Research Article |
Corresponding author: Hideyuki Chiba ( skipper@i.bekkoame.ne.jp ) Academic editor: Carlos Peña
© 2016 Jian-Qing Zhu, Hideyuki Chiba, Li-Wei Wu.
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:
Zhu J-Q, Chiba H, Wu L-W (2016) Tsukiyamaia, a new genus of the tribe Baorini (Lepidoptera, Hesperiidae, Hesperiinae). ZooKeys 555: 37-55. https://doi.org/10.3897/zookeys.555.6144
|
Skippers of the tribe Baorini are evidently a monophyletic group in the subfamily Hesperiinae. In this study, a new Baorini member Tsukiyamaia albimacula gen. n. et sp. n. is described from north Myanmar, southwest China and north Vietnam. Despite its peculiar and striking wing-pattern, this new genus has some important characters of Baorini, such as a broad and bifid uncus and a well-developed gnathos. Based on an analysis of male genitalia and the molecular phylogenies inferred from both mitochondrial and nuclear genes (28 taxa, total aligned length: 2968 bp), it is proposed that the genus Tsukiyamaia is closely related to the genus Polytremis, which has high species diversity in China. This study not only describes a new skipper but also highlights that Tsukiyamaia is important in clarifying phylogenetic relationship of Polytremis and its allies.
Polytremis , new species, cox1 , cox2 , EF-1α
Phylogenetic relationships and higher classifications of Hesperiidae at tribal level were primarily settled by
The members of the tribe Baorini are brown with small semi-hyaline white spots, except for two genera, Zenonia and Prusiana, which have extensive orange markings resembling those of Taractrocerini (
Recently most of newly described Baorini taxa were discovered in the range from the south boundary of Himalayas to South China (
For morphological comparison, eight male and one female specimens of this new taxon were examined. For inferring phylogenetic relationships of tribe Baorini to investigate the position of the new genus, 28 species were sampled (Table
Name | Voucher | Locality | Accession number | Reference | |
---|---|---|---|---|---|
cox1-cox2 | EF-1α | ||||
Calpodes ethlius | 144-ADW | EU364494 | EU364289 |
|
|
Dubiella belpa | 458-ADW | EU364051 | EU364249 |
|
|
Ochlodes bouddha | H1-0635 | Taiwan | KT240162 | KT240144 | this study |
Pyrrhopygopsis crates | 64-ADW | EU364503 | EU364298 |
|
|
Saliana esperi | 514-ADW | EU364501 | EU364296 |
|
|
Suastus gremius | H1-1548 | Taiwan | KT240163 | KT240145 | this study |
Synapte silius | 634-ADW | EU364431 | EU364226 |
|
|
Talides sinois | 512-ADW | EU364457 | EU364252 |
|
|
Thracides phidon | 451-ADW | EU364502 | EU364297 |
|
|
Udaspes folus | H1-1546 | Taiwan | KT240164 | KT240146 | this study |
Baoris farri | H1-0260 | Sichuan, China | KT240165 | KT240147 | this study |
Brobo cinnara | H1-0684 | Fujian, China | KT240166 | KT240148 | this study |
Caltoris bromus | H1-1645 | Taiwan | KT240167 | KT240149 | this study |
Caltoris cahira | H1-1644 | Taiwan | KT240168 | KT240150 | this study |
Iton watsonii | 600-MCZ | EU364490 | EU364285 |
|
|
Parnara guttata | H1-1008 | Sichuan, China | KT240169 | KT240151 | this study |
Pelopidas conjuncta | H1-1565 | Taiwan | KT240170 | KT240152 | this study |
Pelopidas mathias | H1-0617 | Taiwan | KT240171 | KT240153 | this study |
Pelopidas thrax | 570-ADW | EU364492 | EU364287 |
|
|
Polytremis gotama | H1-1019 | Yunnan, China | KT240172 | KT240154 | this study |
Polytremis kiraizana | H1-1437 | Taiwan | KT240173 | KT240155 | this study |
Polytremis lubricans | H1-0052 | Taiwan | KT240174 | KT240156 | this study |
Polytremis matsuii | H1-0982 | Sichuan, China | KT240175 | KT240157 | this study |
Polytremis nascens | H1-0321 | Sichuan, China | KT240176 | KT240158 | this study |
Polytremis pellucida | 234-ADW | EU364493 | EU364288 |
|
|
Polytremis zina | H1-0607 | Taiwan | KT240177 | KT240159 | this study |
Pseudobrobo bevani | H1-0888 | Yunnan, China | KT240178 | KT240160 | this study |
Tsukiyamaia albimacula | H1-1661 | Yunnan, China | KT240179 | KT240161 | this study |
We employed the standard method in Lepidoptera research to examine the male and female genitalia as well as other morphological characters of Tsukiyamaia (
The holotype and one female paratype of the new taxon were deposited in Department of Biology, Shanghai Normal University, China. One male paratype was deposited in the private collection of Jia-Qi Wang. The other paratype from China is in the collection of Kadoorie Conservation China, Kadoorie Farm and Botanic Garden, Hong Kong. The rest of the paratypes are in Hiroshi Tsukiyama’s collection (Chiba-pref., Japan).
Genomic DNA was extracted from the thoracic or leg tissue via using the Purgene DNA Isolation kit (Gentra Systems, Minnesota, USA), following the manufacturer protocol. The primers used for amplifying the mitochondrial cytochrome c oxidase I and II (cox1 and cox2) and nuclear elongation factor 1 alpha (EF-1α) genes were adopted from previous studies (
Molecular sequences of the cox1-cox2 and EF-1α genes were checked and assembled into contiguous arrays using Sequencher 4.8 (GeneCode, Boston, USA). After primer regions were cropped, the sequence dataset was aligned according to amino sequence similarity with the default settings by MUSCLE (
To evaluate species differentiation among Baorini skippers, genetic distance between species was calculated via MEGA 5.1. Pairwise distance with Kimura-2-parameter (
Tsukiyamaia albimacula sp. n.; designated by monotypy.
Antennae: 9.5−10 mm in length, half-length of forewing, nudum 13–14 on apiculus; Labial palpi: Second segment stout and erect, with brown hairs dorsally and yellowish hairs ventrally; third segment short, pointed and erect. Legs: middle tibiae unspined. Wing-shape: Forewing 19−20 mm in length, triangular in shape; costa about 1.4 times as long as dorsum, approximately straight, weakly arched on anterior half; apex angulated; termen lightly curved on anterior half; inner dorsum almost straight. Hindwing nearly triangular in shape; costa slightly longer than dorsum, obviously arched; termen curved on anterior half; tornus concave; dorsum almost straight.
Wing venation (Fig.
Wing markings (Figs
Male genitalia (Figs
The generic name is derived from Hiroshi Tsukiyama, whose outstanding contribution to the taxonomy of Hesperiidae is noteworthy.
Antennae 9.5−10 mm in length, about 1/2 the length of forewing, black brown except club gray dorsally and grayish yellow ventrally; nudum 13-14 on apiculus. Palpi erect, with brown hairs dorsally and yellowish hairs ventrally. Thorax and abdomen covered with brown hairs. Forewing 19−20 mm in length. Both wings ground color black brown at each sides, with white spots and marking; costal area of forewing and entire hindwing covered with brown scales underside; cilia of both wings brown. Upperside forewing: three apical spots in spaces R3−R5, arranged linear; one discal spot present at the middle of the space M2; in space M3, a reduced spot present in the holotype, and absent in two paratypes; cell spots conjoined as trapezium-shaped, which also conjoined with the Cu1 spot. Underside forewing markings same as upperside. Upperside hindwing: only with a cigar-shaped spot in space M1. Underside hindwing: Discal area with a very large, rectangle white marking extending from vein Rs to the middle of space Cu2. Inward margin smooth, upward to the end of the discal cell. Outward margin lightly serrated, and evidently elongated in space M1.
Male genitalia (Figs
Female genitalia (Figs
HOLOTYPE ♂: Phutao, Kachin, N. of MYANMAR, ~1000m, 09-VI-2000, Male genitalia examined by H. Chiba, #HC030511.
PARATYPES: 1♂, the same locality as the holotype, 29-V-2000.; 1♂ ditto, 08-VI-2000.; 1♂ Panglan, ~700m, Kachin, N. of MYANMAR, 02-IX-2002. 1♂ ditto, 04-IX-2002. . 1♂ ditto, 05-IX-2002, 1♂ ditto, 29-IX-2002, 1♂ Mt. Fan Shi Pang, ~1800m, N. VIETNAM, IV-2002. 1♂ Baopo, Dulongjiang, Yunnan, CHINA, 1500m, 29-V-2011, Jia-Qi Wang leg.; 1♀ , Maku, Dulongjiang, Yunnan, CHINA, 1900m, 03-VI-2009, Jian-Qing Zhu, leg; 1♂ CHINA, Yunnan, Tengchong, Gaoligongshan National Nature Reserve, Zhengding, 2200m, 26-IV-2014, LO Yik Fui Philip coll. (YFL140055).
Voltinism. Judging from the collecting data, the species is expected to be multivoltine.
Tsukiyamaia prefers open habitats, such as open field on the hillside, farmland and heavily disturbed shrub land. It is active near the ground and stream under strong sunlight. The female frequents flowers and the male performs padding behavior.
The species is named for its large white marking on underside of the hindwing.
In appearance, Tsukiyamaia is peculiar in Baorini with a large white marking in the center of the hindwing underside. The male genitalia of Tsukiyamaia can be separated from those of Baorini genera by the uncus lacking a pair of basal processes, and the harpe dorsally with a long and straight elongated process and ventrally with a relatively short and small process.
The gene length used in this study included cox1 (1531bp), tRNA-Leu (71 bp), cox2 (141 bp), and EF-1α (1225 bp) genes. Pairwise distance based on mitochondrial sequences showed that the smallest one between Tsukiyamaia albimacula and Polytremis matsuii was 6.8% (Table
Pairwise distance based on the substitution model of Kimura 2-parameter and cox1 – cox2 sequences (aligned length 1743 bp). The dash symbol means the overlap sequence is below 50 bp, the value is excluded.
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 Udaspes folus | ||||||||||||||
2 Suastus gremius | 0.106 | |||||||||||||
3 Synapte silius | 0.122 | 0.101 | ||||||||||||
4 Thracides phidon | 0.123 | 0.116 | 0.113 | |||||||||||
5 Pyrrhopygopsis crates | 0.137 | 0.105 | 0.116 | 0.130 | ||||||||||
6 Talides sinois | 0.121 | 0.065 | 0.097 | 0.109 | 0.118 | |||||||||
7 Ochlodes bouddha | 0.124 | 0.104 | 0.095 | 0.113 | 0.116 | 0.097 | ||||||||
8 Dubiella belpa | 0.132 | 0.111 | 0.112 | 0.105 | 0.133 | 0.118 | 0.096 | |||||||
9 Calpodes ethlius | 0.120 | 0.067 | 0.094 | 0.097 | 0.107 | 0.087 | 0.096 | 0.107 | ||||||
10 Saliana esperi | 0.118 | 0.080 | 0.101 | 0.100 | 0.124 | 0.095 | 0.088 | 0.118 | 0.071 | |||||
11 Parnara guttata | 0.131 | 0.121 | - | - | - | - | 0.123 | - | - | - | ||||
12 Baoris farri | 0.134 | 0.124 | - | - | - | - | 0.113 | - | - | - | 0.110 | |||
13 Pelopidas mathias | 0.113 | 0.111 | 0.106 | 0.096 | 0.112 | 0.104 | 0.103 | 0.115 | 0.097 | 0.102 | 0.109 | 0.090 | ||
14 Pelopidas thrax | 0.121 | 0.091 | 0.115 | 0.112 | 0.118 | 0.100 | 0.100 | 0.121 | 0.098 | 0.096 | - | - | 0.060 | |
15 Pelopidas conjuncta | 0.123 | 0.111 | 0.103 | 0.104 | 0.117 | 0.104 | 0.103 | 0.116 | 0.114 | 0.115 | 0.094 | 0.086 | 0.059 | 0.059 |
16 Brobo cinnara | 0.122 | 0.109 | - | - | - | - | 0.117 | - | - | - | 0.112 | 0.096 | 0.092 | - |
17 Iton watsonii | 0.113 | 0.112 | 0.106 | 0.100 | 0.110 | 0.089 | 0.093 | 0.117 | 0.101 | 0.102 | - | - | 0.074 | 0.084 |
18 Caltoris cahira | 0.126 | 0.098 | 0.111 | 0.115 | 0.127 | 0.109 | 0.106 | 0.117 | 0.101 | 0.101 | 0.116 | 0.117 | 0.101 | 0.100 |
19 Caltoris bromus | 0.125 | 0.113 | 0.116 | 0.113 | 0.129 | 0.105 | 0.114 | 0.119 | 0.096 | 0.103 | 0.127 | 0.137 | 0.110 | 0.095 |
20 Pseudobrobo bevani | 0.126 | 0.099 | - | - | - | - | 0.123 | - | - | - | 0.114 | 0.113 | 0.102 | - |
21 Polytremis lubricans | 0.137 | 0.133 | 0.097 | 0.116 | 0.135 | 0.126 | 0.124 | 0.151 | 0.106 | 0.107 | 0.131 | 0.117 | 0.104 | 0.122 |
22 Polytremis matsuii | 0.127 | 0.129 | - | - | - | - | 0.105 | - | - | - | 0.089 | 0.100 | 0.091 | - |
23 Polytremis kiraizana | 0.123 | 0.116 | 0.107 | 0.103 | 0.128 | 0.101 | 0.103 | 0.115 | 0.109 | 0.102 | 0.104 | 0.107 | 0.094 | 0.092 |
24 Polytremis nascens | 0.116 | 0.115 | - | - | - | - | 0.113 | - | - | - | 0.105 | 0.100 | 0.092 | 0.000 |
25 Polytremis gotama | 0.107 | 0.115 | - | - | - | - | 0.104 | - | - | - | 0.101 | 0.083 | 0.086 | 0.000 |
26 Polytremis zina | 0.116 | 0.115 | 0.108 | 0.123 | 0.142 | 0.100 | 0.107 | 0.117 | 0.092 | 0.109 | 0.093 | 0.091 | 0.082 | 0.089 |
27 Polytremis pellucida | 0.117 | 0.124 | 0.115 | 0.116 | 0.123 | 0.096 | 0.107 | 0.116 | 0.104 | 0.108 | 0.099 | 0.095 | 0.083 | 0.089 |
28 Tsukiyamaia albimacula | 0.120 | 0.119 | 0.114 | 0.121 | 0.123 | 0.114 | 0.108 | 0.128 | 0.121 | 0.109 | 0.105 | 0.098 | 0.087 | 0.085 |
15 | 16 | 17 | 18 | 19 | 20 | 21 | 22 | 23 | 24 | 25 | 26 | 27 | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 Udaspes folus | |||||||||||||
2 Suastus gremius | |||||||||||||
3 Synapte silius | |||||||||||||
4 Thracides phidon | |||||||||||||
5 Pyrrhopygopsis crates | |||||||||||||
6 Talides sinois | |||||||||||||
7 Ochlodes bouddha | |||||||||||||
8 Dubiella belpa | |||||||||||||
9 Calpodes ethlius | |||||||||||||
10 Saliana esperi | |||||||||||||
11 Parnara guttata | |||||||||||||
12 Baoris farri | |||||||||||||
13 Pelopidas mathias | |||||||||||||
14 Pelopidas thrax | |||||||||||||
15 Pelopidas conjuncta | |||||||||||||
16 Brobo cinnara | 0.085 | ||||||||||||
17 Iton watsonii | 0.078 | - | |||||||||||
18 Caltoris cahira | 0.106 | 0.122 | 0.089 | ||||||||||
19 Caltoris bromus | 0.111 | 0.115 | 0.083 | 0.077 | |||||||||
20 Pseudobrobo bevani | 0.098 | 0.096 | - | 0.109 | 0.116 | ||||||||
21 Polytremis lubricans | 0.111 | 0.121 | 0.088 | 0.116 | 0.133 | 0.095 | |||||||
22 Polytremis matsuii | 0.084 | 0.095 | - | 0.119 | 0.124 | 0.107 | 0.105 | ||||||
23 Polytremis kiraizana | 0.087 | 0.084 | 0.072 | 0.105 | 0.112 | 0.098 | 0.105 | 0.061 | |||||
24 Polytremis nascens | 0.092 | 0.091 | - | 0.102 | 0.102 | 0.095 | 0.098 | 0.065 | 0.077 | ||||
25 Polytremis gotama | 0.085 | 0.085 | - | 0.102 | 0.094 | 0.088 | 0.089 | 0.061 | 0.066 | 0.038 | |||
26 Polytremis zina | 0.081 | 0.079 | 0.073 | 0.109 | 0.117 | 0.087 | 0.094 | 0.061 | 0.069 | 0.052 | 0.051 | ||
27 Polytremis pellucida | 0.081 | 0.083 | 0.075 | 0.113 | 0.121 | 0.095 | 0.104 | 0.060 | 0.073 | 0.056 | 0.057 | 0.009 | |
28 Tsukiyamaia albimacula | 0.089 | 0.093 | 0.076 | 0.104 | 0.115 | 0.110 | 0.106 | 0.068 | 0.077 | 0.079 | 0.074 | 0.072 | 0.078 |
The total of eight topologies, inferred by four partitioning datasets and by two tree-reconstructing methods, have similar phylogenetic relationships (summarized in Fig.
Although the monophyly of the tribe Baorini is well-supported by the molecular data, no synapomorphic character in external morphology have been found (
If the key for separation of genera in
We are extremely grateful to Mr Hiroshi Tsukiyama for providing specimens of this peculiar skipper from his collection and his assistance throughout this project. It is our honor to name the new genus after him. We thank Ms Ting-Wei Chen for some DNA sequence work and Mr Yik Fui Philip Lo for the information of the new skipper collected during a joint biodiversity survey project between Kadoorie Conservation China and Tengchong Branch of Gaoligongshan National Nature Reserve.