Research Article |
Corresponding author: Misbah Ullah ( misbah2013@nwsuaf.edu.cn ) Academic editor: Colin Plant
© 2017 Misbah Ullah, Zhaofu Yang, Pingping Qiao, Yalin Zhang.
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:
Ullah M, Yang Z, Qiao P, Zhang Y (2017) A new cryptic species of Nagiella Munroe from China revealed by DNA barcodes and morphological evidence (Lepidoptera, Crambidae, Spilomelinae). ZooKeys 679: 65-76. https://doi.org/10.3897/zookeys.679.11960
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Nagiella occultalis Misbah & Yang, sp. n. from China is described and illustrated. This new species is very similar to N. quadrimaculalis (Kollar, 1844) in general morphological characters of forewing and male genitalia. Molecular evidence shows that these two species diverge in COI barcode region by more than 3.2%. Sequence divergence among the two species is congruent with subtle morphological differences. Wing venation and male genitalia of the two species are compared and illustrated.
COI gene, genitalia, Scopula quadrimaculalis , taxonomy
The subfamily Spilomelinae (Crambidae) is the largest subfamily of pyraloid moths including about 3300 species in more than 300 genera having worldwide distribution (
Recently the integration of DNA barcoding and morphological approaches opened the field for researchers in accelerating species identification and assisted in detecting previously undetected cryptic species (
Three specimens of N. occultalis sp. n. were collected from Taibai Mountain, Shaanxi and Wufeng, Hubei in China and 15 specimens of N. quadrimaculalis were collected from various localities (Table
Identification | BIN | Process ID | Sample ID | Length of sequence (bp) | GenBank Accession | Province | Genitalia slide number |
---|---|---|---|---|---|---|---|
N. occultalis sp. n. | BOLD:AAD8179 | CNPYB439-16 | NAFU Pyr002290 | 658 | KY080696 | Shaanxi | |
N. occultalis sp. n. | BOLD:AAD8179 | CNPYB407-16 | NAFU Pyr002397 | 658 | KY080703 | Shaanxi | NAFU Pyr002065 |
N. occultalis sp. n. | BOLD:AAD8179 | CNPYD499-10 | Pyr000499 | 658 | HM908668 | Hubei | |
N. quadrimaculalis | CNPYA401-10 | NAFU Pyr000401 | 0 | Yunnan | |||
N. quadrimaculalis | CNPYA402-10 | NAFU Pyr000402 | 0 | Sichuan | |||
N. quadrimaculalis | CNPYA403-10 | NAFU Pyr000403 | 0 | Yunnan | |||
N. quadrimaculalis | CNPYA404-10 | NAFU Pyr000404 | 0 | Yunnan | |||
N. quadrimaculalis | CNPYB409-16 | NAFU Pyr002070 | 0 | Shaanxi | NAFU Pyr002070 | ||
N. quadrimaculalis | CNPYB410-16 | NAFU Pyr002261 | 0 | Shaanxi | NAFU Pyr002261 | ||
N. quadrimaculalis | CNPYB411-16 | NAFU Pyr002262 | 0 | Shaanxi | |||
N. quadrimaculalis | BOLD:AAD8178 | CNPYB412-16 | NAFU Pyr002263 | 658 | KY080700 | Shaanxi | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYB413-16 | NAFU Pyr002264 | 658 | KY080702 | Shaanxi | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYB414-16 | NAFU Pyr002265 | 658 | KY080704 | Shaanxi | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYB415-16 | NAFU Pyr002266 | 658 | KY080698 | Shaanxi | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYB416-16 | NAFU Pyr002267 | 658 | KY080694 | Shaanxi | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYB417-16 | NAFU Pyr002268 | 658 | KY080705 | Shaanxi | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYB418-16 | NAFU Pyr002269 | 658 | KY080697 | Shaanxi | |
N. quadrimaculalis | CNPYB419-16 | NAFU Pyr002270 | 0 | Shaanxi | |||
N. quadrimaculalis | CNPYB420-16 | NAFU Pyr002271 | 0 | Shaanxi | |||
N. quadrimaculalis | CNPYB421-16 | NAFU Pyr002272 | 0 | Henan | NAFU Pyr002272 | ||
N. quadrimaculalis | CNPYB422-16 | NAFU Pyr002273 | 0 | Henan | NAFU Pyr002273 | ||
N. quadrimaculalis | CNPYB423-16 | NAFU Pyr002274 | 0 | ||||
N. quadrimaculalis | CNPYB424-16 | NAFU Pyr002275 | 0 | Hunan | |||
N. quadrimaculalis | CNPYB425-16 | NAFU Pyr002276 | 0 | Hunan | |||
N. quadrimaculalis | CNPYB426-16 | NAFU Pyr002277 | 0 | ||||
N. quadrimaculalis | CNPYB427-16 | NAFU Pyr002278 | 0 | Fujian | |||
N. quadrimaculalis | CNPYB428-16 | NAFU Pyr002279 | 0 | Hainan | |||
N. quadrimaculalis | CNPYB429-16 | NAFU Pyr002280 | 0 | Hainan | |||
N. quadrimaculalis | CNPYB430-16 | NAFU Pyr002281 | 0 | ||||
N. quadrimaculalis | CNPYB431-16 | NAFU Pyr002282 | 0 | Zhejiang | |||
N. quadrimaculalis | CNPYB432-16 | NAFU Pyr002283 | 0 | Yunnan | |||
N. quadrimaculalis | CNPYB433-16 | NAFU Pyr002284 | 0 | NAFU Pyr002284 | |||
N. quadrimaculalis | CNPYB434-16 | NAFU Pyr002285 | 0 | ||||
N. quadrimaculalis | CNPYB435-16 | NAFU Pyr002286 | 0 | ||||
N. quadrimaculalis | CNPYB436-16 | NAFU Pyr002287 | 0 | NAFU Pyr002287 | |||
N. quadrimaculalis | CNPYB437-16 | NAFU Pyr002288 | 0 | ||||
N. quadrimaculalis | BOLD:AAD8178 | CNPYB438-16 | NAFU Pyr002289 | 658 | KY080695 | Shaanxi | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYB440-16 | NAFU Pyr002291 | 658 | KY080701 | Shaanxi | NAFU Pyr002291 |
N. quadrimaculalis | BOLD:AAD8178 | CNPYB441-16 | NAFU Pyr002292 | 658 | KY080699 | Shaanxi | |
N. quadrimaculalis | CNPYB408-16 | NAFU Pyr002398 | 0 | Shaanxi | NAFU Pyr002067 | ||
N. quadrimaculalis | BOLD:AAD8178 | CNPYD497-10 | Pyr000497 | 622 | HM908666 | Hubei | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYD498-10 | Pyr000498 | 658 | HM908667 | Hubei | |
N. quadrimaculalis | CNPYD500-10 | Pyr000500 | 0 | Hubei | |||
N. quadrimaculalis | CNPYD501-10 | Pyr000501 | 0 | Hubei | |||
N. quadrimaculalis | CNPYD502-10 | Pyr000502 | 0 | Hubei | |||
N. quadrimaculalis | BOLD:AAD8178 | CNPYD503-10 | Pyr000503 | 658 | HM908669 | Hubei | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYD504-10 | Pyr000504 | 658 | HM908670 | Sichuan | |
N. quadrimaculalis | BOLD:AAD8178 | CNPYD505-10 | Pyr000505 | 658 | HM908671 | Sichuan |
Kimura 2-parameter genetic distances calculated within (in italic) and between three species of Nagiella.
Nagiella occultalis sp. n. | Nagiella quadrimaculalis | Nagiella inferior | Patania ruralis (outgroup) | |
---|---|---|---|---|
Nagiella occultalis sp. n. | 0.0000000 | 0.0072358 | 0.0086344 | |
Nagiella quadrimaculalis | 0.0320975 | 0.000787822 | 0.0101216 | |
Nagiella inferior | 0.0475427 | 0.0598071 | 0.000761036 | |
Patania ruralis (Outgroup) | 0.1156349 | 0.1165689 | 0.1134248 | 0.009202714 |
Genomic DNA was extracted from insect legs by following the method of
Sequence alignment was carried out by using MUSCLE algorithm implemented in MEGA 6.0 (
A total of 18 COI gene sequences of N. occultalis sp. n. and N. quadrimaculalis were obtained. The lengths were from 622–658 bp (mean 656 bp). The genetic distances within and between these two species of Nagiella are given in Table
The specific epithet refers to “cryptic”, as this previously undetected species stood within the N. quadrimaculalis complex.
This species can be distinguished from N. quadrimaculalis by the width and length of the uncus, the proportions of the valva and transtilla, and size of the forewing, as described in Table
(Figs
Male genitalia (Fig.
Female. Unknown
China (Taibai Mountain, Shaanxi; Wufeng, Hubei).
Holotype. ♂: China: Shaanxi, Taibai Mountain, 1051 m, 25 July 2014, Zhou Lin (NWAFU), Specimen ID: NAFU PYR002397. Genitalia slide number: NAFU PYR002397. Paratypes. 1 ♂, same data as the holotype except 24 July 2014; 1 ♂, China, Hubei, Wufeng, Changleping town, 14 July 2008, Zhao Lu.
The genus Nagiella, formerly comprised of three recognized species widespread in Burma, China, Japan and Malaysia (Borneo and Sarawak), is now increased to four with N. occultalis sp. n.
Morphological differences between Nagiella occultalis sp. n. and N. qudrimaculalis.
Characteristics | N. occultalis sp. n. | N. quadrimaculalis |
---|---|---|
Forewing length | 15–16 mm (Fig. |
18–20 mm (Fig. |
Small subdiscal spot on forewing | Proportionally narrower or elongate | Sub-quadrate |
Uncus width and length | 0.4 × 0.6 mm (Fig. |
0.3 × 0.68 mm (Fig. |
Posterior margin of uncus | Slightly notched medially | Evenly rounded |
Valva | Broader, W/L 0.91 × 3.09 mm | Slender, W/L 0.7 mm × 2.08 mm |
Sella with ventral edge | Straight | Slightly incurved |
Subscaphium | Elongate, conical sclerotized | Unsclerotized |
Size of transtilla | Narrower, 0.28 × 0.8 mm | Broadly triangular 0.3 × 0.9 mm |
Phallus | Phallus L/valva L ratio 1.19 (Fig. |
Phallus L/valva L ratio 1.7 (Fig. |
The authors are sincerely thankful to Dr. Jean-François Landry, Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada for assistance with genitalia description and comments on the manuscript. Special thanks are extended to Dr. Christopher H. Dietrich, University of Illinois at Urbana Champaign, USA and Dr. Murray Fletcher, Orange Agricultural Institute, Orange NSW Australia, for reviewing and improving the manuscript. This study was supported by the National Natural Science Foundation of China (31201733), Natural Science Foundation of Shaanxi Province (2016JM3026), The Chinese Universities Scientific Fund (2452015012), The National Key Research and Development Program (2016YFC0501502) and The Ministry of Science and Technology of the People’s Republic of China (2005DKA21402, 2011FY120200).