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DNA Barcodes of Caterpillars (Lepidoptera) from Papua New
Guinea
Author(s): Scott E. Miller , Jan Hrcek , Vojtech Novotny , George D. Weiblen
and Paul D.N. Hebert
Source: Proceedings of the Entomological Society of Washington,
115(1):107-109. 2013.
Published By: Entomological Society of Washington
DOI: http://dx.doi.org/10.4289/0013-8797.115.1.107
URL: http://www.bioone.org/doi/full/10.4289/0013-8797.115.1.107
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PROC. ENTOMOL. SOC. WASH.
115(1), 2013, pp. 107?109
NOTE
DNA barcodes of caterpillars (Lepidoptera) from Papua New Guinea
DOI: 10.4289/0013-8797.115.1.107
This paper provides metadata for DNA
barcode (COI) data in GenBank for a
collection of caterpillar vouchers from
ecological studies inWanang, Papua New
Guinea, sampled as part of long term re-
search on the ecology and taxonomy of
herbivorous insects there (Miller et al.
2003, Craft et al. 2010, Novotny et al.
2010, Whitfeld et al. 2012). This paper
aims to make DNA barcode data avail-
able to document ongoing research, to
contribute to the International Barcode
of Life (iBOL; www.ibol.org) project,
and to encourage enhancement in identi-
?cations, in line with the concept of DNA
barcode data release papers and the Fort
Lauderdale principles for genetic data
(Schindel et al. 2011). Data released on
Genbank (accession numbers HM906186-
HM906529, HQ558240-HQ558310,
KC158225-KC158271) includes the stan-
dard ?elds for the BARCODE data stan-
dard (Benson et al. 2012) and more data,
including images and host plant codes, are
available on BOLD (www.boldsystems.
org; Ratnasingham and Hebert 2007),
accessible from the project CATS using
a DOI (dx.doi.org/10.5883/DS-CATS1).
Field methods and host plants: Trees
and shrubs with stems greater or equal to
5 cm in diameter at breast height (dbh) in
two 100 x 100 m plots near Wanang
(145.182? E, 5.231? S), Madang Province,
Papua New Guinea were destructively
sampled (see Whitfeld et al. 2012 for de-
tails). The two plots were 800 m apart in
a mosaic of primary and secondary rain
forest vegetation at 100?200 m above sea
level in an extensive mixed evergreen
forest on latosols in the Ramu River basin
(Paijmans 1976). The climate is generally
humid and relatively aseasonal. Historical
readings fromMadang (70 km east, 1956?
1970; McAlpine et al. 1983) indicate a
mean annual rainfall of 3500 mm and
mean monthly temperature between
26.2 ?C and 26.7 ?C. Local landowners
practice subsistence agriculture in 0.25?
1.0 ha gardens planted after felling and
burning of primary forest, and we co-
ordinated our sampling with local land-
owners whowere planning to clear the sites
for subsistence gardens. After mapping and
clearing adjacent vegetation, trees were
individually felled and immediately in-
spected for the presence of caterpillars and
leaf miners by a team of eight ?eld
workers. Live caterpillars were hand-
collected and placed in plastic vials for
processing. The live caterpillars were as-
signed to morphospecies (numbered in the
CATX series), documented with photo-
graphs, and, in most cases, with preserved
vouchers. Host plant data are included in the
BOLD records via the plant number in the
form WS2B2315. All plants were vouch-
ered with full data available as part of the
Digital Flora of New Guinea in the Atrium
database (http://ng.atrium-biodiversity.org/
atrium). DNA data (rbcL) for representa-
tives of most plant species are in GenBank
(accession numbers JF738369?JF739166).
Lepidoptera methods: General field
and laboratory methods for Lepidoptera
are described in Miller et al. (2003) and
Novotny et al. (2010). DNA sequencing
(COI barcode) followed standard methods
at the Biodiversity Institute of Ontario,
University of Guelph (Craft et al. 2010,
Wilson 2012), using tissue from the anal
end of the caterpillars. 475 vouchers were
sampled for DNA, resulting in 462 suc-
cessful sequences (including one Coleoptera
larva and three larvae of Hymenoptera
parasitoids accidently sampled with the
caterpillar tissue and preferentially am-
pli?ed from the samples). Caterpillar
vouchers are deposited in the Smithsonian
National Museum of Natural History. The
images in the BOLD records were taken
from live caterpillars to characterize the
morphospecies as they were classi?ed in
the ?eld. In many cases, these images
represent the same specimens that were
sequenced, but not always, and we have
noticed (and removed) several cases of
confused labeling of images, so the
images should be used with care.
Identifications: The 462 successful se-
quences group into 351 barcode clusters.
These were identified using the BOLD
identification tool, primarily against some
18,000 sequences that we have generated
for adult New Guinea Lepidoptera, and
a larger library for adult Australian
Lepidoptera, including more than 30,000
specimens from the Australian National
Insect Collection. Identifications were
made either based on very close matches
at the species level (less than one percent
difference) or using NJ tree topology at
the generic level or above. Some species
remain unidentified, and we welcome
comments on all identifications.
We are currently preparing ecological an-
alyses of the caterpillar communities, their
host plants, and their parasites (e.g., Hrcek
et al. 2011) and continuing taxonomic ana-
lyses of the adult Lepidoptera that can be
linked to these larvae by their DNAbarcodes.
ACKNOWLEDGMENTS
We thank the staff at the New Guinea
BinatangResearchCenter for field assistance,
Wanang landowners for access to field
sites and assistance, Kipiro Damas and
PNG Forest Research Institute for plant
species identification. Karolyn Darrow,
Lauren Helgen, Margaret Rosati, and
many taxonomists have assisted in build-
ing the DNA barcode library used for
Lepidoptera identifications. Field work
was supported by the U.S. National Sci-
ence Foundation grant DEB-0515678,
Czech Science Foundation grant 206/09/
0115, and CzechMinistry of Education &
European Union grant CZ.1.07/2.3.00/
20.0064. DNA sequencing was supported
by a grant from the Government of Canada
through Genome Canada and the Ontario
Genomics Institute in support of the iBOL
project.
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Scott E. Miller, National Museum of
Natural History, Smithsonian Institution,
P.O. Box 37012, MRC 105, Washington,
D.C. 20013-7012, U.S.A. (e-mail: millers@si.
edu)
Jan Hrcek, Faculty of Science, University
of South Bohemia and Biology Center, Czech
Academy of Sciences, Branisovska 31, 37005
Ceske Budejovice, Czech Republic
Vojtech Novotny, Czech Academy of Sci-
ences, Biology Center and University of
South Bohemia, Faculty of Science, Brani-
sovska 31, 37005 Ceske Budejovice, Czech
Republic
George D. Weiblen, University of Min-
nesota, Bell Museum and Department of
Plant Biology, 250 Biological Sciences Cen-
ter, 1445 Gortner Avenue, St. Paul, Minne-
sota 55108-1095, U.S.A.
Paul D.N. Hebert, Department of Integra-
tive Biology and the Biodiversity Institute of
Ontario, University of Guelph, Guelph,
Ontario N1G 2W1, Canada
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