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Broad Polyphyly in Pleurospermum s. l. (Umbelliferae-Apioideae) as Inferred from
nrDNA ITS and Chloroplast Sequences
Author(s) :Carmen M. Valiejo-Roman, Elena I. Terentieva, Michael G. Pimenov, Eugene V. Kljuykov,
Tahir H. Samigullin, and Patricia M. Tilney
Source: Systematic Botany, 37(2):573-581. 2012.
Published By: The American Society of Plant Taxonomists
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Systematic Botany (2012), 37(2): pp. 573–581
©Copyright 2012 by the American Society of Plant Taxonomists
Broad Polyphyly in Pleurospermum s. l. (Umbelliferae-Apioideae) as Inferred from
nrDNA ITS and Chloroplast Sequences
Carmen M. Valiejo-Roman,
Elena I. Terentieva,
Michael G. Pimenov,
Eugene V. Kljuykov,
Tahir H. Samigullin,
and Patricia M. Tilney
Department of Evolutionary Biochemistry, A. N. Belozersky Institute, Moscow State University, Moscow 119991 Russia.
Botanical Garden of Moscow State University, Moscow State University, Moscow 119991 Russia.
Department of Botany and Plant Biotechnology, University of Johannesburg, PO Box 524, Auckland park 2006, South Africa.
Author for correspondence (firstname.lastname@example.org)
Communicating Editor: Rodger Evans
Abstract—Traditionally the genus Pleurospermum and the related genera Aulacospermum, Hymenidium, Hymenolaena, Physospermopsis,
Pseudotrachydium, Pterocyclus, and Trachydium, have been problematic taxa with regard to their circumscription and composition.
Pleurospermum s. s. includes one or two closely related boreal species but more than 40 other species, distributed mainly in the Sino-Himalayan
floristic area, have been attributed to it by different authors in various classifications. Relationships in this taxonomic group are unclear. A
molecular phylogenetic analysis of nuclear (nrITS) and cpDNA (psbA-trnH and trnL-trnF) sequences of representative species of Pleurospermum
s. l. and closely related Hymenidium,Aulacospermum, Trachydium,Physospermopsis, Pseudotrachydium,Sinolimprichtia,Pterocyclus, and
Hymenolaena (55 species in all) was conducted and compared with an analysis of morphological characters. Only two traditional genera were
supported as monophyletic groups in the molecular trees, namely Aulacospermum (including Pseudotrachydium)andHymenolaena. Two stable
groups were revealed within Hymenidium. One group included H. chloroleucum, H. heterosciadium, H. hookeri,andH. delavayi. The second
included H. lindleyanum, H. stellatum, H. huzhihaoi,H. wilsonii, and Trachydium roylei. Various species of Hymenidium and Trachydium were
scattered throughout the tree. The molecular data did not confirm an early divergence between northern and Sino-Himalayan species of
Pleurospermum. This indicates that Pleurospermum s. l. and most of the other genera included in this study are polyphyletic.
Keywords—Apioideae, cpDNA, nuclear markers, phylogeny, Pleurospermum, Umbelliferae.
Pleurospermum has been one of the most problematic
genera of the Umbelliferae subfamily Apioideae with regard
to both monophyly and species composition (Clarke 1879;
Drude 1897–98; Wolff 1925, 1926, 1929; Mukherjee and
Constance 1993; Pimenov and Leonov 1993; Pan ZeHui and
Watson 2005;). Pleurospermum was described by Hoffmann
(1814, 1816) as containing three closely related species,
namely P. austriacum (L.) Hoffm. (nomenclatural type of
the generic name), P. uralense Hoffm., and P. camtschaticum
Hoffm. The latter two species are now treated as conspecific
under the name of P. uralense.Pleurospermum austriacum and
P. uralense are closely related, and were sometimes regarded
as two subspecies of a single species (Horn af Rantzien 1946).
They are distributed in the northern part of the range
(Europe, Siberia and Far East) of Pleurospermum s. l. By
contrast, the majority of species commonly treated under
Pleurospermum are distributed in the southern part of the range,
centered in the Sino-Himalayan floristic region (southwestern
China, adjacent Tibet, and the Indian and Nepali Himalayas).
In total, Pleurospermum s. l. numbers over 40 species.
Morphological investigations of all 80 available species of
Pleurospermum s. l. and closely related genera (Aulacospermum
Ledeb., Hymenolaena DC., Trachydium Lindl., Pseudotrachydium
(Kljuykov, Pimenov & V. N. Tikhom.) Pimenov & Kljuykov,
Physospermopsis H. Wolff, Pterocyclus Klotzsch., and
Keraymonia Farille) based on the distribution of 47 diagnostic
characters (Pimenov and Kljuykov 2000a,b; Pimenov et al.
2000) showed heterogeneity in this genus and called into
question its monophyly. The cluster analysis revealed com-
plex relationships within Pleurospermum s. l. and with the
related genera. Some taxonomic and nomenclatural proposals
were made despite the fact that the morphology of many
species remains poorly investigated due to incomplete collec-
tions. For example, some species are poorly represented in
herbaria, sometimes only by type specimens, while others
were collected without fruits (which are essential for the sys-
tematics of the family) or without flowers. The type speci-
mens of some species have most likely been lost, especially
those of species described by H. Wolff (e.g. P. albimarginatum
H. Wolff, P. longipetiolatum H. Wolff, P. microphyllum H. Wolff,
P. microsciadium H. Wolff, and P. souliei H. Wolff).
On morphological grounds, Pimenov et al. (2000) distin-
guished a large cluster of “southern” species from two
“northern” species and proposed additional modifications to
generic circumscriptions including additions to, or deletions
from, Hymenolaena, Physospermopsis, Pterocyclus, and some
other related genera. The small “northern” cluster corre-
sponds to Pleurospermum s. s. The transfer of the majority of
the “southern” species to Hymenidium was also proposed by
Pimenov et al. (2000).
Hymenidium was originally described by Lindley (1835)
with two species, but these are now treated as conspecific
under the name of the nomenclatural type, H. brunonis Lindl.
The genus had long been neglected until it was enlarged with
the inclusion of most of the former Pleurospermum species
distributed in the Sino-Himalayan region, plus some species
from related genera. At the same time, Pimenov and
Kljuykov (2000b) noted: “It is clear that Hymenidium in our
circumscription is a complex genus, containing probably
some more natural species groups. Unfortunately, many
species in the genus can only be characterized now with
blanks.” Therefore, further splitting did not seem possible,
and the clusters were not treated as taxa only because of
incomplete morphological descriptions. Earlier, Farille et al.
(1985) had made an attempt to partly revise the group and to
define infrageneric taxa in Pleurospermum s. l. He distin-
guished seven subgenera with the type subgenus containing
only the northern species P. austriacum and P. uralense.
Many local high-mountain species of Umbelliferae were
described under the genus Trachydium, which had been
separated concurrently with Hymenidium (Lindley 1835).
Norman (1938) regarded Trachydium as monotypic and
assigned the remaining species to other genera such as
Physospermopsis and Aulacospermum.
As there are different, even polar, viewpoints on the
taxonomy of Pleurospermum and related taxa, all based on
morphology only, we conducted a molecular phylogenetic
analysis of the species of Pleurospermum s. l., Hymenidium,
Aulacospermum,Pseudotrachydium (separately from Aulacospermum),
Hymenolaena, Physospermopsis,Trachydium (including some of
the numerous species variously attributed to this genus),
Sinolimprichtia,andPterocyclus. Our aim was to check the
reliability of morphologically grounded affinities of species
and genera in the group, and to reveal any new affinities.
We regard the molecular phylogenetic analysis as the
main contribution of the present article to the taxonomy
of Pleurospermum s. l. Molecular phylogenetic trees were con-
structed using sequences of the nuclear ribosomal DNA
(nrDNA) ITS. We also investigated the variation within two
cpDNA regions, psbA-trnH intergenic spacer and the trnL-
trnF region (including trnL intron and trnL-trnF intergenic
spacer). We were limited by the availability of material for
the molecular study, using mainly our own collections with
Materials and Methods
Taxon Sampling—Plant material was collected during our expeditions
to China, India, Nepal, and different regions of the former USSR. The
investigated species, their voucher information, and accession num-
bers are listed in Appendix 1. Complete sequences of the nrITS region
were generated for 44 accessions, each representing a distinct species
of Pleurospermum s. l. or related genera. In addition, GenBank data
were downloaded for eight species of Pleurospermum s. l. and related
genera. Sequences of the chloroplast psbA-trnH intergenic spacer and
of the trnL intron,aswellasofthetrnL-F intergenic spacer, were
obtained for 55 accessions. The genera Bupleurum,Heteromorpha,and
Glia were selected as outgroups as they were previously shown to
occupy a basal position in the Apioideae molecular tree (Calvin
2006). We also considered the placement of Pleurospermum s. l. in an
“apioid” analysis (Fig. 1), in which we included representatives of the
main clades and tribes of the recently proposed Apioideae ITS classi-
fication (Downie et al. 2010). That analysis included 100 accessions
from 42 genera.
DNA Extraction, Amplification, Sequencing and Alignment—Total
genomic DNA was isolated from fruit and herbarium leaf material using
a NucleoSpin plant DNA kit (Macherey-Nagel, Du
¨ren, Germany) follow-
ing the manufacturer’s instructions. Polymerase chain reaction (PCR)
amplification was performed on a Biometra T3000 Thermocycler using
Encyclo PCR kit (Evrogen JSC, Moscow, Russia). Details of the PCR ITS
amplifications (including primer locations and characteristics), and
sequencing strategies for ITS were described by Valiejo-Roman et al.
(2002). The sequences of 5.8S rRNA gene were also obtained but excluded
from the analysis due to lack of variation. The psbA-trnH intergenic
spacer was amplified using the primers psbA (Sang et al. 1997) and trnH
(Tate and Simpson 2003). Amplification and sequencing of the psbA-trnH
spacer was conducted in the same way as for the ITS region. The trnL-trnF
region (including trnL intron, and trnL-F intergenic spacer) was amplified
using primers trnL (LSC-40F) and trnF (LSC-40R; Logacheva et al. 2008).
The PCR products were purified using a gel extraction and PCR
cleanup kit (Cytokine Ltd, St. Petersburg, Russia). Automated sequencing
was performed on an ABI 3100 sequencer using the Big Dye Terminator
v.3.1 sequencing kit (ABI, Foster City, California). The nuclear and chlo-
roplast regions were sequenced in their entirety on both strands. The
DNA sequences were aligned using the program MUSCLE (Edgar 2004)
and then manually adjusted using the program BioEdit (Hall 1999).
Molecular-Phylogenetic Analysis—Maximum parsimony analysis of
molecular data involved a heuristic search conducted with PAUP*
version 4.0b8 (Swofford 2003) using TBR (tree-bisection-reconnection)
branch swapping, the multrees option, steepest descent, collapse, and
acctran optimization, with character states specified as unordered and
equally weighted. Five-hundred random addition replicates were per-
formed, saving no more than 1,000 trees per replicate. Bootstrap analyses
(Felsenstein 1985) were performed to assess the degree of support for
particular clades. Bootstrap values were calculated from 500 replicate
analyses using TBR branch swapping and random addition sequence
of taxa, saving 1,000 trees per replicate.
Bayesian analysis of molecular data was performed using the program
MrBayes (Huelsenbeck and Ronquist 2001; Ronquist and Huelsenbeck
2003). The GTR model of substitutions with gamma rate categories was
selected by the Akaike information criterion (Akaike 1974) in Modeltest
(Posada and Crandall 1998). Bayesian inference was performed with two
simultaneous runs with four chains in each; 15,000,000 generations were
performed; trees were sampled every 100 generations. The number of
generations to be discarded was determined by the cold chain log like-
lihood examination using Tracer v. 1.5 (Rambaut and Drummond 2007).
Trees have been submitted to TreeBASE (study number TS2: S12037). To
compare homogeneity of chloroplast and nuclear data, an ILD-test (Farris
et al. 1995) was performed for the combined dataset. In all analyses gaps
were treated as missing data.
Fig. 1. Bayesian tree inferred from the analysis of the 100 samples of
ITS data. Branch lengths are proportional to the amount of character
changes; scale = 0.1 substitutions per character. Maximum parsimony
bootstrap support and Bayesian posterior probabilities are indicated
574 SYSTEMATIC BOTANY [Volume 37
Characteristics of the Nuclear and Chloroplast Sequences—
The length of ITS sequences (100 accessions from 42 genera)
ranged from 442 bp for Scandix stellata to 406 bp for
Heteromorpha arborescens and H. pubescens. An alignment of
100 ITS sequences resulted in a matrix of 508 positions, 72 of
which were excluded because of gap-richness or alignment
ambiguity. Of the remaining 436 positions, 70 were constant,
47 autapomorphic, and 319 parsimony informative.
For comparative purposes, the ITS dataset was sub-
sequently reduced to include the same 55 accessions as
contained in the cpDNA dataset. The length of the ITS region
for the 55 sequences included in our “narrow analysis”
ranged from 441 bp for Hymenidium hookeri to 406 bp for
Heteromorpha arborescens and Heteromorpha pubescens.An
alignment of 55 complete ITS sequences resulted in a matrix
of 486 positions, with 61 excluded because of alignment
ambiguity. Of the remaining 425 positions, 119 were con-
stant, 53 autapomorphic, and 253 parsimony informative.
The length of the psbA-trnH for the 55 sequences ranged
from 416 bp for Hymenolaena badachschanica to 148 bp for
Hymenidium apiolens. The alignment of 55 complete psbA-trnH
sequences resulted in a matrix of 522 positions. A stretch of
231 nucleotide positions in the psbA matrix could not be con-
fidently aligned due to complex patterns of indels, and was
therefore excluded from the phylogenetic analyses. Of the
remaining 291 positions, 128 were constant, 72 autapomorphic,
and 91 parsimony informative.
The length of the trnL-trnF locus for the 55 sequences
ranged from 975 bp for Trachydium variabile to 845 bp for
Hymenidium benthamii. The alignment of these 55 sequences
(which included trnL intron, and trnL-F intergenic spacer)
resulted in a matrix of 1,152 positions, from which 242 posi-
tions were excluded from further consideration due to ambi-
guity of alignment. Of the remaining 910 positions, 705 were
constant, 97 autapomorphic, and 108 parsimony informative.
The combined analysis of 55 nuclear and chloroplast
sequences yielded a matrix of 2,160 aligned positions. After
exclusion of 534 ambiguous positions, 1626 remained, of
which 952 were constant, 222 autapomorphic, and 452 parsi-
mony informative (Table 1).
Comparison of Trees—The BI and MP “apioid” analyses
revealed three well supported clades (Fig. 1, clade A, PP =
0.96, BS = 52%; clade B, PP = 1.00, BS = 100%; clade C, PP =
1.00, BS = 86%). Clade A consisted of subclade 3 (PP = 1.00,
BS = 98%) composed of five Aulacospermum species and three
Pseudotrachydium species, subclade 2 (PP = 1.00, BS = 100%)
with three Physospermopsis species, and subclade 5 (PP = 1.00,
BS = 100%) with two northern, closely related Pleurospermum
species, which corresponds well with traditional taxonomy.
Also in clade A, subclade 1 (PP = 1.00, BS = 98%) included
four Hymenidium species and Trachydium roylei, and subclade
4 (PP = 0.64, BS = 84%) consisted of six Hymenidium species.
Clade B was formed by subclades 6–8. Subclade 6 (PP =
1.00, BS = 81%) consisted of four Trachydium species,
Sinolimprichtia alpina, and Hymenidium virgatum. Subclade 7
(PP = 1.00, BS = 97%) with three Hymenolaena species formed
a monophyletic group, which corresponds well to traditional
taxonomy. Subclade 8 (PP = 1.00, BS = 100%) included two
Pterocyclus species. Pterocyclus angelicoides and P. forrestii
were supported as sister species but P. rivulorum was sup-
ported as a member of Clade C, subclade 9.
Clade C included the representatives of the main
Apioideae clades and two subclades, namely 9 (PP = 1.00,
BS = 67%) and 10 (PP = 1.00, BS = 100%) with Pleurospermum
s. l. species. Subclade 9 (Hymenidium brunonis, Trachydium
subnudum, Pterocyclus rivulorum, Hymenidium apiolens, and
Sinodielsia thibetica) was composed of morphologically dis-
similar species, which is reflected in their attribution to dif-
ferent genera. Subclade 10 (PP = 1.00, BS = 100%) was formed
only by Hymenidium species (H. delavayi, H. heterosciadium,
H. chloroleucum, and H. hookeri).
The dataset of 55 accessions contained plastid data in addi-
tion to nrITS. Analyzed separately, the plastid data analyses
produced poorly resolved trees. Some differences in details
were observed among nuclear and chloroplast data parti-
tions, and between psbA-trnH and trnL-trnF data. The differ-
ences between the ITS and the trnL-trnF trees were
insignificant (p= 0.644), while psbA data were incongruent
with both ITS (p= 0.002) and trnL-trnF (p= 0.004). In the
latter case, major topological conflicts were concentrated in
clade B: subclade 8 became basal in clade C with a moderate
PP value (0.73); one of the Hymenolaena species was sister to
Trachydium simplicifolium (PP = 0.92, BS = 53%). Support for
such relationships does not exceed a “high” threshold (0.95
for PP and 70% for BS), therefore all molecular data were
included in the combined analyses. The resulting trees were
the most resolved and congruent with the results of the
s. l. were compared: 1.) The traditional classification (now
Table 1. Comparison of tree metrics for the maximally parsimonious trees (MPTs) found based on the four different datasets separately and combined.
Dataset morphology ITS1–2 ITS1–2 psbA-trnH trnL-trnF all molecules combined
No. of species 51 100 55 55 55 55
No. of aligned positions 508 486 522 1,152 2,160
No. of characters in analyses 49 436 425 291 910 1,626
No. of aligned positions constant 1 70 119 128 705 952
No. of aligned positions parsimony-informative 44 319 253 91 108 452
No. of aligned positions autapomorphic 4 47 53 72 97 222
No. of MPTs 1,897 2,585 8 497,000 27,002 96
Tree length 361 2,157 959 321 293 1,610
CI 0.288 0.343 0.523 0.673 0.788 0.589
RI 0.58 0.755 0.826 0.824 0.893 0.826
HI 0.712 0.657 0.477 0.327 0.212 0.411
2012] VALIEJO-ROMAN ET AL:BROAD POLYPHYLY IN PLEUROSPERMUM S. L. 575
outdated) of the species in the genera Pleurospermum,
Aulacospermum, Pseudotrachydium, Hymenolaena, Hymenidium,
Physospermopsis, Pterocyclus, Sinolimprichtia, and Trachydium.
2.) Grouping according to nuclear, chloroplast, and combined
DNA sequences (Fig. 1; Fig. 2A-D). The groups obtained (BI
and MP trees) are named below as subclades. 3.) Grouping
according to a broad set of morphological characters
(Pimenov and Kljuykov 2000a,b; Pimenov et al. 2000).
Incongruence of Molecular Trees—In our analyses different
loci resulted in similar, but not congruent, tree topologies.
The ILD-test reveals that the only significantly incongruent
partition is psbA-trnH, while in the ITS and the trnL-trnF trees
the same relationships among the main clades and subclades
were obtained. Despite the fact that the psbA-trnH tree dif-
fers in the placement of subclade 8 and Hymenolaena
badachschanica, we combined all available data. The main
reason for doing this was the increase in resolution and
branch support of trees obtained when the psbA-trnH data
were added to the ITS and the trnL-trnF data (data not
shown). As pointed out above, the branch support for the
groupings using plastid data only is not high so the observed
incongruence may be considered as “soft incongruence”
caused by a lack of sufficient phylogenetic signal in the data
(Mason-Gamer and Kellogg 1996). On the other hand, a large
proportion of missing data may also be the source of the
observed differences in the psbA-trnH tree topology due to
misinterpretation of missing characters (Lemmon et al. 2009).
Indeed, the psbA-trnH spacer length is heterogeneous, so
after exclusion of gap-rich columns and regions of ambigu-
ous alignment, the psbA-trnH data still contain about 50%
missing data for several species. However, deletion of these
“wildcard”-species in a separate psbA-trnH Bayesian analysis
did not alter the relationships of the main clades and
subclades (data not shown). One may therefore conclude that
the topology obtained (Fig. 2C) is due to weak or ambiguous
phylogenetic signals rather than to the effect of missing data.
Well Supported Molecular Clades, Corresponding to the
Morphologically Based Classification—Some relationships
were more or less consistent with both molecular and mor-
Subclade 3 includes all sampled Aulacospermum and
Pseudotrachydium sequences. Both molecular (Figs. 1, 2A–C)
and morphological cluster analyses confirm a relationship
between Aulacospermum and Pseudotrachydium (Pimenov and
Kljuykov 2000a). This corresponds to our earlier classification
of the group (Kljuykov et al. 1976a,b), in which Pseudotrachydium
was regarded as a section within Aulacospermum. The species
of this section have vesiculose fruit surfaces, solid stems, and
yellow petals without claws. Later Pseudotrachydium was
raised to the generic level (Pimenov et al. 2000). Molecular
data support Pseudotrachydium as a monophyletic group within
Aulacospermum. In previous morphological studies, Kljuykov
et al. (1976a,b) found A. stylosum to be the most divergent
among Aulacospermum species, and placed it in a separate sec-
tion Hymenolaenopsis. A set of morphological traits, including
recurved bracts and bractlets, toothed bracts, fistulose stems,
and fruits not covered by wart-like outgrowths, distinguish
A. stylosum from other members of Aulacospermum.Itwas
treated in Hymenolaena by Aitchison (1880) and Lipsky (1900).
In the molecular analyses (Figs. 1, 2A, B), A. stylosum was found
to be sister to Aulacospermum +Pseudotrachydium,andbasalin
Aulacospermum group in the psbA-trnH tree(Fig.2C),butinthe
trnL-trnF tree (Fig. 2D) it is placed outside of subclade 3.
Hymenolaena is the other traditional genus to be supported
by the molecular studies. Three species of Hymenolaena
formed subclade 7 (Figs. 1, 2A, B) with a high support value
(PP = 1.00, BS = 100%). In the chloroplast tree trnL-trnF
(Fig. 2D), the position of these species does not contradict
the ITS but in the cpDNA tree psbA-trnH (Fig. 2C)
H. badachschanica is clustered with T. simplicifolium. The spe-
cies of this genus are clustered together on the basis of a set of
morphological characters in the group under study (Pimenov
and Kljuykov 2000a). All the species have pinnate leaves with
sessile leaflets, petioles almost round in cross-section, and
bracts usually absent. In addition, all Hymenolaena species
have flat petals and 2n= 22 (in contrast to Pleurospermum
and Hymenidium with 2n= 18).
Well Supported Molecular Clades, Not Corresponding to
Traditional Taxonomy—In the molecular trees obtained,
some newly revealed relationships warrant further discus-
sion. Among the molecular subclades, three are formed by
Hymenidium species, which were not recognized as entities
previously. These well-supported subclades are not closely
related to one another in all molecular analyses (Figs. 1,
2A–D; subclades 11, 10, 4). The earliest diverging subclade 11
(Figs. 1, 2A–D) is formed by two species, Hymenidium bicolor
and H. mieheanum, and has strong support (PP = 1.00, BS =
100%) in all molecular trees. Moreover, it is supported by
10 unique, synapomorphic molecular characters. Never-
theless, the two species are morphologically dissimilar. Both
species are unusual representatives of Hymenidium. A signif-
icant difference between H. mieheanum and other species of
the genus has been noted in its protologue (Pimenov and
Kljuykov 2004). In contrast to most Hymenidium species, this
species has long linear or lanceolate calyx teeth, sessile pri-
mary leaf segments, numerous and rigid remains of peti-
oles, and papery bracts. Hymenidium bicolor, with pinnate
leaves, compact umbellets, and numerous broad rigid
bractlets, is probably related to the little-known monotypic
genus Pleurospermopsis C. Norman [P. sikkimense (C. B. Clarke)
C. Norman], not included in the present analysis because
type sheets were not available. A highly supported group
(subclade 10 Figs. 1, 2A–D) containing four Hymenidium spe-
cies sequences, is formed by H. delavayi, H. heterosciadium,
H. chloroleucum, and H. hookeri (three accessions, differing
slightly). These Hymenidium species share common morpho-
logical characters. They have well-developed bracts, dentate,
lanceolate, or linear bractlets, long (longer than stylopods)
calyx teeth, and dorsally compressed mericarps with the mar-
ginal ribs broader than the dorsal ones. Subclade 1 (Figs. 1,
(H. huzhihaoi, H. wilsonii, H. lindleyanum,andH. stellatum)
and Trachydium roylei. This subclade is not formed in the
cpDNA trees (Fig. 2C, D) because the resolution in this part
of clade A is low and may be due to insufficient phylogenetic
information contained in the plastid markers.
The position of Trachydium roylei in this subclade calls for
special comment. In general, Trachydium species are scattered
in the molecular trees. This is not surprising. Norman (1938)
carried out a critical analysis of the genus, suggesting it to be
monotypic. Our molecular analysis further suggests the
demise of Trachydium. The type species, T. roylei, is placed in
subclade 1 (Figs. 1, 2A, B), composed of high-mountain
dwarf (“stemless”) species of Hymenidium (H. stellatum and
its relatives). Therefore, there are no serious objections to
including Trachydium s. s. (i.e. the type species only) within
576 SYSTEMATIC BOTANY [Volume 37
Fig. 2. Bayesian trees inferred from the analysis of the nucleotide data set. A. combined nuclear and chloroplast data. B. ITS data. C. chloroplast
psbA-trnH spacer. D. chloroplast trnL-trnF spacer. Branch lengths are proportional to the amount of character changes; scale = 0.1 substitutions per
character. Maximum parsimony bootstrap support and Bayesian posterior probabilities are indicated above nodes.
2012] VALIEJO-ROMAN ET AL:BROAD POLYPHYLY IN PLEUROSPERMUM S. L. 577
Pleurospermum s. l. or Hymenidium. This position seems to be
natural as T. roylei is similar to these species in life-form and
many other morphological characters (i.e. it is difficult to
find significant differences between them), as well as in plant
geography (mainly endemics of the Western Himalayas).
Trachydium nanum was supported as the closest relative of
Physospermopsis delavayi, which was transfered to Physospermopsis
by Pimenov and Kljuykov (2000b). In the molecular analysis,
Physospermopsis delavayi +Physospermopsis nana (= Trachydium
nanum) formed a separate and well-supported compact
subclade 2 (Figs. 1, 2A–D).
Subclade 4 (Figs. 1, 2A–D) is composed exclusively of six
species of Hymenidium (H. davidii, H. decurrens, H. foetens,
H. linearilobum, H. wrightianum, and H. benthamii). All of these
high-mountain species are similar morphologically how-
ever H. linearilobum, and H. wrightianum form a separate
cluster in morphological analysis. Subclade 5 is sister to
subclade 4 and comprises two species of Pleurospermum s. s.,
viz. P. austriacum and P. uralense. These two species are the
closest in the molecular analysis (Figs. 1, 2A–D). Their posi-
tion in relation to the remaining taxa of Pleurospermum s. l. is
different in the molecular and morphological investigations.
The Pleurospermum s. s. species have a set of unique charac-
ters, inflated mericarp ribs with large secretory ducts (ducts
may sometimes be more than solitary), crushed mesocarp in
mature fruits, flat petals, and puberulent leaf lobes. This was
the basis for the separation of Hymenidium from Pleurospermum
at the generic level. Nevertheless the molecular data did not
confirm an early divergence between northern and Sino-
Himalayan species of Pleurospermum.
Subclades 6 and 9 (Figs. 1, 2A–D) are taxonomically com-
plex groups of closely related taxa. Subclade 6 contains
species from different genera, viz. Hymenidium, Trachydium,
and Sinolimprichtia. Four species of traditional Trachydium
(T. variabile, T. involucellatum, T. simplicifolium, and T. souliei)
are placed in this subclade. Trachydium involucellatum forms
a small subclade with Sinolimprichtia alpina (Figs. 1, 2A, B).
The placement of Sinolimprichtia alpina with Trachydium
involucellatum supports a reclassification of the latter species,
which is obviously not a Trachydium. The molecular data
(only limited) may rekindle a discussion about the generic
position of this species.
The well-supported subclade 9 contains species that are attrib-
uted at present to Hymenidium (H. apiolens and H. brunonis),
Pterocyclus (P. rivulorum), Sinodielsia (S. thibetica), and Trachydium
(T. subnudum). This subclade is composed of morphologi-
cally dissimilar species, which is reflected in their attribution
to different genera. At the same time, all members of the
subclade 9 have a short psbA spacer. Unfortunately, the
subclade includes the nomenclatural type of Hymenidium,
H. brunonis. This means that the separation of the “southern”
group of Pleurospermum s. l. as the genus Hymenidium,isnot
supported by the molecular data. In addition, in the “apioid”
tree (Fig. 1) subclade 9 appears to be distant from all other
subclades of Pleurospermum s. l., and is in clade C formed by
morphologically dissimilar species from other tribes. The
morphological analysis revealed a high degree of similarity
between Sinodielsia thibetica and Trachydium subnudum.Inthe
molecular analysis these species are also clustered together
(Figs. 1, 2A, B, D) with a high support value (PP = 1.00, BS =
100%), except for in the chloroplast psbA-trnH tree (Fig. 2C).
The small subclade 8, with two Pterocyclus species, occu-
pies a basal position in the lineage also containing subclades 6
and 7 (Figs. 1, 2A, B, D). Some other species of Hymenidium,
Trachydium, and Hymenolaena,aswellasKeraymonia cortiformis
and Sinolimprichtia alpina, are placed in the same lineage.
Morphologically, three species of Pterocyclus are clustered
together with P. rivulorum as the most divergent (Pimenov
et al. 2000). In all molecular trees, P. angelicoides and P. forrestii
are also together, but P. rivulorum fell into clade 9. The place-
ment of P. rivulorum in all molecular trees with Hymenidium
apiolens, H. brunonis,Trachydium subnudum,andSinodielsia
thibetica is surprising in light of the comparative morphology
of these species. It differs from the last four species in having
ternate (vs. pinnatisect) leaf blades with cordate, broad, entire
(vs. narrow pinnatifid or bipinnatifid) primary lobes, broad
(vs. narrow) leaf sheaths, and large (8–10 mm long) lanceolate
or lanceolate-linear (vs. small, 2–5 mm long, and ovate to
General Discussion—The groups obtained in the molecu-
lar (Figs. 1, 2A–D) study are evidently polyphyletic, by the
same token they are dissimilar morphologically. There are
no principal arguments to give preference to one type of
evidence to reveal phylogeny and to elaborate taxonomy,
especially if both character types are considered as differ-
ent levels of structural (i.e. morphological) variability
Only a few traditional or recently revised taxa were more
or less supported as clades in the molecular trees. The results
of the molecular phylogenetic analysis of Pleurospermum and
related, or presumably related, taxa are similar to those
obtained previously in molecular phylogenies of some west-
ern North American genera of Apioideae, such as Cymopterus
or Lomatium (Downie et al. 2002; Sun et al. 2004; Sun and
Downie 2010), as well as in our attempt to clarify relation-
ships in the informal generic group Ligusticeae or
Foeniculinae of the Old World (Valiejo-Roman et al. 2006).
These taxa, and Peucedanum s. l., although different in essence,
all belong to complicated genera or tribes, the systematics of
which up to now has not been satisfactorily resolved on a
The present preliminary results could stimulate further
investigation, both molecular and morphological, to include
a search for new material of the described but lost species,
as well as the use of additional molecular markers. Most
probably, such an expanded investigation, with a more
complete set of species and sequences, will lead to further
splitting of traditional Pleurospermum and separate it from
Hymenidium, which is obviously polyphyletic in its presently
accepted form. For instance, two groups in Hymenidium
(subclades 4 and 10), each morphologically homogeneous,
most probably deserve separation as taxa after more
Acknowledgments. The authors thank Prof. Gregory M. Plunkett
and Dr Victoria S. Shneyer for constructive discussion and three anony-
mous reviewers for providing help and advice. The Russian Foundation
of Basic Research (RFBR) for financial support (grants 10-04-00675) and
the National Geographic Society (U. S. A.) for sponsoring the Chinese
expedition are also acknowledged.
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Appendix 1. Species sampled, voucher information (origin; collector,
collector number and herbarium where specimen is deposited), and
GenBank accession numbers for the 55 new accessions of Umbelliferae
examined for ITS (1-2) and cpDNA psbA-trnH intergenic spacers, and the
trnL-trnF region (including trnL intron, and trnL-trnF intergenic spacer).
Aulacospermum anomalum (Ledeb.) Ledeb., cult. Royal Botanic Garden,
Edinburgh, U. K.; no. 19932275: FJ475132, FJ475180. Aulacospermum
popovii (Korovin) Kljuykov, Pimenov & V. N. Tikhom., Kirghizia,
Chatkal distr, 25.VII.1986; Pimenov et al. 364 (MW): - FJ469932, FJ483471,
FJ475133, FJ475181. Aulacospermum simplex Rupr., Kirghizia, cult. Bot.
Gard. MSU, Russia s. n. (MW): FJ475134, FJ475182. Aulacospermum
stylosum (C. B. Clarke) Rech. f. & Riedl, India, Himachal Pradesh, Lahul
and Spiti distr., 28.VIII.2000, Pimenov and Kljuykov,53 (MW): FJ469933,
FJ483472, FJ475135, FJ475183. Bupleurum falcatum L., BG MSU, seeds from
BG Wroclaw (Poland), 1988: HQ246208, HQ246195. Hymenidium hookeri
(C. B. Clarke) Pimenov & Kljuykov, (
Pleurospermum hookeri C. B.
Clarke), India, Jammu and Kashmir, Ladakh, 3.IX.2000; Pimenov,Kljuykov
100 (MW): FJ469946, FJ483485, FJ475150, FJ475197. Hymenidium amabile
2012] VALIEJO-ROMAN ET AL:BROAD POLYPHYLY IN PLEUROSPERMUM S. L. 579
Craib & W. W. Sm.), China, Sichuan, road from Litang to Xiangcheng,
24.IX.1998; Pimenov et al. 396 (MW): FJ469934, FJ483473, FJ475137,
FJ475185. Hymenidium lindleyanum (Klotzsch) Pimenov & Kljuykov
(Hymenolaena lindleyana Klotzsch), India, Jammu and Kashmir,
Ladakh, 30.VIII.2000; Pimenov, Kljuykov 80 (MW): FJ469949, FJ483488,
FJ475153, FJ475200. Hymenidium nanum (Rupr.) Pimenov & Kljuykov
(Hymenolaena nana Rupr.), Kirghizia, E. Tianschan, basin of
Sarydzhaz, 14.VIII.1988; Pimenov and Kljuykov 594 (MW): FJ469952,
FJ483491, FJ475156. Hymenidium apiolens (C. B. Clarke) Pimenov &
Pleurospermum apiolens C. B. Clarke), E. Himalaya, Central
Nepal, Langtang National Park; 31.X.1999; Pimenov and Kljuykov 40
(MW); FJ469935, FJ483474, FJ475138, FJ475186. Hymenidium benthamii
(DC.) Pimenov & Kljuykov (
Pleurospermum benthamii (DC.) C. B.
Clarke), E. Himalaya, Central Nepal, Langtang National Park, 31.X.1999;
Pimenov and Kljuykov 34 (MW): FJ469936, FJ483475, FJ475139,
FJ475187. Hymenidium bicolor (Franch.) Pimenov & Kljuykov (
govanianum (DC.) Benth. ex C. B. Clarke var. bicolor Franch.), China,
Sichuan, between Ranwu and Shanranwu, 25.IX.1998; Pimenov et al. 419
(MW); FJ469937, FJ483476, FJ475140, FJ475188. Hymenidium brunonis (DC.)
Pleurospermum brunonis (DC.) Benth. ex C. B. Clarke), India,
Uttaranchal state, Chamoli distr., W. Himalaya, 8.X.2003; Pimenov and
Kljuykov 43 (MW): FJ469938, FJ483477, FJ475142, FJ475189. Hymenidium
chloroleucum (Diels) Pimenov & Kljuykov (Trachydium chloroleucum
Diels), China, Yunnan, Lijiang Co., 1.X.1998; Pimenov et al. 501 (MW):
FJ469939, FJ483478, FJ475143, FJ475190. Hymenidium davidii (Franch.)
Pimenov & Kljuykov (
Pleurospermum davidii Franch.), China, Sichuan,
Kanding Co., Paoma mountain park, 20.IX.1998; Pimenov et al. 280 (MW):
FJ469940, FJ483479, FJ475144, FJ475191. Hymenidium decurrens (Franch.)
Pimenov & Kljuykov (
Pleurospermum decurrens Franch.), China, Yunnan,
Tali Co., Diancang Shan Mts., 4.X.1998; Pimenov et al. 560 (MW): FJ469941,
FJ483480, FJ475145, FJ475192. Hymenidium delavayi (Franch.) Pimenov &
Kljuykov (Ligusticum delavayi Franch.), China, Yunnan, Lijiang Co., Mts.
Yulongxue Shan, 1.X.1998; Pimenov et al. 50 (MW): FJ469942, FJ483481,
FJ475146, FJ475193. Hymenidium foetens (Franch.) Pimenov & Kljuykov
Pleurospermum foetens Franch.), China, Yunnan, NW part, Degen Co.,
29.IX.1998; Pimenov et al. 481 (MW): FJ469943, FJ483482, FJ475147, FJ475194.
Hymenidium hedinii (Diels) Pimenov & Kljuykov (
Diels), China, Qinghai: Madoi and Bayanka La., 3. IX.1998; G. Miehe
et al.: 98-35626: FJ469944, FJ483483, FJ475148, FJ475195. Hymenidium
heterosciadium (H. Wolff) Pimenov & Kljuykov (
heterosciadium H. Wolff), China, Sichuan, between Xinduqiao and
Yajiange, 22.IX.1998; Pimenov et al. 317 (MW): FJ469945, FJ483484,
FJ475149, FJ475196. Hymenidium huzhihaoi Pimenov & Kljuykov, China,
Sichuan, between Xinduqiao and Yajiang, 22.IX.1998; Pimenov et al. 314
(MW): FJ469947, FJ483486, FJ475151, FJ475198. Trachydium involucellatum
Shan Ren Hwa & Pu Fa Ting, China, Xizang, SE Tibet, 19.VIII.1994;
G. Miehe and Wundisch 94-185-17: FJ469971, FJ483509, HQ246202.
Hymenidium lhasanum Pimenov & Kljuykov, China, Xizang A. R. (Tibet),
NW of Lhasa, 13.VIII.1989; B. Dichore 3917: FJ469948, FJ483487, FJ475152,
FJ475199. Hymenidium linearilobum (W. W. Sm.) Pimenov & Kljuykov
Pleurospermum linearilobum W. W. Sm.), China, Sichuan Xiangcheng
Co., 25.IX.1998; Pimenov et al. 406 (MW): FJ469950, FJ483489, FJ475154.
FJ475201. Hymenidium mieheanum Pimenov & Kljuykov, China, Xizang
A. R. (Tibet), Nyemo, Yarlung Tsangpo Gorge, 5.IX.1998; G. and S. Miehe
China 98-07901: FJ469951, FJ483490, FJ475155, FJ475202. Hymenidium
stellatum (D. Don) Pimenov & Kljuykov (
Pleurospermum stellatum (D. Don)
Benth. ex C. B. Clarke), India, Himachal Pradesh, Kullu distr., 27.VIII.2000;
Pimenov, Kljuykov 26 (MW): FJ469953, FJ483492, FJ475157, FJ475203.
Hymenidium virgatum Pimenov & Kljuykov, China, Sichuan, Kanding Co.,
19.IX. 1998; Pimenov et al. 258 (MW): FJ469954, FJ483493, FJ475158,
FJ475204. Hymenidium wilsonii (H. Boissieu) Pimenov & Kljuykov
Pleurospermum wilsonii H. Boissieu), China, Sichuan, road from Litang
to Xiangcheng, 24.IX.1998; Pimenov et al. 393 (MW): FJ469956, FJ483495,
FJ475159, FJ475206. Hymenidium wrightianum (H. Boissieu) Pimenov &
Pleurospermum wrightianum H. Boissieu), China, Yunnan,
10.IX.1990; KUN N.90-549: FJ469955, FJ483494, FJ475160, FJ475205.
Hymenolaena badachschanica Pissjaukova, Tadzhikistan, Badachschan,
Koitezek Pass., Upaly-Sai, 24.VI.1973; Pimenov and Kljuykov, 1141
(MW): FJ469957, FJ483496, FJ475161. FJ475207. Hymenolaena candollei
Wall. ex DC., India, Jammu and Kashmir, Baralacha La Pass,
28.VIII.2000; Pimenov and Kljuykov, 59 (MW): FJ469958, FJ483497,
FJ475162, FJ475208. Hymenolaena pimpinellifolia Rupr., Kirghizia, Mts.
Kirghiz Alatau, Tuzaschu Pass, 11.VIII.1976; Pimenov 398 (MW):
FJ469959, FJ483498, FJ475163, FJ475209. Keraymonia cortiformis Cauwet &
S. B. Malla, China, Xizang, S Tibet, Central Himalayas, 22.VIII.1993;
G.andS.Miehe:971(FLORA OF TIBET 9537/08): FJ469960, FJ483499,
FJ475164, HQ246197. Physospermum cornubiense (L.) DC, Ukraine, the
Crimea, Alikat-Bogaz Pass, 4.IX.1974; Pimenov and L. P. Tomkovich, s. n.
(MW): FJ475167 FJ475212. Physospermopsis delavayi (Franch.) H. Wolff,
China, Yunnan, Lijiang Co., Mts. Yulongxue Shan; 1.X.1998; Pimenov
et al. 532 (MW): FJ483501; psbA-trnH HQ246209;trnL-trnF HQ246198.
Physospermopsis muktinathensis Farille & S. B. Malla, E. Himalaya, Central
Nepal, south slopes of Annapurna, 23.X.1999; Pimenov and Kljuykov 22
(MW): FJ469961, FJ483500, FJ475165, FJ475210. Physospermopsis nana
(Franch.) Pimenov & Kljuykov (
Pleurospermum nanum Franch.), China,
Yunnan, Tali Co., Diancang Shan Mts., 4.X.1998; Pimenov et al. 557 (MW):
FJ469970, FJ483508, FJ475166, FJ475219. Pleurospermum austriacum (L.)
Hoffm., Romania, Transilvania, distr., Odorhei 6.VIII.1949; E. Ghisa and
E. Topa 2959 (MW): FJ469962, FJ483502, FJ475168, FJ475211. Pleurospermum
uralense Hoffm., Kazakhstan, E. Kazakhstan distr., Kalbin Mts., 29.VII.1972;
Pimenov et al. 242 (MW): FJ475169, FJ475213. Pseudotrachydium dichotomum
(Korovin) Pimenov & Kljuykov, Uzbekistan, S. slope of Seravschan
Mts., 23.VI.1988; Pimenov et al. 30 (MW): HQ246207, HQ342148, FJ475172,
HQ829364. Pseudotrachydium kotschyi (Boiss.) Pimenov & Kljuykov,
Iran, Prov. Hamadan, S. of Hamadan, Alvand Mts., 14.VI.2001; Pimenov
et al. 341 (MW): FJ469963, FJ475170, FJ475214. Pseudotrachydium vesiculoso-
alatum (Rech. f.) Pimenov & Kljuykov, Turkmenia, C. Kopet-Dagh,
13.VI.1978; Pimenov et al. 527 (MW):FJ469964, FJ483503, FJ475171,
HQ246199. Pterocyclus angelicoides (DC.) Klotzsch (
angelicoides Benth. ex C. B. Clarke), India, Uttar Pradesh, Chamoli distr.,
Main Himalayan Range Bhyundar, 18.IX.2000; Pimenov and Kljuykov 233
(MW): FJ469967, FJ483505, FJ475173, FJ475215. Pterocyclus forrestii (Diels)
Pimenov & Kljuykov (Angelica forrestii Diels), China, Yunnan, NW part,
Zhongdian Co., 27.IX.1998; Pimenov et al. 453 (MW): FJ469965, FJ483504,
HQ246200. Pterocyclus rivulorum (Diels) H. Wolff (
rivulorum (Diels) Fu Kun Tsun & Ho Yeh Chi), China, Yunnan, Lijiang
Co., Mts. Yulongxue Shan, 2.X.1998; Pimenov and Kljuykov 540 (MW):
FJ475174, FJ475216. Sinodielsia thibetica (H. Boissieu) Pimenov & Kljuykov
(Vicatia thibetica H. Boissieu), China, Sichuan, Yajiang Co., 22.IX. 1998;
Pimenov and Kljuykov 326 (MW): FJ469969, FJ483507, HQ246211,
HQ246201. Sinolimprichtia alpina H. Wolff, China, Yunnan, NW part,
Degen Co, 28.IX.1998; Pimenov and Kljuykov 487 (MW): FJ475175, FJ475217.
Trachydium roylei Lindl., Pakistan, Hazara, HW Himalaya, 14.IX.1995;
B. Dickore (FLORA OF PAKISTAN 13244): FJ469972, FJ483510, FJ475176,
HQ246203. Trachydium souliei H. Boissieu, China, Yunnan, NW part, Degen
Co., 28.IX.1998; Pimenov et al. 472 (MW): FJ469973, FJ483511, FJ475178,
FJ475218. Trachydium simplicifolium W. W. Smith, China, Yunnan, Lijiang
Co., 2.X.1998; Pimenov et al. 537 (MW):FJ475177, FJ475220. Trachydium
subnudum C. B. Clarke ex H. Wolff, China, Xizang A. R. (Tibet), Nindung
Shang, 7.VIII.1998; G. and S. Miehe; (CHINA 98-03329): FJ469974,
FJ483512, FJ475179, FJ469974. Trachydium variabile H. Wolff, China, Sichuan
25.X.1998; Pimenov et al. 418 (MW): HQ246206, HQ342149, HQ246206,
HQ246205. Heteromorpha arborescens Cham. & Schltdl., Republic of South Africa,
Prov. Limpopo, 05.I.2003; Pimenov et al. 23 (MW): HQ829359, HQ829361.
Heteromorpha pubescens Burtt Davy, Republic of South Africa, Prov.
Mpumalanga, 04.I.2003; Pimenov et al. 10 (MW): HQ829360, HQ829362. Glia
prolifera (Burm. f.) B. L. Burtt, Republic of South Africa, Prov. Western
Cape, 14. I.2003; Pimenov et al. 76 (MW): HQ829358, HQ829363.
Previously published ITS accessions of Apiaceae obtained from
GenBank—Aciphylla congesta Cheeseman; EU886819. Aegopodium
kashmiricum (R. R. Stewart ex Dunn) Pimenov; AF077872. Ammi majus L.;
GQ148786. Angelica sylvestris L.; ASU78414, ASU78474. Angelica tatianae
Bordz.; AF008610, AF009089. Anisosciadium isosciadium Bornm.; EU169244.
Anisotome aromatica Hook. f.; AAU78420, AAU78360. Apium graveolens L.;
GQ379287. Arafoe aromatica Pimenov & Lavrova; AF077874. Aulacospermum
anomalum Ledeb.; AF009120, AF008641. Aulacospermum simplex Rupr.;
AF008640, AF009119. Aulacospermum turkestanicum (Franch.) Schischk.;
GQ379340. Bunium bulbocastanum L.; DQ435210, DQ435249. Bupleurum
falcatum L.; AF077877. Carum carvi L.; AF077878. Cicuta virosa L.; U78372.
Coriandrum sativum L.; HQ377205. Daucus carota L.; AF077780, AF077095.
Echinophora orientalis Hedge & Lamond; EU169267. Elaeosticta paniculata
(Korovin) Kljuykov & Pimenov; DQ422817, DQ422836. Falcaria vulgaris
Bernh.; AF077888. Foeniculum vulgare Mill.; U78385, U78445. Gingidia
trifoliolata (Hook. f.) J. W. Dawson; GTU72367. Glia prolifera (Burm. f.) B. L.
Burtt; DQ368846. Heteromorpha arborescens Cham. & Schltdl.; DQ368849.
Heteromorpha pubescens Burtt Davy; DQ368855. Lecokia cretica DC.;
LCU78358. Nothosmyrnium japonicum Miq.; EU236179. Oenanthe fistulosa L;
AY360249. Physospermopsis delavayi (Franch.) H. Wolff; FJ385056.
Physospermum cornubiense (L.) DC.; AF077904. Pimpinella peregrina L.;
PPRDNA1, PPRDNA2. Pleurospermopsis sikkimensis (C. B. Clarke)
580 SYSTEMATIC BOTANY [Volume 37
C. Norman; GQ379347. Pleurospermum decurrens Franch.; AF164837, AF164862.
Pleurospermum foetens Franch; AF008639, AF009118. Pleurospermum hookeri
C. B. Clarke; AF164838, AF164863. Pleurospermum uralense Hoffm.;
AF008638, AF009117. Pleurospermum wilsonii H. Boissieu; EU236200.
Pleurospermum wrightianum H. Boissieu; EU236201. Pseudotrachydium
dichotomum (Korovin) Pimenov & Kljuykov; GQ379343. Pseudotrachydium
kotschyi (Boiss.) Pimenov & Kljuykov; AY941284, AY941312. Pterocyclus
rivulorum (Diels) H. Wolff; AY038205, AY038219. Pycnocycla aucheriana
Boiss.; AF073533. Scaligeria moreana Engstrand; AF073545S1, AF073545S2.
Scandix stellata Banks & Sol.; AF073629S1, AF073629S2. Selinum carvifolia
(L.) L.; AY328930, AY330496. Seseli krylovii (V. N. Tichom.) Pimenov &
Sdobnina; AF077908. Sinodielsia delavayi (Franch.) Pimenov & Kljuykov;
AY038211, AY038225. Sinolimprichtia alpina H. Wolff; AY328953,
AY330519. Sium latifolium L.; AY360257. Smyrnium olusatrum L.; U30594.
Torilis arvensis (Huds.) Link; AF077801, AF077116. Trachydium simplicifolium
W. W. Sm.; AY038201, AY038215.
2012] VALIEJO-ROMAN ET AL:BROAD POLYPHYLY IN PLEUROSPERMUM S. L. 581