See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/323729283 Petrocodon asterocalyx, a new species of Gesneriaceae from Guangxi, China Article in Phytotaxa · March 2018 DOI: 10.11646/phytotaxa.343.3.6 CITATION READS 1 181 7 authors, including: Long-Fei Fu Yi-Gang Wei Guangxi Institute of Botany, Guangxi Zhuang Autonomous Region and Chinese Ac… Guangxi Institute of Botany, CAS 30 PUBLICATIONS 63 CITATIONS 111 PUBLICATIONS 678 CITATIONS SEE PROFILE SEE PROFILE Stephen Maciejewski Michael Lofurno 6 PUBLICATIONS 11 CITATIONS 2 PUBLICATIONS 1 CITATION SEE PROFILE SEE PROFILE Some of the authors of this publication are also working on these related projects: Urticaceae systematics View project Karst vascular plant diversity View project All content following this page was uploaded by Long-Fei Fu on 23 April 2018. The user has requested enhancement of the downloaded file. Phytotaxa 343 (3): 259–268 ISSN 1179-3155 (print edition) http://www.mapress.com/j/pt/ PHYTOTAXA Copyright © 2018 Magnolia Press Article ISSN 1179-3163 (online edition) https://doi.org/10.11646/phytotaxa.343.3.6 Petrocodon asterocalyx, a new species of Gesneriaceae from Guangxi, China RUI-LI ZHANG1, LONG-FEI FU2, 3, SHU LI2, 3, YI-GANG WEI2, 3, STEPHEN MACIEJEWSKI2, 4, MICHAEL LOFURNO5 & FANG WEN2, 3, * 1College of Landscape Architecture,Beijing University of Agriculture, Beijing Laboratory of Urban and Rural Ecological Environment, Beijing Engineering Research Center of Rural Landscape Planning and Design, CN-102206, Beijing, P.R. China. 2Guangxi Key Laboratory of Plant Conservation and Restoration Ecology in Karst Terrain, Guangxi Institute of Botany, Guangxi Zhuang Autonomous Region and Chinese Academy of Sciences, Guilin, CN-541006, P.R. China. 3Gesneriad Conservation Center of China (GCCC), Guilin Botanical Garden, No. 85, Yanshan Town, Guilin, CN-541006, P.R. China. 4The Gesneriad Society, 2030 Fitzwater Street, Philadelphia, PA 19146, USA. 5Landscape Architect, Philadelphia. PA 19146, USA. *Corresponding author: [email protected] Abstract Petrocodon asterocalyx F.Wen, Y.G.Wei & R.L.Zhang, a new species from the Danxia landform area in Guangxi, South China, is described and illustrated based on molecular and morphological data. The molecular evidence shows that the new species is recovered in a weakly supported clade. Within this clade, the new one is morphologically similar to P. hancei (Hemsl.) A.Weber & Mich.Möller and P. coriaceifolius (Y.G.Wei) Y.G.Wei & Mich.Möller, and it can be distinguished from the former by calyx lobes 20–40 × 2–3 mm, corolla 2.5–3.0 cm long, filaments sparsely erectly pubescent, anthers sparsely pubescent, staminodes 3, and stigmas 2; from latter by leaf blades rhombic-oblong or rhombic, base shallowly cuneate, margin crenulate to serrate, calyx lobes linear, 20–40 × 2–3 cm, and anthers 3.5–3.8 mm long, sparsely pubescent and el- liptical. Key words: Danxia landform, endemism, flora of Guangxi, new taxon, taxonomy Introduction Petrocodon Hance (1883: 167) originally was a small endemic genus in China, with three species and one variety: P. dealbatus Hance (1883: 167), P. dealbatus var. denticulatus (W.T.Wang 1975: 101) W.T.Wang (1990: 420), P. ferrugineus Y.G.Wei (2007: 135) and P. multiflorus F.Wen & Y.S.Jiang in Jiang et al. (2010: 57) before 2011. Wang et al. (2011) preliminarily revised Petrocodon and its relative genera in early 2011. However, only four species from four monospecific genera, Petrocodon, Calcareoboea C.Y.Wu ex H.W.Li (1982: 241), Tengia Chun (1946: 279) and Paralagarosolen Y.G.Wei (2004: 528), were collected and studied. It resulted in the conclusion of revision for previous Petrocodon sensu stricto and its congeners were imperfect because of lacking some necessary and sufficient samples. Soon afterwards, Weber et al. (2011b) revised Petrocodon again late in 2011. The newly defined Petrocodon sensu lato has been swelled to contain at least 31 species and one variety by incorporating some Chinese monotypic genera, i.e. Calcareoboea, Dolicholoma D.Fang & W.T.Wang in Wang (1983: 18), Paralagarosolen and Tengia, some species of two Chinese endemic genera: Lagarosolen W.T.Wang (1984b: 12) (six species), Wentsaiboea D.Fang & D.H.Qin (2004: 533) (one species), four species of Didymocarpus Wallich (1819: 378) and one species of Primulina Hance (1883: 169) (P. guangxiensis Yan Liu & W.B.Xu in Liu et al. (2011: 1)) as well (Xu et al. 2014). It also includes 13 species described after 2011 (Möller et al. 2016, Lu et al. 2017a & 2017b). Almost all known species can be found in limestone areas, with one notable exception, Petrocodon hancei (Hemsley 1890: 299) A.Weber & Mich.Möller in Weber et al. (2011b: 59). It grows on all kinds of substrates from different parent rocks, such as limestone, sandstone, shale and glutenite. When we investigated the transition areas of limestone and Danxia areas in Lechang city, Guangdong (Liu & Huang 1991), we observed many populations of P. hancei from limestone areas, transition zones and Danxia areas. Thus, when we collected some specimens of an unknown species of Petrocodon without flowers from the Danxia area in Ziyuan County, Guangxi in 2005 (Huang et Accepted by Hai He: 11 Feb. 2018; published: 12 Mar. 2018 259 al. 2015a & 2015b), we improperly identified it as P. hancei due to the similar characters. To our surprise, it appeared different from P. hancei when the flowering specimens were collected in May 2012. After consulting the national floras (Wang 1990, Wang et al. 1998), monographs (Li & Wang 2004, Wei et al. 2010) and relevant literature (e.g. Möller et al. 2016, Cen et al. 2017, Lu et al. 2017a) as well as herbarium species from ANU, BJFU, CDBI, CTC, GH, HN, IBK, IBSC, KUN, NU, PH, PE, US and VMN, we considered that it is an undescribed species of Petrocodon. In trying to avoid the misidentification of this new species, we focused on potentially confusing morphological convergence and undertook molecular analysis. Material and methods Morphometric analysis:—Detailed studies on fresh plants and dried specimens of Petrocodon asterocalyx from the herbarium IBK were undertaken. Morphological data used for the description and studies on their variation were also obtained from in situ examination of all living mature plants from the population in Ziyuan county, northern Guangxi, China. For light microscopy (LM), thirty fresh samples (flowers) from 30 mature individuals were fixed in absolute alcohol-acetic acid (3:1 v/v), and herbarium materials were re-hydrated in distilled water with a drop of soap, for 30 min. All observations of specimens and fixed flowers were carried out using the Olympus BH2-RFCA microscope (Tokyo, Japan). Genomic DNA extraction, PCR amplification and sequencing:—To confirm the identity and systematic placement of this new plant, we performed phylogenetic inference of DNA sequence data obtained from the Internal Transcribed Spacer (ITS) region and the plastid trnL-F intron spacer (trnL-F). The new species and its related species were sampled based on previous molecular phylogenetic studies of Gesneriaceae (Möller et al. 2009 & 2011, Weber et al. 2011a & 2011b). Methods used for DNA extraction, PCR amplification and sequencing were carried out as described by (Wei et al. 2013). To elucidate the phylogenetic affinities of the new species, 17 species of Petrocodon including this new species were taken as ingroup, and Primulina dryas (Dunn in Dunn & Tutcher 1912: 195) Mich. Möller & A.Weber in Weber et al. (2011a: 782) and P. pinnata (W.T.Wang (1984a: 25) Y.Z.Wang in Wang et al. (2011: 62) as outgroup based on recent phylogenetic analyses (Möller et al. 2009 & 2011, Weber et al. 2011a & 2011b, Yu et al., 2015). Sequences obtained from this study and NCBI GenBank are listed in Table 1. TABLE 1 Species names, voucher numbers and accession numbers of ITS and trnL-F sequences used in this study. Species name (according to Weber et al., 2011) Voucher number trnL-F ITS Petrocodon ainsliifolius W.H.Chen & Y.M.Shui HQ632941 HQ633038 Petrocodon asterocalyx F.Wen, Y.G.Wei & R.L.Zhang WF120510 KC904957 KC904954 Petrocodon coccineus (C.Y.Wu ex H.W.Li) Y.Z.Wang FJ501516 FJ501341 Petrocodon coriaceifolius (Y.G.Wei)Y.G.Wei & Mich.Möller HQ632943 HQ633040 Petrocodon dealbatus Hance GU350668 GU350636 Petrocodon ferrugineus Y.G.Wei HQ632946 HQ633043 Petrocodon hancei (Hemsl.)A.Weber & Mich.Möller HQ632944 HQ633041 Petrocodon hechiensis (Y.G.Wei, Yan Liu & F.Wen) Y.G.Wei & Mich. HQ632942 HQ633039 Möller Petrocodon hispidus (W.T.Wang) A.Weber & Mich.Möller HQ632939 HQ633036 Petrocodon integrifolius (D.Fang & L.Zeng) A.Weber & Mich.Mölle HQ632940 HQ633037 Petrocodon lithophilus Y.M. Shui, W.H. Chen & Mich. Möller KF202302 KF202295 Petrocodon longgangensis W.H. Wu & W.B. Xu KC765116 KC765114 Petrocodon lui (Yan Liu & W.B.Xu) A.Weber & Mich. Möller HQ632938 HQ633035 Petrocodon niveolanosus (D.Fang & W.T.Wang) A.Weber & Mich. JF697588 JF697576 Möller Petrocodon scopulorum (Chun) Y.Z.Wang HQ632947 HQ633044 Petrocodon tiandengensis (Yan Liu & B.Pan) A.Weber & Mich.Möller HQ632945 HQ633042 Petrocodon viridescens W.H. Chen, Mich. Möller & Y.M. Shui HQ632939 HQ633036 Primulina dryas (Dunn) Mich. Möller & A. Weber FJ501524 FJ501348 Primulina pinnata (W.T. Wang) Y.Z. Wang FJ501526 FJ501349 Sequence alignment and phylogenetic analysis:—Sequence data were edited and assembled using Lasergene Navigator 7.1 (DNAstar, Madison, Wisconsin, USA). Cleaned sequences were aligned with the MEGA 5.1 (Tamura et al. 2011) with additional manual refinements where necessary. Phylogenetic analyses were conducted using maximum 260 • Phytotaxa 343 (3) © 2018 Magnolia Press ZHANG ET AL. parsimony (MP) method implemented in PAUP* 4.0b10. Heuristic searches were performed using a starting tree built from stepwise addition with TBR branch swapping and 1,000 random addition replicates. To assess confidence in clades, bootstrap analyses based on 1,000 replicates with 10 random additions per replicate were used.