antennae, interocellar area and scutum with dense reticulation; clypeus and supraclypeal area shining, frons, paraocular areas and scutum dull. Head black; antennae brown with flagella yellowish ventrally; labial palp segments 1 and 2 dark brown, 3 and 4 ribbon-like, yellowish; all femora and tibiae of middle and hind legs medially dark brown, fore tibiae, all tarsi and remaining parts yellow; gaster dark brown. Marginal zones of metasoma wide and translucent. Forewing with first recurrent vein almost interstitial with first submarginal crossvein. Scattered long white hairs on frons, clypeus, antennal scapes, vertex, mandibles, posterior genae, sides of thorax, venter and gastral tergum 5, pygidial fimbria pale orange.
Male. Length approximately 5.2 mm; wing length approximately 3.6 mm. Head width 1.2 mm. Relative head measurements: width 50; length 60, clypeal length 17; lower interocular distance 22; upper interocular distance 30; interantennal distance 8; antennocular distance 5; interocellar distance 12; ocellocular distance 9. Forewings, labial palps and other characters as in female; inner hind tibial spur roughly pectinate; Terminalia as in Fig.
The specific name means ‘with a furrow’, referring to the deep median furrow in the clypeus.
DNA barcoding has been used to associate the sexes of species. Furthermore, we used integrative taxonomy (
Despite the small size of the present molecular database, both in terms of numbers of species and numbers of nucleotides involved, it provides some insights with respect to validity of the Euhesma groups described by
While
By contrast, and in spite of the fact that the database is far from complete, the COI phylogeny presented here (Suppl. material
Support was also found for some other euryglossine clades. Although results of phylogenetic analyses based on just a small fragment of mitochondrial DNA (used here as the ‘barcoding region’) are often not informative above genus level, some well supported clades (posterior probabilities > 0.8) combine species within the genera Euryglossula, Euryglossina and Pachyprosopis, of which their sister clade appears to be the Euhesma walkeriana species group.
The creation of an open access molecular DNA barcoding project of Australian native bees enhances possibilities for scientists with molecular capabilities to document bee biodiversity (
This paper would not have been completed without the support of the Bush Blitz program provided through ABRS. We thank Jo Harding, Kate Gillespie, Mim Jambrecina and the other members of the Bush Blitz team for their fabulous support in the field. We thank Susan Wright (QM), Nicole Fisher (ANIC), Brian Hanich (WAM) and Terry Houston (WAM) for providing access to the collections, Alexis Tindall for excellent photographic assistance, and Wiebe Hogendoorn for linguistic advice. This work was supported by a Bush Blitz Tactical Taxonomy Grant from the Australian Government through ABRS.
Type localities of new species
Data type: occurence
Explanation note: localities where type specimens where caught.
Phylogenetic relationships of euryglossine species based on CO1 sequence data
Data type: gene tree
Explanation note: tree based on CO1 sequence data.