Journal of
Threatened Taxa | www.threatenedtaxa.org | 26 November 2018 | 10(13):
12817–12820
Dna
barcoding and morphological characterization of moth Antoculeora
ornatissima (Walker, 1858) (Lepidoptera: Noctuidae),
a new range record from western Himalayan region of India
Twinkle Sinha
1, P.R. Shashank 2 & Pratima Chaudhuri Chattopadhyay 3
1,2 Division of Entomology, Indian
Agricultural Research Institute, New Delhi 110012, India
3 Amity Institute
of Biotechnology, Amity University, Noida, Uttar Pradesh 201303, India
1 twinklesinha1901@gmail.com, 2
spathour@gmail.com (corresponding author), 3 pchaudhuri@amity.edu
doi: https://doi.org/10.11609/jott.4127.10.13.12817-12820 | ZooBank: urn:lsid:zoobank.org:pub:7E5BE3DA-2985-412C-A4B6-B31E16AEC95D
Editor: Jatishwor Singh Irungbam,
Biology Centre CAS, České Budějovice,
Czech Republic. Date of publication: 26 November 2018
(online & print)
Manuscript details: Ms
# 4127 | Received 13 March 2018 | Final received 18 September 2018 | Finally
accepted 20 October 2018
Citation: Sinha, T., P.R. Shashank
& P.C. Chattopadhyay (2018).
DNA
barcoding and morphological characterization of moth Antoculeora
ornatissima (Walker, 1858) (Lepidoptera: Noctuidae), a new range record from western Himalayan
region of India. Journal of Threatened Taxa 10(13): 12817–12820; https://doi.org/10.11609/jott.4127.10.13.12817-12820
Copyright: © Sinha et al. 2018.
Creative Commons Attribution 4.0 International License.
JoTT allows unrestricted use of this article in any
medium, reproduction and distribution by providing adequate credit to the
authors and the source of publication.
Funding: DST-SERB
(SB/YS/LS-126/2014).
Competing interests: The authors declare no competing interests.
Acknowledgements: The authors are
grateful to Dr. László Ronkay,
Department of Zoology, Hungarian Natural History Museum, Budapest, H-1431
Hungary for his guidance in noctuids work. We thank
DST-SERB (SB/YS/LS-126/2014) for financial assistance to conduct this work.
Authors are indebted to Head, Division of Entomology, IARI, New
Delhi authorities of ICAR-Indian Agricultural Research Institute, New Delhi for
necessary help.
Abstract: DNA barcoding of Antoculeora
ornatissima (Walker, 1858) was done for the first
time from India. Redescriptions
of genitalia and diagnoses of genus and species are presented with images and
illustrations.
Keywords: Antoculeora ornatissima, DNA barcoding, Lepidoptera,
morphology, new range record, Noctuidae, Plusiinae, western Himalaya.
Noctuidae is one of the
largest families of moths with more than 35,000 known species documented in the
world. The subfamily Plusiinae
(Lepidoptera: Noctuidae) is represented by
approximately 500 species worldwide (Ronkay et al. 2008) of which 59 are reported in India (Shashank & Longjam 2014),
which were grouped in three tribes, Abrostolini Eichlin & Cunningham, 1978, Argyrogrammatini
Eichlin & Cunningham, 1978, and Plusiini Boisduval, 1928. Abrostolini is
represented by two species, Argyrogrammatini by 33
species, and Plusiini by 24 species. Under the subfamily Plusiinae,
the genus Antoculeora is represented by three
species, namely, A. yoshimotoi (Ronkay, 1997), A. locuples
(Oberthür, 1880), and A. ornatissima
(Walker, 1858). Walker (1858) described Plusia ornatissima
on a single female specimen from the southern Himalayan region (northern
Hindustan). Later, Antoculeora
was erected as a subgenus of Erythroplusia
Ichinose, 1962 (Ichinose, 1973) and raised to a full genus by Chou & Lu
(1979). Further, Oberthür
(1880) described P. locuples for the island of
Askold located in the Bay of Vladivostok,
Russia. These two taxa were treated as
synonyms by Staudinger & Rebel (1901), whose opinion was accepted by Hampson (1913) and Warren (1913), followed by all
subsequent authors till Ronkay (1997). Kitching (1987)
expressed his opinion that A. ornatissima
might be a complex of closely related taxa but may also be a single
species. A detailed
history of generic classification and species changes was provided by Kitching (1987), Ronkay (1997),
and Ronkay et al. (2008).
Due
to large intraspecific variation in this species and to resolve a few
ambiguities of the species complex, a need for molecular work was suggested by Ronkay (1997) and Ronkay et al. (2008).
This is the first effort to study the occurrence and DNA barcode of A.
ornatissima from India.
Material and Methods
Sampling
For the present
study, 12 specimens were collected from Chamoli and Katrain, two different localities of northern India. The collection of specimens
were carried out by hand collection of larvae, which were reared to the
adult stage in the Lepidoptera Laboratory, National Pusa
Collection, Indian Agricultural Research Institute, New Delhi (NPC-IARI). Light traps using hanging cloth method were
also used for collection of adult moths.
Further, collected materials were processed by
pinning, spreading, proper labelling, and preparation
of wings and genitalia slides.
All the specimens are preserved in NPC-IARI.
Morphology of adult moths
Genitalia of
both male and female specimens were prepared and images were taken with a Leica
DFC425C digital camera mounted on a Leica M205FA stereozoom
microscope. The terminology used for
male and female genitalia follows Klots (1970). Forewing length was measured from the outer
edge of the tegula to the outer most edge of the
apex.
DNA isolation, PCR amplification, and sequencing
The DNA easy
blood and tissue kit (Quiagen GmbH, Germany) method
was used to extract DNA from one to three legs of each adult. The DNA extraction method provided by Fukova et al. (2009) was followed. For mitochondrial cytochrome c oxidase subunit
I gene analysis was used on the voucher specimens. The genomic DNA was visualized using 0.8% agarose gel and quantified by fluorometer
using standard procedures. Depending
upon the concentration, the DNA samples were diluted with molecular gradient water
to get a working solution of 10–30 ng/µL. A portion of the total DNA was preserved in
glycerol (10%) in -80°C for future reference purposes. The universal barcode primer described by Folmer et al. (1994) (LCO-5’-GGT CAA CAA
ATC ATA AAG ATA TTG G-3’; HCO-5’-TAA ACT TCA GGG TGA CCA AAA AAT CA-3’)
specific to mitochondrial cytochrome c oxidase subunit I gene (COI) was used in
the present study. The optimized PCR
conditions (per 25µL) using Taq DNA polymerase (Fermentas Inc., USA) were 2.5µL of 10 X PCR buffer with 2µL
of 25mM MgCl2, 0.5µL of 10mM dNTPs, 0.5µL
each of forward and reverse primer, IU of Taq, and
17µL of UltraPure water (Invitrogen). Thermocycler
conditions were as follows: initial denaturation for 5min at 94°C followed by
35 cycles of denaturing for 30s at 94°C, annealing for 40s at 54°C and an
extension time of 40s at 72°C, with a final extension for 5min at 72°C. PCR products were visualized on agarose gel after electrophoresis. Single bands were purified using a QIAquick PCR purification kit (Quiagen
GmbH, Germany). Purified PCR products
were sequenced directly in both directions using an automated sequencer (ABI
prism® 3730 XL DNA Analyzer; Applied Biosystems, USA)
at SciGenom Lab, Cochin, India. COI sequences in FASTA format were processed
and submitted to NCBI for GenBank accessions as per Shashank et al. (2014).
Accession numbers for the five specimens are KY886404, KY886405,
KY886406, KY886407, and KY886408.
Results
Redescription of Antoculeora
ornatissima (Walker, 1858)
Systematic accounts
Family: Noctuidae Latreille, 1809
Subfamily: Plusiinae
Boisduval, 1828
Genus: Antoculeora
Ichinose, 1973
Genus: Antoculeora
Ichinose, 1973
Type species: Plusia ornatissima.
The genus Antoculeora is characterized by its large size of 30–42
mm, with broader, acutely pointed forewings.
Head and collar lateritius. Forewing with metallic
sheen present, gamma mark larger and divided into two oval spots. Male genitalia with longer uncus, juxta sclerotised. Asymmetrical valvae with saccular extensions. Female genitalia with longer sacculiform corpus bursae with
strongly ribbed appendix bursae. Ovipositor rather medium sized, weakly sclerotised, papillae anales
rounded.
Antoculeora ornatissima (Walker, 1858) (Images 1A,B)
Plusia ornatissima Walker, 1858: 1786. Type locality: northern Hindostan.
Cerviplusia wukongensis Chou & Lu, 1974: 73. Type locality: China.
Antoculeora ornatissima Goater, Ronkay & Fibiger, 2003.
Redescription: Forewing length of adults 36–40 mm. Spherical rounded medium-sized compound
eyes. Frons and vertex copper-bronze coloured. Frons distinctly exceeding the eyes. Labial palpi
upturned, third segment is pointed and small.
Second segment of labial palpi is densely
covered with reddish scales. Antennae of male and female filiform in
shape. Metathoracic
tuft well developed, looks like a pair of horns and generally reddish-orange coloured. Meso- and metathorax densely
covered with hairs. Foreleg, midleg, and hindleg are similar
in both males and females. Femur and
tibia of foreleg covered with brownish-orange hairs. Tarsi full of spines. Two pairs of spurs present
on hind leg. Abdomen
light brown coloured. Abdominal tuft well developed. Two prominent crest present on abdomen. Anal tuft present. Orange coloured
scales present on the back portion of the entire abdomen. Forewing triangular, subfalcate
and metallic shimmering present on the termen region
and post-medial region. Antemedial line golden coloured
and ends at the stigma. Stigma mark is
like two large oval spots with silver colour filled
into it. Comma-like structure present
above the gamma mark, the best identification feature of this species. Hindwing aeneous brown coloured. Forewing with 12 veins, vein SC and R1 are
free. RS2+RS3 joined by a short vein to discal cell. Vein M2
present near the middle of the discal cell. Veins RS1 and RS2 connected with short vein
to form an areole. Anal
vein separately present. Hindwing with
nine veins present. RS1 joined
with M1 (RS1+M1). M2, M3, and CuA1
originate from the same base stalk. Two
anal veins A1 and A2 freely present.
Male genitalia
(Image 1C,D): Male genitalia robust.
Valvae with longer projections outwards. Fultura inferior rather low, apical part more or less broadly triangular. Tegumen arms are
extremely swollen anteriorly and, in dorsal view, diverge at 180°. The valvae have
projections that can interlock itself when valvae are closed.
Vinculum U-shaped, clavus longer in
shape. Aedeagus
with carina terminated in a broadly half-moon-shaped plate covered with short
but strong teeth at base (and sometimes in medial third); basal part of vesica with two diverticula bearing bundles of short, spiniform cornuti, one of them
usually long and tubular, second much smaller, often without spinules; distal part of vesica
with scobinate walls but without spinulose
field.
Female
genitalia (Image 1E): Corpus bursae
lobular. Proximal third of corpus bursae a large, spacious sac, covered partly with fine,
short, hair-like spiculi. Medial third of corpus bursae
only slightly wrinkled. Ductus bursae smaller and twisted. Diverticule of corpus bursae sclerotised,
proximal papillae anales asymmetrical. Ostium bursae form a heavily sclerotized, double complex. Posterior part smaller, flattened, more or
less rectangular with deeply incised proximal margin; anterior part huge,
axe-head-shaped with pointed postero-lateral
tip. Cervix bursae
elongated, folded.
Material
examined: 1901–1904, 19.v.2015, 2 males, 2 females, Chamoli, Uttarakhand, India, 30.2420N
& 79.6140E, coll. Shashank; 1905,
04.vi.2014, 1 male, Palampur, Himachal Pradesh,
India, 32.1290N & 76.5380E, coll. Pathania;
1906–1908, 05.ix.2007, 2 males, 1 female, Srinagar, Jammu & Kashmir, India,
34.1170N & 74.7760E, coll. Rajesh; 1909–1912,
21.ix.2016, 2 males, 2 females, Katrain, Himachal
Pradesh, India, 32.0970N & 77.1350E, Shashank.
Global
distribution: India (Uttarakhand, Himachal Pradesh,
Jammu & Kashmir, and Sikkim), Pakistan, China, Japan, and Russia.
Conclusions
Our study
highlights the occurrence of species A. ornatissima
from India. Molecular evidence along
with morphology confirm the presence of this
species. There was no molecular data
available for this species from India, which is also its type locality. DNA barcodes provided in this study will help
in the accurate diagnosis of this species from India.
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