Exploring the relationship of Homalosilpha and Mimosilpha (Blattodea, Blattidae, Blattinae) from a morphological and molecular perspective, including a description of four new species

Morphological study

Morphological terminology used in this paper mainly follows McKittrick (1964), Roth (2003) and Li et al. (2018).

The maculae on the pronotum provided effective information for the identification of Homalosilpha species, which could be grouped into three types (Shelford, 1910; Princis, 1966a; Bey-Bienko, 1969; Roth, 1999): (1) white spots scattered on the black pronotum, represented by H. decorata and H. quadrimaculata, (2) one large central dark spot on the pale brown pronotum, represented by H. hanni, H. nigricans and H. gaudens, and (3) the symmetrical multiple spots and stripes on the disk of the pale brown pronotum, represented by the remaining Homalosilpha species. The third type is more complex than the former two, so we herein address the specific part of the maculae for description as shown in Fig. 1.

All specimens were measured by vernier caliper for the body length from the anterior to the posterior, for the body length including tegmina from the anterior to the tip of tegmina, and pronotum length × width at the longest and the widest part. Genital segments of the examined specimens were macerated and photographs (Figs. 1–9) of the specimens were taken as previously described in Liao, Wang & Che (2019). The type materials are deposited in the Institute of Entomology, College of Plant Protection, Southwest University, Chongqing, China (SWU).

DNA extraction, PCR, and sequencing

We sampled seven genes of 13 species (Tables S1 and S2) from Homalosilpha in this study: mitochondrial 12S, 16S, cytochrome c oxidase subunit I (COI) and subunit II (COII), and nuclear 18S, 28S, and histone H3. Total DNA was extracted from the leg tissue of samples according to the Hipure Tissue DNA Mini Kit (Tsingke Biological Technology, Beijing, China). All fragments were amplified using PCR; primers for amplifications are given in Table 1. Reactions were carried out in volumes of 25 µL, containing 22 µL of 1 × 1 T3 supper mix (Tsingke Biological Technology, Beijing, China), 1 µL of each primer and 1 µL of DNA template, except for COII containing 12.5 µL T2 mix (Tsingke Biological Technology, Beijing, China), 8.5 µL of ultrapure water, 1 µL of each primer and 2 µL of DNA template. The amplification conditions were: initial denaturation at 98 °C for 2 min, followed by 35 cycles for 10 s at 98 °C, 10 s at 43−55 °C, and 15 s at 72 °C, with a final extension of 2 min at 72 °C; however for COII: initial denaturation at 94 °C for 5 min, followed by 35 cycles for 45 s at 94 °C, 45 s at 50 °C, and 45 s at 72 °C, with a final extension of 10 min at 72 °C. All sequences were deposited in GenBank (accession numbers in Tables S1 and S2).

Sequence processing and phylogenetic analyses

In this study, a total of 14 COI sequences, whose lengths were 658 bp, were combined with one Protagonista, two Periplaneta, one Rhabdoblatta, one Brephallus and one mantid sequence to infer species delimitation analysis for Homalosilpha and Mimosilpha (Table S1). Intraspecific and interspecific genetic divergence values are quantified based on the Kimura 2-parameter (K2P) distance model (Kimura, 1980), using MEGA 7 (Kumar, Stecher & Tamura, 2016).

To infer the relationship of Homalosilpha and Mimosilpha, we included sequence data from 22 Blattidae taxa (ingroup including seven Homalosilpha and one Mimosilpha species) and 12 outgroup taxa (Table S2). These analyses were performed based on six genes (12S, 16S, 18S, 28S, COII and H3). These data was aligned by online Mafft 7 (https://mafft.cbrc.jp/alignment/server/), and the methods were the same as Wang et al. (2017). Specifically, the Q-INS-i algorithm was selected for non-coding protein genes (12S, 16S, 18S, 28S), the G-INS-i algorithm was selected for coding protein genes (COII, H3) and used with other parameters at their default values. Alignments of sequences were inspected visually and manually adjusted in MEGA 7.0. Poorly aligned characters were removed but these were limited. The remaining bases are as following: 424nt for 12S, 447nt for 16S, 1814nt for 18S, 596nt for 28S, 665nt for COII, 328nt for H3.

The molecular data set was divided into eight partitions (partitioned by gene: 12S, 16S, 18S, 28S, COII_pos1, COII_pos2, H3_pos1, and H3 _pos2). We excluded the third codon positions of the protein-coding genes (COII and H3) because of the high level of mutational saturation. The third codon position (pos3) (I_SS = 0.977) was much more saturated than the first and second codon position (pos12) (I_SS = 0.493) after using Xia’s method implemented in DAMBE 5.0 (Xia, 2013). Phylogenetic analyses were constructed using maximum likelihood (ML) and Bayesian inference methods. ML analyses were performed using RaxML 7.7.1 (Stamatakis, Hoover & Rougemont, 2008), and BI analyses were performed using MrBayes 3.2 (Ronquist et al., 2012). ML analyses was performed using RAxML v.7.7.1 (Stamatakis, Hoover & Rougemont, 2008). We used the GTRGAMMA model for the combined datasets with 1,000 bootstrap replicates. For BI analyses, PartitionFinder v.1.1.1 (Lanfear et al., 2012) was utilized to choose models and model selection was based on BIC. For eight partitions, the best-fitting substitution models are as follows: GTR+I+G: 12S, 16S; TIMef+I+G: 18S, H3_pos1, H3_pos 2; TrN+I+G: 28S, COII_pos1, COII_pos2. Two independent sets of Markov chains were run, each with one cold and three heated chains for 5,000,000 generations, and every 1,000th generation was sampled. Convergence was inferred when a standard deviation of split frequencies < 0.01 was completed. The ESS values are all more than 200.

Species delimitation based onCOIand morphological data

In this study, we acquired 14 COI sequences representing seven Homalosilpha and 1 Mimosilpha species. All of the new sequences have been deposited in GenBank with accession numbers MW201581–MW201594 (Table S1). The COI sequences we acquired had a relatively high AT content (67.2%), with an average nucleotide composition of A = 31%, T = 36.2%, C = 16.5%, and G = 16.3%. Sequence analysis revealed that 109 (16.57%) sites were variable, of which 79 (12%) sites were parsimoniously informative. ML analysis revealed that clades from the same species, constituted monophyletic groups with high support values (Fig. 2). In particular, the male and female of H. clavellata sp. nov., which have different markings on the pronotum, could be matched by COI data.

We observed the lowest and largest K2P interspecies genetic distance (Table 2) among these species. The lowest distance 0.031 was between Homalosilpha sp. and H. clavellata sp. nov., the largest distance 0.084 between H. nigricans and H. arcifera, H. arcifera and H. kryzhanovskii, as well as M. disticha and H. kryzhanovskii. Morphologically, although having a low genetic distance, Homalosilpha sp. and H. clavellata sp. nov. show distinct differences in the maculae on the pronotum, the band in the interocular space (H. clavellata sp. nov.: yellow and straight transverse band, Homalosilpha sp.: yellow and tortuous transverse band) and even in the female genitalia (H. clavellata: anterior of basivalvula with spots, the shape of first valvule uniform, Homalosilpha sp.: surface of basivalvula with fold, the end of the first valvula swollen). Finally, a total of 8 Homalosilpha and Mimosilpha species were recovered after combining the results of molecular data with morphological data, of which 4 species are new to science: H. obtusangula sp. nov., H. recta sp. nov., H. alba sp. nov., and H. clavellata sp. nov. More details about these new Homalosilpha species are provided below.

Relationship of HomalosilphaandMimosilphainferred from two phylogenetic analyses

We acquired 12 12S, 12 16S, 11 18S, 12 28S, 10 COII, 12 H3 sequences with accession numbers MW218556–MW218578, MW219581–MW218604 and MW201809–MW201830 (Table S2). For the concatenate dataset (12S, 16S, 18S, 28S, COII, H3), our likelihood and Bayesian phylogenetic analyses yielded totally identical topologies with generally high node support (Fig. 3 and Fig. S1). All genera of Blattidae clustered together and formed a highly supported monophyletic group (MLB = 100, BPP = 100). Five recognized major lineages of Blattidae from ML and BI inferences were recovered, represented by members of Polyzosteriinae, Archiblattinae and Blattinae. According to our inferred trees, Polyzosteriinae, Archiblattinae and Blattinae were paraphyletic with high support values.

The topology derived from ML and BI analyses shows that all Homalosilpha members cluster together and are recovered as the sister group of Mimosilpha (MLB = 100, BPP = 100). H. clavellata sp. nov. and Homalosilpha sp. were recovered as sister groups with strong support values, which is consistent with the inference based on COI data. And the result is same as the trees of seven genes (12S, 16S, 18S, 28S, COI, COII, H3) (Figs. S2 and S3).

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: [74F6BF88-9FFD-40DB-9C8B-D7650491F0EE]. The online version of this work is archived and available from the following digital repositories: PeerJ, PubMed Central and CLOCKSS.

Taxonomy

Checklist of Homalosilphaworldwide

Key to Homalosilpha species worldwide

• 1. Pronotum mostly black …2

• Pronotum mostly white…3

• 2. Pronotum with two vertical white stripes near the lateral margin …H. decorata

• Pronotum with four white spots respectively at the anterior margin and middle…H. quadrimaculata

• 3. Pronotum with a large black spot at center …4

• Pronotum with multiple black spots or stripes …6

• 4. The large spot connecting both the front and hind margins …H. haani

• The large spot only connected with the hind margin …5

• 5. Pronotum broadest near the hind margin, trapeziform …H. nigricans

• Pronotum broadest near the middle …H. gaudens

• 6. Face pale yellow …H. arcifera

• Face black …7

• 7. Vertex yellow to yellowish brown …8

• Vertex black …10

• 8. Abdomen with yellow spots …H. valida

• Abdomen brownish black to dark …9

• 9. The hind margin of supra-anal plate slightly concave, lateral angles blunt …H. recta sp. nov.

• The hind margin of supra-anal plate round, lateral angles sharp …H. alba sp. nov.

• 10. Interocular space with a yellow straight band …11

• Interocular space with a yellow band …H. ustulata

• 11. The lateral pattern and vertical pattern of pronotum connected with posterior band, spots on both sides of the lateral pattern …H. vicina

• The middle of pronotum with spots or spots and stripes, the both sides of lateral pattern without spots…12

• 12. Pronotum without vertical pattern …H. contraria

• Pronotum with vertical pattern …13

• 13. The vertical pattern of pronotum with one stripe …14

• The vertical pattern of pronotum with spots …16

• 14. The vertical pattern with one short and bold stripe …H. clavellata sp. nov.

• The vertical pattern with one long stripe and posterior vertical pattern bifurcated …15

• 15. The vertical pattern with pinstripe and the anterior pattern connected with lateral pattern …H. obtusangula sp. nov.

• The vertical pattern with black stripe and the central pattern connected with vertical pattern …Homalosilpha sp.

• 16. The hind margin of supra-anal plate with two sharp valves …H. javanica

• The hind margin of supra-anal plate with two round valves …H. kryzhanovskii

Holotype. CHINA: Tibet: male (SWUB125-H) Beibeng Township, Mêdog County, 2013.VII.31, Xinglong Bai & Junsheng Shan leg.

Paratype. 1 female (SWUB125-P1), CHINA: Yunnan: Mt. Fenshuiling, Jinping County, 2016.V, Tianlong He leg.

Etymology. The species epithet comes from the Latin word obtusangulus in reference to the lateral angles of supra-anal plate being obtuse.

Description. Male. Body length 24.4 mm; overall length including tegmen 33.8 mm; pronotum length × width 6.4 × 8.4 mm. Female. Body length 26.5 mm; overall length including tegmen 36.3 mm; pronotum length × width 6.8 × 8.9 mm.

Coloration. Similar in both sexes. Body generally dark brown (Figs. 5A and 5B). Interocular space with a whitish transverse band. Pronotum white, margins black outlined, hind margin black; center of the pronotum with symmetrical blackish markings (Figs. 5C and 5D). Tegmen reddish brown, wing hyaline, brownish except the anal area (Figs. 7A and 7B). Tibiae and tarsi dark reddish brown (Fig. 5B).

Head with vertex unsheltered by pronotum. Interocular distance slightly narrower than the distance between antennae sockets. Pronotum nearly heptagonal, broadest near the middle; the middle of pronotum with black and symmetrical patterns, the anterior pattern connected with lateral pattern, the lateral pattern arc-shaped, central pattern dart-like, the posterior of vertical pattern bifurcated, the posterior band bold (Figs. 5C and 5D). Male supra-anal plate broad and symmetrical, nearly trapezoidal, but with a deep, obtuse-angled invagination at the middle of hind margin; male subgenital plate asymmetrical, left hind corner more protruded than on the right, styli similar, long (Figs. 8A and 8B).

Male genitalia. Left phallomere: L1 fold sclerite; margin of L2d with small serration, posterior of L2v produced as a spiny projection to the end; L3 unciform, the base broad, tapering down, the terminal bifurcate, one branch short, the other long (Fig. 8C). Right phallomere: R1 foot-shaped, left margin with two spines; terminal of R2d sharp, R2v fold sclerite; R3 broad and fold sclerite (Fig. 8C).

Remarks. H . obtusangula sp. nov. resemble H. kryzhanovskii, but can be distinguished from H. kryzhanovskii by the following characters: (1) the pronotum with stripes in the former, while the latter with spots; (2) the hind margin of supra-anal plate deeply concave, while the latter, hind margin slightly concave; (3) the lateral corner sharp in the former, while the latter, the lateral corner rounded

Holotype. CHINA: Yunnan: male (SWUB126-H) Banhong Township, Cangyuan County, 1130 m, 2008.VII.16-18, Jishan Xu & Zhenhua Gao leg.

Etymology. The species epithet ‘rectus’ refers to the hind margin of supra-anal plate and subgenital plate being straight.

Description. Male. Body length 28.3 mm; overall length including tegmen 39.9 mm; pronotum length × width 7.3 × 10.6 mm.

Coloration. Body generally dark brown (Figs. 5E and 5F). Vertex and the area between eyes and ocelli white. Pronotum white, subhyaline, with black outline weak, hind margin blackish weakly, middle of the pronotum with sparse, symmetrical blackish markings (Figs. 5G and 5H). Tegmina yellowish brown, wings hyaline, dark brownish except the anal area (Figs. 7C and 7D). Tibiae and tarsi dark reddish brown.

Head with vertex unsheltered by pronotum. Interocular distance narrower than distance between antennae sockets. Pronotum nearly hexagonal, anterior and hind margins slightly outward, broadest near the middle; the anterior pattern triangular, the lateral pattern and central pattern with two parts, the vertical pattern with only one spot, the posterior band indistinct (Figs. 5E and 5F). Male supra-anal plate broad and symmetrical, nearly trapezoidal, with a very shallow, obtuse-angled invagination at the middle of hind margin, hind lateral angles slightly protruded laterally; subgenital plate of male asymmetrical, quadrate, right lateral hind corner narrowed, styli similar, long (Figs. 8D and 8E).

Male genitalia. Left phallomere: L1 fold sclerite; posterior margin of L2d with small spines, posterior of L2v produced as a spiny projection to the end; L3 unciform, the base broad, tapering down, the terminus bifurcate (Fig. 8F). Right phallomere: R1 foot-shaped, left margin with two unequal spines, the spine on the top longer than the bottom; terminal of R2d unciform, R2v fold sclerite; R3 broad (Fig. 8F).

Remarks. H. recta sp. nov. is similar to H. kryzhanovskii (Figs. 6I–6L, 8M–8O) and H. alba sp. nov. (Figs. 5I–5L, 8G–8I), but can be distinguished from H. kryzhanovskii by the following characters: (1) the composition of black spots in pronotum; (2) vertex white, while in H. kryzhanovskii black; and (3) the hind margin of supra-anal plate slightly concave and with lateral sides curving laterally, however in H. kryzhanovskii the two hind angles nearly round. Furthermore, it can be distinguished from H. alba sp. nov. by the following characters: (1) the composition of black spots with slight difference; and (2) the hind margin of the supra-anal plate slightly concave and blunt, however in H. alba, the lateral angles of the supra-anal plate are sharp.

Holotype. CHINA: Hainan: male (SWUB127-H) Yajiang Scenery Spot, Mt Bawangling, Cangjiang County, 1130 m, 2015.IV.30, Lu Qiu & Qikun Bai leg.

Paratypes. China: Hainan: 1 female (SWUB127-P1). Maogan Township, Baoting County, 2015.IV.11-12, Lu Qiu & Qi-kun Bai leg; 1 female (SWUB127-P2). Mt Diaoluoshan, Lingshui County, 1964.III.28, Si-Kong Liu; 1 female (SWU) Ledong County, 1954.V.4, Keren Huang leg.

Etymology. The species epithet is derived from the Latin word ‘albus’, referring to the vertex being white.

Description. Male. Body length 27.2 mm; overall length including tegmen 33.1 mm; pronotum length × width 6.1 × 8.5 mm. Female. Body length 28.3 mm; overall length including tegmen 36.4 mm; pronotum length × width 7 × 9.9 mm.

Coloration: Similar in both sexes. Body dark brown (Figs. 5I and 5J). Vertex and the area between eyes and ocelli white. Pronotum white, margins black outlined, hind margin black (Figs. 5K and 5L). Tegmina reddish brown, wings hyaline, brown except the anal area (Figs. 7E and 7F). Middle of the abdomen yellowish brown. Tibiae and tarsi reddish brown (Fig. 5L).

Head with vertex unsheltered by pronotum. Interocular distance narrower than the distance between antennae sockets. Pronotum nearly a hexagon, anterior and hind margins slightly outward, broadest near the middle; the anterior pattern and central pattern triangular, the lateral pattern with three parts, the vertical pattern with only one spot, the posterior band bold and with two spots on the band (Fig. 5K). Male supra-anal plate symmetrical, nearly trapezoidal, hind margin widely concave, lateral hind angles sharp and protruded outward; subgenital plate of male asymmetrical, left hind corner angled while right hind corner round, styli similar (Figs. 8G and 8H).

Male genitalia: Left phallomere: L1 lamellar; margin of L2d with small spines, posterior of L2v produced with a spiny projection to the end; L3 with a spiny projection, the basal broad, tapering down, the terminus bifurcate, the lengths equal (Fig. 8I). Right phallomere: R1 foot-shaped, left margin with two equal spines; terminal of R2d unciform, R2v fold sclerite; R3 with complicated and broad sclerite (Fig. 8I).

Remarks. H. alba is similar to H. kryzhanovskii (Figs. 6I–6L, 8M–8O) but can be distinguished from H. kryzhanovskii by the following characters: (1) the black spots smaller and thinner than H. kryzhanovskii in pronotum; and (2) the lateral angles of supra-anal plate sharp, however in H. kryzhanovskii nearly round.

Holotype. CHINA: Yunnan: male (SWUB128-H), Tongbiguan Township, Yingjiang County, 1130 m, 2018.IV.30, Lu Qiu & Wenbo Deng leg.

Paratype. 1 female (SWUB128-P1), CHINA: Yunnan: Tongbiguan Township, Yingjiang County, 1130 m, 2018.IV.30, Lu Qiu & Wenbo Deng leg.

Etymology. This species epithet is derived from the Latin word ‘clavellatus’ referring to the terminal of R2d being produced in a stick shape.

Description. Male. Body length 22.8 mm; overall length including tegmina 29.0 mm; pronotum length × width 4.4 × 7.1 mm. Female. Body length 23.2 mm; overall length including tegmina 29.3 mm; pronotum length × width 5.5 × 8.1 mm.

Coloration: body generally black. Interocular space with a white transverse band, distal half of clypeus white. Pronotum white, margins black outlined; center of the pronotum with symmetrical blackish markings (Figs. 6A and 6B). Tegmen reddish brown, wings hyaline, blackish brown except the anal area (Figs. 7G and 7H). Tibiae and tarsi reddish brown (Fig. 6B).

Head with vertex unsheltered by pronotum. Interocular distance slightly narrower than the distance between antennae sockets. Pronotum nearly hexagonal, broadest on the basal half but near the middle, the anterior pattern connected with lateral pattern, the lateral pattern bold, the vertical pattern connected with posterior pattern, the posterior band bold and with two spots on the band (since the pronotum of the holotype male is deformed, thus this description is only based on the female paratype) (Figs. 6A–6H). Male supra-anal plate broad and symmetrical, nearly trapezoidal, but with a shallow, obtuse-angled invagination at the center of hind margin; male subgenital plate asymmetrical, left hind corner more protruded than that of the right, styli similar, thin (Figs. 8J and 8K).

Male genitalia: Left phallomere: L1 broad and lamellar; margin of L2d with small and dense spines, posterior of L2v produced as an acerose spiny projection to the end; L3 with a spiny projection, the basal broad, tapering down, the terminal bifurcate (Fig. 8L). Right phallomere: R1 foot-shaped, posterior with two spines; terminal of R2d clubbed, R2v fold sclerite and the end with a spine; R3 with broad sclerite (Fig. 8L).

Remarks. H . clavellata sp. nov. is similar to H. arcifera, but can be distinguished by the following characters: (1) different irregular maculae on the pronotum, large in H. clavellata sp. nov., while H. arcifera, small; (2) face and body black in H. clavellata sp. nov., while in H. arcifera, yellowish white. Besides, the pronotum of the male holotype is deformed with posterior and anterior margins deeply concave. We utilized DNA barcoding to pair this species, and the genetic distance between them was 0.00%.

Material examined. 2 females (SWUB129-1, SWUB129-2), CHINA: Yunnan: Yaonan Village, Mt Ailao, Xinping County, 2018.V.24, Lu Qiu & Zhiwei Dong leg.

Description. Female. Body length 22.8–23.5 mm; overall length including tegmen 29.0–30.2 mm; pronotum length × width 4.4–5.0 × 6.9–7.1 mm.

Coloration: Body generally black. Interocular space with a white transverse band. Pronotum white, margins with black outline, hind margin black; center of the pronotum with symmetrical blackish markings. Tegmina brown, wings hyaline. Abdomen brown. Tibiae and tarsi yellow brown.

Head with vertex unsheltered by pronotum. Interocular distance slightly narrower than the distance between antennae sockets. Pronotum nearly hexagonal, broadest near the middle; the anterior pattern polygonal, the lateral pattern with two patterns, the central pattern connected with vertical pattern, the vertical pattern bold and the posterior bifurcated, the posterior band bold. Supra-anal plate broad, symmetrical and with ridges, cerci with obvious segmentation. Subgenital plate symmetrical, divided in two valves.

Natural History. Individuals were captured during night on the village roads (Lu Qiu, personal observation).

Diagnosis. Body small. The diagnoses are similar to Homalosilpha except the outer edge of middle and hind tibiae with two rows of spines (Fig. 4) and the margin of L2d with bigger serration (Fig. 8).

Material examined. 11 males, 7 females (SWUB130-1, SWUB130-2…SWUB130-17, SWU130-18), CHINA: Yunnan: Dadugang, Jinghong City, Xishuangbanna, 2014.IV.28-29, Xinran Li & Hongguang Liu leg; 1 male (SWU130B-19), CHINA: Yunnan: Jinghong City, Xishuangbanna, 545 m, 1974.V.14-15, Io Chou & Feng Yuan leg.

Redescription. Male. Body length 18.2–20.5 mm; overall length including tegmen 22.6–24.9 mm; pronotum length × width 3.7–4.6× 5.4–6.3 mm. Female. Body length 19.2–22.4 mm; overall length including tegmen 21.3–25.7 mm; pronotum length × width 4.3–4.7 × 5.7–6.5 mm.

Coloration. Similar in both sexes. Body dark brown (Figs. 6M and 6N). Pronotum white, with a large central black spot on middle disk, margins with black outline, hind margin black (Figs. 6O and 6P). Tegmen dark brown, wing hyaline, brown except the anal area. Abdomen brown to dark brown. Tibiae and tarsi brown (Fig. 6N).

Head with vertex unsheltered by pronotum. Interocular distance narrower than the distance between antennae sockets. Pronotum nearly hexagonal, broadest near the middle, large black spot connected to the hind margin. Legs with sparse spines. Supra-anal plate of male broad and symmetrical, nearly trapezoidal, posterior margin widely concave, the lateral margin concave, both lateral hind angles sharp; male subgenital plate asymmetrical, left hind corner round, hind margin wavy (Figs. 8P and 8Q).

Male genitalia: Left phallomere: L1, L2 and L3. L1 lamellar; margin of L2d with small spines, posterior of L2v produced a spiny projection to the end; L3 with a spiny projection, the basal broad, tapering down, the terminal bifurcated (Fig. 8R). Right phallomere: R1, R2 and R3. R1 foot-shaped, posterior with two spines; terminal of R2d unciform, R2v fold sclerite; R3 simple and broad (Fig. 8R).