Research Article |
Corresponding author: Zhu-Qing He ( zqhe@bio.ecnu.edu.cn ) Academic editor: Christiane Weirauch
© 2024 Yu-Jiao Guo, Shuang-Qi An, Zhen-Gui Fang, Shu-Fei Wei, Zi-Hao Shen, Zhao-Yang Chen, Zhu-Qing He.
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.
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Species belonging to the genus Goniogryllus Chopard, 1936 are primarily identified based on their morphological characteristics. However, male genitalia provide limited features for classification. In this study, we sequenced the COI gene from 49 samples, and reconstructed a phylogenetic tree. Species were classified by a combination of molecular and morphological data, and some morphological features were re-evaluated. The results indicate that the presence or absence of pubescence on the thorax and abdomen, yellow stripes on the head with or without bifurcation, and the color of the maxillary palpus are interspecific differences. The classification based on these morphological features are consistent with the classification based on COI gene. In contrast, variations in the color of the hind femur, pronotum, and the number of dorsal spurs on the hind tibia are considered to be intraspecific variations. Additionally, Qingryllus striofemorus Chen & Zheng, 1995 and Q. jiguanshanensis Liu, Zhang & Shi, 2017 are recognized as the long-wing forms of G. ovalatus Chen & Zheng, 1996. Therefore, Qingryllus Chen & Zheng, 1995 syn. is a junior synonym of Goniogryllus. Q. striofemorus syn. nov. and Q. jiguanshanensis syn. nov. are junior synonyms of G. ovalatus. Furthermore, G. cirilinearis Xie, 2005 syn. nov. is a junior synonym of G. atripalpulus Chen & Zheng, 1996. G. octospinatus Chen & Zheng, 1995 syn. nov. is a junior synonym of G. chongqingensis Chen & Zheng, 1995. Lastly, Callogryllus yunnanus Wu & Zheng, 1992 is transferred to the genus Goniogryllus, as Goniogryllus yunnanus comb. nov.
COI, morphology, Qingryllus, taxonomy
The genus Goniogryllus Chopard, 1936 belongs to Gryllinae, Gryllidae, Orthoptera. It was established by Chopard in 1936, with G. punctatus Chopard, 1936 from Fujian, China as the type species. Prior to this study, there were 21 species reported from Japan, Assam, and China with 19 species from China (
Goniogryllus is primarily identified based on morphological features, such as the presence or absence of pubescence on the thorax and abdomen, yellow stripes on the head with or without bifurcation, the color of the maxillary palpus, hind femur or pronotum, and the number of dorsal spurs on the hind tibia (
The genus Qingryllus Chen & Zheng, 1995 also belongs to Gryllinae, and its type species, Q. striofemorus Chen & Zheng, 1995, is collected from Xunyangba and Huoditang, Ningshan, Shaanxi, which are also the collecting sites for the holotype of G. ovalatus Chen & Zheng, 1996 and G. atripalpulus (
The holotype of Callogryllus yunnanus Wu & Zheng, 1992 was collected from Yunnan, China. It bears a resemblance to the above two genera. The unique feature of this species is the presence of a pair of short forewing with irregular veins in males.
To clarify the relationship among partial Goniogryllus, Qingryllus and C. yunnanus, and to revise partial species in the genus Goniogryllus, we reconstructed a phylogenetic tree based on molecular markers (COI gene) and estimated the number of molecular operational taxonomic units (MOTUs) by three methods, Automatic Barcode Gap Discovery (ABGD), Assemble Species by Automatic Partitioning (ASAP), and Bayesian Poisson Tree Processes (bPTP). We also combined it with morphological classification to evaluate these morphological features.
The specimens were collected by hand. Photomicrographs were taken by SC2000 digital CMOS Camera, and edited by using Adobe Photoshop. A total of 49 individuals belonging to Goniogryllus, Qingryllus and C. yunnanus were examined for this study. All specimens are preserved in the
Biological History Museum of East China Normal University (
The measurements and indices taken are defined below:
BL – body length, length from the apex of fastigium verticis to the apex of subgenital plate; SZ – body size, length from the apex of fastigium verticis to the end of hindwing (for Qingryllus only); PL – pronotum length, length from anterior margin to posterior margin; FWL – forewing length, length from the base of forewing to the apex; HWL – hindwing length, length from the base of hindwing to the apex; HFL – hind femur length, maximum distance from the base of hind femur to the apex; OL – ovipositor length, straight line distance from the base of ovipositor to the apex.
The terminology used to describe male genitalia followed
The total genomic DNA was extracted from the muscles of one hind leg by AxyPrep Genomic DNA Miniprep Kit (AXYGEN), according to the manufacturer’s instructions. The fragments of the mitochondrial cytochrome-c oxidase subunit I gene (COI, 658 bp) were sequenced by primers COBU TYTCAACAAAYCAYAARGATATTGG and COBL TAAACTTCWGGRTGWCCAAARAATCA (
Genus | Species | No. | voucher | Collection site | GenBank |
---|---|---|---|---|---|
Goniogryllus | punctatus | 1 | H174 | Zhejiang, Hangzhou, Tianmushan Nature Reserve | OR102820 |
2 | 225 | Zhejiang, Hangzhou, Qingliangfeng Nature Reserve | OR102795 | ||
3 | 312 | Zhejiang, Hangzhou, Tianmushan Nature Reserve | OR102798 | ||
4 | 348 | Zhejiang, Hangzhou, Qingliangfeng Nature Reserve | OR102803 | ||
5 | 523 | Zhejiang, Hangzhou, Tianmushan Nature Reserve | OR102829 | ||
6 | 542 | Zhejiang, Hangzhou, Tianmushan Nature Reserve | OR102816 | ||
7 | 632 | Zhejiang, Hangzhou, Qingliangfeng Nature Reserve | OR102817 | ||
8 | 1106 | Zhejiang, Hangzhou, Qingliangfeng Nature Reserve | OR102785 | ||
9 | 2531 | Zhejiang, Hangzhou, Tianmushan Nature Reserve | OR102796 | ||
10 | 2532 | Zhejiang, Hangzhou, Tianmushan Nature Reserve | OR102797 | ||
emeicus | 1 | 675 | Sichuan, Emeishan, Emeishan | OR102818 | |
2 | 679 | Sichuan, Emeishan, Emeishan | OR102819 | ||
3 | 1972 | Sichuan, Emeishan, Emeishan | OR102787 | ||
4 | 1982 | Sichuan, Emeishan, Emeishan | OR102792 | ||
chongqingensis | 1 | 3440 | Chongqing, Beibei, Jinyunshan | OR102799 | |
2 | 3441 | Chongqing, Beibei, Jinyunshan | OR102800 | ||
octospinatus | 1 | 4551 | Hubei, Lichuan, Fubaoshan | OR102802 | |
2 | 4554 | Hubei, Lichuan, Fubaoshan | OR102801 | ||
cirilinearis | 1 | 4535 | Shaanxi, Lan’gao, Dabashan | OR102805 | |
2 | 4536 | Shaanxi, Lan’gao, Dabashan | OR102824 | ||
3 | 4539 | Shaanxi, Lan’gao, Dabashan | OR102807 | ||
atripalpulus | 1 | 4547 | Hubei, Danjiangkou, Wudangshan | OR102811 | |
2 | 4548 | Hubei, Danjiangkou, Wudangshan | OR102812 | ||
glaber | 1 | 1974 | Yunnan, Lushui, Pianma | OR102788 | |
2 | 1976 | Yunnan, Lushui, Pianma | OR102789 | ||
ovalatus | 1 | H007 | Shaanxi, Ningshan, Huoditang | OR102821 | |
2 | H008 | Shaanxi, Ningshan, Huoditang | OR102822 | ||
3 | 1921 | Shaanxi, Huayang, Changqing Nature Reserve | OR102786 | ||
4 | 1981 | Sichuan, Emeishan, Emeishan | OR102791 | ||
5 | 1986 | Sichuan, Emeishan, Emeishan | OR102793 | ||
6 | 1989 | Sichuan, Emeishan, Emeishan | OR102794 | ||
7 | 4532 | Sichuan, Maoxian, Jiudingshan | OR102804 | ||
8 | 4537 | Gansu, Zhouqu, Wuping | OR102806 | ||
9 | 4542 | Chongqing, Wushan, Liziping | OR102809 | ||
10 | 4545 | Hubei, Wufeng, Houhe Nature Reserve | OR102810 | ||
11 | 4550 | Hubei, Lichuan, Fubaoshan | OR102828 | ||
12 | 4552 | Hubei, Lichuan, Fubaoshan | OR102814 | ||
13 | 4555 | Hubei, Lichuan, Fubaoshan | OR102827 | ||
14 | 4556 | Hubei, Lichuan, Fubaoshan | OR102815 | ||
15 | 4558 | Hubei, Lichuan, Fubaoshan | OR102825 | ||
16 | 4559 | Hubei, Lichuan, Fubaoshan | OR102826 | ||
Qingryllus | jiguanshanensis | 1 | 1980 | Sichuan, Emeishan, Emeishan | OR102790 |
striofemorus | 1 | 3882 | Hubei, Shennongjia, Songbai | OR102830 | |
2 | 4540 | Shaanxi, Langao, Dabashan | OR102808 | ||
3 | 4549 | Hubei, Lichuan, Fubaoshan | OR102813 | ||
4 | 4671 | Shaanxi, Ningshan, Huoditang | OR102823 | ||
Callogryllus | yunnanus | 1 | 4224 | Yunnan, Yuxi, Ailaoshan | PP967855 |
2 | 4226 | Yunnan, Yuxi, Ailaoshan | PP967856 | ||
3 | 4228 | Yunnan, Yuxi, Ailaoshan | PP967857 |
A total of 49 COI sequences were obtained from samples of Goniogryllus, Qingryllus and C. yunnanus, which were used for molecular analysis. The COI sequence of Paratrigonidium nitidum Brunner von Wattenwyl, 1893 was used as the outgroup. Additionally, the COI gene of 47 individuals from the Gryllini group were included to assess the monophyly of Goniogryllus (see Table S1 for details). All sequences were aligned in MEGA X following MUSCLE method. We used DAMBE to conduct the saturation test (
When using P. nitidum as the outgroup, a strong clade consisting of 49 individuals from Goniogryllus, Qingryllus, and C. yunnanus was formed, with 7 subclades identified by ABGD and ASAP analysis. The first subclade includes C. yunnanus only. The second subclade consists of G. glaber Wu & Wang, 1992 only. The third subclade includes G. atripalpulus and G. cirilinearis. The fourth subclade includes G. chongqingensis Chen & Zheng, 1995 and G. octospinatus Chen & Zheng, 1995. The fifth subclade includes G. ovalatus, Q. striofemorus, and Q. jiguanshanensis Liu, Zhang & Shi, 2017. The sixth subclade consists of G. emeicus Wu & Wang, 1992 only. The seventh subclade comprises G. punctatus only (Fig.
The relationship among partial Goniogryllus, Qingryllus and C. yunnanus species was inferred from the COI gene. Rooted by P. nitidum, the tree was constructed by Maximum likelihood (ML) with TIM2+I+F model. Topology supports of all major nodes were indicated above branches in this order: the SH-aLRT value, the Bayesian posterior probability and the bootstrap value. The MOTUs was shown by color bars based on ABGD, ASAP, and bPTP analysis in grey, blue, and pink respectively.
Class Insecta Linnaeus, 1758
Order Orthoptera Latreille, 1810
Family Gryllidae Laicharting, 1781
Subfamily Gryllinae Laicharting, 1781
Goniogryllus
Qingryllus
syn. (synonymized by
Goniogryllus punctatus Chopard, 1936.
Average size; black head with yellow stripes extending from occiput, along eyes to lateral ocelli, the yellow stripes divided into two branches from the middle of eyes or no bifurcation; maxillary palpus all black or with yellow stripes on the apex of the 4th joint and the dorsal side of the 5th joint; pronotum all black or with yellow spots or stripes on both sides, glabrous or pubescent; no wings, scaly wings, short wings, or fully developed wings; abdomen glabrous or pubescent; long ovipositor.
The molecular analysis reveals that G. ovalatus, Q. striofemorus, and Q. jiguanshanensis form a cohesive clade with strong support. Morphologically, they share similarities such as processing a pair of yellow-brown longitudinal stripes on each side of the head, and the absence of a tympanum on the fore tibia. Moreover, their male genitalia exhibit resemblances. They are often found in the same collection sites (Fig.
Diagnosis for Goniogryllus species in this study (G. ovalatus with fully developed wings not included).
Feature | G. ovalatus | G. atripalpulus | G. chongqingensis | G. punctatus | G. emeicus | G. glaber | G. yunnanus | ||||||
10♀ | 9♂ | 3♀ | 2♂ | 3♀ | 1♂ | 4♀ | 2♂ | 2♀ | 2♂ | 2♂ | 2♀ | 1♂ | |
Pronotum pubescent | Yes | No | No | No | No | No | No | ||||||
♀ Abdomen pubescent | Yes | Yes | No | No | No | / | No | ||||||
♂ The first three segments of abdomen pubescent | Yes | Yes | No | No | No | No | No | ||||||
Yellow stripes on the head bifurcated | No | Yes | No | Yes | No | No | No | ||||||
Maxillary palpus black | No | Yes | No | Yes | No | Yes | No | ||||||
Apical part of epiphallus with process | Yes | No | Yes | Yes | Yes | / | / |
Goniogryllus ovalatus
Chen & Zheng 1996: 290, 293;
Qingryllus striofemorus
syn. nov. Chen & Zheng 1995: 70, 76;
Qingryllus jiguanshanensis
syn. nov.
G. ovalatus: CHINA • ♂ (1921); Shaanxi Prov., Hanzhong City, Huayang Town, Changqing Nature Reserve (107°33′E, 33°35′N); 8 May. 2019; Zhao-Yang Chen leg. (
No wings, scaly wings, or fully developed wings; pronotum pubescent, if no wings or scaly wings, mesonotum and metanotum pubescent, abdomen pubescent in females, but only the first three segments of abdomen pubescent in males, if fully developed wings, mesonotum, metanotum and abdomen glabrous (Fig.
Dorsal view of body. Aa–Ae female G. ovalatus, Af–Aj male G. ovalatus; Ba–Bc female Q. striofemorus, Bd male Q. striofemorus; C female Q. jiguanshanensis; Da female G. atripalpulus, Db male G. atripalpulus; Ea, Eb female G. cirilinearis, Ec male G. cirilinearis; F female G. chongqingensis; Ga female G. octospinatus, Gb male G. octospinatus; Ha–Hd female G. punctatus, He, Hf male G. punctatus; Ia, Ib female G. emeicus, Ic–Ie male G. emeicus; J male G. glaber; Ka female G. yunnanus, Kb male G. yunnanus. Scale bar = 2mm.
Qingryllus contains two species, Q. striofemorus and Q. jiguanshanensis based on the width of the posterior margin of pronotum and the color of veins of tegmina (Fig.
(mm) ♂: BL 12.28–12.93; SZ 16.97–18.04; PL 2.23–2.65; FWL 9.54; HWL 13.27; HFL 6.83–7.57. — ♀: BL 11.44–13.65; SZ 17.50–17.95; PL 2.34–2.59; FWL 8.4–9.93; HWL 13.70–14.75; HFL 7.49–8.39; OL 9.17–9.94.
China (Chongqing, Gansu, Hubei, Shaanxi, Sichuan).
Goniogryllus atripalpulus
Chen & Zheng 1996: 289, 292;
Goniogryllus cirilinearis
syn. nov.
G. atripalpulus: CHINA • 1♀1♂ (4547, 4548); Hubei Prov., Shiyan City, Danjiangkou County, Wudangshan (111°04′E, 32°28′N); 19 Jun. 2021; Wen-Xuan Bi leg. (
Apterous; mesonotum and metanotum pubescent slightly, abdomen pubescent slightly in females, but only the first three segments of abdomen pubescent slightly in males (Fig.
The main difference between G. cirilinearis and G. atripalpulus is: the occiput of G. cirilinearis is ornamented by two short reddish-brown stripes, while that of G. atripalpulus is not (
(mm) ♂: BL 13.84–14.89; PL 2.79–3.71; HFL 7.93–9.04. — ♀: BL 14.11–15.66; PL 3.14–3.71; HFL 9.37–9.75; OL 11.34–11.79.
China (Gansu, Hubei, Shaanxi).
Goniogryllus chongqingensis
Chen & Zheng 1995: 213, 216;
Goniogryllus octospinatus
syn. nov. Chen & Zheng 1995: 215;
G. chongqingensis: CHINA • 2♀ (3440, 3441); Chongqing City, Beibei District, Jinyunshan (106°23′E 29°50′N); 19 Aug. 2020; Hui-Cong Xie leg. (
Apterous; all glabrous (Fig.
The main difference between G. chongqingensis and G. octospinatus is: the pronotum and the hind femur of G. chongqingensis are black, while there are yellow stripes on both sides of pronotum and a yellow longitudinal stripe on the superior border of the hind femur of G. octospinatus (
(mm) ♂: BL 12.13; PL 2.57; HFL 7.04. — ♀: BL 12.24–13.82; PL 3.24–3.66; HFL 8.27–8.56; OL 9.40–10.99.
China (Chongqing, Hubei, Sichuan).
Callogryllus yunnanus
comb. nov. Wu & Zheng 1992: 95;
CHINA • 1♂2♀ (4224, 4226, 4228); Yunnan Prov., Yuxi City, Xinping County, Ailaoshan (101°26′E 23°58′N); 16 Jul. 2021; Zhu-Qing He leg. (
No wings in females, wings short to the third segment of abdomen in males; all glabrous (Fig.
The molecular result shows that C. yunnanus and Goniogryllus are in the same clade. C. yunnanus and Goniogryllus possess some of the same morphological features such as yellow stripes on each side of the head (
(mm) ♂: BL 9.54; PL 2.38; FWL 2.13; HFL 5.83. — ♀: BL 9.58–10.21; PL 2.45–2.52; HFL 6.94–7.30; OL 8.57–8.96.
Distribution. China (Yunnan).
Goniogryllus punctatus
CHINA • ♀ (312); Zhejiang Prov., Hangzhou City, Lin’an District, Tianmushan Nature Reserve (119°27′E, 30°20′N); 22 Aug. 2016; Jia-Yao Hu leg. (
Apterous; all glabrous (Fig.
(mm) ♂: BL 12.89–12.91; PL 3.02–3.11; HFL 7.92–8.22. — ♀: BL 13.75–14.29; PL 3.07–3.29; HFL 8.85–8.72; OL 10.18–10.82.
China (Fujian, Zhejiang).
Goniogryllus emeicus
Wu & Wang 1992: 230;
CHINA • ♂ (675); Sichuan Prov., Leshan City, Emeishan County, Emeishan (103°21′E, 29°34′N); 31 May. 2017; Wen-Xuan Bi leg. (
Apterous; all glabrous (Fig.
(mm) ♂: BL 10.94–12.37; PL 3.04–3.14; HFL 7.74–8.22. — ♀: BL 13.61–15.59; PL 3.13–3.27; HFL 7.57–8.67; OL 8.97–9.56.
China (Sichuan).
Goniogryllus glaber
Wu & Wang 1992: 227;
CHINA • 2♂ (1974, 1976); Yunnan Prov., Lushui City, Pianma Town (98°37′E 26°00′N); 17 Sep. 2018; Wen-Xuan Bi leg. (
Apterous; all glabrous (Fig.
(mm) ♂: BL 12.13–14.01; PL 2.20–2.37; HFL 6.05–7.08.
China (Yunnan).
The identification of Goniogryllus is primarily based on its morphological characteristics (
The COI gene, also known as DNA barcoding, has proven to be a valuable tool for identifying species, including crickets (
The presence or absence of pubescence on the thorax and abdomen is a classification feature of G. octospinatus and G. pubescens Wu & Wang, 1992 (
Yellow stripes on the head with or without bifurcation is an important feature mentioned by
The color of the maxillary palpus is another classification feature (
Most studies describe the male genitalia of most Goniogryllus species (
The color of the pronotum has been mentioned as a classification feature by
Our molecular analysis revealed that the specimens in this study can be classified into seven species (six species according to the bPTP method), which supports the classification based on the presence or absence of pubescence on the thorax and abdomen, yellow stripes on the head with or without bifurcation, the color of the maxillary palpus, and the shape of male genitalia. However, in Hubei, Shaanxi, and Sichuan, we encountered some “species” that appear different if we rely solely on the color of the hind femur, the color of the pronotum, and the number of dorsal spurs on the hind tibia for identification. When we examined the presence of pubescence on the thorax and abdomen, yellow stripes on the head with or without bifurcation, the color of the maxillary palpus and the shape of male genitalia, these seemingly different “species” showed no discernible variation. Furthermore, our molecular results indicate that they actually belong to the species. Therefore, we consider these features as intraspecific variations.
The presence of specific structures in the male genitalia serves as a mechanism to prevent mating between different species (hybridization), and genital morphology is commonly used for identification purposes in crickets. However, the genitalia of Goniogryllus exhibit only minor differences. Nevertheless, molecular analysis and parts of morphological features provide support for classifying them as distinct species. This may be the first example that genitalia cannot be used for identification in crickets as we know. Our hypothesis is that Goniogryllus species inhabiting high-altitude regions (
G. yunnanus comb. nov. (= C. yunnanus) possesses forewings with irregular veins (Fig.
In conclusion, some morphological features, such as the presence or absence of pubescence on the thorax and abdomen, yellow stripes on the head with or without bifurcation, the color of the maxillary palpus and the shape of male genitalia are interspecific differences, and these can be used as classification features. However, the color of the hind femur and pronotum, and the number of dorsal spurs on hind tibia show intraspecific variations in some species. Therefore, the classification based solely on these morphological features requires verification, particularly for the aforementioned species with only intraspecific variations when published. We recommend that future studies consider utilizing these interspecific differences, increase the sample size, or incorporate molecular data for more robust classification.
We thank Wen-Xuan Bi for providing us with specimens. We also thank Tianmushan Nature Reserve, Qingliangfeng Nature Reserve, Changqing Nature Reserve, Emeishan Nature Reserve, Jinyunshan Nature Reserve, Fubaoshan Nature Reserve, Dabashan Nature Reserve, Wudangshan Nature Reserve, and Houhe Nature Reserve for their cooperation.
Table S1
Data type: .docx
Explanation notes: Description of the Collecting information and COI GenBank accession number of Gryllini.