Acta Arachnologica, 67(1): 23–29, August 31, 2018 Redescription of Chrysilla lauta Thorell 1887 (Araneae: Salticidae) based on the comparison with the holotype, and DNA barcoding Takeshi Yamasaki1*, Marika Yamaguchi1, Luong Thi Hong Phung2, Pao-Shen Huang3 & I-Min Tso3 1 Department of Biological Sciences, Tokyo Metropolitan University, 1–1 Minami-osawa, Hachioji-shi, Tokyo 192–0397, Japan 2 Institute of Ecology and Biological Resources, Vietnam Academy of Science and Technology, Vietnam 3 Department of Life Science, Tunghai University, Taichung, Taiwan *Corresponding author. E-mail: [email protected] Abstract ― Chrysilla lauta Thorell 1887 is the type species of the genus Chrysilla. Zhang & Wang (2017) formally recorded the female of C. lauta for the first time, but it does not include the description of important characters in spider taxonomy such as the palp of males and the epigyne of females. We here redescribe C. lauta in detail, on the basis of the comparison with the holotype, and DNA barcoding using mitochondrial CO1 fragment. Key words ― Chrysillini, taxonomy, mitochondrial DNA, Southeast Asia (THU), and Dr. Booppa Petcharad’s personal collection in Introduction the Thammasat University, Thailand (BP-TU) respectively. The genus Chrysilla Thorell 1887 (Araneae: Salticidae) Their morphology was examined using a Nikon SMZ1270 is currently comprised of ten species, and widely distributed microscope. Multi-focused montage images were produced through South and Southeast Asia (World Spider Catalog using Helicon Focus ver. 4.2.9 from several series of source 2017). The species of Chrysilla show distinct sexual di- images. The source images were obtained by a Canon EOS morphism in proportion of body parts and coloration in the 60D camera attached to a Nikon SMZ1270. adult stage. Therefore, the recognition of conspecific male Specimens used in DNA barcoding are shown in Table and female based on only morphology more or less includes 1. For the methodology of molecular analysis, we followed uncertainty. Recent works based on DNA barcoding have Phung et al. (2016) except PCR experiments. PCR exper- revealed male/female combination in salticids showing dis- iments, with primer combination of CO1-TY-F1 (5’-TGC tinct sexual dimorphism, and subsequently it has been solv- WAT TTT TTC TTT RCA TTT RGC-3’) and CO1-TY-R1 ing synonymies (Suguro & Yahata 2014; Phung et al. 2016). (5’-GCH ACH ACA TAA TAA GTA TCA TG-3’), included The type species of this genus, Chrysilla lauta Thorell an initial 2-min denaturation at 94°C followed by 5 cycles 1887, has been described on the basis of a male specimen of 10 s at 98°C, 30 s at 45°C and 45 s at 68°C, and then 40 from Myanmar. The female has been recorded very recent- cycles of 10 s at 98°C, 30 s at 48°C and 45 s at 68°C, with a ly in Zhang & Wang (2017), however, it does not include final 7-min extension at 68 °C. The sequences obtained were descriptions of taxonomically important characters such as assembled using ChromasPro 1.7.6 (Technelysium Pty Ltd., the palp of males and the epigyne of females. Therefore, the Australia). These sequences, in addition to the homologue detailed description of the both sexes is still necessary. In sequences of Cosmophasis micarioides (L. Koch 1880) the present study, we redescribe C. lauta with detailed mor- (Accesion No.: EU815580) and Phintella versicolor (C. L. phological descriptions based on the holotype and confirm Koch 1846) (Accession No.: LC105656) provided by other the correct male/female combination by DNA barcoding. authors (Maddison et al. 2008; Phung et al. 2016), were aligned using MUSCLE (Edgar 2004) built in MEGA 6.06 Materials and methods (Tamura et al. 2013). The genetic divergence in the K2P The holotype male of Chrysilla lauta was loaned from model (Kimura 1980) was calculated by the pairwise com- the Museo Civico di Storia Naturale “Giacomo Doria”, parison method, and the neighbor-joining tree was construct- Genoa, Italy (MSNG). Additional specimens collected ed using MEGA 6.06. The mitochondrial CO1 sequences in Taiwan and Thailand are deposited in the Department obtained in the present study are deposited in the DNA Data of Life Sciences, Tunghai University, Taichung, Taiwan Bank of Japan (DDBJ). 24 T. Yamasaki, M. Yamaguchi, L. T. H. Phung, P.-S. Huang & I-M. Tso Table 1. Specimens used in the present study. Species Sample code Sex Locality Accession No. Tunghai University, Taic- Chrysilla lauta TW_SAL_20170902B_4 Female LC328902 hung, Taiwan Tunghai University, Taic- Chrysilla lauta TW_SAL_20170902B_5 Female LC328903 hung, Taiwan Forest Recreation Area, Chrysilla lauta TW_SAL_20170906_1 Male Kending National Park, LC328904 Pingtung, Taiwan Tunghai University, Taic- Chrysilla lauta TW_SAL_20170907_1 Male LC328905 hung, Taiwan Tunghai University, Taic- Chrysilla lauta TW_SAL_20170907_2 Male LC328906 hung, Taiwan Nha Ngio, Nakhon Si, Chrysilla lauta TSG_20131009 Male LC328907 Thammarat, Thailand Table 2. Pairwise distances in K2P model (shown below the diagonal) and standard errors (shown above the diagonal). Sample 1 2 3 4 5 6 7 8 1 TW_SAL_20170902B_4 0.000 0.000 0.000 0.005 0.002 0.017 0.018 2 TW_SAL_20170902B_5 0.000 0.000 0.000 0.005 0.002 0.017 0.018 3 TW_SAL_20170906_1 0.000 0.000 0.000 0.005 0.002 0.017 0.018 4 TW_SAL_20170907_1 0.000 0.000 0.000 0.005 0.002 0.017 0.018 5 TW_SAL_20170907_2 0.015 0.015 0.015 0.015 0.005 0.018 0.018 6 TSG_20131009 0.002 0.002 0.002 0.002 0.017 0.016 0.017 7 EU815580 0.145 0.145 0.145 0.145 0.158 0.143 0.017 8 LC105656 0.174 0.174 0.174 0.174 0.182 0.172 0.141 Fig. 1. Neighboor-joining tree generated under K2P distance model. Bootstrap values (1000 replicates) are shown beside nodes. All measurements are given in millimeters. Abbreviations Results and discussion of morphological terms used in the present paper are as fol- lows: ALE, anterior lateral eye; AME, anterior median eye; The genetic divergence among specimens is shown in PLE, posterior lateral eye; PME, posterior median eye; RTA, Table 2, and the constructed neighbor-joining tree is shown retrolateral tibial apophysis on palp. in Fig. 1. Females of C. lauta (TW_SAL_20170902B_4, TW_SAL_20170902B_5) made a cluster with males (TW_ SAL_20170906_1, TW_SAL_20170907_1) identified as Acta Arachnologica, 67(1), August 2018 ⒸArachnological Society of Japan Redescription of Chrysilla lauta 25 Figs. 2–5. Chrysilla lauta, holotype male. 2, habitus, dorsal view; 3, habitus, ventral view; 4, habitus, lateral view; 5, labels. Scales = 1 mm. Figs. 6–8. Chrysilla lauta, holotype male. 6, left palp, ventral view; 7, left palp, retrolateral view; 8, left palp, dorsal view. Scales = 0.5 mm. Acta Arachnologica, 67(1), August 2018 ⒸArachnological Society of Japan 26 T. Yamasaki, M. Yamaguchi, L. T. H. Phung, P.-S. Huang & I-M. Tso Figs. 9–12. Chrysilla lauta, male (TW_SAL_20170906_1). 9, habitus in living condition, dorsal view; 10, habitus, dorsal view; 11, habitus, lateral view; 12, habitus, ventral view. Scales = 1 mm. Figs. 13–15. Chrysilla lauta, male (TW_SAL_20170906_1). 13, left palp, ventral view; 14, left palp, retrolateral view; 15, left palp, dorsal view. Scales = 0.5 mm. C. lauta on the basis of the holotype, without divergence. ing to the locality was not constructed in the present study. From this result, we confirmed the male/female combina- A specimen collected in Tunghai University, Taiwan (TW_ tion proposed in Zhang & Wang (2017) (Fig.1). Although SAL_20170907_2) presented the divergence value of 1.5% the cluster of C. lauta comprised of specimens from Taiwan in K2P model from other specimens collected in the same and Thailand is highly supported, each cluster correspond- locality (Table 2). However, no distinct morphological dif- Acta Arachnologica, 67(1), August 2018 ⒸArachnological Society of Japan Redescription of Chrysilla lauta 27 Figs. 16–19. Chrysilla lauta, female (TW_SAL_20170902B_4). 16, habitus in living condition, dorsal view; 17, habitus, dor- sal view; 18, habitus, lateral view; 19, habitus, ventral view. Scales = 1 mm. ferences were recognized among them. In addition, the mean 9-X-2013, B. Petcharad & T. Suguro leg. TAIWAN: 1 male intraspecific divergence value is considered to be 2.15 % in (THU; TW_SAL_20170906_1), Forest Recreation Area, spiders (Robinson et al. 2009). Therefore, we concluded the Kending National Park, Pingtung; 2 females (THU; TW_ divergence value of 0–1.5 % among our specimens were the SAL_20170902B_4, TW_SAL_20170902B_5), Tunghai intraspecific divergence. University, Taichung, 2-IX-2017, T. Yamasaki leg.; 2 males (THU; TW_SAL_20170907_1, TW_SAL_20170907_2), Taxonomy same locality, 7-IX-2017, T. Yamasaki leg. Chrysilla Thorell 1887 Diagnosis. Males are recognizable by slender body with iridescent bands of scale-like setae (Figs. 9–10); strong leg Remarks. The delimitation of the genus Chrysilla has not I compared with other legs; slender embolus with weakly been defined properly yet. Currently, Chrysilla is comprised curving; tegulum lobe weakly developed and reaching to an- of ten nominal species. However, seven species of them are terior margin of palpal tibia; RTA extending antero-ventrally known only on the basis of one sex (only female: 4 species; with slight curing inward. Females are easily recognizable only male: 3 species). To establish correctly male/female by characteristic markings on abdomen (Figs. 16–17); cop- combination, both morphology and molecular approaches ulatory duct running longitudinally without curving; round are necessary. spermatheca. Measurements (male / female; measurements of the ho- Chrysilla lauta Thorell 1887 lotype in parentheses). Carapace length 1.47–2.10 (1.70) / (Figs. 2–4, 6–24) 1.77–1.82; width 1.08–1.52 (1.30) / 1.20–1.26. ALE–PLE 0.78–1.02 (0.85) / 0.88; ALE–PLE 0.41–0.52 (0.45) / 0.48. Chrysilla lauta Thorell 1887: 378; Prószyński 1976: 154, f. 237; Width of eye row I 1.03–1.45 (1.23) / 1.23–1.25; II 0.97– Prószyński 1983: 44, f 4–6; Żabka 1985: 210, f. 81–82; Song & 1.35 (1.13) / 1.15–1.17; III 1.12–1.51 (1.27) / 1.30–1.35. Chai 1991: 14, f. 2A–B; Song, Zhu & Chen 1999: 507, f. 290N–O; Abdomen length 2.10–3.50 (2.63) / 2.80; width 0.85–1.13 Prószyński & Deeleman-Reinhold 2010: 159, f. 36–37; Zhang & (not measured for the holotype) / 1.28–1.40. Wang 2017: 601, with unlabeled 5 pictures. Cosmophasis longiventris Simon 1903: 732 (synonymized by Żabka Male (Figs. 2–4, 6–15). Carapace shorter and wider than 1985) abdomen (Figs. 2, 9–10). Abdomen slender; almost 3 times longer than wide (Figs. 2, 9–10). Leg I strong compared Type material examined. MYANMAR: Holotype male with other legs. (MSNG), Bhamo, Birmania [Myanmar], 1885, L. Fea. (Label Palp (Figs. 6–8, 13–15). Cymbium strongly tapering to- data is shown in Fig. 5.) ward apex. Bulb anteriorly divided into two portions as with Other material examined. THAILAND: 1 male (BP- other Chrysilla species, slender prolateral and retrolateral TU; TSG_20131009), Nha Ngio, Nakhon Si, Thammarat, portions (Figs. 6, 13); posterior lobe of bulb roundly devel- Acta Arachnologica, 67(1), August 2018 ⒸArachnological Society of Japan 28 T. Yamasaki, M. Yamaguchi, L. T. H. Phung, P.-S. Huang & I-M. Tso Figs. 20–24. Chrysilla lauta, female (TW_SAL_20170902B_4). 20, epigyne, ventral view; 21, 23, clarified epigyne, ventral view; 22, 24, clarified epigyne, dorsal view. Scales = 0.2 mm (20–22); 0.25 mm (23–24). oped, slightly overlapping anterior venter of palpal tibia in pace dorsally covered with white setae and laterally reddish ventral view and projecting ventrally in lateral view (Figs. setae (Figs. 17–18). Abdomen covered with white, black and 6–7, 13–14). Seminal duct beginning from inner margin of reddish setae, and showing complex pattern on dorsum (Figs. retrolateral bulb with strong curving, and running along mar- 17–18). Legs cream. Coloration in living condition shown in gin of bulb toward base of embolus (Figs. 6, 13). Embolus Fig. 16. extending anteriorly from apex of bulb, with slight curve at Distribution. Taiwan (new record), China, Vietnam, tip (Figs. 6, 13). RTA spine-shaped with weakly incurving, Thailand (new record), Myanmar. extending antero-ventrally in retrolateral view (Figs. 6–8, Remarks. Among Chrysilla species, C. volupe (Karsch 13–15). 1879) is very close to C. lauta. Although they are very sim- Coloration and setation in ethanol (Figs. 2–4, 10–12). ilar in the structure of male palp, the coloration in both spe- Carapace densely covered with reddish setae, but sparsely in cies is different from each other (cf. figs. 122–125 in Żabka small specimens; two transversal bands of iridescent scale- 1988; figs. 15–23 in Caleb & Mathai 2014). like setae running on eye row I and III in dorsal view (Fig. Acknowledgements 10); two longitudinal bands of iridescent scale-like setae running on each lateral surface, wider one below ALE to We would like to thank Dr. Maria Tavano (MSNG, Italy) for loan- PLE and narrower one lateral margin of carapace in later- ing the type material, Dr. Noriaki Murakami & Dr. Katsuyuki Eguchi (both, Tokyo Metropolitan University, Japan) for offering laboratory al view (Fig. 11). Abdomen covered with black setae; one equipment, Dr. Booppa Petcharat (Thammasat University, Thailand) longitudinal band of iridescent scale-like setae running me- & Mr. Tatsumi Suguro (Keio Yochisha Elementary School, Japan) for dially on dorsum, and another band extending from anterior offering a valuable specimen and two anonymous referees for their dorsum on each lateral surface (Figs. 10–11). Leg I tinged critical readings of this manuscript. The field works in the Kending National Park were conducted under MOST 106-2621-B-029-004. with black on femur, tibia and metatarsus. Legs II–IV cream. Two of the authors, M. Yamaguchi & P.T.H. Luong, are supported by Coloration in living condition shown in Fig. 9. 25th Fujiwara Natural History Foundation (2017). Female (Figs. 16–24). Carapace longer, and slightly nar- References rower than abdomen (Figs. 16–18). Abdomen oval (Figs. 16–17). Caleb, J. T. D. & Mathai, M. T. 2014. Description of some interesting Epigyne (Figs. 20–24). 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