OPEN ACCESS The Journal of Threatened Taxa is dedicated to building evidence for conservaton globally by publishing peer-reviewed artcles online every month at a reasonably rapid rate at www.threatenedtaxa.org. All artcles published in JoTT are registered under Creatve Commons Atributon 4.0 Internatonal License unless otherwise mentoned. JoTT allows unrestricted use of artcles in any medium, reproducton, and distributon by providing adequate credit to the authors and the source of publicaton. Journal of Threatened Taxa Building evidence for conservaton globally www.threatenedtaxa.org ISSN 0974-7907 (Online) | ISSN 0974-7893 (Print) Short Communication 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 & Pratma Chaudhuri Chatopadhyay 26 November 2018 | Vol. 10 | No. 13 | Pages: 12817–12820 10.11609/jot.4127.10.13.12817-12820 For Focus, Scope, Aims, Policies and Guidelines visit htps://threatenedtaxa.org/index.php/JoTT/about/editorialPolicies#custom-0 For Artcle Submission Guidelines visit htps://threatenedtaxa.org/index.php/JoTT/about/submissions#onlineSubmissions For Policies against Scientfc Misconduct visit htps://threatenedtaxa.org/index.php/JoTT/about/editorialPolicies#custom-2 For reprints contact <[email protected]> Publisher & Host Partners Member Threatened Taxa 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 ISSN 0974-7907 (Online) region of India ISSN 0974-7893 (Print) Communication Short Twinkle Sinha 1 , P.R. Shashank 2 & Pratma Chaudhuri Chatopadhyay 3 OPEN ACCESS 1,2 Division of Entomology, Indian Agricultural Research Insttute, New Delhi 110012, India 3 Amity Insttute of Biotechnology, Amity University, Noida, Utar Pradesh 201303, India 1 [email protected], 2 [email protected] (corresponding author), 3 [email protected] Abstract: DNA barcoding of Antoculeora ornatssima (Walker, 1858) the genus Antoculeora is represented by three species, was done for the frst tme from India. Redescriptons of genitalia namely, A. yoshimotoi (Ronkay, 1997), A. locuples and diagnoses of genus and species are presented with images and illustratons. (Oberthür, 1880), and A. ornatssima (Walker, 1858). Walker (1858) described Plusia ornatssima on a single Keywords: Antoculeora ornatssima, DNA barcoding, Lepidoptera, morphology, new range record, Noctuidae, Plusiinae, western female specimen from the southern Himalayan region Himalaya. (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). Noctuidae is one of the largest families of moths with Further, Oberthür (1880) described P. locuples 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 tll Ronkay (1997). Kitching (1987) expressed his three tribes, Abrostolini Eichlin & Cunningham, 1978, opinion that A. ornatssima might be a complex of closely Argyrogrammatni Eichlin & Cunningham, 1978, and related taxa but may also be a single species. A detailed Plusiini Boisduval, 1928. Abrostolini is represented history of generic classifcaton and species changes was by two species, Argyrogrammatni by 33 species, and provided by Kitching (1987), Ronkay (1997), and Ronkay Plusiini by 24 species. Under the subfamily Plusiinae, et al. (2008). DOI: htps://doi.org/10.11609/jot.4127.10.13.12817-12820 | ZooBank: urn:lsid:zoobank.org:pub:7E5BE3DA-2985-412C-A4B6-B31E16AEC95D Editor: Jatshwor Singh Irungbam, Biology Centre CAS, České Budějovice, Czech Republic. Date of publicaton: 26 November 2018 (online & print) Manuscript details: Ms # 4127 | Received 13 March 2018 | Final received 18 September 2018 | Finally accepted 20 October 2018 Citaton: Sinha, T., P.R. Shashank & P.C. Chatopadhyay (2018). DNA barcoding and morphological characterizaton of moth Antoculeora ornatssima (Walker, 1858) (Lepidoptera: Noctuidae), a new range record from western Himalayan region of India. Journal of Threatened Taxa 10(13): 12817–12820; htps://doi.org/10.11609/ jot.4127.10.13.12817-12820 Copyright: © Sinha et al. 2018. Creatve Commons Atributon 4.0 Internatonal License. JoTT allows unrestricted use of this artcle in any medium, reproducton and distributon by providing adequate credit to the authors and the source of publicaton. Funding: DST-SERB (SB/YS/LS-126/2014). Competng interests: The authors declare no competng 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 fnancial assistance to conduct this work. Authors are indebted to Head, Division of Entomology, IARI, New Delhi authorites of ICAR-Indian Agricultural Research Insttute, New Delhi for necessary help. 12817 DNA barcoding and morphological characterizaton of Antoculeora ornatssima Sinha et al. Due to large intraspecifc variaton in this species and forward and reverse primer, IU of Taq, and 17µL of UltraPure to resolve a few ambiguites of the species complex, a water (Invitrogen). Thermocycler conditons were as need for molecular work was suggested by Ronkay (1997) follows: inital denaturaton for 5min at 94°C followed by and Ronkay et al. (2008). This is the frst efort to study 35 cycles of denaturing for 30s at 94°C, annealing for 40s the occurrence and DNA barcode of A. ornatssima from at 54°C and an extension tme of 40s at 72°C, with a fnal India. extension for 5min at 72°C. PCR products were visualized on agarose gel afer electrophoresis. Single bands were Material and Methods purifed using a QIAquick PCR purifcaton kit (Quiagen Sampling GmbH, Germany). Purifed PCR products were sequenced For the present study, 12 specimens were collected directly in both directons using an automated sequencer from Chamoli and Katrain, two diferent localites of (ABI prism® 3730 XL DNA Analyzer; Applied Biosystems, northern India. The collecton of specimens were carried USA) at SciGenom Lab, Cochin, India. COI sequences in out by hand collecton of larvae, which were reared to the FASTA format were processed and submited to NCBI adult stage in the Lepidoptera Laboratory, Natonal Pusa for GenBank accessions as per Shashank et al. (2014). Collecton, Indian Agricultural Research Insttute, New Accession numbers for the fve specimens are KY886404, Delhi (NPC-IARI). Light traps using hanging cloth method KY886405, KY886406, KY886407, and KY886408. were also used for collecton of adult moths. Further, collected materials were processed by pinning, spreading, Results proper labelling, and preparaton of wings and genitalia Redescripton of Antoculeora ornatssima (Walker, 1858) slides. All the specimens are preserved in NPC-IARI. Systematc accounts Family: Noctuidae Latreille, 1809 Morphology of adult moths Subfamily: Plusiinae Boisduval, 1828 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 Genus: Antoculeora Ichinose, 1973 microscope. The terminology used for male and female Type species: Plusia ornatssima. 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 lateritus. Forewing with metallic sheen present, gamma mark larger and divided into two oval DNA isolaton, PCR amplifcaton, and sequencing spots. Male genitalia with longer uncus, juxta sclerotsed. The DNA easy blood and tssue 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 extracton method strongly ribbed appendix bursae. Ovipositor rather provided by Fukova et al. (2009) was followed. For medium sized, weakly sclerotsed, 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 quantfed Antoculeora ornatssima (Walker, 1858) (Images 1A,B) by fuorometer using standard procedures. Depending Plusia ornatssima Walker, 1858: 1786. Type locality: upon the concentraton, the DNA samples were diluted northern Hindostan. with molecular gradient water to get a working soluton Cerviplusia wukongensis Chou & Lu, 1974: 73. Type of 10–30 ng/µL. A porton of the total DNA was preserved locality: China. in glycerol (10%) in -80°C for future reference purposes. Antoculeora ornatssima Goater, Ronkay & Fibiger, The universal barcode primer described by Folmer et 2003. al. (1994) (LCO-5’-GGT CAA CAA ATC ATA AAG ATA TTG Redescripton: