A peer-reviewed open-access journal PhytoKeys 80: 113–120 (2017) Macrosolen bidoupensis (Loranthaceae), a new species... doi: 10.3897/phytokeys.80.13338 113 RESEARCH ARTICLE http://phytokeys.pensoft.net Launched to accelerate biodiversity research Macrosolen bidoupensis (Loranthaceae), a new species from Bidoup Nui Ba National Park, southern Vietnam Shuichiro Tagane1, Van Son Dang2, Nguyen Van Ngoc1,3, Hoang Thi Binh1,3, Natsuki Komada4, Jarearnsak Sae Wai5,6, Akiyo Naiki7, Hidetoshi Nagamasu8, Hironori Toyama1, Tetsukazu Yahara1 1 Center for Asian Conservation Ecology, Kyushu University, 744 Motooka, Fukuoka, 819-0395, Japan 2 The VNM Herbarium, Institute of Tropical Biology, Vast, 85 Tran Quoc Toan Street, District 3, Ho Chi Minh City, Vietnam 3 Department of Biology, Dalat University, 01 – Phu Dong Thien Vuong, Dalat, Vietnam 4 Laboratory of Forest Resources and Society, Graduate School of Agriculture Kyoto University, Kitashirakawaoiwakecho, Sakyo-ku, Kyoto, 606-8502, Japan 5 Prince of Songkla University, Hat Yai, Thailand 6 Institute of Ecology and Evolutionary Biology, National Taiwan University, Taipei, Taiwan 7 Iriomote Station, Tropical Biosphere Research Center, University of the Ryukyus, 870 Uehara, Taketomi-cho, Yaeyama-gun, Okinawa, 907-1541, Japan 8 The Kyoto University Museum, Kyoto University, Yoshida Honmachi, Sakyo-ku, Kyoto, 606-8501, Japan Corresponding author: Shuichiro Tagane (stagane29@gmail.com) Academic editor: D. Nickrent | Received 21 April 2017 | Accepted 15 May 2017 | Published 5 June 2017 Citation: Tagane S, Dang VS, Ngoc NV, Binh HT, Komada N, Wai JS, Naiki A, Nagamasu H, Toyama H, Yahara T (2017) Macrosolen bidoupensis (Loranthaceae), a new species from Bidoup Nui Ba National Park, southern Vietnam. PhytoKeys 80: 113–120. https://doi.org/10.3897/phytokeys.80.13338 Abstract Macrosolen bidoupensis Tagane & V.S.Dang, sp. nov. (Loranthaceae) is newly described from Bidoup Nui Ba National Park in Lam Dong Province, southern Vietnam. The new species is characterized by small broadly elliptic to circular leaves, sessile to short petioles, slightly cordate to rounded leaf bases, 4–5 pairs of lateral veins and a basally green corolla tube. An illustration, a summary of DNA barcoding of the plastid genes rbcL and matK, and a key to the species of Macrosolen in Vietnam are provided. Keywords Bidoup Nui Ba National Park, DNA barcoding, Loranthaceae, Macrosolen, new species, Vietnam Copyright Shuichiro Tagane 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. 114 Shuichiro Tagane et al. / PhytoKeys 80: 113–120 (2017) Introduction Macrosolen (Blume) Rchb. is a small parasitic shrub in the family Loranthaceae. The genus is characterized by being totally glabrous with 6-merous flowers subtended by one bract and two bracteoles (often connate), reflexed corolla lobes at anthesis, and 4-locular anthers. The genus comprises ca. 40 species widely distributed in tropical South and Southeast Asia. In Vietnam, it has been recorded by de Loureiro (1790, as Loranthus), Lecomte (1915, as Elytranthe), Danser (1938) and Hô (2003), and recently a taxonomic revision of Loranthaceae in the country by Han (2014) enumerated seven species of Macrosolen: M. annamicus Danser, M. avenis Danser, M. bibracteolatus (Hance) Danser, M. cochinchinensis (Lour.) Tiegh., M. dianthus Danser, M. robinsonii (Gamble) Danser and M. tricolor (Lecomte) Danser. During botanical surveys in Bidoup Nui Ba National Park, Lam Dong Province, southern Vietnam in 2016, we encountered an undescribed species of Macrosolen. We here describe this new species as Macrosolen bidoupensis Tagane & V.S.Dang, and provide illustrations, and a key to the species of Macrosolen in Vietnam. In addition to the morphological examination, DNA sequences are extremely helpful for delimiting species (Hebert and Gregory 2005, Dick and Webb 2012). We sequenced two DNA barcode regions of rbcL and matK following the recommendation of CBOL Plant Working Group (2009). Materials and methods Morphological observations To verify the validity of the new species, we undertook morphological comparisons to closely related species based on herbarium specimens (ANDA, BKF, BO, BRUN, FU, HN, KYO, SAR, TNS, VNM), specimen images on the web (e.g. JSTOR Global Plants, http://plants.jstor.org/ and Muséum National d’Histoire Naturelle, http:// www.mnhn.fr/), and consulting the relevant literature (e.g. Barlow 1995, 1997, 2002, Qiu and Gilbert 2003, Hô 2003, Han 2014). DNA barcoding DNA amplification and sequencing of portions of the chloroplast genes rbcL and matK followed established protocols (Kress et al. 2009, Dunning and Savolainen 2010) as in Toyama et al. (2015). Macrosolen bidoupensis (Loranthaceae), a new species... 115 Taxonomy Macrosolen bidoupensis Tagane & V.S.Dang, sp. nov. urn:lsid:ipni.org:names:77163237-1 Figures 1, 2 Diagnosis. Macrosolen bidoupensis is similar to Macrosolen tricolor Danser of China, Laos and Vietnam in leaf shape and size and by possession of a 2-flowered umbel, but differs in having a sessile to very short petiole (up to 0.7 mm long in M. bidoupensis vs. 2–3 mm long in M. tricolor), more lateral veins (4 or 5 vs. 2 or 3 pairs), slightly cordate to rounded leaf bases (vs. cuneate) and a basally green corolla tube (vs. reddish around the basal 1/3). The leaf shape is also somewhat similar to Macrosolen platyphyllus Danser of Thailand, Peninsular Malaysia and Borneo, but easily distinguished in having much smaller leaves and flowers, and very short petioles. Type. VIETNAM. Lam Dong Province, Bidoup Nui Ba National Park, in lower montane evergreen forest, 12°10'34.7"N, 108°41'08.4"E, alt. 1533 m, 21 Feb. 2016, with flowers, Tagane S., Nagamasu H., Naiki A., Dang V. Son, Ngyuen V. Ngoc, Binh T. Hoang & Wai J. V4083 (holotype-KYO!, fl. spirit collection; isotypes NTU!, the herbarium of Bidoup Nui Ba National Park!, VNM!). Description. Hemi-parasitic shrub, 25–40 cm tall, totally glabrous. Branches terete, grayish to grayish brown, lenticellate. Leaves opposite; blade elliptic, broadly elliptic, broadly ovate-elliptic, or circular, 1.2–4.8 × 1.2–3.5 cm, coriaceous, apex broadly obtuse to rounded, base slightly cordate to rounded, margin entire, grayish green and slightly lustrous adaxially, dull olive abaxially, midrib prominent at basal 1/3 to 1/2 abaxially, secondary veins 4 or 5 pairs, obscure or visible adaxially, obscure to faintly visible abaxially; petiole to 0.7 mm long. Inflorescences axillary, sometimes at older leafless nodes, 2-flowered umbels; peduncle ca. 0.9 mm long. Pedicel 1.1–1.5 mm long; central bract suborbicular, ca. 1 mm long, bracteoles suborbicular, connate, ca. 1 mm long; mature buds 2.8–3.2 cm long; calyx limb (calyculus) annular, ca. 0.7 mm long; corolla green tinged with red at top of tube and lobes, symmetrically 6-winged at about two thirds the length, tube 2.2–2.4 cm long in anthesis, gradually dilated, inflated, slightly curved, lobes lanceolate, 6–8 mm long, reflexed in anthesis; stamens 6, anthers ca. 2.3 mm long, filaments basally adnate to corolla tube, free part 2.3–3.3 mm long; ovary 2.8–3.3 mm long; style 2.8–3.2 cm long, stigma capitate. Berry reddish orange, subglobose to slightly depressed-barrel-shaped, ca. 8 mm high, ca. 6 mm wide, ovoid when dry, apex beaked by calyx limb. Seed 1, ellipsoidal, ca. 5 mm high, ca. 3 mm wide, longitudinally 6-grooved. The measurements of the flower characters are derived from the spirit collection. Other specimen examined in Vietnam. Bidoup Nui Ba National Park; in lower montane evergreen forest, 12°10'34.9"N, 108°41'04.4"E, alt. 1533 m, 22 Feb. 2016, with fruits, Tagane et al.V4169 (FU, NTU, the herbarium of Bidoup Nui Ba National Park, VNM). 116 Shuichiro Tagane et al. / PhytoKeys 80: 113–120 (2017) Figure 1. Macrosolen bidoupensis Tagane & V.S.Dang. A Branch with flower buds B 2-flowered inflorescence C Flower with corolla tube opened D Fresh fruit E Dried fruit F Seed G seed (left) and longitudinal section of seed (right). Materials: Tagane et al. V4083 (KYO). Drawn by N. Komada. Macrosolen bidoupensis (Loranthaceae), a new species... 117 Figure 2. Macrosolen bidoupensis Tagane & V.S.Dang. A Branch with flower buds B Flower C Fruits. Photos by Tagane et al. V4083. 118 Shuichiro Tagane et al. / PhytoKeys 80: 113–120 (2017) Phenology. Flowering and fruiting specimens were collected in February. Distribution and habitat. Vietnam (so far known only from Bidoup Nui Ba National Park, Lam Dong Province). In lower montane evergreen forest, ca. 1500 m altitude. GenBank accession no. Tagane et al. V4083: LC259010 (rbcL), LC259011 (matK). The partial rbcL sequence of M. bidoupensis was identical to M. tricolor (GenBank accession no. HQ317771) of the total 517 bp and differed at 3 bases of the 518 total from M. cochinchinensis (KP094775 and HQ317768). The partial matK sequence of M. bidoupensis differed at 23 bases from M. cochinchinensis (EU544439) of the 813 total. Etymology. The specific epithet bidoupensis is derived from its type locality. Conservation status. Least Concern (LC). Bidoup Nui Ba National Park is located on Central highland of Vietnam. It covers ca. 700.38 km2 and is the core zone of the Langbiang biosphere reserve (Bidoup-Nui Ba National Park 2016). Macrosolen bidoupensis is commonly found in the lower montane evergreen forest in the national park. Since the forest is widely seen and well-protected, we suggest this status as LC according to IUCN (2012). Note. A similar species, Macrosolen tricolor is known from lower elevations (below 100 m, Qiu and Gilbert 2003), and the two species do not occur sympatrically. Key to the species of Macrosolen in Vietnam 1 – 2 – 3 – 4 – 5 – 6 – 7 – Leaf blade 1.2–5.5 × 1.2–3.5 cm, apex rounded to broadly obtuse .............2 Leaf blade (3–)4–15 × (1.2–)1.5–7 cm, apex acute to acuminate, or obtuse .... 3 Petiole to 0.7 mm long; lateral veins 4 or 5 pairs .................. M. bidoupensis Petiole 2–3 mm long; lateral veins 2 or 3 pairs ............................ M. tricolor Mature flower bud 1–1.5(–2.3) cm long .....................................................4 Mature flower bud 3–8.5 cm long ..............................................................5 Petiole (1–)5–10 mm long; inflorescences racemose, (2–)4–8-flowered; peduncle (5–)15–20 mm long ........................................... M. cochinchinensis Petiole 2–4 mm long; inflorescences umbellate, 2- or 3-flowered; peduncle 1–2.5 mm long .......................................................................M. robinsonii Flowers subsessile, pedicel less than 1 mm long; bract 2–3 mm long; anther 5–8 mm long ............................................................................ M. dianthus Flowers distinctly pedicellate, pedicel 2–6 mm long; bract 1–1.5 mm long; anther 2–5 mm long ...................................................................................6 Mature flower bud more than 4.5 cm long; anthers 4–6 mm long ............... ..............................................................................................M. annamicus Mature flower bud 3–4 cm long; anthers 2–3 mm long ..............................7 Corolla tube curved near the middle, asymmetrically winged........ M. avenis Corolla tube curved above the middle, symmetrically winged ....................... ........................................................................................ M. bibracteolatus Macrosolen bidoupensis (Loranthaceae), a new species... 119 Acknowledgements Authors thank the staff of Bidoup Nui Ba National Park for their kind assistance in the field survey. We are grateful to Keiko Mase for her help with DNA barcoding, and to the curators and staff of the following herbaria: ANDA, BKF, BO, BRUN, FU, HN, KYO, SAR, TNS, VNM for allowing us to examine their collections. We also thank the editors of the journal and two anonymous reviewers and for their help and advice. The present study was supported by the Ministry of the Environment, Japan (4-1601) and was partially supported by JSPS KAKENHI Grant Number 15H02640 and 24255007. References Barlow BA (1995) New and noteworthy Malesian species of Loranthaceae. Blumea 40: 15–31. Barlow BA (1997) Loranthaceae. Flora Malesiana (ser. I) 13: 209–442. Barlow BA (2002) Loranthaceae. Flora of Thailand 7(4): 665–706. Bidoup Nui Ba National Park, (2016–onwards) official website, http://bidoupnuiba.gov.vn/en/ introduction-menu.html [Accessed 14 May, 2017] CBOL Plant Working Group (2009) A DNA barcode for land plants. Proceedings of the National Academy of Sciences of the United States of America 106: 12794–12797. https:// doi.org/10.1073/pnas.0905845106 Danser BH (1938) The Loranthaceae of French Indo-China and Siam. Bulletin du Jardin Botanique de Buitenzorg, Sér. 3, 16: 1–63. Dick CW, Webb CO (2012) Plant DNA barcodes, taxonomic management, and species discovery in tropical forests. In: Kress WJ, Erickson DL (Eds) DNA Barcodes. Springer, New York, NY. Methods and Protocols 858: 379–393. https://doi.org/10.1007/978-1-61779-591-6_18 Dunning LT, Savolainen V (2010) Broad-scale amplification of matK for DNA barcoding plants, a technical note. Botanical Journal of the Linnean Society 164: 1–9. https://doi. org/10.1111/j.1095-8339.2010.01071.x Han LN (2014) Taxonomy of Loranthaceae in Vietnam. Master thesis, Published by author, Ha Noi, 161 pp. [In Vietnamese] Hebert PDN, Gregory TR (2005) The promise of DNA barcoding for taxonomy. Systematic Biology 54: 852–859. https://doi.org/10.1080/10635150500354886 Hô PH (2003) Cay Co Viet Nam: An Illustrated Flora of Vietnam Vol. 2. Published by the author, Montreal. IUCN (2012) IUCN Red List Categories and Criteria: version 3.1. second edition. IUCN, Gland and Cambridge. Kress WJ, Erickson DL, Jones FA, Swenson NG, Perez R, Sanjur O, Bermingham E (2009) Plant DNA barcodes and a community phylogeny of a tropical forest dynamics plot in Panama. Proceedings of the National Academy of Sciences of the United States of America 106(44): 18621–18626. https://doi.org/10.1073/pnas.0909820106 120 Shuichiro Tagane et al. / PhytoKeys 80: 113–120 (2017) Lecomte H (1915) Loranthaceae. In: Lecomte PH, Gagnepain F (Eds) Flore Générale de l’Indo-Chine 5, Masson, Paris, 185–210. de Loureiro J (1790) Flora Cochinchinensis 1. Typis et expensis Academicis, Ulyssepone. Qiu HX, Gilbert MG (2003) Loranthaceae. In: Wu ZY, Raven PH, Hong DY (Eds) Flora of China 5. Beijing, Science Press and Missouri Botanical Garden Press, St. Louis, 220–239. http://www.efloras.org Toyama H, Kajisa T, Tagane S, Mase K, Chhang P, Samreth V, Ma V, Sokh H, Ichihasi R, Onoda Y, Mizoue N, Yahara T (2015) Effects of logging and recruitment on community phylogenetic structure in 32 permanent forest plots of Kampong Thom, Cambodia. Philosophical Transactions of the Royal Society B: Biological Sciences 370(1662): 20140008. https://doi.org/10.1098/rstb.2014.0008