DOI: 10.5061/dryad.rn8pk0p5g
title: Data from: Genetic polymorphism in Chondrilla (Asteraceae) in the South of European Russia and the nature of Chondrilla juncea L.

description: We studied genetic diversity in 54 populations of nine species of the
 genus Chondrilla (C. acantholepis, C. ambigua, C. brevirostris, C.
 canescens, C. graminea, C. juncea, C. laticoronata, C. latifolia, and C.
 pauciflora) from SE European Russia and the neighboring territories of NW
 Kazakhstan. Analyses of the trnT–trnF region of plastid DNA and the
 internal transcribed spacer of ribosomal DNA (ITS1–5.8S–ITS2) using
 Statistical Parsimony, Maximum Likelihood and Neighbor Net methods
 revealed two major evolutionary lineages roughly corresponding to two
 subgenera traditionally recognized within Chondrilla in the region. Within
 the first evolutionary lineage (subgenus Brachyrhynchus) sexual diploid C.
 ambigua and its putatively hybrid apomictic derivatives C. brevirostris,
 C. laticoronata and C. pauciflora can be recognized. Their identity is
 confirmed by analyses of ISSR markers. The second evolutionary lineage
 (subgenus Chondrilla) is comprised comprises by C. juncea, C.
 acantholepis, C. canescens, C. graminea, and C. latifolia in European
 Russia. Analyses of morphological variability and the genealogy of plastid
 and nuclear markers genealogy favor their treatment as a single species C.
 juncea.    
description: Samples were collected in 2014 to 2016 from 54 natural
 populations of 9 <i>Chondrilla</i> species (<i>C.
 acantholepis</i>, <i>C. ambigua</i>, <i>C.
 brevirostris</i>, <i>C. canescens</i>, <i>C.
 graminea</i>, <i>C. juncea</i>, <i>C.
 laticoronata,</i> <i>C. latifolia </i>and <i>C.
 pauciflora</i> Ledeb.) in the Astrakhanskaya, Volgogradskaya,
 Penzenskaya, Samarskaya, Saratovskaya and Ulyanovskaya Provinces of
 Russia, Krasnodarsky Krai, the Republic of Kalmykia, the Crimea, and
 Western Kazakhstan. Three plants were sampled from each population as
 herbarium specimens (kept at the herbarium SARBG) and silica gel dried
 floret ligules. From sympatric populations of <i>C.
 juncea</i>, <i>C. graminea</i> and <i>C.
 latifolia</i> with numerous morphologically intermediate plants,
 three plants were randomly chosen and designated as <i>C. juncea /
 C. graminea</i> or <i>C. juncea / C. latifolia,</i>
 respectively. DNA was extracted from silica gel dried
 ligules (connate petals of ligulate florets) using commercially available
 NucleoSpin® Plant II DNA extraction kit (MACHEREY-NAGEL GmbH, Düren,
 Germany) according to the manufacturer’s protocol. The
 plastid <i>trn</i>T<i>−trn</i>L and
 <i>trn</i>L−<i>trn</i>F intergenic spacers (IGS)
 and the <i>trn</i>L intron were amplified using standard
 primers <i>a</i> and <i>b</i>,
 <i>c</i> and <i>d</i>, <i>e</i> and
 <i>f</i> (Taberlet et al., 1991). The nuclear internal
 transcribed spacer regions of ribosomal DNA
 (<i>ITS</i>1<i>−</i>5.8S<i>−ITS</i>2)
 were amplified using primers ITS 1, ITS2, ITS3 and ITS 4 (White et al.
 1990). Polymerase chain reactions (PCR) were carried
 out in 20 µl reaction volumes in a Mastercycler gradient thermocycler
 (Eppendorf, Germany). The reaction mixture contained 4 µl of ready-to-load
 PCR mix 5X MasTaq<sup>DD</sup>Mix-2025 (200 µM of each dNTP, 2
 mM MgCl<sub>2</sub>, 1.5 U “hot-start” SmarTaq DNA-polymeraze,
 reaction buffer, stabilizer and two inert dyes; Dialat Ltd., Moscow,
 Russia), 15 µl of deionized water, 3.4 pmol of each primer and 1 µl of
 template DNA of unknown final concentration. The PCR conditions were as
 follows: preliminary denaturation for 5 minutes at 95°C; 35 cycles of 30
 seconds at 95°C, 30 seconds at 55°C and 2 minutes at 72°C, followed by
 final elongation for 10 minutes at 72°C. PCR products were purified with a
 DNA Gel Extraction Miniprep Kit (BioSilica Ltd., Novosibirsk, Russia)
 according to the manufacturer’s instructions.<br> Direct sequencing
 was performed in both directions on the ABI PRISM 3130 XL genetic analyzer
 (Applied Biosystems, Foster City, CA, USA), using ABI Prism BigDye
 Terminator Cycle Sequencing Ready Reaction Kit v. 3.1 for cycle sequencing
 reactions at the facilities of Sintol Ltd. (Moscow, Russian
 Federation).