DOI: 10.5061/dryad.d2547d88m
title: Data from: Oxyrrhynchium hians (Brachytheciaceae, Bryophyta) includes several morphologically distinct and cryptic species in northwestern Europe

description: A study of the variable species Oxyrrhynchium hians s.l. in NW Europe
 based on nuclear ITS, and plastid rpl16 and trnL‑trnF, as well as
 morphology, revealed unsuspected species level diversity. Three taxa are
 distinguishable by morphology: O. distichum with complanate or
 sub-complanate branch leaves and long and narrow leaf lamina cells, O.
 hians with cordate or broadly ovate, concave leaves that are evenly
 arranged around the stems and branches, and O. swartzii with mostly
 complanate or sub-complanate branch leaves and compared with O. distichum
 relatively short and wide leaf lamina cells. In Sweden O. distichum grows
 almost exclusively on base-rich or calcareous rocks and has been recorded
 from a belt stretching from the Baltic Sea islands of Öland and Gotland to
 Dalarna and southernmost Norway, whereas the other two species grow on
 various substrates and have wider distributions. Oxyrrhynchium hians grows
 in more nutrient-rich habitats than O. swartzii and is therefore absent
 from regions with relatively poor soils. Oxyrrhynchium swartzii occurs
 northwards to Sør-Trøndelag in Norway and Jämtland and Medelpad in Sweden
 and includes two semi-cryptic species that differ slightly in size and may
 have relatively more western and eastern distributions, respectively, in
 Fennoscandia.
description: For the molecular portion of this study, 32 specimens of
 <em>O. hians </em>s.l. from Sweden, six from Norway, and three
 from Switzerland were included. One specimen each of the two closely
 related species <em>O. schleicheri</em> (R. Hedw.) Röll and
 <em>O. speciosum</em> (Brid.) Warnst., two other unambiguous
 members of <em>Oxyrrhynchium</em> (Ignatov and Huttunen 2002,
 Ignatov and Isoviita 2003), were included as outgroup.
 Total DNA was extracted using the NucleoMag® Plant kit for DNA
 isolation from plant tissue (Macherey-Nagel) with the KingFisher Duo
 magnetic particle processor. Double stranded DNA templates were prepared
 by polymerase chain reaction (PCR). PCR was performed using IllustraTM Hot
 Start Mix RTG (GE Healthcare) in a 25 µl reaction volume according to the
 manufacturer’s instructions. In all cases, the
 specified PCR programs were initiated by a denaturation step of 5 min at
 95º C and followed by a final extension period of 10 min at 72º C. The PCR
 programs were, for the nuclear internal transcribed spacers 1 and 2 (ITS),
 4 cycles of 30 sec at 95º C, 40 sec at 55º C, and 1 min at 72º C, 4 cycles
 of 30 sec at 95º C, 30 sec at 53º C, and 1 min at 72º C, 35 cycles of 30
 sec at 95º C, 30 sec at 50º C, and 1 min at 72º C, with the primers
 ‘ITS4bryo’ (Stech 1999) and ‘ITSbryoR’ (Hedenäs 2014). For the plastid
 <em>rpl</em>16 G2 intron (<em>rpl</em>16), 43
 cycles of 30 sec at 95º C, 30 sec at 58º C, and 1 min 15 sec at 72º C,
 with<em> </em>the primers ‘F71’ (Jordan, et al. 1996) and
 ‘rpl16-antR2’ (Hedenäs 2008), and for the plastid
 <em>trn</em>LUAA intron plus
 <em>trn</em>LUAA-<em>trn</em>FGAA spacer
 (<em>trn</em>L-<em>trn</em>F), 4 cycles of 30 sec
 at 95º C, 40 sec at 57º C, and 1 min at 72º C, 4 cycles of 30 sec at 95º
 C, 30 sec at 55º C, and 1 min at 72º C, 35 cycles of 30 sec at 95º C, 30
 sec at 52º C, and 1 min at 72º C, with the primers ‘trnC’ and ‘trnF’
 (Taberlet, et al. 1991). The amplified PCR products
 were purified from excess primers and nucleotides by adding 1µl of
 Exonuclease I (20U/µl) and 4µl of FastAP Thermosensitive Alkaline
 Phosphatase (1U/µl) (Thermo Scientific™) and incubating at 37° C for 30
 min followed by an enzyme inactivation step at 80° C for 15 min. The
 purified PCR products, together with the same primers used for PCR
 amplification, were subsequently sent to Macrogen Europe B.V (Amsterdam,
 The Netherlands) for single-stranded sequencing on an Applied Biosystems
 3730XL sequencer. Nucleotide sequence fragments were
 edited and assembled for each DNA region using PhyDE® 0.9971 (http://www.phyde.de/index.html; accessed 17 February 2023). The assembled sequences were aligned manually in PhyDE®. Regions of partially incomplete data in the beginning and end of the sequences were identified and were excluded from subsequent analyses. Gaps were coded using the simple indel coding of Simmons and Ochoterena (2000) in SeqState (Müller 2005).
description: # Data from: Oxyrrhynchium hians (Brachytheciaceae, Bryophyta) includes
 several morphologically distinct and cryptic species in northwestern
 Europe ## Description of
 the data and file structure Sequence alignments for ITS, rpl16, and
 trnL-trnF Date of data collection: 2022-2023 Geographic range: Europe,
 mainly Scandinavia Files: hians_ITS.txt (ITS); hians rpl16.txt (rpl16);
 hians_trnLF.txt (trnL-trnF); these are sequence alignments in FASTA format
 Usage notes: The positions in the files correspond with ITS, rpl16, and
 trnL-trnF, as follows: File: hians_ITS.txt ITS positions: 1-753 ITS indel
 coding positions: 755-773 File: hians rpl16.txt rpl16 positions: 1-723
 rpl16 indel coding positions: 725-730 File: hians_trnLF.txt trnL-trnF
 positions: 1-447 trnL-trnF indel coding positions: 449-450 ##
 Sharing/Access information Specimen data linked to the sequence numbers
 can be found in the paper (when published) at: *
 Data was derived from herbarium
 material located in S and TRH (Index Herbariorum abbreviations): *