Fig. 4 in Microphysogobio luhensis Huang, Chen, Zhao & Shao, 2018, n. sp.

© 2018 Academia Sinica, Taiwan Open Access The Octocorals of Dongsha Atoll (South China Sea): An Iterative Approach to Species Identification Using Classical Taxonomy and Molecular Barcodes Yehuda Benayahu1,*, Leendert Pieter van Ofwegen2, Chang-feng Dai3, Ming-Shiou Jeng4, Keryea Soong5, Alex Shlagman1, Samuel W. Du6, Prudence Hong6, Nimrah H. Imam6, Alice Chung6, Tiana Wu6, and Catherine S. McFadden6 1School of Zoology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv, 69978, Israel. E-mail: [email protected]. ac.il (Shlagman) 2Naturalis Biodiversity Center, P.O. Box 9517, 2300 RA Leiden, The Netherlands. E-mail: [email protected] 3Institute of Oceanography, National Taiwan University, Taipei, Taiwan. E-mail: [email protected] 4Biodiversity Research Center, Academia Sinica, Taipei, Taiwan. E-mail: [email protected] 5Institute of Marine Biology, National Sun Yat-sen University, Kaohsiung, Taiwan. E-mail: [email protected] 6Department of Biology, Harvey Mudd College, Claremont, CA 91711-5990, USA. E-mail: [email protected] (Du); [email protected] (Hong); [email protected] (Imam), [email protected] (Chung), [email protected] (Wu), [email protected] (McFadden) (Received 7 August 2018; Accepted 16 September 2018; Published 3 December 2018; Communicated by Benny K.K. Chan) Citation: Benayahu Y, van Ofwegen LP, Dai C-f, Jeng MS, Soong K, Shlagman A, Du SW, Hong P, Imam NH, Chung A, Wu T, McFadden CS. 2018. The octocorals of Dongsha Atoll (South China Sea): an iterative approach to species identification using classical taxonomy and molecular barcodes. Zool Stud 57:50. doi:10.6620/ZS.2018.57-50. Yehuda Benayahu, Leendert Pieter van Ofwegen, Chang-feng Dai, Ming-Shiou Jeng, Keryea Soong, Alex Shlagman, Samuel W. Du, Prudence Hong, Nimrah H. Imam, Alice Chung, Tiana Wu, and Catherine S. McFadden (2018) Surveys of octocorals from Dongsha Atoll, Taiwan were conducted during 2011, 2013 and 2015 by SCUBA at a depth range of 6-25 m. The collections yielded ~540 specimens, encompassing the variety of taxa occurring in the explored sites; estimates of their abundances were also recorded. Dongsha features a highly diverse octocoral fauna, and octocorals are the dominant benthic organisms in the surveyed reef sites, often covering the majority of the hard substratum. Specimens were identified to the genus and species levels based on an iterative approach that integrates classical taxonomy with character-based molecular barcodes. A total of 51 nominal species representing 20 genera belonging to seven families were recorded, plus ~30 colonies that could only be assigned to a genus. Members of the family Alcyoniidae were the most abundant and diverse taxa, with 27 nominal species plus at least one potentially new, undescribed species of Sinularia, and 5-7 species each of Cladiella, Lobophytum and Sarcophyton. Problems with the taxonomic identification and phylogenetic relationships of species in these genera are discussed. The peculiarity of the Dongsha octocoral species composition is noted, and the composition is also compared to the other Taiwanese reef systems. Key words: Alcyonacea, Coral reefs, DNA barcoding, Pure characteristic attributes, Taiwan, Species diversity, New records. *Correspondence: E-mail: [email protected] BACKGROUND The Taiwanese reef systems at the junction of the Philippine-Japan islands arc include a variety of well-developed reef communities at the northern edge of the South China Sea and additionally Zoological Studies 57: 50 (2018) doi:10.6620/ZS.2018.57-50 1 © 2018 Academia Sinica, Taiwan around the adjacent Pacific offshore islands (e.g. Spalding et al. 2001; Ribas-Deulofeu et al. 2016). The octocoral fauna of these reef communities in Taiwan has been a subject of taxonomic research since the pioneering studies carried out by Utinomi (1950a) along the northeastern coast of Taiwan, as well as at its southern end (Utinomi 1950b 1951 1959). Later studies have addressed the octocoral diversity of the southern Taiwanese reefs at Kenting National Park, the Pacific Green Is. (Lyudao) (Benayahu and Perkol-Finkel 2004; Benayahu et al. 2004), the Penghu Archipelago (Benayahu and McFadden 2011; Benayahu and Ofwegen 2011; Ofwegen and Benayahu 2012; Benayahu et al. 2012), as well as the abundance and ecological significance of octocorals on the Taiwanese reefs (e.g. Dai 1990 1991a 1991b 1993; Fan et al. 2005). Of special interest is the reef-building function played by Sinularia octocorals in some coral reefs there (Jeng et al. 2011a). This record of publications from the region clearly indicates the high diversity and ecological importance of octocorals on the various reef systems in Taiwan, in both the East China Sea and Pacific Ocean. Dongsha Atoll is located in the Pratas Islands at the edge of the continental shelf in the northern South China Sea (20°41'N, 116°48'E). It has a diameter of about 25 km, and its lagoon, reef flats and fore-reef cover ca. 500 km2 in total area. The reef flats are ca. 1 m deep, and the lagoon is 12-21 m deep (Soong et al. 2002). These reefs are affected by the northeast monsoon during October-April, by the typhoon season between June-September, and by SW winds at other times of the year. The atoll experiences two contrasting physical phenomena: repetitive anomalies in the sea-surface temperature that exceed the coral bleaching threshold (DeCarlo et al. 2015); and regular effects of the world’s strongest internal waves that result in the rhythmic upwelling of cold deep waters on the outer reef slopes of the atoll (Hsu and Liu 2000; Dai 2005; Liu et al. 2013; Wang 2016). This combination leads to a clear spatial separation between ‘thermally-susceptible’ stony coral genera, which mainly inhabit the forereef, and ‘thermally-resistant’ genera, which mainly reside in the lagoon. Consequently, it has been suggested that Dongsha Atoll constitutes a potential thermal refuge for reef-building corals in the northern South China Sea, facilitating the development of resilience and resistance to bleaching in coral communities of the lagoon (Tkachenko and Soong 2017). Estimates of the number of species of stony corals found at Dongsha Atoll have steadily increased over time: 45 species were reported by Yang et al. (1975), 63 by Fang et al. (1990), and 137 by Dai et al. (1995). More recent surveys have revealed 257 stony coral species, 56 octocorals, six hydrozoans, and two antipatharians (Jeng et al. 2008 2011b). A comprehensive field guide to the octocorals of Dongsha Atoll was published recently and presents the diverse species composition there, comprising ten families (Dai and Chin 2017). These surveys have also indicated that fleshy octocorals, such as those of the families Alcyoniidae, Briareidae, Nephtheidae and Xeniidae, are abundant on the northern, eastern, and southern reef slopes of the atoll, but are relatively rare on the western reef slope (5-25 m) and almost absent in the lagoon. The current study addresses the octocoral diversity at Dongsha based on the comprehensive collections conducted there. We use an iterative, integrative approach to identify species based on the reconciliation of morphological features with character-based molecular barcodes. In addition to a systematic list of the octocoral taxa found at Dongsha Atoll, we discuss aspects of their phylogeny and note their abundance and geographic distribution in relation to other Taiwanese reef systems. MATERIALS AND METHODS Material was collected, using SCUBA, during three field trips conducted in July 2011 (3 days, 6 dives), September 2013 (4 days, 7 dives), and June 2015 (6 days, 12 dives). All collections were made on the reef slope at a depth range of 6-25 m (Fig. 1). These collections yielded ~540 specimens of octocorals, encompassing the variety of taxa occurring in the explored sites. During the field work underwater abundance estimates of the different morphospecies were made visually and divided into four qualitative categories: rare, sporadic, abundant, and dominant (see also Benayahu et al. 2004). Most of the colonies were photographed in situ prior to collection, which assisted in determining the abundance of the identified species. All samples were preserved in 70% ethanol and subsamples were removed and preserved in absolute ethanol and salt-saturated DMSO buffer for molecular studies. Specimens were identified at the species level based on an iterative approach that integrated page 2 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan classical taxonomy using morphological characters with molecular barcodes. Species identifications based on morphological characters were compared to identifications made independently by matching barcode sequences to an existing reference database (McFadden et al. 2006 2009 2014; Benayahu et al. 2012). When species identifications based on morphology vs. molecular barcodes were in disagreement we iteratively confirmed and reconsidered both sources of information and sought to resolve the discrepancy. Morphological species identification In order to identify morphospecies, sclerites from different parts of the studied colony were obtained by dissolving the tissues in 10% sodium hypochlorite, followed by careful rinsing in distilled water. Particular care was taken to examine polyp sclerites when applicable, as these play a major role in the taxonomy of the speciose genus Sinularia (e.g. McFadden et al. 2009; Ofwegen and Benayahu 2012). When necessary, sclerites were prepared for scanning electron microscopy as follows: they were carefully rinsed with double-distilled water, dried at room temperature, coated with gold, and examined with a Jeol 6480LV electron microscope operated at 10 kV. Identification of species was facilitated by comparisons with permanent sclerite preparations of type material (when available) kept in the Steinhardt Museum of Natural History, Tel Aviv University, Israel (SMNH) and the Netherlands Center for Biodiversity, Naturalis, Leiden (RMNH). The identified specimens are deposited at SMNH (collection numbers preceeded by ZMTAU) (Table 1). Molecular species identification Extraction of DNA from ethanol-preserved tissue samples, PCR amplification, and sequencing of the mtMutS, COI (including the adjacent intergenic region, igr1) and 28S rDNA barcoding regions followed the protocols published in McFadden et al. (2011 2014). The Fig. 1. Collection sites of octocorals at Dongsha Atoll. N page 3 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan Table 1. List of Octocorallia species of the orders Helioporacea Bock, 1938 and Alcyonacea (Lamouroux, 1816) from Dongsha Atoll with indication of the Museum inventory numbers (ZMTAU Co), their collection site, depth of occurrence, abundance estimate on the reef, and Distribution in Taiwan (southern Taiwan and Penghu Archipelago) Classification Family Helioporidae Moseley, 1876 Heliopora de Blainville, 1830 Heliopora coerulea (Pallas, 1766) Occurrence: ZMTAU Co 36388, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013. Field notes: Rare. Distribution in Taiwan: Southern Taiwan. Family Alcyoniidae Lamouroux, 1812 Aldersladum Benayahu & McFadden, 2011 Aldersladum jengi Benayahu & McFadden, 2011 Occurrence: ZMTAU Co 35392, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011. Field notes: Rare. Distribution in Taiwan: Penghu. Cladiella Gray, 1869 Cladiella australis (Macfadyen, 1936) Occurrence: ZMTAU Co 35324, Co 35333, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 35344, Dongsha, (20°46.697'N; 116°51.051'E), 7-10 m, 8 July 2011; ZMTAU Co 35366, Dongsha, (20°46.700'N; 116°53.137'E), 10-11 m, 9 July 2011; ZMTAU Co 36277, Co 36279, Co 36284, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 6-10 m, 11 September 2013; ZMTAU Co 36313, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-12 m, 14 September 2013; ZMTAU Co 36344, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 36912, Dongsha, (20°44.422'N; 116°54.865'E), 19-21 m, 12 June 2015; ZMTAU, Co 36987, Dongsha, (20°41.994'N; 116°55.370'E), 4-6 m, 15 June 2015. Field notes: Common. Distribution in Taiwan: Southern Taiwan and Penghu Archipelago. Cladiella conifera (Tixier-Durivault, 1943) Occurrence: ZMTAU Co 35376, Dongsha, (20°46.689'N; 116°47.932'E), 5m, 9 July 2011. Field notes: Rare. Distribution in Taiwan: New record. Cladiella krempfi Hickson, 1919 Occurrence: ZMTAU Co 35347, Dongsha, (20°46.697'N; 116°51.051'E), 7-10 m, 8 July 2011; ZMTAU Co 35389, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 36289, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013. Field notes: Rare. Distribution in Taiwan: Penghu Archipelago. Cladiella latissima (Tixier-Durivault, 1948) Occurrence: MTAU Co 35320, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 36336, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013. Field notes: Rare. Distribution in Taiwan: Penghu Archipelago. Cladiella pachyclados (Klunzinger, 1877) Occurrence: MTAU Co 35377, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 35409, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 36314, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-12 m, 14 September 2013; ZMTAU 36352, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013. Field notes: Sporadic. Distribution in Taiwan: Southern Taiwan and Penghu Arcipelago. page 4 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan Table 1. (Continued) Cladiella sp. Occurrence: ZMTAU Co 35323, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 35425, (20°46.778'N; 116°48.533'E), 5-6 m, 10 July 2011; ZMTAU Co 36901, Dongsha, (20°44.316'N; 116°54.827'E), 12-14 m, 12 June 2015; ZMTAU Co 36947, Dongsha, (20°43.448'N; 116°42.520'E), 4-5 m, 13 June 2015; ZMTAU Co 36963, Dongsha, (20°41.509'N; 116°55.379'E), 6-8 m, 14 June 2015; ZMTAU Co 37012, Dongsha, (20°35.859'N; 116°51.380'E), 5-8 m, 16 June 2015. Field notes: Sporadic. Klyxum Alderslade, 2000 Klyxum sp. Occurrence: MTAU Co 35404, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 36278, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 6-10 m, 11 September 2013; ZMTAU Co 36348, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 36380, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013; ZMTAU Co 36968, Dongsha, (20°41.994'N; 116°55.370'E), 4-6 m, 15 June 2015; ZMTAU Co 37008, Dongsha, (20°35.859'N; 116°51.380'E), 5-8 m, 16 June 2015. Field notes: Sporadic. Distribution in Taiwan: Southern Taiwan: K. simplex and Penghu: K. utinomii. Lobophytum von Marenzeller, 1886 Lobophytum catalai Tixier-Durivault, 1957 Occurrence: ZMTAU Co 35340, Dongsha, (20°46.697'N; 116°51.051'E), 7-10 m, 8 July 2011; ZMTAU Co 35396, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 36285, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 6-10 m, 11 September 2013. Field notes: Rare. Distribution in Taiwan: New record. Lobophytum crassum von Marenzeller, 1886 Occurrence: ZMTAU, Co 35321, Co 35325, Co 35334 Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; Co 35354, Dongsha, (20°46.697'N; 116°51.051'E), 7-10 m, 8 July 2011; Co 35370, Dongsha, (20°46.700'N; 116°53.137'E), 10-11 m, 9 July 2011; Co 35383, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; Co 35398, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 36276, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 6-10 m, 11 September 2013; ZMTAU Co 36290, Co 36304, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36321, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-12 m, 14 September 2013; ZMTAU Co 36346, Co 36349, Co 36364, Co 36369, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013. Field notes: Dominant. Distribution in Taiwan: Southern Taiwan and Penghu Archipelago. Lobophytum hirsutum Tixier-Durivault, 1956 Occurrence: ZMTAU Co 36385, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013. Field notes: Rare. Distribution in Taiwan: New record. Lobophytum hsiehi Benayahu & Ofwegen, 2011 Occurrence: ZMTAU Co 35319, Co 35326, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 35365, Dongsha, (20°46.700'N; 116°53.137'E), 10-11 m, 9 July 2011; ZMTAU Co 35399, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 35421, Dongsha, (20°46.778'N; 116°48.533'E), 5-6 m, 10 July 2011; ZMTAU Co 36282, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 6-10 m, 11 September 2013; ZMTAU Co 36320, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 7-8 m, 14 September 2013; ZMTAU Co 36342, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 36387, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013. Field notes: Dominant. Distribution in Taiwan: Penghu Archipelago. Lobophytum legitimum Tixier-Durivault, 1970 Occurrence: ZMTAU Co 36311, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-12 m, 14 September 2013. Field notes: Rare. Distribution in Taiwan: New record. Lobophytum pauciflorum (Ehrenberg, 1834) Occurrence: ZMTAU Co 35380, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011. page 5 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan Table 1. (Continued) Field notes: Rare. Distribution in Taiwan: Southern Taiwan. Lobophytum rigidum Benayahu, 1995 Occurrence: ZMTAU Co 35329, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 36306, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36934, Dongsha, (20°43.448'N; 116°42.520'E), 3-4 m, 13 June 2015; ZMTAU Co 37015, Dongsha, (20°35.859'N; 116°51.380'E), 5-8 m, 16 June 2015. Field notes: Rare. Distribution in Taiwan: New record. Paraminabea Williams & Alderslade, 1999 Paraminabea aldersladei (Williams, 1992) Occurrence: ZMTAU Co 36949, Dongsha, (20°43.448'N; 116°42.520'E), 4-5 m, 13 June 2015. Field notes: Rare. Distribution in Taiwan: Southern Taiwan. Sarcophyton Lesson, 1834 Sarcophyton cinereum (Tixier-Durivault, 1946) Occurrence: ZMTAU Co 35413, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 36354, Co 36366, Co 36372, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013.ZMTAU Co 36275, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 6-10 m, 11 September 2013. Field notes: Rare. Distribution in Taiwan: New record. Sarcophyton crassocaule Moser, 1919 Occurrence: ZMTAU Co 35330, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 36294, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013. Field notes: Rare. Distribution in Taiwan: Southern Taiwan. Sarcophyton ehrenbergi von Marenzeller, 1886 Occurrence: ZMTAU Co 35313, Co 35337, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 35356, Dongsha, (20°46.697'N; 116°51.051'E), 7-10 m, 8 July 2011; ZMTAU Co 35359, Dongsha, (20°46.700'N; 116°53.137'E), 10-11 m, 9 July 2011; ZMTAU Co 35382, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 35403, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 36280, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 6-10 m, 11 September 2013; ZMTAU Co 36293, Co 36295, Co 36299, Co 36303, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36308, Co 36317, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-12 m, 14 September 2013; ZMTAU Co 36331, Co 36338, Co 36339, Co 36340, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013; ZMTAU Co 36351, Co 36353, Co 36362, Co 36363, Co 36368, Co, Co 37370, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 36382, Co 36398, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-12 m, 14 September 2013; ZMTAU Co 37020, Co 37022, Dongsha, (20°35.941'N; 116°51.632'E), 7-9 m, 16 June 2015. Field notes: Dominant. Distribution in Taiwan: Southern Taiwan and Penghu Archipelago. Sarcophyton elegans Moser, 1919 Occurrence: ZMTAU Co 36318, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-12 m, 14 September 2013; ZMTAU Co 36966, Dongsha, (20°41.509'N; 116°55.379'E), 6-8 m, 14 June 2015. Field notes: Rare. Distribution in Taiwan: New record. Sarcophyton nanwanensis Benayahu & Perkol-Finkel, 2004 Occurrence: ZMTAU Co 35346, Co 35350, Dongsha, (20°46.697'N; 116°51.051'E), 7-10 m, 8 July 2011; ZMTAU Co 35386, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 35407, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 35415, Dongsha, (20°46.778'N; 116°48.533'E), 5-6 m, 10 July 2011; ZMTAU Co 36310, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-12 m, 14 September 2013. Field notes: Common. Distribution in Taiwan: Southern Taiwan. page 6 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan Table 1. (Continued) Sarcophyton trocheliophorum von Marenzeller, 1886 Occurrence: ZMTAU Co 35317, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 35363, Dongsha, (20°46.700'N; 116°53.137'E), 10-11 m, 9 July 2011; ZMTAU Co 35379, Co 35388, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 35418, Dongsha, (20°46.778'N; 116°48.533'E), 5-6 m, 10 July 2011; ZMTAU Co 36302,Co 36291, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36335, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013; ZMTAU Co 36350, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 36996, Dongsha, (20°41.994'N; 116°55.370'E), 5-7 m, 15 June 2015. Field notes: Dominant. Distribution in Taiwan: Southern Taiwan and Penghu Archipelago. Sinularia May, 1898 Sinularia abrupta Tixier-Durivault, 1970 Occurrence: ZMTAU Co 35331, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 36305, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36332, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013; ZMTAU Co 36341, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013. Field notes: Sporadic. Distribution in Taiwan: Penghu Archipelago. Sinularia acuta Manuputty & van Ofwegen, 2007 Occurrence: ZMTAU Co 35393, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 36376, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013; ZMTAU Co 36897, Dongsha, (20°44.316'N; 116°54.827'E), 12-14 m, 12 June 2015; ZMTAU Co 36943, Dongsha, (20°43.448'N; 116°42.520'E), 4-5 m, 13 June 2015; ZMTAU Co 36952, Co 36957, Dongsha, (20°41.509'N; 116°55.379'E), 6-8 m, 14 June 2015; ZMTAU Co 36984, Co 36988, Dongsha, (20°41.994'N; 116°55.370'E), 4-8 m, 15 June 2015; ZMTAU Co 36992, Co 36997, Co 37003, Dongsha, (20°41.994'N; 116°55.370'E), 5-7 m, 15 June 2015; ZMTAU Co 37009, Co 37016, Dongsha, (20°35.859'N; 116°51.380'E), 5-8 m, 16 June 2015; ZMTAU Co 37028, Dongsha, (20°35.941'N; 116°51.632'E), 7-9 m, 16 June 2015. Field notes: Dominant. Distribution in Taiwan: Penghu Archipelago. Sinularia brassica May, 1898 Occurrence: ZMTAU Co 36328, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013; ZMTAU Co 35349,Co 35352, Dongsha, (20°46.697'N; 116°51.051'E), 7-10 m, 8 July 2011; ZMTAU Co 35360, Co 35368, Dongsha, (20°46.700'N; 116°53.137'E), 10-11 m, 9 July 2011; ZMTAU Co 35381, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 35427, Co 35428, Dongsha, (20°46.778'N; 116°48.533'E), 5-6 m, 10 July 2011; ZMTAU Co 36345, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 36908, Dongsha, (20°44.316'N; 116°54.827'E), 12-14 m, 12 June 2015; ZMTAU Co 36911, Dongsha, (20°44.422'N; 116°54.865'E), 19-21 m, 12 June 2015; ZMTAU Co 37030, Dongsha, (20°35.941'N; 116°51.632'E), 7-9 m, 16 June 2015. Field notes: Common. Distribution in Taiwan: Southern Taiwan. Sinularia capillosa Tixier-Durivault, 1970 Occurrence: ZMTAU Co 35410, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 35417, Dongsha, (20°46.778'N; 116°48.533'E), 5-6 m, 10 July 2011. Field notes: Rare. Distribution in Taiwan: New record. Sinularia ceramensis Verseveldt, 1977 Occurrence: ZMTAU Co 35351, Co 35343, Dongsha, (20°46.697'N; 116°51.051'E), 7-10 m, 8 July 2011; ZMTAU Co 36326, Co 36334, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013; ZMTAU Co 36355, Co 36357, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 35390, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 36333, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013; ZMTAU Co 36379, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013; ZMTAU Co 36938, Dongsha, (20°43.448'N; 116°42.520'E), 4-5 m, 13 June 2015; ZMTAU Co 37429, Dongsha, (20°43.448'N; 116°42.520'E), 3-4 m, 13 June 2015; ZMTAU Co 37001, Dongsha, (20°41.994'N; 116°55.370'E), 5-7 m, 15 June 2015; ZMTAU Co 37010, Dongsha, (20°35.859'N; 116°51.380'E), 5-8 m, 16 June 2015. Field notes: Dominant. Distribution in Taiwan: New record. page 7 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan Table 1. (Continued) Sinularia curvata Manuputty & van Ofwegen, 2007 Occurrence: ZMTAU Co 35338, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 35367, Dongsha, (20°46.700'N; 116°53.137'E), 10-11 m, 9 July 2011. Field notes: Rare. Distribution in Taiwan: New record. Sinularia densa (Whitelegge, 1897) Occurrence: ZMTAU Co 35373, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 36902, Co 36907, Dongsha, (20°44.316'N; 116°54.827'E), 12-14 m, 12 June 2015; ZMTAU Co 36935, Dongsha, (20°43.448'N; 116°42.520'E), 3-4 m, 13 June 2015; ZMTAU Co 36965, Dongsha, (20°41.509'N; 116°55.379'E), 6-8 m, 14 June 2015; ZMTAU Co 36977, Co 36985, Dongsha, (20°41.994'N; 116°55.370'E), 4-6 m, 15 June 2015; ZMTAU Co 37005, Dongsha, (20°41.994'N; 116°55.370'E), 5-7 m, 15 June 2015. Field notes: Sporadic. Distribution in Taiwan: Southern Taiwan. Sinularia erecta Tixier-Durivault, 1945 Occurrence: ZMTAU Co 36329, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013. Field notes: Rare. Distribution in Taiwan: Southern Taiwan and Penghu Archipelago. Sinularia exilis Tixier-Durivault, 1970 Occurrence: ZMTAU Co 36360, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 36898, Dongsha, (20°44.316'N; 116°54.827'E), 12-14 m, 12 June 2015; ZMTAU Co 36940, Dongsha, (20°43.448'N; 116°42.520'E), 4-5 m, 13 June 2015; ZMTAU Co 36994, Dongsha, (20°41.994'N; 116°55.370'E), 5-7 m, 15 June 2015; ZMTAU Co 37006, Dongsha, (20°35.859'N; 116°51.380'E), 5-8 m, 16 June 2015. Field notes: Common. Distribution in Taiwan: Southern Taiwan. Sinularia flexibilis (Quoy & Gaimard, 1833) Occurrence: ZMTAU Co 35405, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 36301, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36325, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013. Field notes: Common. Distribution in Taiwan: Southern Taiwan. Sinularia heterospiculata Verseveldt, 1970 Occurrence: ZMTAU Co 36281, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 6-10 m, 11 September 2013. Field notes: Rare. Distribution in Taiwan: New record. Sinularia hirta (Pratt, 1903) Occurrence: ZMTAU Co 36298, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36392, Co 36383, Co 36394, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013; ZMTAU Co 36941, Co 36945, Dongsha, (20°43.448'N; 116°42.520'E), 4-5 m, 13 June 2015. Field notes: Rare. Distribution in Taiwan: Penghu Archipelago. Sinularia humesi Verseveldt, 1971 Occurrence: ZMTAU Co 36309, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-12 m, 14 September 2013; ZMTAU Co 36395, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013; ZMTAU Co 36917, Dongsha, (20°44.422'N; 116°54.865'E), 19-21 m, 12 June 2015; ZMTAU Co 36899, Dongsha, (20°44.316'N; 116°54.827'E), 12-14 m, 12 June 2015. Field notes: Sporadic. Distribution in Taiwan: Southern Taiwan. Sinularia humilis van Ofwegen, 2008 Occurrence: ZMTAU Co 35316, Co 35328, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 36330, Dongsha, (20°46'25.8"N; 116°46'52.1"E), 10-13 m, 14 September 2013; ZMTAU Co 36390, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 page 8 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan Table 1. (Continued) September 2013; ZMTAU Co 36900, Dongsha, (20°44.316'N; 116°54.827'E), 12-14 m, 12 June 2015; ZMTAU Co 36916, Dongsha, (20°44.422'N; 116°54.865'E), 19-21 m, 12 June 2015; ZMTAU Co 36920, Co 36930, Dongsha, (20°43.448'N; 116°42.520'E), 3-4 m, 13 June 2015; ZMTAU Co 36942, Co 36944, Dongsha, (20°43.448'N; 116°42.520'E), 4-5 m, 13 June 2015; ZMTAU Co 36986, Dongsha, (20°41.994'N; 116°55.370'E), 4-6 m, 15 June 2015. Field notes: Common. Distribution in Taiwan: New record. Sinularia lochmodes Kolonko, 1926 Occurrence: ZMTAU Co 35318, Co 35332, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 35391, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 35423, Co 35426, Dongsha, (20°46.778'N; 116°48.533'E), 5-6 m, 10 July 2011; ZMTAU Co 36292, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36356, Co 36375, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 36377, Co 36386, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013; ZMTAU Co 36903, Dongsha, (20°44.316'N; 116°54.827'E), 12-14 m, 12 June 2015; ZMTAU Co 36936, Co 36937, Dongsha, (20°43.448'N; 116°42.520'E), 4-5 m, 13 June 2015. Field notes: Dominant. Distribution in Taiwan: Southern Taiwan and Penghu Archipelago. Sinularia maxima Verseveldt, 1971 Occurrence: ZMTAU Co 35384, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 35414, Dongsha, (20°46.778'N; 116°48.533'E), 5-6 m, 10 July 2011; ZMTAU Co 36283, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 6-10 m, 11 September 2013; ZMTAU Co 36300, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36381, Co 36384, Co 36391, Co 36399, Dongsha, (20°43'23.6"N; 116°42'31.5"E), 7-8 m, 16 September 2013; ZMTAU Co 36924, Dongsha, (20°43.448'N; 116°42.520'E), 3-4 m, 13 June 2015. Field notes: Dominant. Distribution in Taiwan: Southern Taiwan and Penghu Archipelago. Sinularia ovispiculata Tixier-Durivault, 1970 Occurrence: ZMTAU Co 35353, Dongsha, (20°46.697'N; 116°51.051'E), 7-10 m, 8 July 2011; ZMTAU Co 37584, Dongsha, (20°46.767'N; 116°48.393'E), 10 m, coll. Ming-Shion Jeng, 29 May 2015; ZMTAU Co 36995, Dongsha, (20°41.994'N; 116°55.370'E), 5-7 m, 15 June 2015. Field notes: Rare. Distribution in Taiwan: Southern Taiwan. Sinularia pavida Tixier-Durivault, 1970 Occurrence: ZMTAU Co 35374, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 35408, Dongsha, (20°45.609'N; 116°44.525'E), 7-11 m, 10 July 2011; ZMTAU Co 36946, Dongsha, (20°43.448'N; 116°42.520'E), 4-5 m, 13 June 2015. Field notes: Rare. Distribution in Taiwan: Penghu Archipelago. Sinularia penghuensis van Ofwegen & Benayahu, 2012 Occurrence: ZMTAU Co 35322, Co 35314, Dongsha, (20°46.703'N; 116°50.324'E), 8-10 m, 8 July 2011; ZMTAU Co 35387, Dongsha, (20°46.689'N; 116°47.932'E), 5 m, 9 July 2011; ZMTAU Co 35422, Dongsha, (20°46.778'N; 116°48.533'E), 5-6 m, 10 July 2011; ZMTAU Co 36287, Co 36296, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36343, Dongsha, (20°46'50.2"N; 116°48'38.5"E), 7-8 m, 15 September 2013; ZMTAU Co 36914, Dongsha, (20°44.422'N; 116°54.865'E), 19-21 m, 12 June 2015; ZMTAU Co 36919, Co 36921, Dongsha, (20°43.448'N; 116°42.520'E), 3-4 m, 13 June 2015; ZMTAU Co 36974, Dongsha, (20°41.994'N; 116°55.370'E), 4-6 m, 15 June 2015; ZMTAU Co 37000, Dongsha, (20°41.994'N; 116°55.370'E), 5-7 m, 15 June 2015; ZMTAU Co 37025, Dongsha, (20°35.941'N; 116°51.632'E), 7-9 m, 16 June 2015. Field notes: Dominant. Distribution in Taiwan: Penghu Archipelago. Sinularia querciformis (Pratt, 1903) Occurrence: ZMTAU Co 36297, Dongsha, (20°35'23.5"N; 116°46'52.1"E), 12-17 m, 11 September 2013; ZMTAU Co 36928, Dongsha, (20°43.448'N; 116°42.520'E), 3-4 m, 13 June 2015. Field notes: Rare. Distribution in Taiwan: Southern Taiwan. page 9 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan 0.02 S.ehrenbergi Co37022 L.hirsutum RMNH Coel.40955 S.tortuosum Co34684* Sinularia brassica Co36345 L.hirsutum Co36385 S.trocheliophorum Co35388 L.pauciflorum Co35380 L.legitimum Co36311 S.ehrenbergi Co34705* S.ehrenbergi Co37020 L.rigidum Co36934 S.nanwanensis Co35346 Sinularia humesi Co36395 L.crassum RMNH 40945 L.crassum Co35354 S.trocheliophorum Co36996 S.trocheliophorum Co35317 L.crassum Co36304 S.nanwanensis Co35407 S.ehrenbergi Co35382 S.ehrenbergi Co36338 S.ehrenbergi Co35356 S.elegans Co36318 S.elegans RMNH Coel.41011 L.crassum Co35325 L.crassum Co36364 S.ehrenbergi Co36362 S.nanwanensis Co35415 S.crassocaule Co36294 L.patulum NTM C13965 S.nanwanensis Co36310 S.trocheliophorum Co35363 S.ehrenbergi Co36351 S.trocheliophorum Co36291 S.crassocaule RMNH Coel.33076 L.catalai Co35340 S.ehrenbergi Co36363 L.crassum RMNH Coel.40950 S.ehrenbergi Co35337 S.ehrenbergi Co35313 L.pauciflorum UF2856 S.ehrenbergi RMNH Coel.40963 S.tumulosum Co34698* L.crassum Co35383 L.crassum Co35398 L.rigidum Co35329 L.crassum Co36321 S.cinereum Co36372 L.hsiehi Co34694* L.rigidum Co36306 L.hsiehi Co33593* L.catalai RMNH Coel.40957 S.trocheliophorum Co36302 L.crassum RMNH Coel.33061 S.trocheliophorum Co36335 S.trocheliophorum Co35418 S.trocheliophorum Co33621* Sinularia maxima Co36300 L.catalai Co36285 L.crassum Co35370 S.crassocaule Co35330 L.crassum Co35321 S.elegans Co36966 S.cinereum Co36366 S.nanwanensis Co35386 L.crassum RMNH Coel.40954 S.nanwanensis Co33078* L.crassum Co36369 L.catalai Co35396 S.ehrenbergi Co36295 L.rigidum Co37015 L.crassum Co36346 S.ehrenbergi RMNH Coel.40960 S.cinereum RMNH Coel.41004 S.nanwanensis Co35350 S.trocheliophorum Co35379 L.crassum Co36290 L.hsiehi Co34717* S.cinereum Co35413 Sinularia flexibilis Co36325 L.crassum Co35334 100 100 100 100 100 100 100 99 91 80 M S L C 1.0 1.0 1.0 1.0 1.0 1.0 ns 1.0 ns 1.0 However, six colonies identified as S. crassocaule and L. rigidum (Fig. 3F) belonged to a wellsupported, genetically distinct fourth clade that was sister to the [‘Sarcophyton’ + ‘Lobophytum’] clade; a reference specimen of S. crassocaule from Indonesia also belonged to this fourth clade, Fig. 4. Maximum likelihood tree based on concatenated mtMutS and igr1 + COI sequences (1552 bp) for the soft coral genera Lobophytum and Sarcophyton. Circled letters indicate clades: M, “mixed” clade; C, “Crassocaule” clade; S, “Sarcophyton” clade; L, “Lobophytum” clade (see McFadden et al. 2006). Numbers above nodes: ML bootstrap values; below nodes: Bayesian posterior probabilities. Boldface taxon labels: reference sequences from other studies; *specimen from Penghu Archipelago (Benayahu et al. 2012). Co: ZMTAU accession number. page 16 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan although it was genetically distinct from the S. crassocaule in Dongsha. Four species belonging to the ‘Sarcophyton’ clade (S. cinereum: Fig. 5A, S. elegans, S. nanwanensis: Fig. 5B, S. trocheliophorum: Fig. 5C, D) were morphologically and phylogenetically distinct from one another and had unique barcode sequences that closely matched reference material of those species. Within the ‘Lobophytum’ clade, five species were identified based on morphology: L. catalai, L. Fig. 5. Underwater photographs of Dongsha Atoll octocorals: (A) Sarcophyton cinereum, (B) S. nanwanensis, (C) S. trocheliophorum, (D) S. trocheliophorum, (E) Lobophytum crassum and (F) L. pauciflorum. (A) (C) (E) (B) (D) (F) page 17 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan crassum: Fig. 5E, L. hirsutum, L. legitimum, and L. pauciflorum: Fig. 5F. Colonies identified as L. crassum, however, belonged to four genetically distinct sub-clades, each of which also included reference material previously identified as L. crassum as well as some other species (Fig. 4). Two ‘mixed’ clade species were identified, S. ehrenbergi (Fig. 6A) and L. hsiehi (Fig. 6B). Barcode sequences for S. ehrenbergi varied slightly among individuals but most haplotypes Fig. 6. Underwater photographs of Dongsha Atoll octocorals: (A) Sarcophyton ehrenbergi, (B) Lobophytum hsiehi, (C) Litophyton sp., (D) Sinularia ceramensis, (E) S. brassica, (F) S. flexibilis. (A) (C) (E) (B) (D) (F) page 18 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan matched existing reference material of that species; we were unable to obtain sequence data from any specimens of L. hsiehi. Other taxa All other taxa were identified to the genus level based on a congruence of morphological characters and molecular barcodes. Species-level identifications were not possible, however, for the alcyoniid genera Klyxum and some Cladiella and for the majority of Nephtheidae genera as a result of the need for taxonomic revisions coupled with a lack of reference sequences with confirmed species IDs. Previous work has demonstrated that neither the mtMutS nor COI barcodes resolve species boundaries adequately in Klyxum and Cladiella (Benayahu et al. 2012). Although the 28S rDNA barcode provides better resolution for species in those genera (Benayahu et al. 2012), we encountered some incongruence between morphological and molecular identifications (data not shown) that limited our ability to assign species names with confidence to some specimens. Both molecular and morphological data support the conclusion that the Nephtheidae genera Capnella, Lemnalia, Litophyton (Fig. 6C), Paralemnalia, and Stereonephthya are each represented by just a single species at Dongsha. No or only minimal (< 0.2%) variation in mtMutS + igr1 + COI haplotypes were observed among specimens within any of those genera. Although all colonies of Dendronephthya also shared an identical mitochondrial haplotype, morphological differences suggested that 3-4 species may be present at Dongsha. Previous molecular work has shown that mitochondrial barcode markers may not exhibit sufficient variation to distinguish morphospecies of that genus (Park et al. 2012). DISCUSSION Diversity and biogeography of Dongsha Atoll octocorals The octocoral fauna of the Taiwanese reef systems, including southern Taiwan and the Penghu Archipelago, has been studied extensively (e.g. Benayahu and Perkol-Finkel 2004; Benayahu et al. 2004; Benayahu and McFadden 2011; Benayahu and Ofwegen 2011; Ofwegen and Benayahu 2012; Benayahu et al. 2012). Dongsha Atoll shares a number of common species with both of these other reef systems (Table 1). This resemblance is in particular indicated by the shared occurrence of species whose type locality is southern Taiwan (Sarcophyton nanwanensis) and the Penghu Archipelago (Aldersladum jengi, Lobophytum hsiehi, Sinularia penghuensis, S. soongi, and S. wanannensis). Octocoral diversity is higher at Dongsha Atoll than at Penghu, however, comprising 15 genera in seven families compared to 11 genera in just three families, respectively. Notably, the family Xeniidae is absent at Penghu while at Dongsha xeniids are present although rather rare. The alcyoniid genera Cladiella, Lobophytum, Sarcophyton and Sinularia are the most speciesrich and numerically dominant octocorals in all three Taiwanese reef systems, although the species richness of alcyoniids at Dongsha is about twice that of the more northerly reef system of Penghu. Dongsha has 7, 5 and 27 species of Lobophytum, Sarcophyton and Sinularia, respectively, compared to 4, 3 and 16 species of those same genera at Penghu (Benayahu et al. 2012). Three species of Sinularia encountered most frequently at Penghu (S. molesta, S. ornata, S. peculiaris) have not been reported from Dongsha, whereas several of the most common species found at Dongsha are not present at Penghu (S. ceramensis: Fig. 6D, S. brassica: Fig. 6E, S. flexibilis: Fig. 6F, S. tumulosa: Fig. 7A) or are rare there (S. acuta: Fig. 7B, S. maxima; Fig. 7C, S. lochmodes: Fig. 3A, S. slieringsi). Species such as S. penghuensis: Fig. 3D and S. wanannensis: Fig. 7D are, however, common at both locations, as are Cladiella australis: Fig. 7E, Lobophytum crassum: Fig. 5C, L. hsiehi: Fig. 6B, and Sarcophyton trocheliophorum: Fig. 5D. The reefs of southern Taiwan (Kenting National Park) are reported to have a species richness similar to that of Dongsha and to share many of these same common taxa (Benayahu et al. 2004). However, because studies there were conducted prior to the use of molecular barcoding markers to facilitate species identifications, further studies and re-examination of material from Kenting using such an integrative approach might yield different estimates of both species richness and identity of the taxa present. Character-based DNA barcodes as a tool for octocoral identification The identification of shallow-water IndoPacific octocorals to the species level is extremely page 19 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan challenging, particularly within speciose genera such as Sinularia (e.g. Benayahu et al. 2012; McFadden et al. 2014). Morphospecies are distinguished primarily on the basis of differences in colony growth form and the distribution and shape of sclerites within the tissues (e.g. Fabricius and Alderslade 2001; McFadden et al. 2009). A high level of taxonomic expertise is required to evaluate the latter, and the degree to which such morphological characters may vary intraspecifically Fig. 7. Underwater photographs of Dongsha Atoll octocorals: (A) Sinularia tumulosa, (B) S. acuta, (C) S. maxima, (D) S. wanannensis, (E) Cladiella australis and (F) Cladiella sp. (A) (C) (E) (B) (D) (F) page 20 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan has been assessed for only a few Sinularia species (e.g. S. brassica: Benayahu et al. 1998; S. leptoclados: Verseveldt 1980; Ofwegen et al. 2013; S. polydactyla: Verseveldt 1980; Ofwegen et al. 2016). Moreover, most of the earlier studies were based on limited numbers of specimens, did not examine the entire range of intraspecific variation in morphological characters, and lack supporting molecular data. Although the use of molecular barcode markers facilitates the identification of taxa without the need for extensive taxonomic expertise, the single-gene markers currently available are inadequate to discriminate closelyrelated species within most genera (McFadden et al. 2006 2009 2011 2014). For instance, in a previous survey of octocorals in Palau, application of genetic distance thresholds using the mtMutS DNA barcode discriminated 80% of Sinularia morphospecies, while the overall concordance between identifications based independently on morphology vs. barcode sequences was 77.6% (McFadden et al. 2014). When the molecular barcode was combined with assessment of colony growth form; however, 96% of specimens were identified to the species level. In the present study we used a similar combined approach, reconciling morphological characters with barcode sequences in order to discriminate taxa and assign appropriate binomials. When intraspecific and interspecific genetic distance values are both low and overlap extensively, as they often do in Sinularia and other octocoral genera (McFadden et al. 2011 2014), methods of species discrimination that rely on arbitrary threshold values may fail when even a small amount of intraspecific variation or sequencing error is present. Characterbased barcodes, however, are more robust to such noise, as species-diagnostic nucleotide characters (pure characteristic attributes, PuCAs) can be distinguished from characters that vary intraspecifically, and only the former can be used to identify species (DeSalle et al. 2005; Rach et al. 2008; Zou et al. 2011; Bergmann et al. 2013; Paknia et al. 2015). For example, Sinularia ceramensis from Dongsha Atoll exhibited intraspecific variation at 8 of 735 nucleotide positions in the mtMutS barcode sequence, leading to intraspecific genetic distances that ranged from 0-0.7% and overlapped with some interspecific genetic distances. All individuals of S. ceramensis, however, shared diagnostic PuCAs at two nucleotide positions in mtMutS (T at position 204 and C at position 294) (Fig. 2), allowing them to be discriminated unequivocally from all other species regardless of the noise contributed by intraspecific variation. Unlike threshold-based methods; however, application of character-based barcodes requires that a sufficient number of individuals of a species be sequenced in order to distinguish species-diagnostic PuCAs from nucleotides that vary intraspecifically. Although application of a characterbased rather than a genetic distance-based barcode allowed better discrimination of species of Sinularia that differed from one another by only 1-2 diagnostic nucleotides (genetic distances < 0.3%), we nonetheless encountered distinct morphospecies that shared identical mtMutS sequences and therefore could not be distinguished using either a characteror distance-based approach. Among the species belonging to the confusing 5C (“leptoclados”) sub-clade of Sinularia, S. acuta, S. abrupta, S. penghuensis, and S. slieringsi shared identical mtMutS haplotypes, and S. wanannensis differed from them at only a single nucleotide position (Fig. 2). Further investigation of those morphospecies using multilocus next-generation RADseq methods supports the genetic distinctions among them, however, and also suggests that a nuclear gene barcode marker such as 28S rDNA—perhaps in combination with mtMutS—may better discriminate closely-related Sinularia species (unpub. data). Regardless of whether a characteror threshold-based approach is used to discriminate taxa, the use of molecular barcodes to assign binomials relies on the establishment of a database that links sequence data to validated reference specimens (Puillandre et al. 2011 2012). A limitation to the establishment of such a database for octocorals is that sequences of type colonies along with their appropriate taxonomic descriptions (i.e. including SEM images of their entire suite of sclerites) are usually only available for recently described species. For most taxa, any reference sequences that are currently available derive from freshly collected (non-type) material, as old museum specimens often fail to yield DNA of a quality suitable for sequencing. Even if the newly collected reference material was identified by a skilled taxonomist, it has not always been compared directly to the respective type material and may not have been obtained from the type locality. Such practices introduce sources of error into the sequence database, and may result in molecular barcodes being assigned to reference specimens that have been mis-identified. These page 21 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan taxonomic errors are propagated further when the barcode sequence is then used to assign the incorrect binomial to additional material. Unfortunately, such errors can be difficult to detect and correct, especially when type material is not available for comparison, because it has either been lost or destroyed or its depository is not known. For most octocoral taxa, taxonomic revisions accompanied by barcoding of type material or other validated exemplars (including neotypes) will be necessary before it will be possible to use molecular barcodes alone to assign bionomials with a high degree of confidence. Phylogenetic relationships among genera of the family Alcyoniidae Molecular barcodes and morphological analyses both discriminated > 5 species in each of the alcyoniid genera Cladiella, Lobophytum, and Sarcophyton at Dongsha Atoll. The problems of species identification in these genera are similar to those in Sinularia, with the additional complication of a lack of congruence between morphologically defined genera and molecular clades (McFadden et al. 2006; Benayahu et al. 2012). McFadden et al. (2006) first recognized that Sarcophyton and Lobophytum comprise three morphologically and phylogenetically distinct clades, one of which includes nominal species of both genera (the ‘mixed’ clade). Suggestions that this third, ‘mixed’ clade should be considered a separate genus have been further complicated by the subsequent recognition that some nominal species of both genera fall outside of those three main clades (Benayahu et al. 2012), including S. crassocaule, S. tumulosum and L. rigidum (Fig. 4). Reconciliation of the genus-level taxonomy with the observed phylogenetic relationships among species will require either recognizing each distinct clade (i.e. clades M, C, S and L in Fig. 4) as a separate genus or, alternatively, synonymizing Lobophytum with Sarcophyton in order to recognize a single genus that encompasses a range of distinct morphologies. The current taxonomic division between Cladiella and Klyxum also does not reflect the phylogenetic relationships among species assigned to those genera (Benayahu et al. 2012). Molecular evidence suggests that C. australis (Fig. 7E), the species of Cladiella most commonly encountered at Dongsha, actually belongs to the genus Klyxum. Indeed, both C. australis and C. kashmani (an Indian Ocean species) bear some morphological resemblance to Klyxum spp. as their colonies are rather flaccid in contrast to most typical Cladiella spp. (personal observations). In addition, both C. australis and C. kashmani feature elongated sclerites that differ from the usual dumbbells found in Cladiella (Benayahu and Schleyer 1996: figs. 4, 8-11). At both of the mitochondrial barcode loci we sequenced (mtMutS and igr1+COI), C. australis and the unidentified species of Klyxum collected at Dongsha shared identical haplotypes, which differed from other Cladiella species by 1.5%. At the nuclear 28S rDNA locus, C. australis and Klyxum sp. differed from one another by 1.5% compared to a 7% average distance of both species from other Cladiella. Reassignment of C. australis (and C. kashmani) to Klyxum will necessitate revising the diagnosis of that genus to accommodate their different sclerite morphologies. In addition to the observed incongruence between morphologically defined genera and molecular clades in Sarcophyton, Lobophytum, Cladiella, and Klyxum, species in these genera also exemplify the previously discussed challenges of using molecular barcodes to assign binomials when reference sequences are not derived from type material. For example, specimens identified in this study as Sarcophyton crassocaule (based on morphology) are not a close genetic match to reference material collected at a different geographic location and identified as that same species by a different taxonomic expert (Fig. 4). No sequence data are currently available for type material of S. crassocaule, and the high degree of intraspecific morphological variation exhibited by this species has been noted previously, indicating some confusion regarding its morphology-based diagnosis (Verseveldt 1982). Further in-depth study is necessary to reconcile the morphological and genetic variation in this species, as well as its apparently close relationship to L. rigidum (Fig. 3F). The only morphological description of L. rigidum is presented in the original description of its type colony (Benayahu 1995), and there are no data on intraspecific variability in that species. Similarly, specimens identified in other studies as Lobophytum crassum exhibit considerable sequence variation, and some haplotypes match reference sequences attributed to different morphospecies (Fig. 4). Interestingly, this species features a bewildering array of colony and sclerite variation (e.g. Verseveldt 1983: 25-32), which might also explain the noted sequence variation. Without further examination and sequencing of type material it is not clear if all of these genetically page 22 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan different forms are indeed L. crassum, or if the observed genetic polymorphisms are indicative of cryptic species. The same is true for Cladiella pachyclados (see Benayahu et al. 2012). Until sequences can be obtained from type specimens, or from newer material collected from type localities that is a close morphological match to the original types (e.g. Ofwegen et al. 2016), the use of molecular barcodes to assign Latin binomials in these genera will continue to be fraught with uncertainty. CONCLUSIONS Dongsha Atoll features a highly diverse octocoral fauna. Octocorals are the dominant benthic organisms at the surveyed sites, and members of the family Alcyoniidae—mainly the genera Lobophytum, Sarcophyton and Sinularia— may constitute 60-70% live coverage on some of the reefs (personal observations). Our photographic records also reveal the competitive abilities of certain octocorals to dominate reef areas and successfully overgrow some stony corals (Figs. 7F; 8A-C) or to hamper their growth when they closely interact (Fig. 8D). Undoubtedly, the exact nature of such competitive interactions should be further studied along with the adaptive features of Dongsha octocorals in order to understand their success there. Integrative approaches that combine morphological, molecular, and other lines of evidence to distinguish among species have greatly facilitated our ability to assess the specieslevel diversity of octocorals and to compare community compositions among diverse locations (McFadden et al. 2014). As shown in the current Fig. 8. Underwater photographs of Dongsha Atoll octocorals: (A) Litophyton sp. colonies densely occupy reef space, (B) colony of Sarcophyton ehrenbergi overgrowing Acropora sp., (C) colonies of Sinularia penghuensis surrounding colony of the stony coral Symphyllia agaricia and (D) colony of S. ehrenbergi inhibiting growth of Acropora sp. (A) (C) (B) (D) page 23 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan study and elsewhere (McFadden et al. 2011), the use of character-based barcodes rather than genetic distance thresholds to distinguish taxa can overcome some – but not all – of the limitations imposed by the low levels of sequence variation that characterize the octocoral mitochondrial genome in general, and the most commonly used barcoding loci in particular. Although integrated approaches facilitate the recognition of cryptic species and have led to the correction of longstanding taxonomic errors (e.g. Ofwegen et al. 2013 2016), assignment of binomials to octocorals still remains a considerable challenge. Until reference sequences are available for a majority of museum types or taxonomically validated exemplars (e.g. recently collected material from type localities that has been compared directly to types, if they are available), the use of molecular barcodes to assign binomials will remain impossible in many genera, such as most of the Nephtheidae obtained at Dongsha. In those genera in which many barcodes are associated with reference specimens whose identity has not been validated by direct comparison to type material (i.e. most Alcyoniidae), the assignment of binomials remains susceptible to the inadvertent propagation of taxonomic errors. Although molecular barcoding is already a valuable approach to discriminating taxa and quantifying biodiversity in octocorals, most genera will require taxonomic revisions that include molecular characterization of species before it will be possible to use barcodes to assign Latin binomials with any certainty. Acknowledgments: This study was made possible by a grant to YB from the Taiwanese Ministry of Science (MOST) to conduct octocoral surveys in Dongsha. We thank the staff members of Dongsha Atoll National Park, Dongsha Atoll Research Station (DARS), and Biodiversity Research Center, Academia Sinica (BRCAS) for assistance during the field work. We thank V. Wexler for assistance with graphics, M. Weis for technical assistance, and A. Gonzalez, L. Mattson and M. Blevins for assistance with DNA sequencing. N. Paz is acknowledged for skillful editorial assistance. SWD, PH, NHI, AC and CSM were supported in part by the Howard Hughes Medical Institute Undergraduate Science Education Program award #52007544 to Harvey Mudd College. Authors’ contributions: The contributions of the coauthors are as follows: YB field work, taxonomy and writing of MS, LPO taxonomy, CfD writing, MSJ facilities and field work, KS writing and facilities, AS curatorial skills, SWD, PH, NHI, AC, TW molecular work, CSM field work, phylogeny, writing of MS. Competing interests: YB, LPO, CfD, MSJ, KS, AS, SWD, PH, NHI, AC, TW and CSM have no conflicts of interest. Availability of data and materials: Sequences are available in the GenBank and material in the Steinhardt Museum of Natural History, Tel Aviv University, Israel. Consent for publication: Not applicable. Ethics approval consent to participate: Not applicable. REFERENCES Benayahu Y. 1995. Species composition of soft corals (Octocorallia, Alcyonacea) on the coral reefs of Sesoko Island, Ryukyu Archipelago, Japan. Galaxea 12:103-124. Benayahu Y, Jeng MS, Perkol-Finkel S, Dai CF. 2004. Soft corals (Octocorallia, Alcyonacea) from southern Taiwan. II. Species diversity and distributional patterns. Zool Stud 43:548-560. Benayahu Y, McFadden CS. 2011. A new genus of soft coral of the family Alcyoniidae (Cnidaria, Octocorallia) with redescription of a new combination and description of a new species. ZooKeys 84:1-11. Benayahu Y, Perkol-Finkel S. 2004. Soft corals (Octocorallia: Alcyonacea) from southern Taiwan. I. Sarcophyton nanwanensis sp. n. (Octocorallia: Alcyonacea). Zool Stud 43:537-547. Benayahu Y, Schleyer MH. 1996. Corals of the South-West Indian Ocean III. Alcyonacea (Octocorallia) of Bazaruto Island, Mozambique: a redescription of Cladiella australis (Macfadyen, 1936) and description of Cladiella kashmani spec. nov. Oceanogr Res Inst Invest Rep 69:1-22. Benayahu Y, van Ofwegen LP. 2011. New species of octocorals (Coelenterata: Anthozoa) from the Penghu Archipelago, Taiwan. Zool Stud 50:350-362. Benayahu Y, van Ofwegen LP, Alderslade P. 1998. A case study of variation in two nominal species of Sinularia (Coelenterata: Octocorallia), S. brassica May, 1898, and S. dura (Pratt, 1903), with a proposal for their synonymy. Zool Verhandel 323(23):277-309. Benayahu Y, van Ofwegen LP, Dai CF, Jeng MS, Soong K, Shlagman A, Hsieh HJ, McFadden CS. 2012. Diversity, distribution, and molecular systematics of octocorals (Coelenterata: Anthozoa) of the Penghu Archipelago, Taiwan. Zool Stud 51:1529-1548. Bergmann T, Rach J, Damm S, DeSalle R, Schierwater B, Hadrys H. 2013. The potential of distance-based thresholds and character-based barcoding for defining taxonomic problematic entities by CO1 and ND1. Mol Ecol Res 13:1069-1081. doi:10.1111/1755-0998.12125. page 24 of 26Zoological Studies 57: 50 (2018) © 2018 Academia Sinica, Taiwan Dai CF. 1990. Interspecific competition in Taiwanese corals with special reference to interactions between alcyonacean and scleractinian corals. Mar Ecol Progr Ser 60:291-297. doi:10.3354/meps060291. Dai CF. 1991a. Reef environment and coral fauna of southern Taiwan. Atoll Res Bull 354:1-28. doi:10.5479/ si.00775630.354.1. Dai CF. 1991b. Distribution and adaptive strategies of alcyonacean corals in Nanwan Bay, Taiwan. Hydrobiologia 216:241-246. doi:10.1007/BF00026469. Dai CF. 1993. Patterns of coral distribution and benthic space partitioning on the fringing reefs of southern Taiwan. Marine Ecology 14(3):185-204. doi:10.1111/j.14390485.1993.tb00479.x. Dai CF. 2005. Coral reefs in Dongsha Island: past, present and future. In Proc. 10th Coral Reef Symp. Okinawa. Dai CF, Chin CH. 2017. Octocoral Fauna of Dongsha Atoll. Marine National Park Heaquarters, Kaohsiung, Taiwan. Dai CF, Fan TY, Wu CS. 1995. Coral fauna of Tungsha Tao (Pratas Islands). Acta Oceanographica Taiwanica 34(3):116. Darriba D, Taboada GL, Doallo, Posada D. 2012. jModelTest 2: more models, new heuristics and parallel computing. Nat Methods 9:772. doi:10.1038/nmeth.2109. DeCarlo TM, Karnauskas KB, Davis KA, Wong GT. 2015. Climate modulates internal wave activity in the Northern South China Sea. Geophys Res Lett 42(3):831-838. doi:10.1002/2014GL062522. DeSalle R, Egan MG, Siddall M. 2005. The unholy trinity: taxonomy, species delimitation and DNA barcoding. Phil Trans R Soc B 360:1905-1916. doi:10.1098/ rstb.2005.1722. Fabricius K, Alderslade P. 2001. Soft corals and sea fans: a comprehensive guide to the tropical shallow-water genera of the Central West-Pacific, the Indian Ocean and the Red Sea. Australian Institute of Marine Science, Townsville. Fan TY, Chou YH, Dai CF. 2005. Sexual reproduction of the alcyonacean coral Lobophytum pauciflorum in southern Taiwan. Bull Mar Sci 76(1):143-154. Fang LS, Shao KT, Liu L, Li JJ. 1990. A survey of the marine ecological resources of Dong-Sha Atoll. Fishery Management Department, Kaohsiung City government, Taiwan, 61 pp. (in Chinese with English inventory) Hsu MK, Liu AK. 2000. Nonlinear internal waves in the South China Sea. Can J Remote Sens 26(2):72-81. doi:10.1080 /07038992.2000.10874757. Jeng MS, Dai CF, Chen JP. 2011b. The assessment of ecological status and change trends of coral reefs at Dongsha Atoll. Marine National Park Headquarters, Construction and Planning Agency, Ministry of the Interior, R.O.C. (in Chinese with English abstract) Jeng MS, Dai CF, Chen JP, Wang WL, Meng PJ. 2008. Coral reef resource surveys and monitoring of Dongsha Atoll. Marine National Park Headquarters, Construction and Planning Agency, Ministry of the Interior, R.O.C. (in Chinese with English abstract) Jeng MS, Huang HD, Dai CF, Hsiao YC, Benayahu Y. 2011a. Sclerite calcification and reef-building in the fleshy octocoral genus Sinularia (Octocorallia: Alcyonacea). Coral Reefs 30:925-933. Katoh K, Kuma K, Toh H, Miyata T. 2005. MAFFT version 5: improvement in accuracy of multiple sequence alignment. Nucleic Acids Res 33:511-513. doi:10.1093/nar/gki198. Liu AK, Su FC, Hsu MK, Kuo NJ, Ho CR. 2013. Generation and evolution of mode-two internal waves in the South China Sea. Cont Shelf Res 59:18-27. doi:10.1016/ j.csr.2013.02.009. Maddison DR, Maddison WP. 2005. MacClade 4. Sinauer Associates Inc., Sunderland, MA. McFadden CS, Alderslade P, van Ofwegen LP, Johnsen H, Rusmevichientong A. 2006. Phylogenetic relationships within the tropical soft coral genera Sarcophyton and Lobophytum (Anthozoa, Octocorallia). Invertebr Biol 125:288-305. doi:10.1111/j.1744-7410.2006.00070.x. McFadden CS, van Ofwegen LP, Beckman EJ, Benayahu Y, Alderslade P. 2009. Molecular systematics of the speciose Indo-Pacific soft coral genus, Sinularia (Anthozoa: Octocorallia). Invertebr Biol 128:303-323. McFadden CS, Benayahu Y, Pante E, Thoma JN, Nevarez PA, France SC. 2011. Limitations of mitochondrial gene barcoding in Octocorallia. Mol Ecol Resour 11:19-31. doi:10.1111/j.1755-0998.2010.02875.x. McFadden CS, Brown AS, Brayton C, Hunt CB, van Ofwegen LP. 2014. Application of DNA barcoding to biodiversity studies of shallow-water octocorals: molecular proxies agree with morphological estimates of species richness in Palau. Coral Reefs 33:275-286. doi:10.1007/s00338-0131123-0. van Ofwegen LP, Benayahu Y. 2012. Two new species and a new record of the genus Sinularia (Octocorallia, Alcyonacea) from the Penghu Archipelago, Taiwan. Zool Stud 51:283-398. van Ofwegen LP, Benayahu Y, McFadden CS. 2013. Sinularia leptoclados (Ehrenberg, 1834) (Cnidaria, Octocorallia) re-examined. ZooKeys 272:29. doi:10.3897/ zookeys.272.4406. van Ofwegen LP, McFadden CS, Benayahu Y. 2016. Sinularia polydactyla (Ehrenberg, 1834) (Cnidaria, Octocorallia) reexamined, with the description of a new species. ZooKeys 581:71. doi:10.3897/zookeys.581.7455. Paknia O, Bergmann T, Hadrys H. 2015. Some ‘ant’swers: application of a layered barcode approach to problems in ant taxonomy. Mol Ecol Resour 15:1262-1274. doi:10.1111/1755-0998.12395. Park E, Hwang DS, Lee JS, Song JI, Seo TK, Won YJ. 2012. Estimation of divergence times in cnidarian evolution based on mitochondrial protein-coding genes and the fossil record. Mol Phylogenet Evol 62:329-345. doi:10.1016/j.ympev.2011.10.008. Puillandre N, Bouchet P, Boisselier-Dubayle MC, Brisset J, Buge B, Castelin M, Chagnoux S, Cristophe T, Corbari L, Lambourdiere J, Lozouet P, Marani G, Rivasseau A, Silva , Terryn Y, Tillier S, Utge J, Samadi S. 2012. New taxonomy and old collections: integrating DNA barcoding into the collection curation process. Mol Ecol Resour 12:396-402. doi:10.1111/j.1755-0998.2011.03105.x. Puillandre N, Macpherson E, Lambourdiere J, Cruaud C, Boisselier-Dubayle MC, Samadi S. 2011. Barcoding type specimens helps to identify synonyms and an unnamed new species in Eumunida Smith, 1883 (Decapods: Eumunididae). Invertebr Syst 25:322-333. Rach J, DeSalle R, Sarkar IN, Schierwater B, Hadrys H. 2008. Character-based DNA barcoding allows discrimination of genera, species and populations in Odonata. P R Soc B 275:237-247. doi:10.1098/rspb.2007.1290. Ribas-Deulofeu L, Denis V, De Palmas S, Kuo CY, Hsieh HJ, Chen CA. 2016. Structure of benthic communities along the Taiwan latitudinal gradient. PLoS ONE page 25 of 26Zoological Studies 57: 50 (2018)