Research Article |
Corresponding author: Quan-Ru Liu ( liuquanru@bnu.edu.cn ) Academic editor: Hugo de Boer
© 2024 Li-Hua Wang, Shuo Li, De-Ning Zhang, Quan-Ru Liu.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Wang L-H, Li S, Zhang D-N, Liu Q-R (2024) A new combination and synonym in Bupleurum (Apiaceae, Apioideae), based on morphological, molecular and cytological evidence. PhytoKeys 239: 239-253. https://doi.org/10.3897/phytokeys.239.116877
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Specimen examinations and field observations revealed that Bupleurum smithii var. parvifolium was distinctly different from B. smithii var. smithii in umbel, leaf, and fruit morphology, but was very similar to B. commelynoideum var. flaviflorum. Based on these morphological evidences, the present study re-examined the taxonomic status of these taxa through morphological, cytological, and phylogenetic analyses. The results showed distinguishable features in the width of middle leaves and bracteoles of B. smithii var. parvifolium compared to B. smithii var. smithii. Morphological variation between B. smithii var. parvifolium and B. commelynoideum var. flaviflorum was continuous and overlapping. Notably, the chromosome number of B. smithii var. parvifolium was 2n = 14 (x = 7), consistent with B. commelynoideum var. flaviflorum, whereas B. smithii var. smithii was 2n = 64 (x = 8). Additionally, phylogenetic analyses revealed B. commelynoideum var. flaviflorum nested within B. smithii var. parvifolium, and that both were distant from the B. smithii var. smithii and B. commelynoideum var. commelynoideum. Based on the evidence above, the differences between B. smithii var. parvifolium and B. smithii var. smithii extend beyond the level of intraspecific variation, and B. commelynoideum var. flaviflorum is considered to be identical with B. smithii var. parvifolium. Hence. A new combination and status, B. parvifolium (Shan & Y.Li) Q.R.Liu & L.H.Wang, comb. et stat. nov., is proposed. Furthermore, B. commelynoideum var. flaviflorum should be treated as a synonym of B. parvifolium.
Apiaceae, chloroplast genome, chromosome counts, new combination, synonymy
The genus Bupleurum (Apiaceae, Apioideae), comprising 180–220 species (
Bupleurum smithii Wolff was classified into three varieties: B. smithii var. smithii, B. smithii var. parvifolium Shan & Y.Li, and B. smithii var. auriculatum Shan & Y.Li, based on leaf morphology (
Upon a thorough examination of the type specimens, it was discovered that the isotype specimen of B. commelynoideum var. flaviflorum was identified as B. commelynoideum var. flaviflorum (WUK0423353) and B. smithii var. parvifolium (WUK0033909). Such instances are common during specimen reviews, highlighting the need for meticulous morphological and phylogenetic analyses to elucidate the taxonomic placement and phylogenetic position of these taxa.
Our study involved the examination of collections and digital images of B. smithii var. smithii, B. smithii var. parvifolium, B. smithii var. auriculatum, B. commelynoideum var. flaviflorum, B. commelynoideum var. commelynoideum from the Chinese Virtual Herbarium (http://www.cvh.ac.cn/) and the Global Biodiversity Information Facility (https://www.gbif.org/). The images were sourced from specimens deposited at BJFC, BM, BNU, CDBI, HNWP, HSIB, KUN, NAS, P, PE, PEY, SZ, and WUK. The specimens with well-preserved leaves and flowers were selected, covering all districts of the distribution area. A total of 43 specimens of B. smithii var. smithii, 50 of B. smithii var. parvifolium, 29 of B. commelynoideum var. commelynoideum, and 39 of B. commelynoideum var. flaviflorum were examined, including 55 sheets of specimens collected by our team (Suppl. material
The examination of characters was conducted using ImageJ (
The materials used in the cytological studies were sourced from Xiaowutai Mountain (for B. smithii var. smithii), Qilian Mountain (for B. smithii var. parvifolium), Luya Mountain (for B. smithii var. auriculatum) and Min Mountain (for B. commelynoideum var. flaviflorum). The voucher specimens are detailed in Table
Voucher information and GenBank accession numbers of newly sequenced plastome sequences, asterisks for cytology.
Taxa | Location | Voucher information | Accession |
---|---|---|---|
B. baimaense | Deqincountry, Yunnan, China | YNNU-19-302 (KUN) | OR778864 |
B. commelynoideum var. commelynoideum | Kangding, Sichuan, China | BNU2023WLH0174 (BNU) | OR778865 |
B. smithii var. smithii *[1] | Xiaowutai Mountain, Hebei, China | BNU2020DT007 (BNU) | OR387522 |
B. smithii var. smithii [2] | Dongling Mountain, Hebei, China | DL023-3 (BNU) | OR811239 |
B. smithii var. auriculatum* | Luya Mountain, Shanxi, China | BNU2021SX017 (BNU) | OR811240 |
B. commelynoideum var. flaviflorum* | Min Mountain, Gansu, China | BNU2023WLH190 (BNU) | OR778866 |
B. smithii var. parvifolium [1] | Hualong County, Qinghai, China | BNU2022WLH061 (BNU) | OR778870 |
B. smithii var. parvifolium [2] | Haiyan County, Qinghai, China | ZZU2021QH004 (BNU) | OR778869 |
B. smithii var. parvifolium* [3] | Tianjun county, Qinghai, China | BNU2022WLH030 (BNU) | OR778871 |
In this study, the whole chloroplast (cp) genome of Bupleurum was sequenced to investigate its phylogeny and evolution. Fresh leaves from nine Bupleurum specimens were field-collected and rapidly desiccated using Silica Gel. Voucher specimens were deposited at Herbarium of Beijing Normal University (BNU), and listed in Table
Phylogenetic analyses were conducted using the maximum likelihood (ML) and Bayesian inference (BI) methods with IQ-TREE (
The Principal Component Analysis (PCA) revealed that traits such as width of middle leaves, width of bracteoles, length of bracteoles, and the number of rays were effective for interspecific classification (Fig.
Comparison of morphological characteristics and geographic distribution.
Morphology | B. smithii var. smithii | B. smithii var. parvifolium | B. commelynoideum var. flaviflorum | B. commelynoideum var. commelynoideum |
---|---|---|---|---|
Basal leaves | 7–15 × 0.8–1.5 cm | 6–8 × 0.3–0.7 cm | 6–8 × 0.3–0.5 cm | 8–15 × 0.25–0.4 cm |
Middle stem leaves | 7–13 × 0.8–1.5 cm | 4–7 × 0.4–0.7 cm | 6–9 × 0.3–0.5 cm | 8–11 × 0.25–0.4 cm |
Umbels | Numerous | Numerous | Numerous | Single |
Rays | 4–9 | 4–9 | 4–9 | 3–4 (6) |
Bracteole | (6)7–9, 5–7 × 3–4.5 mm | (5) 6–7, 3–4.5 × 2–2.5 mm | (5) 6–7, 3–5 × 2–2.5 mm | 7–9, 5–6.5 × 3–5 mm |
Petal color | Yellow or abaxially purplewish-tinged | Yellow or abaxially purplewish-tinged | Yellow or abaxially purplewish-tinged | Purple or yellowish-tinged |
Fruit | Rectangular, 3.0–3.6 × 1.2–1.4 | Ovoid, 1.9–2.5 × 1.1–1.3 | Ovoid, 2.0–2.4 × 1.1–1.3 | Tapered, 2–2.5 × 1.5 |
Vittae in each furrow | 4 | 6 | 6 | 4 |
Vittae on commissure | 3 | 3 | 3 | 3 |
Distribution | Shanxi, Hebei, Beijing and N Henan | E Qinghai, Gansu, Ningxia, E Xizang, and NW Sichuan | S Gansu, S Qinghai, and W Sichuan. | W Sichuan, Xizang and NW Yunnan |
PCA and box plots showed intermittent distinctions in leaf shape, bracteoles, and fruit size between B. smithii var. smithii and B. smithii var. parvifolium, providing reliable evidence for interspecific differentiation. Cluster diagram results corroborated the cohesion of B. smithii var. parvifolium and B. commelynoideum var. flaviflorum into a single group, providing robust support for considering them as the same taxon. Box plots visually depicted continuous and overlapping variations between these two taxa.
Cytological analysis reveals that the chromosome number of B. smithii var. parvifolium is 2n = 14 (x = 7) (Fig.
Chromosome base diversity is high within the genus Bupleurum, including several cases with x = 4, 5, 6, 7, 8, 11, and 13 (
In recent years, researchers have gradually emphasized the role of chromosomes in species delimitation. For example, B. komarovianum was once treated as a variety of B. chinense (
The phylogenetic analysis based on chloroplast genome sequences reveals a consistent topology between the maximum likelihood (ML) tree and the Bayesian inference (BI) tree. The genus Bupleurum was divided into two clades with high support (BS = 100%, PP = 1), with all Chinese Bupleurum species belonging to Subg. Bupleurum. Bupleurum commelynoideum var. flaviflorum was nested within B. smithii var. parvifolium (BS = 100%, PP = 1), forming an individual clade. This clade was further related to B. sikangense X.J.He & C.B.Wang (Fig.
In this study, the reconstruction of the phylogenetic tree utilizing the chloroplast genome yielded results consistent with the topological structure presented by
In terms of distribution patterns, B. smithii var. parvifolium is a widespread taxon mainly in northwest China, including Gansu, Ningxia, Qinghai, eastern Xizang, and western Sichuan. On the other hand, B. commelynoideum var. flaviflorum is distributed in SW China, including S Gansu, S Qinghai, and W Sichuan. The distribution of these two taxa overlaps in northwest Sichuan and southern Gansu, precisely where the type specimens of B. commelynoideum var. flaviflorum were collected. In contrast, B. smithii var. smithii primarily occur in the Taihang Mountains and Yanshan Mountains in E China. Bupleurum commelynoideum is a species that occurs throughout the Hengduan Mountains region. The distribution areas of B. smithii var. parvifolium and B. smithii var. smithii are separated by the Qinling Mountains and the Loess Plateau (Fig.
≡ B. smithii var. parvifolium Shan & Y.Li, Acta Phytotax. Sin. 12(3): 273. 1974. Type. CHINA. Gansu: Tianzhuxian, Wushao Mountain ca. 2800 m, 22. 07. 1959, Y.Q. He 4267 (Holotype: WUK0389736!). Basionym.
= B. commelynoideum var. flaviflorum Shan & Y.Li, Acta Phytotax. Sin. 12(3): 276. 1974. syn. nov. ≡ B. smithii var. flaviflorum (Shan & Y.Li) X.J.He & C.B.Wang J. Syst. Evol. 49 (6): 586. 2011.Type. CHINA. Gansu: Min County, Min Mountain, ca. 3500 m, 10. 08. 1937, T.P. Wang 7535 (Holotype: PE00935517! Isotypes: WUK0033909! WUK0423353!).
Bupleurum parvifolium is morphologically similar to B. smithii, can be distinguished from the latter by its cauline leaves sessile, 4–9 × 0.3–0.7 cm; bracteoles 6–9, ovate or broad-ovate, 3–5 × 2–2.5 mm; fruit ovoid, brown, 2.0–2.5 × 1.1–1.3 mm; vittae 3 in each furrow, 6 on commissure.
Plant 15–40 cm, perennial. Rhizome brown, usually branched. Stems many, tufted, base without fibrous remnant sheaths. Basal leaves narrowly lanceolate, 6–8 × 0.3–0.7 cm, thickly papery, base tapered into petiole, not embracing. Cauline leaves sessile, 4–9 × 0.3–0.7 cm. Apical leaf long-ovate, 1.5–7.5 × 1–1.7 cm, base rounded, sometimes auriculate, clasping, apex acuminate. Bracts 0 or 1–2, broadly ovate, 7–18 × 4–11 mm, unequal; rays 4–9, 0.5–4 cm, unequal, angled; bracteoles 6–9, ovate or broad-ovate, 3–5 × 2–2.5 mm, equal, acute, apiculate, slightly exceeding flowers; umbellules 0.8–1.1 cm across. Petals yellow, occasionally abaxially purplish-red. Stylopodium low-conic, discoid, dark yellow or purple-brown. Fruit ovoid, brown, 2.0–2.5 × 1.1–1.3 mm; ribs acute, prominent; vittae 3 in each furrow, 6 on commissure (Fig.
Flowering from July to August and fruiting from August to September.
Qinghai, Gansu, Ningxia, Sichuan, Xizang. It grows on mountains at elevations of 2700–700 m.
China. Gansu: Min County, 02 July 1936, T.P. Wang 4852 (KUN), 19 August 1937, T.P. Wang 7535 (WUK), 19 August 1937, T.P. Wang 7535 (PE), 01 July 1936, T.P. Wang 4742 (PE), 02 July 1936, T.P. Wang 4852 (PE), 30 June 1936, T.P. Wang 4699 (PE); Tianzhu County, Y.Q. He 4267 (WUK), 12 July 1959, Y.Q. He 4628 (WUK); Yuzhong County, 09 August 1959, Y.Q. He 5981 (WUK), 04 August 2000, X.G. Sun et al. 2126 (PE), 2 September 2023, L.H. Wang & J.L. Li BNU2023-WLH242 (BNU); Xiahe County, 10 July 1937, T.P. Wang 7171 (WUK), 29 July 1937, K.T. Fu 1438 (PE); Minle County, 29 August 1934, C.W. Yao 336 (NAS); Hezuo County, 09 September 2011, X. Yin et al. LiuJQ-GN-2011-278 (KUN); Maqu County, 04 August 2011, X. Yin et al. LiuJQ-GN-2011-280 (KUN); Shandan County, 10 July 1959, Y.Q. He 4052 (WUK); Qinghai: Tianjun County, 05 August 2022, L.H. Wang et al. BNU2022-WLH030 (BNU); Gangcha County, 30 July 2022, L.H. Wang et al. BNU2022-WLH017 (BNU); Huzhu County, 05 August 2022, Q.Y. Zhang ZQY2022003 (BNU); Xunhua County, 06 August 2022, L.H. Wang et al. BNU2022-WLH039 (BNU), 06 August 2022, L.H. Wang et al. BNU2022-WLH051 (BNU); Hulong County, 07 August 2022, LH Wang et al. BNU2022-WLH061 (BNU); Xining City, 11 September 2022, S.B. Zhang BNU2022-0911 (BNU); Haeyan County, 04 July 1958, B.Q. Zhong 8410 (KUN); Angqian County, 11 August 1972, Zangyao team 1283 (KUN); Qilian County, 27 July 1958, Gan & Qing, BQ Zhong 8573 (WUK), 12 August 2013, X.C. Chen et al. 4487 (HNWP); Xinghai County, 09 August 1919, T.N. He 416 (WUK); Datong County, 17 August 1962, Fan & Liang 00495 (HNWP), B.W. Li 72-070 (HNWP); Ghindu County, 10 August 1983 X.J. Xun 83-144 (HNWP), 15 August 1996, T.N. Ho et al. 1887 (PE), 15 August 1996, T.N. Ho et al. 1887 (HNWP); Zeku County, 21 August 1967, L.H. Zhou 1628 (HNWP); Tungrin County, 24 July 1970, S.W. Liu et al. 1412 (HNWP), 07 August 2010, S.L. Chen et al. ChenSL0915 (KUN); Menyuan County, 10 July 1970, L.H. Zhou 1036 (HNWP); Zhidoi County, 10 September 1966, L.H. Zhou 454 (HNWP); Yushu County, 30 August 1996, T.N. Ho et al. 2698 (PE), 12 August 1964, Qinghai Plant Team 620 (WUK); Nangqen County, 05 September 199, T.N. Ho et al. 2913 (PE); Huangzhong County, 25 July 2014, Y.H. Wu 050810 (HNWP), 25 July 2014, Y.H. Wu 050644 (HNWP); Ningxia: Jingyuan County, 14 August 1942, T.P. Wang 13561 (KUN); Tongxin County, 12 August 1981, Y.P. Xu et al. 1701 (WUK); Longde County, 10 July 1942, T.P. Wang 13052 (WUK); Guyuan County, 07 August 1953, T.P. Wang 17175 (WUK); Sichuan: Ruoergai, County, X.J. He et al. SCU-20080522 (KUN), 15 July 1993, Z.M. Tan 93-88 (PE), 06 August 1961, S. Jiang 6833 (PE); Hongyuan County, 18 September 2012, Y.D. Gao et al. GaoXF-12-029 (KUN); Pingwu County, H.L. Tsising 10904 (NAS); Daofu County, 06 September 1960, Sichuan Team 16217 (NAS); Songpan County, 3 October 1983, F.D. Pu et al. 021 (CDBI), 02 August 1984, J. He et al. 140 (CDBI); Daoge County, 01 August 1980, Vegetation group 28404 (CDBI); Baxoi County, 27 August 1973, Qinghai-Tibet Team 73-1267 (PE), 14 September 2008, T. Zhang et al. 08CS701 (KUN), 15 August 2014, X.C. Chen et al. 032-2 (HNWP), 15 August 2014, X.C. Chen et al. 032 (HNWP); Wuqi County, 27 August 1976, Tibet Expedition Team 12951 (PE).
We are grateful to the curators of the herbaria of WUK, PE, HNWP and KUN for access to the specimens for our study. We thank Associate Researcher Wen-Jun Li for the field assistance and Dan-Hui Liu for the modification of English.
The authors have declared that no competing interests exist.
No ethical statement was reported.
This research was funded by the Biodiversity survey and assessment project of the Ministry of Ecological Environment (No. 2019HJ2096001006).
Conceptualization: LHW. Data curation: SL. Funding acquisition: QRL. Investigation: SL, DNZ, LHW. Methodology: LHW. Supervision: QRL. Writing - original draft: LHW. Writing - review and editing: LHW, QRL.
Quan-Ru Liu https://orcid.org/0000-0003-4270-4746
All of the data that support the findings of this study are available in the main text or Supplementary Information.
Taxa | Accession number | Taxa | Accession number |
---|---|---|---|
B. boissieuanum | NC036017 | B. longiradiatum | MT261186 |
B. candollei | MT261183 | B. marginatum | MN968501 |
B. chinense | MT083933 | B. rockii | MT261189 |
B. densiflorum | MT261184 | B. shanianum | MW135452 |
B. euphorbioides | NC053888 | B. sibiricum | MT261190 |
B. falcatum | NC027834 | B. sikangense | NC056803 |
B. fruticosum | NC060426 | B. thianschanicum | MT261192 |
B. gibraltaricum | NC060427 | P. astrantioideum | NC071798 |
B. hamiltonii | NC056298 | P. szechenyii | NC071808 |
Specimens for measurements and initial measurements of traits data
Data type: xlsx
Traits selected for subsequent principal component and cluster analysis after the first PGA
Data type: xlsx