A peer-reviewed open-access journal ZooKeys | 112: 161-178 (2022) SL ae RET #ZooKeys https:/ / ZOO keys. pensoft.net Launched to accelerate biodiversity research New species of the genus Molanna Curtis, 1834 (Trichoptera, Molannidae) in China inferred from morphology and DNA barcodes Xin-yu Ge', Lang Peng’, Jie Du’, Chang-hai Sun’, Bei-xin Wang' | Department of Entomology, College of Plant Protection, Nanjing Agricultural University, 210095, Nanjing, China 2 Jiuzhaigou Administration Bureau, 623402, Jiuzhaigou County, Aba Prefecture, Sichuan Province, China Corresponding authors: Chang-hai Sun (chsun@njau.edu.cn), Bei-xin Wang (wangbeixin@njau.edu.cn) Academic editor: Ana Previsi¢ | Received 30 March 2022 | Accepted 16 June 2022 | Published 14 July 2022 https://zoobank. org/2923FC2A-DA45-46 D2-BB8 B-F3B32BBD0448 Citation: Ge X-y, Peng L, Du J, Sun C-h, Wang B-x (2022) New species of the genus Molanna Curtis, 1834 (Trichoptera, Molannidae) in China inferred from morphology and DNA barcodes. ZooKeys 1112: 161-178. https:// doi.org/10.3897/zookeys.1112.84475 Abstract The male adult of Molanna truncata Ge, Peng & Sun sp. nov. is described and illustrated based on material col- lected in Si-chuan, China. It could be diagnosed by the subtriangular superior appendages when viewed dorsal- ly, and by the mesal appendages each having a slender thorn and inferior appendages with a tiny inner process. Based on morphology of genitalia, we provide a dichotomous key to adult males of Molanna from the Oriental region. The DNA barcodes (partial mt COI sequences) of M/. truncata sp. nov. are generated and compared with existing sequences of Molanna species from Oriental and Palearctic regions. The mean intraspecific diver- gence of Molanna was 1.58% with a maximum of 8.50% in M. moesta. The Automatic Barcode Gap Discovery (ABGD) analysis of Molanna inferred 9 OTUs and thresholds of interspecific divergence of 10%. Divergence of M. truncata sp. nov. haplotypes from all other Molanna haplotypes ranged from 10.1% to 18%. We discuss distribution and potential groups of species within the Oriental Molanna species based on morphology. Keywords Caddisflies, COI sequence, integrative taxonomy, morphology, Oriental region, wing venation Introduction Molannidae Wallengren, 1891 is a small family of Holarctic and Oriental biogeo- graphic regions, with two genera, Molanna Curtis, 1834 and Molannodes McLachlan, Copyright Xin-yu Ge et al. 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. 162 Xin-yu Ge et al. / ZooKeys 1112: 161-178 (2022) 1866 (Morse 2021). Curtis (1834) erected the genus Molanna with Molanna angustata Curtis, 1834 as the type species by original designation. At present, the genus includes 25 extant species and three fossil species worldwide (Morse 2021). Among them, 12 species were reported from the Oriental region, six were restricted to the Nearctic re- gion, four from the Palaearctic region and two from the East Palaearctic region; only M. moesta Banks, 1906 occurs in both the East Palaearctic and Oriental regions (Banks 1906; Malicky and Chantaramongkol 1989; Nebois 1993; Malicky 1994; Ito 2006). At present, three species were reported from China, among them, MZ. moesta, distribut- ed from northeast to southwest in China (i.e. Hei-long-jiang, Jiang-xi, Guang-dong, Si- chuan, Gui-zhou and Yun-nan), while 1. kunmingensis Hwang, 1957 and M. xiaguana Malicky, 1994 are reported from the Yunnan Province exclusively (Yang et al. 2016). Adults of Molanna live around lakes or running waters and are easy to recognize because in repose the adults look like short branch segments (Schmid 1998). Larvae mostly occur in lakes or slower currents of streams, and inhabit sandy substrates (Wig- gins 1996). Larvae of Molanna flavicornis Banks, 1914 were found to inhabit the pro- fundal zone (up to 20 m deep; Neave 1933). DNA barcodes, the 658 bp fragment of the mitochondrial gene cytochrome c oxidase I unit (CO]), has provided important evidence to confirm new species and has proved to be useful for association between larvae and adults (Hebert et al. 2003; Zhou et al. 2007; Xu et al. 2018; Hu et al. 2019; Ge et al. 2020). But existing DNA barcodes of Molanna species from China are very few, and only one barcode has, to date, been recorded in Barcode of Life Data Systems (BOLD) (Ratnasingham and Hebert 2013). This lack of barcode resources greatly restricts accurate monitoring of Molanna species for metabarcoding of environmental DNA (Lin et al. 2021). In this study, we describe a new Chinese species of Molanna and provide DNA bar- codes of this species. The variation in male hind wing venation and DNA barcodes among species from the Palaearctic and Oriental regions are discussed. Finally, a key and map with distribution to the known adults of Mo/anna from the Oriental region are given. Materials and methods Sample collection Adult specimens were collected into 95% alcohol using pan traps with 15-W ultraviolet light tubes in the Jiu-zhai-gou County, Si-chuan Province, PR China, during July. The specimens were then sorted and stored in 95% alcohol at -20 °C temperature. Morphological study The methods used for preparation of genitalia followed Peng et al. (2020). For wing venation studies, Right wings were cut from the body, mounted in a microscope slide with glycerin and covered with a coverslip to ensure that the wings were fully flattened. A new species of genus Molanna in China 163 Wing and genitalia structures were traced in pencil using a Nikon Eclipse 80i mi- croscope and an Olympus SZX10 stereomicroscope equipped with a camera lucida. Pencil drawings were scanned with an Epson Perfection (V30 SE) scanner, then placed as templates in Adobe Photoshop (Version: 13.0) and inked digitally to produce illus- trations. The illustrations were then arranged using Adobe Photoshop (version 13.0). Genitalia, wings and the remainder of each specimen were stored in a microvial in 95% alcohol. Type and voucher specimens were deposited at the Insect Collection, Nanjing Agricultural University (NJAU), Nanjing, Jiangsu Province, PR China. Terminology The terminology for male genitalia follows those of Wiggins (1968) and Ito (2006). The terminology for wing venation follows that of Schmid (1998). DNA analysis The right hindleg of two adults was removed for genomic DNA extractions. DNA extraction, PCR amplification, fragment sequencing, and analysis followed the pro- cedures of Xu et al. (2018). The universal primers LCO1490 and HCO2198 (Folmer et al. 1994) were used to amplify the 658 bp fragment of the mitochondrial (mt) cytochrome c oxidase I unit (COI). Raw sequences were assembled and edited in Se- quencher 4.5 (Gene Codes Corporation, Ann Arbor, Michigan, USA). Neighbor-join- ing (NJ) tree of eight species within the genus Mo/anna was constructed using MegaX (Kumar et al. 2018), with the following parameters: Kimura 2-parameter substitution model (K2P), pairwise gap deletion, and others as defaults. The same software was used to calculate the K2P corrected p-distance of the 658 bp COI fragment among all Molanna sequences available (Table 1). COI sequences of Molanna species were also applied to the Automatic Barcode Gap Discovery (ABGD) tool (Puillandre et al. 2012) to compare the operational taxonomic unit (OUT) number with the Barcode Index Numbers (BINs). COI sequences of new species were uploaded to GenBank. Accession numbers of the analyzed Molanna specimens are shown in Table 1. Results Taxonomy Molanna truncata Ge, Peng & Sun, sp. nov. https://zoobank.org/82EBA03B-56EC-42F7-83E4-3CD0F8D4D3BF Fig. LA-F Type material. Holotype: 13, PR. Cutna, Si-chuan Province, Aba Prefecture, Jiu-zhai- gou County, Jiu-zhaigou National Nature Reserve, Xi-niu-hai (Fig. 2), 33°11'42"N, 164 Xin-yu Ge et al. / ZooKeys 1112: 161-178 (2022) 103°53'46"E, alt. 2348 m, 7 VII 2019, leg. X.Y. Ge & Y. Wang (NJAU). Paratypes: 2 4, same data as holotype (NJAU). Other specimens. 1d PR. Cura, Si-chuan Province, Aba Prefecture, Jiu- zhaigou County, Jiu-zhaigou National Nature Reserve, Wu-hua-hai, 33°9'32"N, 103°52'55"E, alt. 2377 m, 20 VII 2014, leg. Y. Cao (NJAU). 24 PR. China, Si- chuan Province, Aba Prefecture, Jiu-zhaigou County, Jiu-zhaigou National Nature Reserve, Lu-wei-hai, 33°13'18"N, 103°54'48"E, alt. 2299 m, 10 VII 2019, 19 VII 2014 lee -CO¥sOin NIA), Diagnosis. The new species is similar to M. yaeyamensis Ito 2006 from Japan (Ishigaki and Iriomote islands), but can be differentiated by the following characters: (1) superior appendages in MV. truncata sp. nov. in dorsal view have subrhomboid shape and basally bearing a slender thick seta, whereas M. yaeyamensis have subsquare superior appendages with no such thick setae in dorsal view; (2) in MZ. truncata sp. nov. mesal appendages have a subrectangular shape in lateral view, whereas in MZ. yaeyamensis mesal appendages have an ellipsoidal shape in lateral view; (3) inferior appendages with tiny triangular inner processes in M. truncata sp. nov., compared to long digitate inner processes (about 2/3 as long as dorsal process) in M. yaeyamensis, and 4) ventral processes of inferior appendages in M. truncata sp. nov. in ventral view are somewhat stub-like and the lateral margin has a distally bristled, tiny process, which is both absent in M. yaeyamensis. Description. Specimens in alcohol with compound eyes black, thorax, abdomen and legs black to grayish white, without patterns. Body medium-sized, length 7.3— 7.7 mm (NV = 3). Head 0.95 mm long, about 1.75 times wider than length, ocelli absent. Front of vertex with subquadrate setal wart, posterolateral portion with two pairs of setal warts. Pronotum nearly trapezoidal, Pronotum anterior margin slightly sinuous, slightly concave anteromesad, posterior margin slightly concave, with one pair of setal warts. Male genitalia: Abdominal segment IX in lateral view (Fig. 1A), irregularly pentag- onal, convex anteriorly and posteriorly. Superior appendages in lateral view (Fig. 1A), trapezoidal, covered with many long and short setae, posterior margin straight; in dorsal view (Fig. 1B), subrhomboid, posterior margin irregularly serrated, each with slender thick setae at ventromesal base. In lateral view (Fig. 1A) mesal appendages sub- rectangular shape with narrow base and wider distal part, distal end slightly produced ventrad, with each side having 3—4 thick long setae, setose apically; in dorsal view, somewhat tubular, with base slightly thickened, with longitudinal membranous lobe mesally from base to apex. In lateral view, inferior appendages one-segmented, slightly shorter than mesal appendages, each divided into dorsal and ventral processes exter- nally, and with a tiny triangular inner process (Fig. 1B); dorsal processes each with apex curved upwards in lateral view (Fig. 1B), sparsely setose, apical and lateral margins each sinuate, inner margins each arc-shaped in dorsal view; ventral processes in lateral view halfway shorter than dorsal processes, in ventral view (Fig. 1C) somewhat stub-like, lateral margin with distally bristled, tiny process. Phallus arched, with one pair of thin, long sclerites on apical half of dorsal surface (Fig. 1D). Male wings: Forewings (Fig. 1E): Venation fairly complete, typical for the genus, without obvious marks. R, very short jointed with R,, M 3-branched. M and Cu, A new species of genus Molanna in China 165 mem lob thick setae Figure |. Molanna truncata sp. nov. Male adult, holotype A genitalia, lateral view B genitalia, dorsal view C genitalia, ventral view D phallus, lateral view E forewing F hind wing. Abbreviations: sup app, superior appendage. mes app, mesal appendage; mem lob, membranous lobe; dor pro, ven pro, inn pro, dorsal, ventral, and inner processes of inferior appendage, respectively; pha, phallus. Sc, Subcosta; R, Radius; M, Media; Cu, Cunitus; A, Anal. Scale bars: 200 pm. 166 Xin-yu Ge et al. / ZooKeys 1112: 161-178 (2022) fused at very base, Cu, not extended to margin, 1A and 2A merged at base and running to posterior margin and then curved anteriad to Cu,,. Hind wings (Fig. 1F): Venation of male hind wing much reduced, with band of dark setae running near midline. Etymology. The Latin adjective truncatus, -a, um refers to the truncate shape of the superior appendages posterior margin in lateral view. Distribution. China (Si-chuan). Key to the adult males of Molanna Curtis, 1834 from Oriental region 1 In lateral view, mesal appendages distal end furcated...... ice eeeeeeeeeeeeeeeee 2 = In lateral view, mesal appendages distal end unfurcated...... eee eee esos 5 2, Distal end of mesal appendages dorsum with 3 large spines ........ceeeeeeeeees Ee oR E EAE ry Peay Carr tec ee Be rr RAE YT es VET nD M. gamdaha (Fig. 3A) Distal end of mesal appendages dorsum without 3 large spines.............00 3 3 In lateral view, superior appendages triangular, with posterior margins con- GA re centessse dag cen cde deen a sev Os Gus Poncen eevee duh vetedueee M. crinitaa (Fig. 3B) In lateral view, superior appendages trapezoidal 0... eee ceeeeseeseeeeeeeecseeeee A 4 In lateral view, mesal appendages with upper and lower lobes pointing to and neatly contacting Gach other: cco ticespernsecenntaes teen M. saetigera (Fig. 3C) In lateral view, mesal appendages with upper and lower lobes divided widely Bry pe Ba Ped roc IRA na er De eProp M. oglamar (Fig. 3D) 5 Inferior appendages without ventral processes .........sssssesseseceeeeeseeeeeeseeeeees 6 — Inferior appendages withevertral processes, sis.uc.3rsceboatalhtetuaubhnekocs eoncdhuand 10 6 Superior appendages in lateral view leaf-shaped or trapezoid... eee 7, -. Superior appendages in lateral view finger-shaped ........ cess eseeseeeeeeeeeeeeee 9 7 Superior appendages in lateral view, leaf shape... M. kunmingensis (Fig. 3E) — Superior appendages in lateral view, trapezoid shape ....... sce eeeseeseeeeeeeeeee 8 8 In lateral view, mesal appendages finger-like... M. moesta (Fig. 3F) = In lateral view, mesal appendages inflated, hammer-like..... eee UM aie ees cte Meth seg Macend.s Nace Maye lead: Bulan Mayetnsde: bel olen sne BRS M. paramoesta (Fig. 3G) 9 In lateral view, mesal appendages tapering from base to apex, with distal half curved downwards at 90 degree oo... ee eeeeeeeeseeseeeee M. taprobane (Fig. 3H) - In lateral view, mesal appendages not as above........... M. xiaguana (Fig. 31) 10 Inferior appendages without inner Processes .........cseeseeseeseceeeeeseeeseeaeeeees i — Inferiorappendages: with inner Processes’ vo. cccoseeveccouvornrscesqorsvwsecgernvecsecseace 12 11 Ventral processes of inferior appendage with thorn distally wo... eee B svapentushorcetemse justi vent aoe w aahaue eras wad aacopananekn neve M. jolandae (Fig. 3J) - Ventral processes of inferior appendage without thorn distally... SR cere 233k rok sir Deen ar LLM nee RE Ae Ae fe M. cupripennis (Fig. 3K) 12 Inferior appendages with tiny triangular inner processes .........sceseseseeseeees ARE re oa Ee AAR ei ali Soa i dead «tee M. truncata sp. nov. 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ABMION Aueulian AUBULIOD) AayUNod DISOOU ‘WP DISOOU PW ‘Aou ‘ds pJvoUNAL We ‘Aou ‘ds DJvoUNAL WW pIsaoul WW pIsaoul “Wy pISaoul “Wy SUuDIIG|D ‘W SUDIIGID ‘We supdIg|0 W DSOALaU J DSOALAU JY pIsaoul We pIsaoul “JW pIsaoul We ‘ds'7x W SUDIIGID “We subdIg]0 ‘W SUuDIIg|O W subaig|o ‘W supdIg]0 ‘W DASU DAS A DASIU DASIU ‘We ‘ds w ADUD]8O JW DIDISNSUD PW DIDISNSUD ‘PW DIVISNSUD PW DIVISNBUD DIVISNSUD sored ‘ponunuo’) *| aque 170 Xin-yu Ge et al. / ZooKeys 1112: 161-178 (2022) Figure 2. Type locality of Molanna truncata sp. nov., Xi-niu-hai in the Jiu-zhaigou National Nature Reserve, China. 70° E 80° E 90° E 100°E 110°E 120°E 130°E 140°E 30°N 20°N 20°N 10°N 10°N Q° A M. gamdaha H oO M. xiaguana A M. erinitara I oO M. taprobane 0° Ly M. saetigera J O M. jolandae Ay M. oglamar K (@) M. cupripennis HM. kummingensisi L * M. truncata sp.nov. - 1) M. moesta 10°S WM. paramoesta =M * M. yaeyamensis 10°S 0 195 390 780 1,170 1,560, Miles 80° E 90° E 100° E 110°E 120°E Figure 3. Distribution of Molanna species in the Oriental region. A new species of genus Molanna in China 171 DNA barcodes analyses The aligned 32 sequences ranged from 634 to 658 base pairs, including 29 sequences with a full barcode length of 658 base pairs. As some morphospecies showed comparably high intraspecific divergence, no definite “barcode gap” was observed based on pairwise distance (Fig. 4A). The ABGD analysis of the genus Molanna recognized 9 OTUs with a prior intraspecific divergence of P_ = 3.59% (Fig. 4B). The NJ tree based on 32 COI Molanna haplotypes conducted in the current study does not provide clear information on species differentiation, except for . angustata and M. albicans (Zetterstedt, 1840) (Fig. 5), both distributed in the Palaearctic region. Even though each of the species was clustered in separate clades, division into clades is not supported by the presented NJ analysis (Fig. 5). However, the NJ analysis suggests relatively high differentiation in populations of M. moesta (Fig. 5). Two Molanna truncata sp. nov. haplotypes were clustered; however, their differentiation from the “Molanna XZ.sp.(KX102865)” haplotype is not supported by the NJ tree. The interspecific divergence (K2P p value) ranged from 10.1% to 19.5% (Table 1), with the mean divergence of 16.16%. The lowest intraspecific divergence was observed for haplotypes from European populations of M. angustata and M. nigra (Zetterstedt, 1840) and M. albicans, as well as Russian and Japanese populations of M. moesta (Table 1). Highest intraspecific divergence was observed in M. moesta (Table 1). The mean intraspecific divergence of all species was 1.58% with a maximum of 8.50% in M. moesta. In addition, one unnamed species VM. sp. (BOLD: AAP1029) was associated with MM. moesta clades. Divergence of M. truncata nov. sp. haplotypes from all other Molanna haplotypes ranged from 10.1% (from the “Molanna XZ.sp. haplotype) to 18% (from M. angustata, Table 1). = Recursive Partition » Initial Partition Frequency ak 2 2 So 5 = ¢ we 0100 "0 FIOO “0 6100 “0 Lz00 “0 2€00 “0 &L00 “0 O10 “O £6100 8920 "0 £2600 stso 0 i=) i) =] Prior intraspecific divergence COI divergence Figure 4. Histogram of pairwise K2P distances and number of the OTUs of 32 aligned sequences A the histogram was created using the K2P model in the Automatic Barcode Gap Discovery (ABGD) analysis. The horizontal axis shows the pairwise K2P-distance, and the vertical axis shows the number of pairwise sequence comparisons B the number of the OTUs by the prior intraspecific divergence calculated with the ABGD online-tool. 172 Xin-yu Ge et al. / ZooKeys 1112: 161-178 (2022) 99 5 Molannamoesta\|KX 107440 99 Molannamoesta\|KX 105642 Molanna moesta\|KX 103332 100 Molanna sp.|LC619232 100 Molannamoesta\|KX291805 100 Molanna moesta|\|KX295053 Molannamoesta|\|KX292654 100 pr Molanna moesta\HQ958937 Molannamoesta|K-X291103 Molannanigra|K-X291834 — Molanna nigra\|HM422029 Molanna nigra\KX 143012 Molannanigra\|KX291651 Molannaoglamar|KX295021 100 pm Molannanervosa|KX103890 Molannanervosa\|KX105001 Molanna XZ sp.|KX102865 100 5 Molanna truncata sp.nov. Molanna truncata sp.nov. Molanna angustata|KX293662 Molanna angustata\|KX104560 92 100 Molannaangustata\|GU713189 Molanna angustata|CKX29 1340 Molanna angustata|KX295460 85 99 | Molannaalbicans|KX103592 Molannaalbicans|KX105019 100 Molannaalbicans|KX103901 Molannaalbicans|KX 104075 7 | Molannaalbicans|KX106945 Molannaalbicans|KX106945 Molannaalbicans|KX 104075 Molannaalbicans|KX103901 0.020 Figure 5. Neighbor-joining cladogram based on the 658 bp long mtCOI sequence of eight Molanna species. Numbers on branches represent bootstrap support (>70%) based on 1000 replicates; scale equals K2P genetic distance. A new species of genus Molanna in China 173 Discussion In order to verify the description of Molanna truncata sp. nov., we compared the il- lustrations of male genitalia of all available species and wing venations of most species (Curtis 1834; Banks 1906; Hwang 1957; Wiggins 1968; Malicky and Chantaramong- kol 1989; Nebois 1993; Malicky 1994; Ito 1998, 2006; Olah and Johanson 2010). The male genitalia varied greatly among Molanna species in the superior appendages, the mesal appendages and in the inferior appendages. However, some species showed similarities in these characters, as well as in the hindwing venation. Considering the above-mentioned similarities and differences in morphology, we could argue for par- ticular groups of species within the Oriental Molanna species. The first group is characterised by the superior appendages with the height at least twice as long as its length or approximately equal to its length, and the media veins unbranched or occasionally 2-branched in hindwings. This group could be further di- vided into three subgroups according to morphology of the mesal appendages, inferior appendages and venation degeneration in the hindwings. The first subgroup consists of M. paramoesta Wiggins, 1968 and M. moesta, in which the posterior margins of the superior appendages are concave, the mesal appendages are oblique and unfurcated at the distal end when viewed laterally; and the inferior appendages are without any ven- tral processes. Although these two species both have a wide distribution in the Oriental region, but their ranges do not overlap. The second subgroup consists of M. gamdaha Olah & Johanson, 2010, M. crinita Wiggins, 1968, M. saetigera Wiggins, 1968 and M. oglamar Malicky & Chantaramongkol, 1989. In this subgroup, the mesal append- ages are furcated at the distal end, the inferior appendages divided into dorsal, ventral and inner processes. Furthermore, media veins are usually unbranched and fused with cubitus at the base or at the distal end. Unlike MZ. paramoesta and M. moesta, these four species are regional endemics, also with non-overlapping ranges. ‘The relatively localized dispersal of adults and the disjunct distribution of adequate habitats in some cases lead to small scale allopatric speciation (Vitecek et al. 2015; Thomas et al. 2020). Molanna saetigera and M. paramoesta, on the contrary, have overlapping ranges. The third subgroup is composed of M. truncata sp. nov. and M. yaeyamensis in which the mesal appendages are unfurcated at the distal end in lateral view. The inferior append- ages and media veins are, however, the same as in the latter group. The second group is characterised by the digitate superior appendages and the vari- able mesal appendages, which are either hammer-like or with their distal ends curved ventrad. The hindwings have relatively complete venation. We divided the group into two subgroups based on the shape of inferior appendages. ‘The first subgroup consists of M. taprobane Flint, 1973 and M. xiaguana, having elongate-triangular inferior ap- pendages. The second subgroup consists of M. jolandae Neboiss, 1993 and M. cuprip- ennis Ulmer, 1906, with bifurcated inferior appendages. both of which show rather unique distribution patterns in Indonesia (i.e., restricted allopatric distribution on Sulawesi; Fig. 3). They exhibit similar morphology, however, genetic data were not available for the current study, thus their relationship and evolutionary history remain 174 Xin-yu Ge et al. / ZooKeys 1112: 161-178 (2022) A Cu, Cu, ; A A. Molanna flavicornis B. Molanna uniophila D. Molanna taprobane De Sc+R, SctR, pea se ee —<—<——————— F. Molanna saetigera H. Molanna jolandae 2a eae he, S2ceaer o> M+Cu I. Molanna gamdaha J. Molanna nervosa Figure 6. Male hind wings of eight known Molanna species. A M. flavicornis B M. uniophila C M. moestaD M. taprobane E M. cupripennis F M. saetigera G M. yaeyamensis H M. jolandae\ M. nervosa J M. gamdaha. Scale bar: 200 um. to be investigated. Sulawesi island is however, known as a biodiversity hotspot due to its complex geological history (e.g., Tanzler et al. 2016). Wings are one of the most important organs of insects, and venation modifications may reflect successful adaptation to different environmental conditions. Based on A new species of genus Molanna in China KS A. Molanna truncata sp. nov. B. Molanna crinita Figure 7. Male hind wings of Molanna truncata sp. nov. and M. crinita. A M. truncata sp. nov. B ™. crinita. Scale bar: 200 um. Schmid’s terminology (Schmid 1998) for the hindwing venation of male M. flavicornis, we compared the venation of nine known males of Molanna, and found that venation showed a trend of degradation. Molanna flavicornis and M. uniophila Vorhies, 1909 had the most complete venation in the hindwing, i.e., subcosta was free, radius had 4 branches, media had 2 branches, and cubitus 2 branches. The hindwing venation of these two species could represent the primitive form (Fig. 6A, B). Other species showed venation of hindwings more or less reduced (Figs 6C-—J, 7A, B). Regarding the evolution of hindwings venation in the genus, further support from molecular data could help clarify the true evolutionary pattern. In previous barcode studies on Trichoptera, thresholds of intraspecific divergence (as uncorrected pairwise differences in the COI region) have been estimated to reach at most 11.7% in Hydropsychidae Curtis, 1835 (Zhou et al. 2007) and 11% in Drusus Stephens, 1837 (Kudini¢é et al. 2016). In this study, the ABGD analysis of Molanna inferred 9 OTUs and thresholds of interspecific divergence of 10%; however, it seems that MZ. moe- sta has diverged into three geographic populations (MZ. moesta from Laos, Russia and Ja- pan were recognized as two OTUs; Table 1), with a threshold of intraspecific divergence ranging between 0.30%-8.50% (Table 1). Molanna truncata sp. nov. is morphologically most similar to M. yaeyamensis, but molecular data for the latter were not available. The minimum interspecific divergence was 10.10% between M. truncata sp. nov. and M. XZ sp. As the M. XZ_sp. was collected in close proximity to the known range of M. yaeya- mensis, it is highly likely that M/. XZ_sp. is indeed MZ. yaeyamensis. However, in order to check this, we would need to examine the specimen in detail. The suggested clustering of Oriental Molanna into groups and subgroups based on the structure of male genitalia and the hindwing venation should be further evaluated using an integrated approach, ie., more detailed morphological analysis encompassing more specimens, a multigene phylogeny including all Molanna species and detailed species distribution (Johanson et al. 2012; Jiang et al. 2021). Such an approach would enable reconstruction of the history of the genus in the Oriental region; however, at present, data are not available. Molanna truncata sp. nov. is endemic to Jiuzhaigou Natural Reserve. Apart from this species, we have collected an additional 24 species of Trichoptera (belonging to 22 genera and 14 families) during 2014—2019 in the Reserve (Cao et al. 2016), however, it is the only Molanna species occurring in the area. The area harbours high diversity of freshwater habitats (114 alpine lakes, 17 groups of waterfalls, 47 springs, and 11 sec- tions of rapids; Deng 2012), thus, high diversity of Trichoptera can be expected. The 176 Xin-yu Ge et al. / ZooKeys 1112: 161-178 (2022) Reserve is an example of typical karst geology, with a high amount of travertine calcite deposits in freshwater habitats (Wang et al. 2018). The majority of the lakes are oligo- trophic, with low concentrations of total suspended solids and low turbidity (Li et al. 2020). Water temperature is relatively low in the lakes, whereas conductivity, alkalinity, and pH are relatively high (Cao et al. 2016). Adults of M. truncata sp. nov. were col- lected from one of such alpine lakes located at altitudes ranging from 2299 to 2377 m. Acknowledgements We are grateful to the editor and the two anonymous reviewers for useful recommen- dations to improve the manuscript. 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