A new genus Vittaliana belonging to the tribe Opsiini (Hemiptera: Cicadellidae) from India and its molecular phylogeny

For morphological studies

Data was collected as previously described in Meshram, Shashank & Sinha (2017) specifically, in and around ICAR research institutes, Vittal, Kasargod (Kerala: India), with a mercury vapor lamp. Hence, no specific permissions were required for any of the collection localities/activities. Specimens were processed by a series of steps like sorting, cleaning, and mounting. Male genitalia dissections were carried out as described by Oman (1949) and Knight (1965) as follows, the abdomen was removed by inserting a sharp pin between the abdomen and thorax with gentle piercing. The abdomen was treated in 10% KOH for 2∼4 h to remove unsclerotized material by gently prodding the abdomen with the head of a pin. Afterward, the abdomen was rinsed thoroughly in water. The internal structures were then removed by a hooked pin, before being stored in glycerol vials for study.

Photographs were taken with a Leica DFC 425C digital camera on the Leica M205FA stereo zoom automontage microscope.

Repository of the Material: The holotype and paratypes are deposited in National Pusa Collection (NPC), Indian Council of Agricultural Research-Indian Agricultural Research Institute (IARI)-New Delhi, India (with repository number: Holotype: RRS2; Paratypes: RRS3, RRS4, RRS5)

New Taxon LSID. Vittaliana: urn:lsid:zoobank.org:act:51DA3683-0359-444F-8C11-F630518D8506, Vittaliana reticulata: urn:lsid:zoobank.org:act:E690A7AF-7FCD-4460-93BC-1274D932C5F4.

The electronic version of this article in Portable Document Format (PDF) will represent a published work according to the International Commission on Zoological Nomenclature (ICZN), and hence the new names contained in the electronic version are effectively published under that Code from the electronic edition alone. This published work and the nomenclatural acts it contains have been registered in ZooBank, the online registration system for the ICZN. The ZooBank LSIDs (Life Science Identifiers) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix http://zoobank.org/. The LSID for this publication is: urn:lsid:zoobank.org:pub:228D17FC-590C-41C7-9434-60554F753DBA. The online version of this work is archived and available from the following digital repositories: PeerJ, PubMed Central, and CLOCKSS.

Molecular studies

Alignment and phylogenetic analyses

For phylogenetic analysis, the majority of species sequences were taken from Zahniser & Dietrich (2013) and Zhaniser & Dietrich (2010) and Du, Liang & Dai (2019). A dataset consisting of the newly sequenced taxa and 76 sequences of Deltocephalinae species. The outgroups consist of two species from Aphrodinae and one species from Euacanthellinae (Table 1).

The histone H3 and 28S rDNA sequences aligned separately with the MUSCLE application in MEGA 6 (Tamura et al., 2013a; Tamura et al., 2013b). The aligned sequences of the two gene regions were concatenated into one dataset using Sequence Matrix 1.7.8 (Vaidya, Lohman & Meier, 2011) and obtained NEXUS data block for combined data set as follows: #NEXUS begin data; dimensions ntax = 80 nchar = 6974; format datatype = dna; gap = −; missing = ?; matrix; end;

The NEXUS file used in the phylogenetic analysis deposited in public repository TreeBASE (Study ID S26664; https://treebase.org).

Maximum likelihood (ML) analysis of combined gene region (H3, 28S rDNA) were constructed in IQtree v1.4.1 (Nguyen et al., 2015) using the best-fit substitution model automatically selected by the software according to the Bayesian information criterion scores and weights (BIC) with partitions. An ultrafast bootstrap (UFB) (Minh, Nguyen & von Haeseler, 2013) with 1,000 replicates and the SH-like approximate likelihood ratio test (SH-aLRT) (Guindon et al., 2010) and Bayesian-like transformation of aLRT (aBayes) (Anisimova et al., 2011) were used in the analysis to assess branch support and obtained tree was visualized in FigTree v1.4.2.

Taxonomy

Type species: Vittaliana reticulata sp. nov. Sunil and Meshram [Figs. 1A–1J, 2A–2G, 3A–3H]

Diagnosis

This genus is placed in the subtribe Opsiina of Opsiini based on all these characters macropterous; ovipositor not protruding far beyond pygofer apex and subgenital plates with a lateral row of macrosetae; aedeagal shafts divided near to base. The new genus can be differentiated from all related genera in this subtribe by a combination of the following characters: Body and face whitish with yellow, and brown mottling; anterior margin of crown with five white patches, slightly produced, slightly concave; pronotum with concave posterior margin; aedeagal shafts arising from near to base, aedeagus distinctive outward curved apically, without processes arising from base, with a pair of medial processes arising from mid-length of shafts, apex without processes.

Description

Colour. Body and face whitish with yellow and brown mottling. Crown anterior margin with five white patches (Fig. 1D). Pronotum brown mottled with white patches. Scutellum yellow with an orange basal triangle (Fig. 1D). Eyes red; forewing white speckled with brown patches (Fig. 1B).

Body length. Male 3.6 mm long; 1.4 mm wide across eyes. Female 3.8 mm long; 1.46 mm wide across eyes.

Head. Anterior margin of head slightly produced, head in dorsal view as wide as pronotum; crown length 2/3rd as long as median length of the pronotum, anterior margin produced with concave posterior margin; face with brown and white irregular mottling; ocelli small, close to eyes on anterior margin of crown; clypellus 2.8x as long as wide; gena obtusely incised laterally (Fig. 1C).

Thorax. Pronotum anterior margin convex, 2x as broad as long, hind margin slightly concave; scutellum 1.5x as broad as long, and 0.6x as long as width of pronotum, with distinct scutoscutellar suture (Fig. 1D). Forewing elongate, veins raised, three subapical and four apical cell, claval vein raised with 2 crossveins, appendix extended around the apex (Fig. 1I). Hind wing veination complete, appendix broad (Fig. 1J).

Legs. Prothoracic femur with AM1 seta only; intercalary row with one row of more than 5 fine setae; AV row with 2–3 macrosetae, AV1 seta long; AD setae small and sparsely arranged. Prothoracic tibia on dorsal surface rounded, AD row with 4–5 setae long, distributed widely; AV setae moderately dense and long (Fig. 1F). Mesothoracic femur with AD setae small; AV row with basal half short setae and rest with macrosetae; AM seta present; intercalary row with one row of more than 5 fine setae (Fig. 1G). Metathoracic femur with setal formula 2+2+1; lateral surface area broadened distally; metathoracic tibia flattened, tibial row AD setae long and densely arranged, PD with long macrosetae placed equidistantly, AV setae small and densely arranged, PV with macrosetae moderately arranged, Metatarsomere I length equals to tarsomere II and tarsomere III combined (Fig. 1H).

Male genitalia. Pygofer longer than wide with macrosetae in midlateral region, with anal tube long, 3/4th membranous from the base (Fig. 2A). Valve triangular with broad base (Fig. 2B). Subgenital plate triangular, basally broad, posterior half gradually narrowed towards apex, with 7–8 submarginal macrosetae, 7 microsetae medially on distal 2/3rd (Fig. 2F). Style broadly bilobed basally, subapical angle not prominent with few setae (Fig. 2C). Connective Y-shaped, stem 1.3x as long as arms (Fig. 2G). Aedeagus with well-developed dorsal apodeme, with a pair of medial processes arising from mid-length of shafts, gonopores subapical (Fig. 2D).

Female genitalia. Female seventh sternite trapezoid in shape, sternite 2x as wide as median length, hind margin with sinuate with shallow notch medially (Fig. 3I); first pair of valvulae wider beyond the base and narrowed at apex, with irregular sculpture on apical 1/2th, dorsal hyaline area restricted to basal half (Fig. 3F); second pair of valvulae, with small teeth and sculpting on apical half (Fig. 3H).

Etymology

This genus was named after the place of collection, Vittal in Kerala, India.

Distribution

Kerala, Karnataka, India

Diagnosis. In addition to generic character, the specific characters for this species are: anterior margin of crown slightly produced with five white patches (Fig. 1D), face with brown and white irregular marking (Fig. 1C). Pygofer with a group of macrosetae confined to the mid-lateral region (Fig. 2A). Style with beak-like apophysis, directed posteriorly (Fig. 2C). Connective Y- shaped, stem longer than arms (Fig. 2G). Aedeagal gonopore opens subapically on ventral margin with constriction (Fig. 2D). Seventh sternite 2x as wide as median length, hind margin with shallow notch medially (Fig. 3I).

Description.

Colour. Body and face whitish with yellow and brown mottling; anterior margin of crown with five white patches (Fig. 1D). Pronotum brown mottled with white patches. Scutellum yellow with an orange basal triangle. (Fig. 1D); fore wing white speckled with brown patches (Fig. 1B).

Anterior margin of head slightly produced, head in dorsal view as wide as pronotum (Fig. 1D). Ocelli small, close to eyes on anterior margin of crown; clypellus 2.8x as long as wide. Gena obtusely incised laterally (Fig. 1C). Pronotum anterior margin convex, 2x as broad as long; scutellum 1.5x as broad as long with distinct scutoscutellar suture (Fig. 1D). Fore wing macropterous, veins raised, appendix expanded around the apex (Fig. 1I).

Male genitalia. Pygofer longer than wide with a group of macrosetae confined to mid-lateral region (Fig. 2A). Valve 2x as wide at base as long medially (Fig. 2B). Subgenital plate triangular, broad at base slightly tapering towards the apex with 7–8 submarginal macrosetae (Fig. 2F). Style bilobed basally, subapical angle not prominent with beak-like apophysis, directed posteriorly (Fig. 2C). Connective Y- shaped, stem longer than arms (Fig. 2G). Aedeagus with a pair of medial processes arising from mid-length of shafts, gonopore opens subapically on ventral margin with constriction (Fig. 2D).

Female genitalia. Seventh sternite 2x as wide as median length, hind margin with shallow notch medially (Fig. 3I). First pair of valvulae with an irregular sculpture on apical 1/2th (Fig. 3F). Second pair of valvulae with small teeth and sculpting on apical half (Fig. 3F).

Type material

Molecular analysis

Maximum likelihood (ML) analysis using IQtree v1.4.1 of the 80 taxa and 6074bp aligned nucleotide position of combined Histone H3 and 28S rDNA (D2 & D9-D10 region) yielded maximum likelihood phylogenetic tree (Fig. 4) with good SH-like approximate likelihood ratio test (SH-aLRT), ultrafast bootstrap (UFB) and Bayesian-like transformation of aLRT (aBayes). Our new species Vittaliana reticulata is sister to clade Opsius versicolor (Distant), Opsiini gen. sp., Libengaia sp., Hishimonus phycitis (Distant) and Yinfomibus menglaensis Du, Liang & Dai. Among them V. reticulata closely associated with Opsius versicolor with good SH-aLRT score (77.7), UFB (89) and moderate aBayes (0.689), indicates that the new genus belongs to the tribe Opsiini under subfamily Deltocephalinae. All included 11 species of Opsiini including new genus, form two clades in the phylogenetic tree resolve as paraphyletic with respect to clade Pendarini + Athysanini + Scaphytopiini in one clade and with Hecalini in another clade with SH-aLRT (>90), low UFB (>50) and aBayes (1).