Lepidoptera: Noctuidae), a New Range Record from Western Himalayan Region of India

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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, P.R. Shashank & Praꢀma Chaudhuri Chaꢁopadhyay

26 November 2018 | Vol. 10 | No. 13 | Pages: 12817–12820 10.11609/joꢁ.4127.10.13.12817-12820

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Dna barcoding and morphological characterization of moth Antoculeora ornatissima (Walker, 1858) (Lepidoptera: Noctuidae), a new range record from western Himalayan region of India

ISSN 0974-7907 (Online) ISSN 0974-7893 (Print)

Twꢄꢇkle Sꢄꢇꢊꢀ¹, P .R. Sꢊꢀsꢊꢀꢇk²& Pꢉꢀꢆꢈꢀ Cꢊꢀꢃdꢊꢃꢉꢄ Cꢊꢀꢋꢂpꢀdꢊyꢀy³

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^1,2Division of Entomology, Indian Agricultural Research Insꢁtute, New Delhi 110012, India ³Amity Insꢁtute of Biotechnology, Amity University, Noida, Uꢀar Pradesh 201303, India ¹[email protected], ²[email protected] (corresponding author), ³[email protected]

Absꢁꢉꢀꢅꢁ: DNA barcoding of Antoculeora ornaꢀssima (Walker, 1858)

the genus Antoculeora is represented by three species,

namely, A. yoshimotoi (Ronkay, 1997), A. locuples

(Oberthür, 1880), and A. ornaꢀssima (Walker, 1858). Walker (1858) described Plusia ornaꢀssima 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).

was done for the ﬁrst ꢁme from India. Redescripꢁons of genitalia and diagnoses of genus and species are presented with images and illustraꢁons.

Keywꢂꢉds: Antoculeora ornaꢀssima, DNA barcoding, Lepidoptera,

morphology, new range record, Noctuidae, Plusiinae, western Himalaya.

Noctuidae is one of the largest families of moths with Further, Oberthür (1880) described P . l ocuples for the more than 35,000 known species documented in the island of Askold located in the Bay of Vladivostok, Russia. world. The subfamily Plusiinae (Lepidoptera: Noctuidae) These two taxa were treated as synonyms by Staudinger is represented by approximately 500 species worldwide & Rebel (1901), whose opinion was accepted by Hampson (Ronkay et al. 2008) of which 59 are reported in India (1913) and Warren (1913), followed by all subsequent (Shashank & Longjam 2014), which were grouped in authors ꢁll Ronkay (1997). Kitching (1987) expressed his three tribes, Abrostolini Eichlin & Cunningham, 1978, opinion that A. ornaꢀssima might be a complex of closely Argyrogrammaꢁni Eichlin & Cunningham, 1978, and related taxa but may also be a single species. A detailed Plusiini Boisduval, 1928. Abrostolini is represented history of generic classiﬁcaꢁon and species changes was by two species, Argyrogrammaꢁni by 33 species, and provided by Kitching (1987), Ronkay (1997), and Ronkay Plusiini by 24 species. Under the subfamily Plusiinae, et al. (2008).

DOI: hꢀp s ://doi.org/10.11609/joꢀ.4127.10.13.12817-12820 | ZꢂꢂBꢀꢇk: urn:lsid:zoobank.org:pub:7E5BE3DA-2985-412C-A4B6-B31E16AEC95D

Edꢄꢁꢂꢉ: Jaꢁshwor Singh Irungbam, Biology Centre CAS, České Budějovice, Czech Republic.

Dꢀꢁe ꢂf pꢃblꢄꢅꢀꢆꢂꢇ: 26 November 2018 (online & print)

Mꢀꢇꢃsꢅꢉꢄpꢁ deꢁꢀꢄls: Ms # 4127 | Received 13 March 2018 | Final received 18 September 2018 | Finally accepted 20 October 2018 Cꢄꢁꢀꢆꢂꢇ: Sinha, T., P.R. Shashank & P.C. Chaꢀopadhyay (2018). DNA barcoding and morphological characterizaꢁon of moth Antoculeora ornaꢀssima (Walker, 1858) (Lepidoptera: Noctuidae), a new range record from western Himalayan region of India. Journal of Threatened Taxa 10(13): 12817–12820; hꢀps://doi.org/10.11609/

joꢀ.4127.10.13.12817-12820

Cꢂpyꢉꢄgꢊꢁ: © Sinha et al. 2018. Creaꢁve Commons Aꢀribuꢁon 4.0 Internaꢁonal License. JoTT allows unrestricted use of this arꢁcle in any medium, reproducꢁon and distribuꢁon by providing adequate credit to the authors and the source of publicaꢁon.

Fꢃꢇdꢄꢇg: DST-SERB (SB/YS/LS-126/2014). Cꢂꢈpeꢆꢇg ꢄꢇꢁeꢉesꢁs: The authors declare no compeꢁng interests. Aꢅkꢇꢂwledgeꢈeꢇꢁs: 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 authoriꢁes of ICAR-Indian Agricultural Research Insꢁtute, New Delhi for necessary help.

12817

DNA bꢀꢉꢅꢂdꢄꢇg ꢀꢇd ꢈꢂꢉpꢊꢂlꢂgꢄꢅꢀl ꢅꢊꢀꢉꢀꢅꢁeꢉꢄzꢀꢆꢂꢇ ꢂf Aꢇtꢀꢁꢂꢃꢄꢀꢅꢆ ꢀꢅꢇꢆꢈꢉꢉꢊꢋꢆ

Sꢄꢇꢊꢀ eꢁ ꢀl.

Due to large intraspecific variaꢁon in this species and forwardandreverseprimer,IUofTaq,and17µLofUltraPure to resolve a few ambiguiꢁes of the species complex, a water (Invitrogen). Thermocycler condiꢁons were as need for molecular work was suggested by Ronkay (1997) follows: iniꢁal denaturaꢁon for 5min at 94°C followed by and Ronkay et al. (2008). This is the first effort to study 35 cycles of denaturing for 30s at 94°C, annealing for 40s the occurrence and DNA barcode of A. ornaꢀssima from at 54°C and an extension ꢁme of 40s at 72°C, with a final

India.

extension for 5min at 72°C. PCR products were visualized

on agarose gel aﬅer electrophoresis. Single bands were purified using a QIAquick PCR purificaꢁon kit (Quiagen GmbH, Germany). Purified PCR products were sequenced

Material and Methods

Sampling
For the present study, 12 specimens were collected directly in both direcꢁons using an automated sequencer from Chamoli and Katrain, two diﬀerent localiꢁes of (ABI prism® 3730 XL DNA Analyzer; Applied Biosystems, northern India. The collecꢁon of specimens were carried USA) at SciGenom Lab, Cochin, India. COI sequences in out by hand collecꢁon of larvae, which were reared to the FASTA format were processed and submiꢀed to NCBI adult stage in the Lepidoptera Laboratory, Naꢁonal Pusa for GenBank accessions as per Shashank et al. (2014). Collecꢁon, Indian Agricultural Research Insꢁtute, New Accession numbers for the ﬁve specimens are KY886404, Delhi (NPC-IARI). Light traps using hanging cloth method KY886405, KY886406, KY886407, and KY886408. were also used for collecꢁon of adult moths. Further, collected materials were processed by pinning, spreading, Results proper labelling, and preparaꢁon of wings and genitalia Redescripꢁon of Antoculeora ornaꢀssima (Walker, 1858)

slides. All the specimens are preserved in NPC-IARI.

Systemaꢁc accounts

Family: Noctuidae Latreille, 1809

Subfamily: Plusiinae Boisduval, 1828

Morphology of adult moths

Genitalia of both male and female specimens were Genus: Antoculeora Ichinose, 1973 prepared and images were taken with a Leica DFC425C digital camera mounted on a Leica M205FA stereozoom Geꢇꢃs: Antoculeora Iꢅꢊꢄꢇꢂse, 1973 microscope. The terminology used for male and female

Type species: Plusia ornaꢀssima.

genitalia follows Klots (1970). Forewing length was
The genus Antoculeora is characterized by its large size measured from the outer edge of the tegula to the outer of 30–42 mm, with broader, acutely pointed forewings.

most edge of the apex.

Head and collar lateriꢁus. Forewing with metallic sheen

present, gamma mark larger and divided into two oval

spots. Male genitalia with longer uncus, juxta scleroꢁsed.

DNA isolaꢁon, PCR ampliﬁcaꢁon, and sequencing

The DNA easy blood and ꢁssue kit (Quiagen GmbH, Asymmetrical valvae with saccular extensions. Female
Germany) method was used to extract DNA from one genitalia with longer sacculiform corpus bursae with to three legs of each adult. The DNA extracꢁon method strongly ribbed appendix bursae. Ovipositor rather provided by Fukova et al. (2009) was followed. For medium sized, weakly scleroꢁsed, papillae anales mitochondrial cytochrome c oxidase subunit I gene rounded. analysis was used on the voucher specimens. The genomic DNA was visualized using 0.8% agarose gel and quanꢁﬁed Aꢇtꢀꢁꢂꢃꢄꢀꢅꢆ ꢀꢅꢇꢆꢈꢉꢉꢊꢋꢆ (Wꢀlkeꢉ, 1858) (Images 1A,B)

by ﬂuorometer using standard procedures. Depending

Plusia ornaꢀssima Walker, 1858: 1786. Type locality:

upon the concentraꢁon, the DNA samples were diluted northern Hindostan.

with molecular gradient water to get a working soluꢁon

Cerviplusia wukongensis Chou & Lu, 1974: 73. Type

of 10–30 ng/µL. A porꢁon of the total DNA was preserved locality: China. in glycerol (10%) in -80°C for future reference purposes. The universal barcode primer described by Folmer et 2003. 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’) mm.

Antoculeora ornaꢀssima Goater, Ronkay & Fibiger,

Redescripꢁon: Forewing length of adults 36–40
Spherical rounded medium-sized compound specific to mitochondrial cytochrome c oxidase subunit I eyes. Frons and vertex copper-bronze coloured. Frons gene (COI) was used in the present study. The opꢁmized disꢁnctly exceeding the eyes. Labial palpi upturned, third PCR condiꢁons (per 25µL) using Taq DNA polymerase segment is pointed and small. Second segment of labial (Fermentas Inc., USA) were 2.5µL of 10 X PCR buffer with palpi is densely covered with reddish scales. Antennae 2µL of 25mM MgCl₂, 0.5µL of 10mM dNTPs, 0.5µL each of of male and female filiform in shape. Metathoracic tuﬅ

12818

Journal of Threatened Taxa | www.threatenedtaxa.org | 26 November 2018 | 10(13): 12817–12820

B
A

C

E
D

Iꢈꢀge 1. Aꢇtꢀꢁꢂꢃꢄꢀꢅꢆ ꢀꢅꢇꢆꢈꢉꢉꢊꢋꢆ (Wꢀlkeꢉ, 1858): A - ꢀdꢃlꢁ ꢈꢀle, B - ꢀdꢃlꢁ feꢈꢀle, C - ꢈꢀle geꢇꢄꢁꢀlꢄꢀ, D - ꢀedeꢀgꢃs, E - feꢈꢀle geꢇꢄꢁꢀlꢄꢀ

well developed, looks like a pair of horns and generally present. Orange coloured scales present on the back reddish-orange coloured. Meso- and metathorax densely porꢁon of the enꢁre abdomen. Forewing triangular, covered with hairs. Foreleg, midleg, and hindleg are subfalcate and metallic shimmering present on the similar in both males and females. Femur and ꢁbia of termen region and post-medial region. Antemedial line foreleg covered with brownish-orange hairs. Tarsi full of golden coloured and ends at the sꢁgma. Sꢁgma mark is spines. Two pairs of spurs present on hind leg. Abdomen like two large oval spots with silver colour ﬁlled into it. light brown coloured. Abdominal tuﬅ well developed. Comma-like structure present above the gamma mark, Two prominent crest present on abdomen. Anal tuﬅ the best idenꢁﬁcaꢁon feature of this species. Hindwing

Journal of Threatened Taxa | www.threatenedtaxa.org | 26 November 2018 | 10(13): 12817–12820

12819

aeneous brown coloured. Forewing with 12 veins, vein SC from India, which is also its type locality. DNA barcodes and R1 are free. RS2+RS3 joined by a short vein to discal provided in this study will help in the accurate diagnosis of cell. Vein M2 present near the middle of the discal cell. this species from India. 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.

References

Bꢂꢄsdꢃvꢀl, A.J.A. (1829). Europaeorum Lepidopterorum Index

Methodicus. 103pp.

Cꢊꢂꢃ, I. & T. Lꢃ (1974). Studies on Chinese Plusiinae (Lepidoptera:

Noctuidae). Acta Entomologica Sinica 17(1): 66–82.

Cꢊꢂꢃ, I. & T. Lꢃ (1979). Eight new species of Plusiinae and revision of some of its known species (Lepidoptera: Noctuidae). Acta Entomologica

Sinica 22(1): 61–72.

Eꢄꢅꢊlꢄꢇ, T.D. & H.B. Cꢃꢇꢇꢄꢇgꢊꢀꢈ (1978). The Plusiinae (Lepidoptera:

Noctuidae) of America north of Mexico, emphasizing genitalic and

larval morphology. United States Department of Agriculture Technical Bulleꢀn 1567: 1–222.

Fꢂlꢈeꢉ, O., M. Blꢀꢅk, W. Hꢂeꢊ, R. Lꢃꢁz & R. Vꢉꢄjeꢇꢊꢂek (1994). DNA

primers for ampliﬁcaꢁon of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine

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Fꢃkꢂvꢀ, I., L.G. Neveꢇ, N.M. Bꢀꢉꢅeꢇꢀs, N.A. Gꢃꢇd, M. Dꢀlꢄkꢂvꢀ & F.

Mꢀꢉeꢅ (2009). Rapid assessment of the sex of codling moth Cydia pomonella (Linnaeus) (Lepidoptera: Totricidae) eggs to larvae. Journal

of Applied Entomology 133(4): 249–261.

Male genitalia (Image 1C,D): Male genitalia robust.
Valvae with longer projecꢁons 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 projecꢁons 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 someꢁmes in medial third); basal part of vesica with two diverꢁcula bearing bundles of short, spiniform cornuꢁ, one of them usually long and tubular, second much smaller, oﬅen 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. Diverꢁcule of corpus bursae scleroꢁsed, proximal papillae anales asymmetrical. Osꢁum bursae form a heavily scleroꢁzed, double complex. Posterior part smaller, flaꢀened, more or less rectangular with deeply incised proximal margin; anterior part huge, axe-head-shaped with pointed postero-lateral ꢁp. Cervix bursae elongated, folded.

Gꢂꢀꢁeꢉ, B., L. Rꢂꢇkꢀy & M. Fꢄbꢄgeꢉ (2003). Catocalinae and Plusiinae,

Noctuidae Europaeae,10, Sorø, 1: 452pp.

Hꢀꢈpsꢂꢇ, G.F. (1913). Catalogue of the Lepidoptera Phalaenae in the Collecꢀon of the Briꢀsh Museum. Catocalinae Conꢀnued, Mominae,

Phytometrinae, Vol. 13. Briꢁsh Museum Trustees, London, 609pp. Iꢅꢊꢄꢇꢂse, T. (1962). Studies on the Noctuid subfamily Plusiinae of Japan.

Bulleꢀn of Faculty of Agriculture, Tokyo University of Agriculture and Technology 6: 1–127.

Iꢅꢊꢄꢇꢂse, T. (1973). A revision of some genera of the Japanese Plusiinae, with descripꢁons of a new genus and two new subgenera (Lepidoptera, Noctuidae). Kontyu 41(2): 135–140. Kꢄꢁꢅꢊꢄꢇg, I.J. (1987). SpectaclesandsilverYs:asynthesisofthesystemaꢁcs, cladisꢁcs & biology of the Plusiinae (Lepidoptera: Noctuidae). Bulleꢀn