Research Article |
Corresponding author: Menglin Wang ( wangmenglin123@126.com ) Academic editor: Mike Wilson
© 2019 Songping Zhao, Thierry Bourgoin, Menglin Wang.
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:
Zhao S, Bourgoin T, Wang M (2019) The impact of a new genus on the molecular phylogeny of Hemisphaeriini (Hemiptera, Fulgoromorpha, Issidae). ZooKeys 880: 61-74. https://doi.org/10.3897/zookeys.880.36828
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A new genus, Retaldar gen. nov. of the family Issidae (Hemisphaeriinae, Hemisphaeriini) is described from Guangxi Province of China. A revised molecular analysis for the Hemisphaeriini based on partial sequences of 18S, 28S, COXI and Cytb, provides evidence for a new lineage within the subtribe Mongolianina. With two subgroups of genera now identified, the monophyly of Mongolianina is discussed from both a morphological and a molecular basis.
Hemisphaeriinae, molecular, morphology, new species, Oriental, planthoppers
Recently
In this paper, we describe and sequence the genes (18S, 28S (D3–D5) and (D6–D7), COXI and Cytb) of a new species of Hemisphaeriini from Guangxi Province of China, and provide several sequences for other known genera. These new data allow the reassessment of the subtribal division of Hemisphaeriini proposed by
The specimens were collected by net capture and stored in alcohol. The genitalia were separated from the insect body using micro-scissors, and then boiled in 10% NaOH solution for few minutes until muscles were completely dissolved leaving tegumentary structures. After rinsing in distilled water several times to clean the residual NaOH solution, the abdomen was subsequently transferred to glycerine for final dissection and observation. Genitalia were finally conserved under the specimen in genital vials. Photographs for external morphology and genitalia characters were taken using Leica DFC camera attached to Leica M205FA stereomicroscope and further refined with LAS X software. Morphological interpretations and subsequent terminologies follow
Total genomic DNA was extracted from the legs of type specimens using a Sangon Ezup column animal genomic DNA purification kit. The genes (18S, 28S, COXI, Cytb) were amplified using the same primers and amplification procedure as in
Taxa sampling used for the phylogeny tree and corresponding sequence numbers registered in GenBank. The symbol * denotes new added sequences in this study.
Taxa name | Collecting location | Gene 18S | Gene 18S | Gene 18S | Gene 28S | Gene 28S | COXI | Cytb |
---|---|---|---|---|---|---|---|---|
(1F–5R) | (3F–Bi) | (A2–9R) | (D3–D5) | (D6–D7) | ||||
Hemisphaeriini | ||||||||
Ceratogergithus pseudotessellatus (Che, Zhang & Wang, 2007) | China | KX761574 | – | KX761576 | KX761444 | KX702806 | KX702919 | KX702906 |
Ceratogergithus spinosus (Che, Zhang & Wang, 2007) | China | KX761491 | KX761491 | KX761491 | KX761532 | KX761521 | KX761502 | KX761513 |
Choutagus longicephalus Zhang, Wang & Che, 2006 | China | KX650620 | KX650620 | KX650620 | KX761450 | KX702810 | KX761460 | – |
Clypeosmilus centrodasus Gnezdilov & Soulier-Perkins, 2017 | Vietnam | – | – | KX761575 | – | – | KX761470 | KX761474 |
Eusudasina nantouensis Yang, 1994 | China | JX196136 | – | – | – | – | HM052838 | HM452266 |
Euxaldar lenis Gnezdilov, Bourgoin & Wang, 2017 | Vietnam | KX761573 | – | KX761565 | KX761412 | – | – | – |
Gergithoides carinatifrons Schumacher, 1915 | China | – | KX761538 | KX761538 | – | KX702805 | KX761555 | KX702905 |
Gergithoides rugulosus (Melichar, 1906) | China | JX196163 | – | – | – | – | HM052835 | HM452279 |
Gergithus yunnanensis Che, Zhang & Wang, 2007 | China | KX702831 | KX702831 | KX702831 | KX761456 | MN381848* | KX702924 | KX702915 |
Hemisphaerius palaemon Fennah, 1978 | China | KX761486 | KX761486 | KX761486 | KX761526 | KX761517 | KX761497 | KX761508 |
Hemisphaerius rufovarius Walker, 1858 | China | KX702825 | KX702825 | KX702825 | KX761454 | KX702812 | KX702923 | KX702913 |
Hemisphaerius lysanias Fennah, 1978 | Vietnam | KX702833 | KX702833 | KX702833 | KX761404 | KX702860 | KX702933 | KX702883 |
Hemisphaerius coccinelloides (Burmeister, 1834) | Philippines | KX702834 | KX702834 | KX702834 | KX761405 | KX702861 | KX702934 | KX702884 |
Hemisphaerius sp. | Laos | KX702835 | KX702835 | KX702835 | KX761406 | KX702862 | KX761556 | KX702885 |
Hemisphaerius testaceus Distant, 1906 | China | JX196135 | – | – | – | – | HM052831 | HM452258 |
Macrodaruma pertinax Fennah, 1978 | Vietnam | KX702832 | KX702832 | KX702832 | KX761402 | KX702859 | KX702931 | KX702882 |
Macrodaruma sp. | China | KX702828 | KX702828 | KX702828 | KX761399 | KX702857 | KX702927 | KX702881 |
Maculergithus multipunctatus (Che, Zhang & Wang, 2007) | China | KX702816 | KX702816 | KX702816 | KX761443 | KX702804 | KX702918 | KX702904 |
Maculergithus nonomaculatus (Meng & Wang, 2012) | China | KX761492 | KX761492 | KX761492 | KX761533 | KX761522 | KX761503 | KX761514 |
Mongoliana triangularis Che, Wang & Chou, 2003 | China | – | – | KX761561 | KX761528 | – | – | KX761510 |
Mongoliana sinuata Che, Wang & Chou, 2003 | China | KX702820 | KX702820 | KX702820 | KX761448 | – | KX761459 | KX702908 |
Mongoliana sp.1 | Thailand | – | – | MN422135* | MN381854* | – | – | MN332233* |
Mongoliana sp. 2 | China | KX761572 | MN422136* | KX761566 | KX761534 | MN381849* | – | – |
Mongoliana serrata Che, Wang & Chou, 2003 | China | JX196160 | – | – | – | – | HM052830 | HM452272 |
Neogergithoides tubercularis Sun, Meng & Wang, 2012 | China | KX702822 | KX702822 | KX702822 | KX761451 | MN381845* | KX761558 | KX702910 |
Ophthalmosphaerius trilobulus (Che, Zhang & Wang, 2006) | China | KX702826 | KX702826 | KX702826 | KX761455 | KX702813 | KX761462 | KX702914 |
Retaldar yanitubus sp. nov. | China | MN381856* | MN381856* | MN381856* | MN381853* | MN381851* | MN381857* | MN332232* |
Thioniini | ||||||||
Thionia sp. | Panama | KX761539 | KX761539 | KX761407 | KX702935 | KX702886 | ||
Hysteropterini | ||||||||
Celyphoma quadrupla Meng & Wang, 2012 | China | KX702815 | KX702815 | KX702815 | KX761442 | KX702803 | KX702917 | KX702903 |
Mulsantereum maculifrons (Mulsant & Rey, 1855) | France | KX761569 | KX761569 | KX761569 | KX761400 | MN381847* | KX702928 | KX761551 |
Issini | ||||||||
Issus coleoptratus (Fabricius, 1781) | France | KX761568 | KX761568 | KX761568 | KX761403 | KX761560 | KX702932 | KX761550 |
Kodaianellini | ||||||||
Kodaianellissus intorqueus Wang, Bourgoin & Zhang, 2017 | China | KX761476 | KX761476 | KX761476 | KX761480 | KX761482 | KX761472 | |
Kodaianella bicinctifrons Fennah, 1956 | China | KX702814 | KX702814 | KX702814 | KX761441 | KX702802 | KX761458 | KX702902 |
Sarimini | ||||||||
Sarima bifurca Meng & Wang, 2016 | China | KX702819 | KX702819 | KX702819 | KX761447 | KX702808 | KX702921 | KX761552 |
Tetrica sp. | China | KX702821 | KX702821 | KX702821 | KX761449 | KX702809 | KX702922 | KX702909 |
Parahiraciini | ||||||||
Flavina hainana (Wang & Wang, 1999) | China | KX702824 | KX702824 | KX702824 | KX761453 | MN381846* | KX702912 | |
Fortunia sp. | China | KX761487 | KX761487 | KX761487 | KX761527 | KX761518 | KX761498 | KX761509 |
Neodurium hamatum Wang & Wang, 2011 | China | KX702818 | KX702818 | KX702818 | KX761446 | MN381844* | KX702920 | |
Rhombissus sp. | China | MN381855* | MN381855* | MN381855* | MN381852* | MN381850* | MN332231* |
Retaldar yanitubus sp. nov., here designated.
Genus name masculine from the free combination of the latin word ‘rete’ meaning network as for the reticulated forewings and the suffix ‘-aldar’ from the genus Euxaldar Fennah, 1978.
This new genus is similar to the genus Clypeosmilus Gnezdilov & Soulier-Perkins, 2017 in general appearance, but differs by: 1) a more complex and obscure reticular venation of the forewing (Fig.
Retaldar yanitubus sp. nov., holotype. 6 male genitalia, lateral view 7 male anal tube, dorsal view 8 gonostylus, lateral view 9 phallic complex, left lateral view 10 phallic complex, right lateral view 11 apical half of phallic complex, ventral view. The arrows on Fig.
Head with compound eyes slightly wider than pronotum, almost same width as mesonotum (Fig.
Retaldar yanitubus sp. nov., paratype. 12 female anal tube, dorsal view 13 gonoplacs, lateral view 14 gonoplacs, dorsal view 15 gonapophysis IX and gonaspiculum bridge, dorsal view 16 gonapophysis IX and gonaspiculum bridge, lateral view 17 sternite VII 18 gonocoxa VIII and gonapophysis VIII, ventral view.
Anal tube in lateral view long and curved (Fig.
Anal tube in dorsal view a little longer than wide (Fig.
China (Guangxi).
Specific epithet built by the arbitrary combination of the alphabet letter “Y” and “anal tube” latinised into “anitubus”, referring to the Y-shaped male anal tube in dorsal view.
The species is close to Clypeosmilus centrodasus Gnezdilov & Soulier-Perkins, 2017, from which it differs by its generic characters (complex reticular venation (Fig.
Holotype: ♂, CHINA: Guangxi Province, Jinxiu, Dayaoshan natural reserve, Hekou, 24°14'11"N, 110°14'11"E, 689.9 m, 23 vii 2018, Feilong Yang & Kun Zhao leg.
Paratypes: 2♀♀, same data as for holotype.
Length: male (including forewings) (N =1): 3.1 mm; female (including forewings) (N =2): 3.3–3.4 mm.
Vertex tawny, disc with two dark brown circular markings; anterior, lateral and posterior margins tawny (Figs
Head and thorax.
Vertex 2.5 times wider than long in midline, without median carina; anterior margin straight; posterior margin roundly concaved (Fig.
Male genitalia.
Anal tube in lateral view arc-shaped, gradually narrowing from the base to the end, apical part conical (Fig.
Female genitalia.
Anal tube in dorsal view ovate, widest near middle, 1.2 times longer in midline than widest part, apical margin and lateral margins rounded (Fig.
Genes sequences were registered in GenBank with accession numbers as following: MN381856 (whole 18S), MN381853 (28S D3–D5), MN381851 (28S D6–D7), MN381857 (COXI), MN332232 (Cytb). The COXI sequence of this species differs respectively by 87 bp (over 601 bp: 14.5%) and 103 bp (over 681 bp: 15.1%) from Eusudasina nantouensis Yang, 1994 (Genbank accession number: HM052838) and Clypeosmilus centrodasus (Genbank accession number: KX761470).
As in the genus Euxaldar (
In-group sampling comprised 14 Hemisphaeriini genera and 27 species while 12 other issid genera were used as outgroups (Fig.
According to their results,
In the phylogeny of
Within Hemisphaeriinae, Hemisphaeriini sec.
Hemisphaeriina remains the most diverse subtribe with, at least, 11 described genera, eight having being sequenced and providing the following topology: ((Ophthalmosphaerius + Gergithus) + (Hemisphaerius + (Ceratogergithus + (Maculergithus + (Neogergithoides + (Choutagus + Gergithoides)))))). Three genera remain unsequenced: Epyhemisphaerius Chan & Yang, 1994, Neohemisphaerius Chen, Zhang & Chang, 2014 and Rotundiforma Meng, Wang & Qin, 2013. Moreover, Gergithus yunnanensis Che, Zhang & Wang, 2007 probably belongs to another new genus and Gergithus s.s. should remain in an incertae sedis position within Hemisphaeriini. According to
According to our analysis, Mongolianina should be now restricted to the following genera: (Mongoliana + (Euxaldar + Macrodaruma)), separated from a third new lineage (Retaldar + (Clypeosmilus + Eusudasina)). The support of this last lineage is high (BS= 94) and higher than the Mongolianina lineage itself. Because the monophyly of the Mongolianina lineage itself remains weak and unstable, we prefer to keep provisionally a paraphyletic Mongolianina, including in it this new lineage. When more genera will be sequenced, a better and stronger topology will probably appear. Genera such as Bruneastrum Gnezdilov, 2015, Tapirissus Gnezdilov, 2014 and Neotapirissus Meng & Wang, 2016 should also join this group, but, as for Hemisphaeriina, new morphological analyses are still needed to better identify these lineages without molecular sequencing.
The new species Retaldar yanitubus gen. & sp. nov. has a protruded clypeus in lateral view. In the tribe Hemisphaeriini, several other genera also display such a protruded clypeus: Euxaldar, Clypeosmilus and Eusudasina. They also share a plesiomorphic clearly visible CuP on the forewing while this vein is not visible in Hemisphaeriina (
Including more sequenced taxa in the molecular analysis, revisiting morphological characteristics of Hemisphaeriini and investigating the etho-ecology of these fascinating ladybug-like planthoppers is now urgently needed.
We sincerely thank Feilong Yang and Kun Zhao for collecting specimens for this study and Dr M. Wilson for useful comments and English review. This study was supported by Sichuan Science and Technology Program (2019YFH0086) and Sichuan Education Program (18ZA0481) of China, the Doctoral Scientific Research Foundation (17E070), and the Fundamental Research Funds (17B009) of China West Normal University, China.