Print
New insights into the phylogeny and taxonomy of Chinese Physospermopsis (Apiaceae)
expand article infoXin-Rui Xu, Xian-Lin Guo, Megan Price, Xing-Jin He, Song-Dong Zhou
‡ Sichuan University, Chengdu, China
Open Access

Abstract

Physospermopsis (Apiaceae) comprises about 10 species, but its taxonomy and phylogeny are disputed. The genus is mostly distributed in the Himalayas and Hengduan Mountains at high elevation. Earlier molecular studies involving six species of Physospermopsis indicated that this genus is not monophyletic and is nested in the East Asia Clade. Therefore, the aims of this study were to re-assess the phylogenetic position of, and interspecific relationships within, Physospermopsis based on two chloroplast loci (rpl16, rps16) and one nuclear region, the internal transcribed spacers of ribosomal DNA (ITS). Eight species involving 13 populations of Physospermopsis were collected. These were sequenced and analyzed with the sequences of 31 other Apiaceae species obtained from the NCBI to determine phylogenetic relationships using Bayesian inference (BI) and Maximum likelihood (ML). Our study found that Physospermopsis is monophyletic, nested in Pleurospermeae of Apiaceae, sister to Pleurospermum. And we propose that the Physospermopsis clade should be replaced by the East Asia Clade. However, the interspecific relationships within Physospermopsis were not well resolved and the positioning of species was unclear. Diagnostic characteristics to distinguish Physospermopsis species in the field and laboratory are provided for future Physospermopsis phylogenetic studies.

Keywords

Apiaceae, morphology, phylogeny, Physospermopsis, taxonomy

Introduction

Physospermopsis H. Wolff (1925: 276) has been reported to contain about 10 species, with eight species distributed in China (Pan and Watson 2005). However, 11 species were reported by Pimenov and Kljuykov (Pimenov and Kljuykov 2000a, b, c; Pimenov 2017). There has been difficulty in interpretation of diverse morphology to diagnose species and even limits of the genus. Most species of this genus occur in the Himalayas and Hengduan Mountains, and of these, four are endemic to the Hengduan Mountains (Wang and Pu 1992). In China, most species of Physospermopsis grow in open forests, scrubs, grasslands and alpine meadows at elevations of 2250–4800 m (Wolff 1929; Mukherjee 1982; Farille and Malla 1985; Pan and Watson 2005). Physospermopsis is characterized by having a long, conic taproot, ribbed stem, pinnate, rarely entire leaf blade, prominent, leaf-like bracts, variable bracteoles, minute calyx teeth, emerald young fruits, ovoid to broadly ovoid mature fruit with slightly cordate base (Wolff 1925; Pan and Watson 2005). Based on an analysis of previous research (Wang and Pu 1992; Pu and Liu 2005, 2006), Physospermopsis species usually possess a pericarp with wavy stria or reticulate ornamentation, prominent or inconspicuous fruit ribs, diverse carpoderms and endosperms, and pollen morphology showing a trend from rhomboidal type to rectangular type.

Previous studies on Physospermopsis have been extensive, including on micromorphology, anatomy and pollen morphology (Wang and Pu 1992; Pu and Liu 2005, 2006). However, previous molecular phylogenetic analyses of Physospermopsis have only involved a small number of taxa mostly using internal transcribed spacer (ITS) sequences (Downie et al. 2000; Calviño et al. 2006; Zhou et al. 2008, 2009; Downie et al. 2010; Valiejo-Roman et al. 2012). Phylogeny of Physospermopsis has been disputed with Downie et al. (2000) placing P. kingdon-wardii (H.Wolff) C.Norman (1938: 231) and P. rubrinervis (Franchet) C.Norman in the Komarovia clade based on the materials collected from Yunnan, China. Then, Calviño et al. (2006) provisionally positioned Physospermopsis in the Physospermopsis clade based on more comprehensive maximum parsimony (MP) analyses of ITS sequences, which arose as a weakly supported sister group to the Komarovia clade. Later, Zhou et al. (2008) studied five species of Physospermopsis and concluded that Physospermopsis was not a monophyletic group. Zhou et al. (2008) placed P. kingdon-wardii and P. rubrinervis in the East Asia clade and referred them to Trachydium J.Lindley (1835: 232). Zhou et al. (2008) also concluded that P. cuneata H.Wolff (1929: 126) was nested in Pimpinelleae and should be close to Pimpinella C.Linnaeus (1753: 263), while P. muliensis R.H.Shan & S.L.Liou (1979: 105) and P. shaniana C.Y.Wu & F.T.Pu (1993: 1285) were allied within Pleurospermeae. Additionally, the East Asia clade was proposed as the synonym of Physospermopsis clade due to almost all of its species being primarily distributed in East Asia (Zhou et al. 2008). The following year, Zhou et al. (2009) added P. delavayi (the nomenclatural type of Physospermopsis) to their previous analyses and placed it in Pleurospermeae. Downie et al. (2010) decided that Physospermopsis should be placed in the Physospermopsis clade (East Asia clade) and was not monophyletic, but did not include the type species in their analyses potentially influencing their conclusions. Valiejo-Roman et al. (2012) conducted a molecular phylogenetic analysis of the genus Pleurospermum G.F.Hoffmann (Hoffmann 1814) and its related genera, including three Physospermopsis species.

Physospermopsis is a taxonomically complex genus whose generic limits with Pleurospermum, Tongoloa H.Wolff (1925: 279), and Trachydium are problematic (Pan and Watson 2005). Therefore, misidentification was common due to the absence of convincing morphological evidence, limitations of collected materials and examinations of type specimens. Additionally, until now there has been no comprehensive analysis using molecular phylogenetics and morphology within the one study. Therefore, we aimed to determine an accurate phylogeny of Physospermopsis and infrageneric relationships within Physospermopsis based on molecular, morphology data and combined analysis linking phylogeny and morphology. We acquired accurate data by collecting field specimens of eight Physospermopsis species involving 13 populations from their type localities and adjacent areas. Species were identified by field observations, validation with herbarium specimens and primary literature.

Material and methods

Specimen examinations, field investigations and morphology observations

The taxonomic identification of Physospermopsis species was by field investigations and specimen examinations from herbaria BM, BNU, CDBI, CVH, E, HITBC, ILL, K, KUN, LBG, LE, MW, NAS, NHW, P, PE, PEY, SABG, SM, SZ, UC, WU, WUK.

In the field investigations, we sampled three populations of P. delavayi, two populations of P. rubrinervis, two populations of P. shaniana, one population of P. obtusiuscula (1938: 231) and one population of P. nana (2000: 538) in Yunnan Province. We sampled one population of P. kingdon-wardii and one population of P. obtusiuscula in Tibet. One population of P. alepidioides (H.Wolff & Hand.-Mazz.) R.H.Shan (1941: 187) and one population of P. muliensis were sampled in Sichuan Province. All populations were collected from the type locality and adjacent regions, and the features of every species were closely matched with the types and original descriptions (de Candolle 1830; Franchet 1894; Diels 1912; Wolff 1929; Wolff et Handel-Mazzetti 1933; Shan et Liou 1979). The specific collection information are listed in Appendix 1.

Fruits, leaf segments and specimens from these eight species of Physospermopsis were collected in the field for morphological study. Morphological analyses of leaves and fruits based on herbarium specimens or formaldehyde-acetic acid-alcohol (FAA) preserved material were photographed by a stereomicroscope Nikon SMZ25 (Japan). The morphological data were measured using KaryoType (Altnordu et al. 2016).

Taxon sampling

We sampled 13 populations, representing eight species of Physospermopsis in our phylogenetic analysis, and obtained 31 sequences of other Apiaceae species from the NCBI (Appendix 1). Based on previous research (Zhou et al. 2009), Bupleurum krylovianum B.K.Schischkin (1935: 2010) and Bupleurum rockii H. Wolff (1929: 187) were selected as the outgroup for studying the phylogenetic position of Physospermopsis. We chose Pl. franchetianum W.B.Hemsley (1892: 307) and Pl. wrightianum H.Boissieu (1903: 847) as the outgroup for studying interspecific relationships within Physospermopsis. The DNA sequences of two chloroplast loci (rpl16, rps16) and one nuclear region, the internal transcribed spacers of ribosomal DNA (ITS), were used for phylogenetic analyses. According to the research to date (Zhou et al. 2008, 2009; Downie et al. 2010; Guo et al. 2018; Panahi et al. 2018), these three markers should be sufficient to obtain the general information about relationships within the genus and its phylogenetic position within the family Apiaceae.

DNA extraction and sequencing

The fresh leaves were collected from field specimens in Yunnan, Sichuan and Tibet, China. Voucher specimens were deposited in the Herbarium of Sichuan University (SZ). Total genomic DNA was extracted from silica-dried leaves with plant genomic DNA kit (Cwbio Biosciences, Beijing, China). The universal primers ITS4 (5’-TCC TCCGCT TAT TGA TAT GC-3’) and ITS5(5’-GGA AGT AAA AGT CGT AAC AAG G-3’; White et al. 1990) were used to amplify the entire internal transcribed sequences. The rpl16 intron region was amplified using primers F71(5’-GCT ATG CTT AGT GTG TGA CTC GTT G-3’) and R1516 (5’-CCC TTC ATT CTT CTA TGT TG-3’) (Jordan et al. 1996; Kelchner and Clark 1997). The rps16 sequences were amplified with primers rps16 3’exon (5’-CCT GTA GGY TGN GCN CCY TT-3’) and rps16 5’exon (5’-AAA CGA TGT GGN AGN AAR CA-3’)(Downie and Katz-Downie 1999). PCR amplification was implemented in a 30 μL volume reaction, including 3 μL total DNA, 1.5 μL forward primer, 1.5 μL reverse primer, 15 μL 2×Taq MasterMix (Cwbio, Beijing, China), and 9 μL ddH2O. The amplification of the ITS region was obtained by initial denaturation for 3 min at 94 °C, followed by 30 cycles of 45 s at 94 °C, 70 s at 54 °C, and 90 s at 72 °C, then final extension of 10 min at 72 °C. Amplification of cpDNA intron regions was obtained by initial denaturation for 3 min at 94 °C, followed by 36 cycles of 45 s at 94 °C,70 s at 58.5 °C, and 90 s at 72 °C, then final extension of 10 min at 72 °C. All PCR products were separated using a 1.5% (w/v) agarose TAE gel and sent to Sangon (Shanghai, China) for sequencing. New sequences obtained for this study have been deposited in GenBank. GenBank accession numbers are provided in the Appendix 1.

Data analysis

We used SegMan7 (Burland 2000) to assemble ITS and cpDNA sequences. ClustalX (Jeanmougin et al. 1998) was used to align DNA sequences with manual adjustment. We then used MEGA7 (Kumar et al. 2016) to manually adjust and obtain ITS and cpDNA datasets. Gaps were positioned to minimize nucleotide mismatches. Bayesian inference (BI) and Maximum likelihood (ML) methods were used for phylogenetic analyses, using MrBayes v3.2 (Ronquist et al. 2012) and RAxML v8.2.4 (Stamatakis 2014), respectively. Before undertaking BI analyses, MrModeltest version 2.2 (Nylander 2004) was used to determine the best model of nucleotide substitution and the GTR+G model under the Akaike Information Criterion (Akaike 1974) was selected. Bayesian analyses were performed over 20 million generations with a variant of Markov Chain Monte Carlo (MCMC) method and the trees were saved to a file every 1,000 generations. The first 20% trees were discarded as “burn-in” and the remaining 80% trees were used to build a majority-rule consensus tree based on analysis of the program Tracer v1.4 (Drummond and Rambaut 2007). ML analyses were performed using RAxML v8.2.4 with the GTR+G model and 1,000 bootstrap replicates. We constructed the BI tree with ITS data from all 44 taxa to test the systematic position of Physospermopsis. And we mapped some valuable morphological characteristics of Physospermopsis on phylogenetic tree, including leaves, bracts and bracteoles, ribs of fruits. The BI and ML trees were constructed for analysis of interspecific relationships within Physospermopsis using ITS and plastid datasets from the 13 Physospermopsis populations we sampled, one Physospermopsis species and the two Pleurospermum species downloaded from NCBI. Detailed information on the investigated taxa can be found in the Appendix 1.

Results

Morphological characteristics of Physospermopsis

Through observations in the field, the most important characteristic to identify Physospermopsis species was prominent bracts and bracteoles. Physospermopsis shaniana, P. nana, P. muliensis, P. rubrinervis, P. obtusiuscula and P. kingdon-wardii usually have leaf-like bracts and bracteoles (Fig. 1A3–F3). While P. alepidioides and P. delavayi possess lanceolate or oblong bracts and bracteoles with a 2–3-lobed apex and dark purple margin (Fig. 1G3, H3). Furthermore, leaf shape varies with species and can be obovate-lanceolate (e.g. P. alepidioides), triangular (e.g. P. rubrinervis), obovate-orbicular (e.g. P. delavayi) or linear-lanceolate (e.g. P. nana) segments (Fig. 1B2, D2, G2, H2). Besides, the leaves of P. kingdon-wardii and P. obtusiuscula are 2-pinnate and ovate-oblong, and have 2–6 pairs of ovate pinnae with pinnatisect margin. Physospermopsis muliensis and P. shaniana possess 3–5 pairs pinnae with pinnatifid margin, narrowly winged petioles and narrow and purple-red sheaths. Fruit morphology was recorded prior to alcohol preservation because the alcohol altered the color slightly (as is seen in photographs). The fruits of Physospermopsis were emerald green or chartreuse, ovoid to broadly ovoid, and typically had a slightly cordate base, a gradually narrowed and laterally flattened apex, with filiform or prominent ribs. Fruit shape and size of all Physospermopsis species were similar except that P. kingdon-wardii had fruit half the size of other species and very prominent and sinuate ribs. Physospermopsis nana and P. muliensis fruits had relatively prominent and filiform ribs, but P. muliensis fruits had scattered warts especially on the ribs and P. nana had smaller fruit. Physospermopsis obtusiuscula fruits were ovoid with narrowly winged and sinuolate ribs. The fruit of P. delavayi had an obvious cordate base, and filiform and less prominent ribs. Physospermopsis alepidioides, P. rubrinervis and P. shaniana had ovoid, verucose fruits with prominent ribs, but P. alepidioides did not have a cordate base, while the other two species had a slightly cordate base. The fruit of P. shaniana had many small warts distinguishing it from P. rubrinervis. For easy reading and comparison, the main morphological characteristics were listed in Table 1.

Figure 1. 

Morphological characters of Physospermopsis A1–H1 habit A2–H2 basal leaf A3–H3 umbel A4–H4 mericarps A1 habit of P. shaniana B1 habit of P. nana C1 habit of P. muliensis D1 habit of P. rubrinervis E1 habit of P. obtusiuscula F1 habit of P. kingdon-wardii G1 habit of P. delavayi H1 habit of P. alepidioides A2 basal leaf of P. shaniana B2 basal leaf of P. nana C2 basal leaf of P. muliensis D2 basal leaf of P. rubrinervis E2 basal leaf of P. obtusiuscula F2 basal leaf of P. kingdon-wardii G2 basal leaf of P. delavayi H2 basal leaf of P. alepidioides A3 umbel of P. shaniana B3 umbel of P. nana C3 umbel of P. muliensis D3 umbel of P. rubrinervis E3 umbel of P. obtusiuscula F3 umbel of P. kingdon-wardii G3 umbel of P. delavayi H3 umbel of P. alepidioides A4 mericarps of P. shaniana B4 mericarps of P. nana C4 mericarps of P. muliensis D4 mericarps of P. rubrinervis E4 mericarps of P. obtusiuscula F4 mericarps of P. kingdon-wardii G4 mericarps of P. delavayi H4 mericarps of P. alepidioides.

Table 1.

The morphological characteristics of eight Physospermopsis species.

Taxa Bracts Bracteoles Fruits Leaf shape Ribs Stems Umbels
Physospermopsis delavayi lanceolate or oblong lanceolate broadly ovoid, with obvious cordate base winged, obovate-orbicular filiform branched above 7–13
P. alepidioides lanceolate or oblong ovate-lanceolate,entire ovoid, with obscure cordate base entire, obovate-lanceolate prominent branched above 5–14
P. muliensis leaf-like lanceolate, entire broadly ovoid or ovoid narrowly winged, pinnatifid relatively prominent, with scattered warts branched above, slender 7–18
P. rubrinervis leaf-like leaf-like, with purplish margin ovoid, with slightly cordate base triangular, with purple-red nerves prominent branched above, dark purple 6–17
P. shaniana leaf-like leaf-like, with purplish margin ovoid, with slightly cordate base narrowly winged, pinnatifid prominent,with small warts branched at the base, reduced 6–15
P. obtusiuscula leaf-like ovate-oblong ovoid to broadly ovoid ovate-oblong, pinnatisect with narrowly winged, sinuolate branched at the base, dark purple-green 5–20
P. kingdon-wardii leaf-like leaf-like, with purplish margin broadly ovoid ovate-oblong, pinnatisect prominent, sinuate, with sparse minute warts reduced, often acaulescent 5–11
P. nana leaf-like leaf-like, with membranous margin broadly ovoid linear-lanceolate prominent, narrowly sinuolate-winged reduced, slender 4–13

Phylogenetic analyses

Through comprehensive sampling, the ITS analyses indicated that the 13 populations of Physospermopsis we sampled and P. muktinathensis M.A.Farille & S.B.Malla (1985: 512) formed an individual clade. Physospermopsis was confirmed to be a monophyletic group and nested in Pleurospermeae. Trachydium roylei Lindl. (1835: 232) and Pl. wilsonii H.Boissieu (1906: 433) were the closest relatives of Physospermopsis (Fig. 2).

Figure 2. 

Bayesian tree inferred from the analysis of the 44 samples of ITS data. Branch lengths are proportional to the amount of character changes, scale = 0.02 substitutions per character. The tree is rooted with Bupleurum. The names of the clades identified are those of Zhou et al. (2008, 2009).

The ITS dataset tree topologies generated from BI and ML analyses were consistent. Therefore, only the BI tree with posterior probabilities (PP, 0–1) and bootstrap support values (BS, 0–100%) is illustrated in Fig. 3A. The first to differentiate from Physospermopsis was P. muktinathensis, which is distributed in Nepal. Three populations of P. delavayi and one of P. alepidioides united as a strongly supported (BIPP = 1; MLBS = 100%) group. Physospermopsis obtusiuscula was supported as a sister group to P. kingdon-wardii (BIPP = 1; MLBS = 100%). Physospermopsis rubrinervis, P. muliensis, P. nana and P. shaniana were allied in all trees (BIPP = 1; MLBS = 100%). However, clear interspecific relationships between P. rubrinervis, P. muliensis, P. nana and P. shaniana were not strongly supported by ML or BI analyses.

Figure 3. 

Bayesian trees of Physospermopsis and its related genus inferred from ITS (A) and plastid rpl16+rps16 (B). Values on the branches indicate their support (Bayesian posterior probability/ Maximum-likelihood bootstrap). Branch lengths are proportional to the amount of character changes, scale = 0.01 (A), 0.005 (B) substitutions per character.

The cpDNA dataset tree topologies inferred by BI and ML analyses were consistent (Fig. 3B). However, results of a partition homogeneity test for the ITS and cpDNA datasets indicated that these genomes provide significantly different phylogenetic estimates. The taxa involved in this conflict are highlighted in Fig. 3. There was no chloroplast data for P. muktinathensis. The first to differentiate were P. obtusiuscula and P. kingdon-wardii (BIPP = 1; MLBS = 99%). The relationships of the three P. delavayi populations differed from the ITS dataset tree topology, although this cpDNA dataset relationship was not strongly supported. The cpDNA dataset tree topologies indicated that LYG population was closer to the HB population (BIPP = 0.33; MLBS = 47%), while LYG was closer to LGH in the ITS dataset tree topologies (BIPP = 0.34; MLBS = 70%). Additionally, the relationships between P. rubrinervis, P. muliensis, P. nana were not consistent with the ITS tree, where P. nana allied with P. rubrinervis in the cpDNA tree (BIPP = 1; MLBS = 72%), whereas P. nana allied with P. muliensis in the ITS tree (BIPP = 1; MLBS = 94%).

Discussion

The Phylogenetic position of Physospermopsis and relationship between Physospermopsis and Pleurospermum

Physospermopsis is monophyletic. The reasons for previous designations as a polyphyletic genus were likely attributable to the misidentification of several species (e.g. P. rubrinervis, P. kingdon-wardii, P. cuneata). Besides, P. cuneata is a poorly known species and unusual within the genus for its lack of conspicuous bracts and bracteoles, and therefore the phylogenetic placement of it is highly controversial. However, the most recent consensus is that P. cuneata should not be placed in Physospermopsis (Zhou et al. 2008; Zhou et al. 2009; Pimenov 2017). So previous molecular studies only involved five physospermopsis species which were widely accepted; we added another three physospermopsis species in this study, including P. alepidioides, P. obtusiuscula, and P. nana. Evidence obtained through more precise checking of generic type, infrageneric types and extensive herbarium specimens, literature and field investigations, analyzing morphological characters, and ITS and cpDNA evidence. This comprehensiveness allows us to be confident that Physospermopsis is monophyletic and nested in Pleurospermeae. In addition, we propose that the Physospermopsis clade should be replaced by the East Asia Clade.

The molecular results indicated that Physospermopsis is closest to Pleurospermum. Morphologically, Pleurospermum usually possess numerous bracts and bracteoles with white scarious margins, conspicuous or obsolete calyx teeth, white or purple-red petals with clawed base and narrow apex, prominent, acute ribs (Pan and Watson 2005). However, we found that Physospermopsis differed from Pleurospermum by less prominent and even inconspicuous fruit ribs, and the bracts and bracteoles did not have white scarious margins, resulting in an obvious, diagnostic boundary between Pleurospermum and Physospermopsis. The closeness of the two genera is also evidenced in pollen morphology. Wang and Pu (1992) found P. alepidioides and P. muliensis pollen to be rhomboidal and similar to several Pleurospermum species whereas other Physospermopsis species (P. rubrinervis and P. delavayi) have more advanced rectangular types. In addition, Pan and Watson (2005) identified several Physospermopsis species (e.g. P. obtusiuscula) with morphological similarities to Pleurospermum species, including having long fruit ribs and bulgy fruit walls, while other species had flattening of fruit and reduced wall thickness. Consequently, Physospermopsis is sister to Pleurospermum.

Interspecific relationships within Physospermopsis

The morphological characteristics mapped on the phylogenetic tree indicated that most closely related species have similar morphological characteristics. For example, P. rubrinervis, P. muliensis and P. shaniana are highly consistent on leaves, bracts and bracteoles, ribs on fruits (Fig. 2). Similarly, these species are the geographically sympatric species (Fig. 4). Resolution of the relationships between these species will only be achieved through continued studies, which may be difficult due to their geographic and morphological similarities. However, we can learn that P. nana, P. rubrinervis, P. muliensis and P. shaniana are the more advanced species in Physospermopsis. The morphological characters of P. nana are the most particular; these might be caused by hybridization with Pleurospermum species.

Figure 4. 

Geographic distribution of the eight Chinese Physospermopsis species in China. The altitude, scale, name of provinces and provincial capitals are also showed on the map.

The interspecific relationships between certain species within Physospermopsis are evident based on the consistencies between ITS and cpDNA trees. For instance, P. alepidioides showed a close affinity to P. delavayi in phylogenetic tree, and they have similar bracts and bracteoles (entire or 2–3-lobed at apex, with dark purple margin) (Figs 1, 2). However, differing leaf shapes can be used to easily distinguish these two species because P. alepidioides has an undivided leaf with sparsely serrated margin (Fig. 1H2) and P. delavayi has a pinnate leaf (Fig. 1G2). Physospermopsis kingdon-wardii is sister to P. obtusiuscula, which is consistent with their geographic closeness. Physospermopsis kingdon-wardii appears more morphologically similar to P. obtusiuscula (including the leaves, bracts and bracteoles), but differs in its reduced stem, small stature and small fruits with prominent and sinuate ribs (Fig. 1).

The topologies of the ITS and cpDNA trees differed in the positioning of P. delavayi, P. muliensis and P. nana (Fig. 3). This inconsistency between nrDNA ITS and cpDNA data has been reported in some studies of Apiaceae (Lee and Downie 2006; Zhou et al. 2008, 2009; Spalik et al. 2009; Bone et al. 2011; Yi et al. 2015; Panahi et al. 2018). This difference generally has been caused by incomplete lineage sorting, hybridization, homoplastic substitutions and introgression. Since we did not sample by lineage and execute gene flow analysis, what caused the inconsistency cannot be determined. Previous studies have indicated that Pleurospermeae occupies a relative position in the base of the Apioideae (Zhou et al. 2008, 2009; Downie et al. 2010), the differentiation time should be earlier. Thus, for Physospermopsis, we infer the more effective reason for the inconsistency between nrDNA ITS and cpDNA data is hybridization. A further study based on widely sampling and deeper analysisis needed. However, several diagnostic characteristics can be utilized in the field and laboratory to separate them. P. rubrinervis can easily be recognized by purple-red nerves on the leaves, bracts and bracteoles with purple-red margin (Fig. 1). Physospermopsis muliensis possesses a slender, branched stem and narrowly winged basal petioles with narrow sheaths (Fig. 1). Physospermopsis nana has bracts and bracteoles with white scarious margins and linear-lanceolate segments with membranous-margined sheaths (Fig. 1). The stem of P. shaniana was reduced and branched at the base, and had prominent bracts 1–2-pinnate with developed, broad sheaths (Fig. 1).

Taxonomy

Chinese Physospermopsis species

Physospermopsis H.Wolff (1925: 276)

Type

Physospermopsis delavayi H.Wolff (1925: 278)

Key to the Chinese Physospermopsis species

1 Leaves entire, margin sparsely serrate P. alepidioides
Leaves pinnate or pinnatifid 2
2 Stems reduced, sometimes acaulescent 3
Stems developed 5
3 Bracteoles margin membranous P. nana
Bracteoles margin purplish 4
4 Fruits ovoid, with slightly cordate base; ribs prominent, with small warts P. shaniana
Fruits broadly ovoid; ribs prominent and sinuate, with sparse minute warts P. kingdon-wardii
5 Nerves of leaves purple-red P. rubrinervis
Nerves of leaves green 6
6 Bracts lanceolate or oblong; ribs filiform P. delavayi
Bracts leaf-like; ribs prominent 7
7 Bracteoles ovate-lanceolate,entire; ribs with scattered warts P. muliensis
Bracteoles ovate-oblong; ribs with narrowly winged, sinuolate P. obtusiuscula

Physospermopsis alepidioides (H.Wolff et Hand.-Mazz.) R.H.Shan, 1941: 187

Haploseseli alepidioides H.Wolff et Hand.-Mazz., 1933: 722

Type

China. Sichuan: Yanyuan County, 2700–2800 m, 7 Oct 1914, Handel-Mazzetti 5562 (holotype: WU [WU0060774]).

Diagnostic characters

Physospermopsis alepidioides usually possesses an entire leaf blade with a sparsely serrated margin. The shape of the entire leaf segment is an obvious diagnostic characteristic to distinguish it from other Physospermopsis species. The stem of it is velutinous.

Distribution

Endemic to China, Sichuan (Fig. 4).

Habitat

Physospermopsis alepidioides usually occurs in open forests and grasslands.

Additional specimens examined

China. Sichuan Province: Muli County, Hetaowan, 2300 m, 8 Aug 2019, X.R.Xu XXR2019080801 (SZ); Muli County, Hetaowan, 2250 m, 23 Dec 1982, Y.B.Yang & Y.L.Cao 400 (CDBI); Muli County, Liziping, 21 Sep 2011, X.G.Ma m11092101 (SZ); unknown locality, 2650 m, 19 Jul 1983, Anonymous 22 (HITBC); Yanyuan County, Mt. Huolu, 3950 m, 22 Jul 1983, Anonymous 25 (HITBC).

Physospermopsis rubrinervis (A.R.Franchet) C.Norman, 1938: 231

Trachydium rubrinerve A.R.Franchet, 1894: 112

Pleurospermum rubrinerve (A.R.Franchet) M.Hiroe, 1979: 747

Type

China. Yunnan: Eryuan County, Mt. Luoping, 3200 m, 31 Aug 1888, Delavay 3235 (holotype: P [P00245453]; lectotype, designated by Pimenov 2017, pg. 188: P [P00245453] ; isolectotypes: K [K001235378], P [P00245454, P00834665]).

Diagnostic characters

Physospermopsis rubrinervis usually possesses dark purple, sparsely branching stems. The basal blade is ovate to broadly ovate in outline, having almost purple-red nerves.

Distribution in China

Sichuan, Yunnan (Fig. 4).

Distribution outside China

India, Nepal.

Habitat

This species grows in the forest edge or rhododendron shrubs at an elevation of 2800–4800 m.

Additional specimens examined

China. Sichuan Province: Yanbian County, Yankou xiang, 3150 m, 20 Sep 2002, X.F.Gao, Y.L.Peng & G.Sun 3753 (PE); Meigu County, Ligou xiang, 3600 m, 5 Aug 1959, 1591 (SM); Butuo County, Wuke pasture, 3500 m, 1 Jul 1976, Vegetation expedition 13827 (CDBI); Dukou County, Mt. Dahei, 1400 m, 18 Jun 1983, Qinghai-Tibet Expedition 11231 (KUN); Puge County, 9 Aug 1960, Anonymous 25099 (SM); Yunnan Province: Eryuan County, Mt. Luoping, 3200 m, 17 Aug 2019, X.R.Xu XXR2019081701 (SZ); Lushui County, 3000 m, 18 Oct 2019, X.L.Guo G19101802 (SZ); unknown locality, 21 Sep 1959, S.G.Wu 2715 (KUN).

Physospermopsis kingdon-wardii (H.Wolff) C.Norman, 1938: 231

Trachydium kingdon-wardii H.Wolff, 1929: 124

Pleurospermum kingdon-wardii (H.Wolff) M.Hiroe,1979: 747

Type

China. Yunnan: A-tun-tsi, screes, turf, 14000 ft. (ca. 4267 m), 7 Aug 1913, Kingdon-Ward 992 (lectotype: E [E00000221]).

Diagnostic characters

P. kingdon-wardii is similar to P. obtusiuscula in shape of basal leaves, but the stem of P. kingdon-wardii is reduced. The fruits are smaller than other species, and the immature fruits sometimes have sparse minute warts. Additionally, the ribs are prominent, often sinuate.

Distribution in China

Tibet, Yunnan (Fig. 4).

Distribution outside China

Bhutan, Nepal, Sikkim.

Habitat

Physospermopsis kingdon-wardii usually grows in alpine meadows or scrubs at about 3900 m elevation.

Additional specimens examined

China. Tibet Province: Nyalam County, 4000 m, 24 Aug 2019, X.L.Guo G19082407 (SZ); Bainang County, 4580 m, 24 Aug 1988, Anonymous 8 (CDBI); Zayü County, 4180 m, 27 Sep 1982, Qinghai-Tibet Expedition 10772 (PE); Zayü County, 4370 m, 31 Aug 2003, X.F.Gao, W.G.Tu, H.He & Y.K.Qiao 6745 (CDBI); Yunnan Province: Dêqên County, 4300 m, 28 Sep 1981, L.R.Xu 129 (WUK); Dêqên County, 3900 m, 18 Aug 1940, K.M.Feng 6746 (PE); Dêqên County, 4500 m, 23 Sep 1986, H.Sun & Z.G.Qian 751 (KUN); Zhongdian County, 4300 m, 2 Oct 1986, H.Sun & Z.G.Qian 0980 (KUN).

Physospermopsis obtusiuscula (DC.) C.Norman, 1938: 231

Hymenolaena obtusiuscula DC., 1830: 246

Trachydium obtusiusculum (DC.) C.B.Clarke, 1879: 673

Pleurospermum obtusiusculum (DC.) M.Hiroe, 1979: 741

Aulacospermum obtusiusculum (DC.) A.R.Naqshi, U.Dhar et P.N.Kachroo, 1995: 107

Type

Nepal. “Ad Gossain-Than Nepalensium, Wallich [543]” (lectotype: G-DC; isolectotypes: BM [BM000622303, BM000944782], K [K000697363], K-WALLICH, LE).

Diagnostic characters

Physospermopsis obtusiuscula sometimes is flushed. The stems are dark purple-green, simple, and occasionally branched at the base. The fruit ribs are narrowly winged and sinuolate, which is a unique character in Physospermopsis.

Distribution in China

Sichuan, Tibet, Yunnan (Fig. 4).

Distribution outside China

Bhutan, India, Nepal, Sikkim.

Habitat

Physospermopsis obtusiuscula grows in shrubs or grassland at an elevation of ca. 4000 m.

Additional specimens examined

China. Sichuan Province: Xiangcheng County, 3900 m, 9 Aug 1981, Qinghai-Tibet Expedition 3986 (PE); Xiangcheng County, 9 Aug 1981, Qinghai-Tibet Expedition 3942 (PE); Tibet Province: Yadong County, 3500 m, 20 Aug 2019, X.L.Guo G19082009 (SZ); Nyingchi County, 3400 m, 8 Aug 1983, B.S.Li & S.Z.Cheng 6199 (PE); Nyingchi County, Mt. Shergyla, 3346 m, 13 Oct 2009, J.Luo, S.L.Wang & G.Y.Wang LiuJQ-09XZ-388 (KUN); Nyingchi County, Mt. Shergyla, 3346 m, 13 Oct 2009, J.Luo, S.L.Wang & G.Y.Wang LuoJian-ZX-0938 (PE); Yadong County, 3980 m, 14 Sep 1974, Qinghai-Tibet Expedition 74-2505 (PE); Yadong County, 4000 m, 12 Sep 1974, Qinghai-Tibet Expedition 2416 (PE); Nyalam County, 3800 m, 2 Sep 1972, 1736 (PE); Zayü County, 4300 m, 26 Sep 1982, Qinghai-Tibet Expedition 10635 (PE); Yunnan Province: Dêqên County, Baimaxueshan, 4100 m, 15 Aug 2019, X.R.Xu XXR2019081502 (SZ); Gongshan County, Dulongjiang, 2900 m, 15 Sep 1938, T.T.Yü 20274 (PE); Gongshan County, Dulongjiang, 3800 m, 9 Aug 1938, T.T.Yü 19829 (PE).

Physospermopsis muliensis R.H.Shan et S.L.Liou, 1979: 105

Type

China. Sichuan: Muli County, 4000 m, 20 Oct 1937, T.T.Yu 14579 (holotype: PE [P01432306]).

Diagnostic characters

Physospermopsis muliensis usually possesses branching stems, ovate-oblong leaf blades, narrow sheaths, leaf-like bracts, lanceolate bracteoles, and ovoid fruits with filiform ribs with sparse scattered warts. Basal and lower petioles are narrowly winged.

Distribution

Endemic to China, Sichuan (Fig. 4).

Habitat

Physospermopsis muliensis usually grows in wet grasslands at 2500–4100 m elevation.

Additional specimens examined

China. Sichuan Province: Muli County, Kangwuliangzi, 3800 m, 9 Aug 2019, X.R.Xu XXR2019080903 (SZ); Muli County, 3900 m, 20 Oct 1981, L.R.Xu 0187 (WUK); unknown locality, 3600 m, 29 November 2005, Anonymous 416 (PE); unknown locality, 2580 m, 28 November 2005, Anonymous 418 (PE); unknown locality, 3650 m, 29 November 2005, Anonymous 419 (PE); Muli County, 3800 m, 21 Sep 1955, Anonymous 267 (PE); Muli County, 3620 m, 15 Oct 1982, Y.B.Yang & G.Yao 174 (CDBI); Muli County, 3200 m, 3 Sep 1978, Q.S.Zhao, K.H.Mou & Y.B.Yang 8395 (CDBI); Muli County, 3650 m, 25 Oct 1986, Y.J.Li et al. 987 (CDBI); Muli County, 3150 m, 25 Oct 1982, Vegetation expedition 29926 (CDBI); Muli County, 3900 m, 22 Aug 1983, 39 (HITBC); Dukou County, 21 Sep 1978, 412 (SM); Jinyang County, Mt. Shizi, 4000 m, 18 Aug 1978, 683 (SM); Mianning County, Juexingou, 3000 m, 11 Oct 1978, Mianning expedition 667 (SM); Ningnan County, 3700 m, 24 Aug 1978, Ningnan expedition 547 (SM); Dukou County, Yanbian, 28 Sep 1978, Anonymous 435 (SM); Meigu County, 3700 m, 18 Aug 1979, Anonymous 594 (SM); Yanbian County, 28 Sep 1978, Anonymous 475 (SM); Zhaojue County, 4040 m, 13 Jul 1979, Plants census expedition 179 (SM); Xide County, 29 Jun 1979, Anonymous 495 (SM); Mabian County, 3400 m, 1 Jul 1978, Mabian expedition 798 (SM); Muli County, 3900 m, 2 Aug 1978, Muli expedition 577 (SM); Muli County, Changhaizi, 3669 m, 7 Oct 2009, Z.L.Nie, Y.Meng & T.Deng SunH-07ZX-2330 (KUN).

Physospermopsis shaniana Z.Y.Wu et F.D.Pu, 1993: 1285

Trachydium forrestii Diels, 1912: 291

Physospermopsis forrestii (Diels) C.Norman, 1938: 231

Pleurospermum forrestii (Diels) M.Hiroe, 1958: 123

Type

China. Yunnan: Lijiang range, shady, grassy openings in pine forests on the eastern flank, Aug 1906, Forrest 2855 (lectotype, designated by Pimenov 2017, pg. 188: E [E00000219]; isolectotype: P [P00245432]).

Diagnostic characters

Physospermopsis shaniana usually possesses 2-pinnate/pinnatifid, ovate-oblong leaf blades, broad sheaths, broadly ovoid fruits, white petals, and leaf-like, 2-pinnate bracts. The pinnae are subsessile with pinnatifid margin. The stems of P. shaniana are reduced and branched at the base. The branches are longer than the main stem.

Distribution in China

Sichuan, Tibet, Yunnan (Fig. 4).

Distribution outside China

Myanmar.

Habitat

Physospermopsis shaniana usually grows in pasture and grassy slopes.

Additional specimens examined

China. Sichuan Province: Meigu County, Kongmingzhai, 3600 m, 5 Aug 1959, Anonymous 1591 (KUN); Zhaojue County, Jiefanggou, 3200 m, 9 Jul 1976, Vegetation expedition 12928 (PE); Yunnan Province: Qiaojia County, Yaoshan, 3200 m, 14 Jul 2019, X.R.Xu XXR2019071404 (SZ); Luquan County, Jiaozixueshan, 3700 m, 17 Jul 2019, X.R.Xu XXR2019071701 (SZ); Dongchuan County, Huizedahai, 3400 m, 30 Jul 1964, Northeast Yunnan Expedition 434 (KUN); Qiaojia County, G.M.Yang SCSB-W-1237 (KUN); Zhenkang County, 8 Jul 1938, T.T.Yu 17120 (PE); Zhenkang County, snow range, 3450 m, 4 Aug 1938, T.T.Yu 17174 (PE); Dongchuan County, Mubanghai, 3360 m, 16 Aug 1964, Northeast Yunnan Expedition 822 (LBG,KUN); Zhaotong County, 24Gang, 2300 m, 10 Aug 1974, Anonymous 234 (KUN); Qiaojia County, Yaoshan, 3100 m, 16 Jul 1973, B.X.Sun 1017 (KUN).

Physospermopsis delavayi (A.R.Franchet) H.Wolff, 1925: 278

Arracacia delavayi A.R.Franchet, 1894: 115

Pleurospermum delavayi (A.R.Franchet) M.Hiroe, 1958: 120

Type

China. Yunnan: Mosuoying, Yangyushan, 15 Sep 1885, Delavay 2017 (lectotype, designated by Pimenov and Kljuykov 2000c, pg. 537: P [P00245424]).

Diagnostic characters

P. delavayi usually possesses a conspicuously winged rachis and yellow-green, round fruits. The bracts are usually smaller than other species of Physospermopsis. The basal leaves are obovate to obovate-orbicular with incised-serrate or lobed margin or cuneate with partite margin.

Distribution

Endemic to China, Hunan, Sichuan, Yunnan. (Fig. 4).

Habitat

Physospermopsis delavayi prefers to grow in the pine forest or open grasslands.

Additional specimens examined

China. Sichuan Province: Yanyuan County, Lugu Lake, 3200 m, 11 Aug 2019, X.R.Xu XXR2019081102 (SZ); Yunnan Province: Lijiang County, Mt. Yulong, 3000 m, 13 Aug 2019, X.R.Xu XXR2019081305 (SZ); Shangri-La, 3200 m, 18 Jul 2018, X.L.Guo G18071802 (SZ); Lijiang County, Ganhaizi, 3100 m, 14 Feb 1968, Anonymous s.n. (HITBC); Lijiang County, Baishuihe, 2980 m, 4 Aug 1962, Anonymous s.n. (HITBC); Shuangbai County, Tuodian, 6 Oct 1958, S.Q.Huang 0217 (LBG); Chuxiong County, Baomanjie, 19 Sep 1958, S.Q.Huang 035 (LBG); Jianshui County, Yangjieba, 17 Mar 1941, S.E.Liu 018312 (PE); Eryuan County, Chaijiaying, Mt. Longtou, 21 Jul 1929, R.C.Qin 23338 (PE); unknown locality, Anonymous 3365 (PE); unknown locality, M.Chen 2460 (KUN); W.R.He 2381 (KUN); Fumin County, Mt. Laoqing, 2300 m, 19 Oct1964, B.Y.Qiu 596086 (KUN); Yiliang County, Qixingcun, 27 Aug 1975, B.Y.Qiu 60727 (KUN); Xicheng County, Qingshuihe, 1900 m, 12 Oct1982, B.T.Yue 2088 (KUN); Shangri-La County, Habacun, 2800 m, 3 Aug 1962, A.L.Zhang 100750 (KUN); Lijiang County, Mt. Yulong, Baishuihe, 3000 m, 11 Jul 1962, A.L.Zhang, S.W.Yu 100919 (KUN); Lijiang County, Yuhucun, 29 Aug 2010, Z.X.Wang WZX2010082911 (SZ); Lijiang County, Yuhucun, 31 Aug 2010, Z.X.Zhang WZX2010083102 (SZ); unknown locality, 29 Aug 2010, P.Gao, S.Liu 10829-6 (SZ); unknown locality, 17 Sep 1938, 12366 (PEY); 22 Sep 1919, K.K.TSOONG 387 (PEY).

Physospermopsis nana (A.R.Franchet) M.G.Pimenov et E.V.Kljuykov, 2000c: 538

= Pleurospermum nanum A.R.Franchet, 1894: 140. Type: CHINA. Yunnan: Dali County, Mt. Cang, 25 Sep 1884, Delavay 197 (syntypes: P [P00834544, P00834545]).

Type

China. Yunnan: Mt. Cang, 4000 m, 30 Aug 1889, Delavay 4066 (lectotype, designated by Pimenov and Kljuykov 2000c, pg. 538: P [P00834546]; isolectotype, designated by Pimenov and Kljuykov 2000c, pg. 538: P [P00834547]).

Diagnostic characters

Physospermopsis nana usually possesses reduced stem, membranous-margined sheaths, and leaf-like bracts. The bracteoles are pale green with whitish margin in lower half. The ultimate segments are linear-lanceolate. The characters mentioned above are sufficient to distinguish it from other Physospermopsis species.

Distribution

Endemic to China, Sichuan, Tibet, Yunnan. (Fig. 4).

Habitat

Physospermopsis nana usually grows on marshy meadows, under shrubs.

Additional specimens examined

China. Sichuan Province: Yajiang County, 3800 m, 9 Aug 1979, Yajiangdui 293 (SM); Muli County, Sanqu, 3600 m, 13 Sep 1983, Qinghai-Tibet Expedition 14043 (KUN); Tibet Province: Cona County, 4561 m, 10 Aug 2015, L.Wei & J.C.Hao 15544 (BNU); Lahsa County, 5200 m, 3 Sep 1965, G.C.Xia & T.K.Mi 2610 (KUN); Gê’gyai County, 5200 m, 21 Aug 1976, Qinghai-Tibet Expedition 8710 (KUN); Lahsa County, 6000 m, 1 Sep 1965, Y.T.Zhang & K.Y.Lang 2412 (KUN); Gê’gyai County, 5185 m, 9 Sep 2017, Y.He BNU2017XZ325 (BNU); Shigatse, 4645 m, 23 Aug 2017, Y.He & D.H.Liu BNU2017XZ064 (BNU); Gar County, 4360 m, 31 Aug 2008, J.H.Chen et al YangYP-Q-0122 (KUN). Yunnan Province: Lanping County, 4200 m, 18 Oct 2019, X.L.Guo G19101802 (SZ); Lijiang County, Mt. Yulong, 4400 m, 18 Aug 1976, Y.Q.He 049 (WUK); Dêqên County, Baima snow range, 4400 m, 14 Jul 1981, Qinghai-Tibet Expedition 2774 (HITBC); Gongshan County, Mt. Nanwan, 3400 m, 22 Sep 1997, 9604 (KUN); Zhongdian County, 4190 m, 2 Oct 2005, Z.D.Fang et al PL-130 (SABG); Lijiang County, Yuhu, 4000 m, 8 Sep 1955, G.M.Feng 21475 (KUN); Lijiang County, 4100 m, 31 Aug 1963, C.Z.Bao 20229 (KUN); Zhongdian County, 4200 m, 2 Aug 1986, H.Sun & Z.G.Qian 991 (KUN); Lijiang County, snow range, 15 Sep 1940, R.C.Qin 31043 (KUN); Zhongdian County, Mt. Haba, 4000 m, 31 Aug 1962, Zhongdian Expedition 1687 (KUN); Zhongdian County, Haba snow range, 26 Aug 1939, G.M.Feng 2215 (KUN); Lijiang County, Yulong, 4000 m, 31 Aug 1963, C.Z.Bao 20226 (KUN); Lijiang County, 2600 m, 27 Jul 1937, D.J.Yu 15374 (KUN); Lijiang County, Mt. Yulong, 3200 m, 26 Aug 1961, R.L.Xiong & Y.F.Qi 612715 (KUN); Dali County, 20 Aug 1945, H.C.Wang 4507 (KUN); unknown locality, 1963, J.S.Yang 2374 (KUN); Zhongdian County, 3400 m, 28 Jun 2009, Z.D.Fang G-297 (SABG); Dali County, Mt. Cang, 3460 m, 15 Oct 1990, CLD-90 (PE).

Conclusion

Physospermopsis is monophyletic and nested in Pleurospermeae, sister to Pleurospermum. Although the interspecific relationships within Physospermopsis were not well resolved and the positioning of species was unclear, the relationships of P. alepidioides and P. delavayi, P. kingdon-wardii and P. obtusiuscula are close. Diagnostic characteristics for distinguishing the species in the field and laboratory are provided for necessary morphological and molecular research in future Physospermopsis phylogenetic studies.

Acknowledgements

We thank W. Gou, H.Y. Zheng, D.F. Xie for their help in preparing this paper, curators and staff of the following herbaria: E, K, P, BM, LE, PE, SM, SZ, UC, WU, BNU, CVH, KUN, LBG, PEY, WUK, CDBI, SABG, HITBC who allowed the authors to examine their specimens or get digital information via the Internet. This work was supported by the National Natural Science Foundation of China [Grant No. 31872647], and the Chinese Ministry of Science and Technology through the “National Science and Technology Infrastructure Platform” project [Grant No. 2005DKA21403-JK], the fourth national survey of traditional Chinese medicine resources [Grant No. 2019PC002].

References

  • Boissieu H (1903) Pleurospermum Hoffm. Bulletin de l’Herbier Boissier, sér 2 3(10): 847–848.
  • Bone TS, Downie SR, Affolter JM, Spalik K (2011) A phylogenetic and biogeographic study of the genus Lilaeopsis (Apiaceae tribe Oenantheae). Systematic Botany 36(3): 789–805. https://doi.org/10.1600/036364411X583745
  • Calviño CI, Tilney PM, Wyk BEV, Downie SR (2006) A molecular phylogenetic study of southern African Apiaceae. American Journal of Botany 93(12): 1828–1847. https://doi.org/10.3732/ajb.93.12.1828
  • Clarke CB (1879) Umbelliferae. In: Hooker JD (Ed.) The Flora of British India. L Reeve & Co, London, 673 pp.
  • de Candolle AP (1830) Cxlix. Hymenolaena. Prodromus Systematis Naturalis Regni Vegetabilis 4: e246.
  • Diels FLE (1912) Umbelliferae. Notes from the Royal Botanic Garden Edinburgh 5(25): e291.
  • Downie SR, Katz-Downie DS (1999) Phylogenetic analysis of chloroplast rps16 intron sequences reveals relationships within the woody southern African Apiaceae subfamily Apioideae. Canadian Journal of Botany 77(8): 1120–1135. https://doi.org/10.1139/b99-086
  • Downie SR, Watson MF, Spalik K, Katz-Downie DS (2000) Molecular systematics of Old World Apioideae (Apiaceae): Relationships among some members of tribe Peucedaneae sensu lato, the placement of several island-endemic species, and resolution within the apioid superclade. Canadian Journal of Botany 78(4): 506–528. https://doi.org/10.1139/b00-029
  • Downie SR, Spalik K, Katz-Downie DS, Reduron JP (2010) Major clades within Apiaceae subfamily Apioideae as inferred by phylogenetic analysis of nrDNA ITS sequences. Plant Diversity and Evolution 128(1–2): 111–136. https://doi.org/10.1127/1869-6155/2010/0128-0005
  • Farille MA, Malla SB (1985) Apiaceae Himalayenses, III. Candollea 40: 512–520.
  • Franchet AR (1894) Notes sur quelques Ombellifères du Yunnan. Bulletin de la Société Philomathique de Paris 8(6): 106–146.
  • Guo XL, Wang CB, Wen J, Zhou SD, He XJ (2018) Phylogeny of Chinese Chamaesium (Apiaceae: Apioideae) inferred from ITS, cpDNA and morphological characters. Phytotaxa 376(1): 1–16. https://doi.org/10.11646/phytotaxa.376.1.1
  • Hiroe M (1958) Umbelliferae of Asia (excluding Japan). Eikodo (Akira Imagawa), Kyoto, 123–219.
  • Hiroe M (1979) Umbelliferae of World. Ariake Book Company, Tokyo, 741–747.
  • Hoffmann GF (1814) Genera plantarum Umbelliferarum eorumque characters naturales secundum numerum, figuram, situm et proportionem fructificationis et fructus partium. Mosquae.
  • Jordan WC, Courtney MW, Neigel JE (1996) Low levels of intraspecific genetic variation at a rapidly evolving chloroplast DNA locus in North American Duckweeds (Lemnaceae). American Journal of Botany 83(4): 430–439. https://doi.org/10.1002/j.1537-2197.1996.tb12724.x
  • Kelchner SA, Clark LG (1997) Molecular evolution and phylogenetic utility of the chloroplast rpl16 intron in Chusquea and the Bambusoideae (Poaceae). Molecular Phylogenetics and Evolution 8(3): 385–397. https://doi.org/10.1006/mpev.1997.0432
  • Kumar S, Stecher G, Tamura K (2016) Mega7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33(7): 1870–1874. https://doi.org/10.1093/molbev/msw054
  • Lee C-S, Downie SR (2006) Phylogenetic relationships within Cicuta (Apiaceae tribe Oenantheae) inferred from nuclear rDNA ITS and cpDNA sequence data. Canadian Journal of Botany 84(3): 453–468. https://doi.org/10.1139/b06-016
  • Lindley J (1835) Illustrations of the Botany and Other Branches of the Natural History of the Himalayan Mountains. Royle, John Forbes, London, 232 pp.
  • Linnaeus C (1753) Species Plantarum (Vol. 1). Laurentii Salvii, Holmiae, 263 pp.
  • Mukherjee PK (1982) Resume of Indian umbellifers. Monographs in Systematic Botany from the Missouri Botanical Garden 6: e59.
  • Naqshi AR, Dhar U, Kachroo PN (1995) Central Asia and Kashmir Himalaya-Archaeobotany and Floristics. Scientific Publishers, India, 107 pp.
  • Norman C (1938) Physospermopsis H. Wolff. Journal of Botany. British and Foreign 76: e231.
  • Nylander JAA (2004) MrModelTest 2.2. Program Distributed by the Author. Evolutionary Biology Centre, Uppsala University.
  • Pan ZH, Watson MF (2005) Physospermopsis H. Wolff. In: Wu ZY, Raven RH (Eds) Flora of China (Vol. 14). Science Press, Beijing and Missouri Botanical garden Press, Saint Louis, 31–33.
  • Panahi M, Banasiak Ł, Piwczyński M, Puchałka R, Kanani MR, Oskolski AA, Modnicki D, Miłobędzka A, Spalik K (2018) Taxonomy of the traditional medicinal plant genus Ferula (Apiaceae) is confounded by incongruence between nuclear rDNA and plastid DNA. Botanical Journal of the Linnean Society 188(2): 173–189. https://doi.org/10.1093/botlinnean/boy055
  • Pimenov MG, Kljuykov EV (2000a) Taxonomic revision of Pleurospermum HOFFM. and related genera of Umbelliferae I. General part. Feddes Repertorium 111(7–8): 499–515. https://doi.org/10.1002/fedr.20001110717
  • Pimenov MG, Kljuykov EV (2000b) Taxonomic revision of Pleurospermum HOFFM. and related genera of Umbelliferae II. The genera Pleurospermum, Pterocyclus, Trachydium, Keraymonia, Pseudotrachydium, Aulacospermum, and Hymenolaena. Feddes Repertorium 111(7–8): 517–534. https://doi.org/10.1002/fedr.20001110718
  • Pimenov MG, Kljuykov EV (2000c) Taxonomic revision of Pleurospermum HOFFM. and related genera of Umbelliferae III. The genera Physospermopsis and Hymenidium. Feddes Repertorium 111(7–8): 535–552. https://doi.org/10.1002/fedr.20001110719
  • Pu GZ, Liu QX (2006) The micromorphological features of pericarp surface of Physospermopsis and Trachydium (Apiaceae) in China and its taxonomic significance. Journal of Plant Resources and Environment 15: 1–6. https://doi.org/10.3969/j.issn.1674-7895.2006.03.001
  • Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, Hohna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice Across a Large Model Space. Systematic Biology 61(3): 539–542. https://doi.org/10.1093/sysbio/sys029
  • Schischkin BK (1935) Flora Zapadnoi Sibiri. Tomsk State University, Tomsk, 2010 pp.
  • Shan RH (1941) On reduction of the genus Haplesoseseli WOLFF et HANDEL MAZZETTI. Sinensia 12(1–6): 185–187.
  • Shan RH, Liou SL (1979) Physospermopsis Wolff Apiaceae. In: Shan RH, She ML (Eds) Flora Reipublicae Popularis Sinicae. Science Press, Beijing, 105 pp.
  • Spalik K, Downie SR, Watson MF (2009) Generic delimitations within the Sium alliance (Apiaceae tribe Oenantheae) inferred from cpDNA rps16-5` trnK(UUU) and nrDNA ITS sequences. Taxon 58(3): 735–748. https://doi.org/10.1002/tax.583004
  • Valiejo-Roman CM, Terentiev EI, Pimenov MG, Kljuykov EV, Samigullin TH, Tilney PM (2012) Broad Polyphyly in Pleurospermum s. l. (Umbelliferae-Apioideae) as Inferred from nrDNA ITS and Chloroplast Sequences. Systematic Botany 37(2): 573–581. https://doi.org/10.1600/036364412X635593
  • Wang PL, Pu FD (1992) Pollen morphology diversity and evolution trend of the genus Physospermopsis in the Hengduan Mountains of China. Yunnan Zhi Wu Yan Jiu 4: 413–417.
  • White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (Eds) PCR Protocols: A Guide to Methods and Applications. Academic Press Inc, San Diego, California, 315–322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
  • Wolff H (1925) Neue umbelliferen-gattungen aus ostasien. Notizblatt Des Königl Botanischen Gartens Und Museums Zu Berlin-Dahlem 9(84): 275–280. https://doi.org/10.2307/3994550
  • Wolff H, Handel-Mazzetti H (1933) Umbelliferae. Symbolae Sinicae 7(3): e722.
  • Wu CY, Pu FT (1993) Vascular Plants of the Hengduan Mountains. Science Press, Beijing, 1285 pp.
  • Yi T-S, Jin G-H, Wen J (2015) Chloroplast capture and intra- and inter- continental biogeographic diversification in the Asian-New World disjunct plant genus Osmorhiza (Apiaceae). Molecular Phylogenetics and Evolution 85: 10–21. https://doi.org/10.1016/j.ympev.2014.09.028
  • Zhou J, Peng H, Downie SR, Liu ZW, Gong X (2008) A molecular phylogeny of Chinese Apiaceae subfamily Apioideae inferred from nuclear ribosomal DNA internal transcribed spacer sequences. Taxon 57: 402–416. https://doi.org/10.2307/25066012
  • Zhou J, Gong X, Downie SR, Peng H (2009) Towards a more robust molecular phylogeny of Chinese Apiaceae subfamily Apioideae: Additional evidence from nrdna its and cpDNA intron (rpl16 and rps16) sequences. Molecular Phylogenetics and Evolution 53(1): 56–68. https://doi.org/10.1016/j.ympev.2009.05.029

Appendix 1

Voucher details and GenBank accession numbers of taxa used in this study. A n-dash (–) indicates unavailable information; new sequences are in bold.

Taxa Voucher Locality Genbank accession numbers
ITS rpl16 rps16
Angelica archangelica Downie 79 (ILL) cult. University of Joensuu Botanical Garden, Finland AH003539 AF094362 AF110536
Angelica likiangensis 200421 (NHW) Lijiang, Yunnan, China DQ263587 FJ385074 FJ385172
Angelica maowenensis ZJ0582 (KUN) Mt. Gongga, Sichuan, China EU236157 FJ385075 FJ385173
Aulacospermum anomalum 19932275 (E) cult. Royal Botanic Garden, Edinburgh, United Kingdom AF008641, AF009120 AF094440 AF110558
Aulacospermum simplex Dingelstedt & Sovetkina 367 23-VII-1927 (LE) Kazakhstan GQ379339 AF110557
Bupleurum krylovianum ZJ0726 (KUN) KaNaSi Lake, Xinjiang, China FJ385035 FJ385082 FJ385180
Bupleurum rockii J059 (KUN) Ninglang, Yunnan, China FJ385036 FJ385083 FJ385181
Chamaesium paradoxum ZJ0560 (KUN) Daocheng-Litang, Sichuan, China EU236161 FJ385085 FJ385184
Chamaesium thalictrifolium ZJ0607 (KUN) Zhangla-Caowan, Sichuan, China EU236162 FJ385086 FJ385185
Chamaesium wolffianum ZJ0525 (KUN) Shudu Lake, Yunnan, China EU236163 FJ385087 FJ385186
Changium smyrnioides J101 (KUN) Jiangsu Institute of Botany, China DQ517340 FJ385088 FJ385187
Chuanminshen violaceum J105 (KUN) Cangxi, Sichuan, China FJ385040 FJ385089 FJ385188
Conioselinum chinense Raiche 30046 (UC) California, America U78374 AF094421 GU395135
Conioselinum vaginatum ZJ0731 (KUN) KaNaSi Lake, Xinjiang, China FJ385041 FJ385091 FJ385190
Haplosphaera phaea ZJ0521 (KUN) Shudu Lake, Yunnan, China EU236167 FJ385096 FJ385194
Hansenia weberbaueriana ZJ0697 (KUN) KIB nursery, Yunnan, China EU236180 FJ385115 FJ385212
Heracleum bivittatum ZJ0611 (KUN) MaoCountyg, Sichuan, China EU236168 FJ385098 FJ385196
Heracleum forrestii ZJ091032 (KUN) Shangri-La, Yunnan, China EU236169 FJ385099 FJ385197
Korshinskya bupleuroides Pimenov et al. 106 (MW) FJ489360, FJ489391
Korshinskya olgae Pimenov et al. 228 (MW) FJ489359, FJ489390
Physospermopsis alepidioides XXR2019080801 (SZ) Muli, Sichuan, China MT533355 MT542144 MT561011
Physospermopsis delavayi (HB) G18071802 (SZ) Shangri-La, Yunnan, China MN658653 MN786490 MN786487
Physospermopsis delavayi (LGH) XXR2019081102 (SZ) Lugu Lake, Sichuan, China MN658656 MN786488 MN786486
Physospermopsis delavayi (LYG) XXR2019081305 (SZ) Lijiang, Yunnan, China MN658657 MN786489 MN786485
Physospermopsis kingdon-wardii G19082407 (SZ) Nyalam, Tibet, China MN659655 MN786491 MN786484
Physospermopsis muktinathensis Pimenov & Kljuykov 22 (MW) Annapurna, Nepal FJ469961, FJ483500
Physospermopsis muliensis XXR2019080903 (SZ) Muli, Sichuan, China MT533356 MT542145 MT561012
Physospermopsis nana G19101802 (SZ) Lanping, Yunnan, China MT542694 MT561018 MT561017
Physospermopsis obtusiuscula (DQ) XXR2019081502 (SZ) Dêqên, Yunnan, China MT533361 MT542149 MT561016
Physospermopsis obtusiuscula (YD) G19082009 (SZ) Yadong, Tibet, China MT533360 MT542148 MT561015
Physospermopsis rubrinervis (EY) XXR2019081701 (SZ) Eryuan,Yunnan, China MN658654 MN786492 MN786483
Physospermopsis rubrinervis (LS) G19101802 (SZ) Lushui, Yunnan, China MT533359 MT542143 MT561010
Physospermopsis shaniana (LQ) XXR2019071701 (SZ) Luquan, Yunnan, China MT533357 MT542146 MT561014
Physospermopsis shaniana (QJ) XXR2019071404 (SZ) Qiaojia, Yunnan, China MT533358 MT542147 MT561013
Pleurospermum austriacum Ghisa and Topa 2959 (MW) FJ469962, FJ483502
Pleurospermum foetens Chungtien 1181 (E) Yunnan, China FJ483482, FJ469943 AF094438 AF110559
Pleurospermum franchetianum ZJ0573 (KUN) Kangding, Sichuan, China EU236198 FJ385137 FJ385232
Pleurospermum uralense LQX031 (NAS) Liaoning, China JF977839 AF094439 AF110560
Pleurospermum wilsonii ZJ0624 (KUN) Hongyuan, Sichuan, China EU236200 FJ385139 FJ385234