Research Article |
Corresponding author: Chun-Lei Xiang ( xiangchunlei@mail.kib.ac.cn ) Academic editor: Eberhard Fischer
© 2024 Yue Zhao, Fei Zhao, Alan J. Paton, Jin-Fei Xiao, Ya-Ping Chen, Chun-Lei Xiang.
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:
Zhao Y, Zhao F, Paton AJ, Xiao J-F, Chen Y-P, Xiang C-L (2024) Using scanning electron microscopy and molecular data to discover a new species from old herbarium collections: The case of Phlomoides henryi (Lamiaceae, Lamioideae). PhytoKeys 238: 127-146. https://doi.org/10.3897/phytokeys.238.117180
|
Phlomoides is one of the largest genera of Lamiaceae with approximately 150–170 species distributed mainly in Eurasia. In this study, we describe and illustrate a new species, P. henryi, which was previously misidentified as P. bracteosa, from Yunnan Province, southwest China. Molecular phylogenetic analyses revealed that P. henryi is found within a clade in which most species lack basal leaves. In this clade, the new species is morphologically distinct from P. rotata in having an obvious stem and, from the rest, by having transparent to white trichomes inside the upper corolla lip. In addition, micro-features of trichomes on the calyx and leaf epidermis can differentiate the new species from other species grouped in the same clade and a key, based on trichome morphology for these species, is provided. The findings demonstrate that the use of scanning electron microscopy can reveal inconspicuous morphological affinities amongst morphologically similar species and play an important role in the taxonomic study of the genus Phlomoides.
Micromorphology, Phlomoides, Phlomis, Phlomideae, taxonomy, trichomes
The tribe Phlomideae (Lamiaceae, Lamioideae) was originally established by
As currently defined, Phlomoides consists of approximately 150–170 species and ranks the second largest genus within subfamily Lamioideae (
During the past ten years, phylogenetic and taxonomic studies have focused on Phlomoides from China (
In total, we sampled 49 out of 58 (84.48%) Chinese species of Phlomoides for molecular phylogenetic analyses. Sampling is primarily based on previous molecular phylogenetic studies of Phlomoides (
In addition, six species from the subclade comprising the potential new species, as well as P. bracteosa, were sampled to investigate macro-micro-features of trichomes on flora bracts and leaves. The list of sampled species and their origins are given in Table
List of sampled Phlomoides species to investigate macro/micro features of trichomes and their voucher information.
Taxon | Geographical origin | Voucher information | Collection date |
---|---|---|---|
P. henryi | China, Yunnan Province, Honghe Hani and Yi Autonomous Prefecture, Jianshui County, on the forest edge, 23°57′52.54″N, 102°59′49.47″E, alt. 1279 m. | F. Zhao et al. XCL2222 (KUN) | 3 Sep. 2022 |
P. bracteosa | India, Uttarakhand, Bhyundar Valley, alt. 3430 m. | Anonymous 6583 (PE) | 16 Aug. 1975 |
P. breviflora | China, Tibet, Nielamu County, Qu Town, 28°4′44″N, 86°0′2.109″E, alt. 3246 m | Y.P. Chen et al. EM1139 (KUN) | 12 Sep. 2019 |
P. macrophylla | China, Xizang Province, Yadong County, on the way from Yadong to Nathu La Pass. | Y.P. Chen et al. EM1094 (KUN) | 9 Sep. 2019 |
P. nyalamensis | China, Xizang Province, Nyalam County, Zhangmu Town, on the way from Lixin to Xuebugang, 27°56′37.0356″N, 85°58′28.1712″E, alt. 2896 m. | Y.P. Chen et al. EM1145 (KUN) | 13 Sep. 2019 |
P. tibetica | China, Xizang Province, Linzhi City, Shergyla Mountain. | C.L. Xiang et al. XCL1458 (KUN) | 15 Sep. 2016 |
P. milingensis | China, Xizang Province, Linzhi City, Miling County, Lilong Town, Lilonggou, 29°1′45.6″N, 93°53′34.7″E, alt. 3188 m. | C.L. Xiang et al. XCL1469 (KUN) | 16 Sep. 2016 |
P. rotata | China, Xizang Province, Changdu City, Zuogong County, Dondara Mountain, 29°42′59.9″N, 98°1′7.3″E, alt. 5034 m. | C.L. Xiang et al. XCL1419 (KUN) | 12 Sep. 2016 |
Total genomic DNA was extracted using the CTAB method (
The combined dataset of nine plastid DNA regions was analysed using Bayesian Inference (BI) and Maximum Likelihood (ML). Three species of Phlomis were selected as outgroup, based on previous studies (
Species concept, definitions of characters and depiction generally follow
Micro-features of leaf epidermis and floral bracts were investigated using Light Microscopy (LM) and Scanning Electron Microscopy (SEM). Photographs and morphological observations were taken using a Leica DM2500 light microscope (Leica Microsystems GmbH, Wetzlar, Germany). Mature leaves and floral bracts were taken from our collection (Table
A total of 18 sequences were newly sequenced in the present study and they were submitted to GenBank under accession nos. OR674852–OR674869. The aligned length of the combined plastid dataset was 9259 bp (2380 bp for atpB-rbcL, 421 bp for psbA-trnH, 1361 bp for rpl16, 681 bp for rpl32-trnL, 967 bp for rps16, 958 bp for trnK, 868 bp for trnL-trnF, 831 bp for trnS-trnG and 792 bp for trnT-L), respectively. The topologies of the BI and ML trees were consistent with each other, only the Bayesian 50% majority-rule consensus tree being presented, with the posterior probabilities (PP) and Bootstrap support (BS) and values being superimposed near the nodes (Fig.
Monophyly of the genus Phlomoides was recovered (Fig.
As shown in Fig.
Two basic types of trichomes can be observed in Phlomoides: eglandular hairs and glandular hairs, as reported by
Different types of trichomes of Phlomoides A short simple non-glandular trichomes (P. macrophylla) B short simple non-glandular trichomes (P. breviflora) C long simple non-glandular trichomes (P. henryi) D symmetrically non-glandular stellate (P. breviflora) E non-glandular stellate with central long branch (P. bracteosa) F bi- or trifurcate non-glandular stellate (P. nyalamensis) G sub-sessile/ sessile glandular trichomes (P. macrophylla) H simple glandular trichomes of (P. bracteosa) I branched glandular trichomes (P. breviflora).
Figs
Photos of bracts, SEM of bracts of Phlomoides henryi and related species A, B P. henryi C, D P. bracteosa E, F P. breviflora G, H P. macrophylla I, J P. nyalamensis K, L P. tibetica M, N P. milingensis O, P P. rotata. A, C, E, G, I, K, M, O photos of bracts B, D, F, H, J, L, N, P SEM of bracts.
SEM of both sides of leaves of Phlomoides henryi and related species A, B P. henryi C, D P. bracteosa E, F P. breviflora G, H P. macrophylla I, J P. nyalamensis K, L P. tibetica M, N P. milingensis O, P P. rotata A, C, E, G, I, K, M, O SEM of adaxial leaves B, D, F, H, J, L, N, P SEM of abaxial leaves.
Distribution of different types of trichome in the examined Phlomoides spp.
Tissue | Species | Eglandular hairs | Glandular hairs | ||||||
---|---|---|---|---|---|---|---|---|---|
Simple | Branched | Simple | Branched | ||||||
Short | Long | Symmetrically stellate | Stellate with central long branch | Bi- or trifurcate | Sub-sessile/ sessile | Short stalked | |||
Adaxial leaf | P. henryi | + | + | – | – | – | + | – | – |
P. bracteosa | + | – | + | + | + | + | – | – | |
P. breviflora | + | – | – | – | – | + | – | – | |
P. macrophylla | + | – | – | – | – | + | – | – | |
P. nyalamensis | + | – | – | – | – | + | – | – | |
P. tibetica | + | + | – | – | – | + | – | – | |
P. milingensis | + | + | – | – | – | + | – | – | |
P. rotata | + | + | – | – | – | + | – | – | |
Abaxial leaf | P. henryi | + | + | – | – | – | + | – | – |
P. bracteosa | + | – | + | + | + | + | + | – | |
P. breviflora | + | – | + | + | + | + | + | + | |
P. macrophylla | + | – | + | + | + | + | + | – | |
P. nyalamensis | – | – | – | – | + | + | – | – | |
P. tibetica | + | + | – | + | – | + | + | – | |
P. milingensis | + | – | – | + | + | + | + | – | |
P. rotata | + | – | + | + | + | + | – | – | |
Bract | P. henryi | + | + | – | – | – | + | – | – |
P. bracteosa | + | + | + | + | + | + | – | – | |
P. breviflora | + | – | + | + | – | + | + | – | |
P. macrophylla | + | + | – | – | – | + | – | – | |
P. nyalamensis | + | + | – | – | – | + | – | – | |
P. tibetica | + | + | – | – | – | + | – | – | |
P. milingensis | + | + | – | + | + | + | – | – | |
P. rotata | + | + | – | – | – | + | – | – |
Simple short eglandular trichomes were observed in every species on leaf and bract surface, but were missing in the abaxial leaf of Phlomoides nyalamensis, since it was nearly glabrous (Fig.
Trichomes were transparent to white or brown to black in Phlomoides. Trichomes inside the upper corolla lip of the new species (P. henryi), P. bracteosa and P. rotata were transparent to white, while the other five species were brown to black. Bract trichomes of P. tibetica and P. milingensis were brown to black (Fig.
Herbaria house millions of specimens that embody the plant diversity on the Earth. Many new species have been lurking in herbaria for many years before being published.
The resulting phylogenetic tree of Phlomoides in this study was similar to that in previous study (
As above mentioned, we believe that the differences merit recognition of the new species and we describe it below.
Phlomoides is a morphologically diverse and taxonomically difficult group with many characters used for traditional taxonomy being highly variable. In this study, we investigated trichome micro-morphology on bracts and leaves of Phlomoides henryi and related species. We found that trichomes are a useful character to distinguish some morphologically similar species. Based on the colour of trichomes, we can separate two groups of those species. Phlomoides nyalamensis, P. macrophylla, P. tibetica, P. milingensis and P. breviflora have brown to black trichomes on the upper corolla lip, while the other species (P. rotata, P. bracteosa and the new species described here, P. henryi) have transparent to white trichomes on the upper corolla lip. Trichome density and bract trichome colour can separate P. tibetica from the similar P. milingensis. Both species are distributed in Xizang at an altitude from 3500–4500 m and
Phlomoides henryi Y.Zhao & C.L.Xiang A habitat B plant with linear-tuberous roots C inflorescence D verticillaster E flowers F dissected flower G appendages at base of posterior filaments H fruiting calyces I dissected calyces J bracts K floral leaves L stem leaves. Photographs by Yue Zhao, except C by Li Chen.
In order to distinguish those species grouped with the new species in the phylogenetic tree (Fig.
1 | Upper corolla lip with trichome transparent to white | 2 |
– | Upper corolla lip with trichomes brown to black | 4 |
2 | No branched trichomes on leaf and bract | P. henryi |
– | With branched trichomes on leaf and bract | 3 |
3 | Floral bracts with branched trichomes, bracts linear to lanceolate | P. bracteosa |
– | Floral bracts with simple trichomes only, bracts needle-like to subulate | P. rotata |
4 | Floral bracts with trichomes brown to black | 5 |
– | Floral bracts with trichomes transparent to white | 6 |
5 | Floral bracts with black simple trichomes, no branched trichomes | P. tibetica |
– | Floral bracts with brown simple and branched trichomes | P. milingensis |
6 | Floral bracts less than 1 cm long, with branched trichomes | P. breviflora |
– | Floral bracts more than 1 cm long, with only simple trichomes | 7 |
7 | Flower purple | P. nyalamensis |
– | Flower white | P. macrophylla |
China, Yunnan Province, Honghe Hani and Yi Autonomous Prefecture, Jianshui County, on the forest edge, 23°57′52.54″N, 102°59′49.47″E, alt. 1279 m, 3 Sep 2022, F. Zhao, Y. Zhao & C.L. Xiang XCL2222 (holotype: KUN!; isotypes: KUN!, CSH!).
Within the subclade, Phlomoides henryi is morphologically similar to P. rotata for having transparent to white trichomes inside the upper corolla lip rather than brown to black and is distinct from all other species by lacking branched hairs. P. bracteosa has similar transparent to white trichomes inside the upper corolla lip, but with branched trichomes on both sides of leaves and floral bracts. The differences between P. henryi, P. rotata and P. bracteosa are listed in Table
Morphological comparisons amongst Phlomoides henryi, P. rotata and P. bracteosa.
Characters | P. henryi | P. rotata | P. bracteosa |
---|---|---|---|
Height | 100–150 cm | 2.5–10 cm | 20–50 (–100) cm |
Basal leaves | absent | rosette basal leaves | absent |
Floral leaf shape | narrowly lanceolate | lanceolate, oblanceolate, or linear | Ovate to lanceolate |
Floral leaf petiole length | 5–35 mm | Lack obvious petiole | Upper floral leaves sessile, lower floral leaves with petiole 5–20 mm long |
Floral bracts shape | Subulate | Needle-like to subulate | Linear to lanceolate, often with enlarged bracts |
Branched trichomes | No branched trichomes | With branched trichomes on abaxial leaves | With branched trichomes on both sides of leaves and bracts |
Roots robust, linear-tuberous. Stems 1–1.5 m tall, subquadrangular, robust, simple pilose. Basal leaves absent; stem leaves with petioles 4–15 cm long, with simple trichomes, broadly ovate to ovate-oblong, papery, 10–18 × 15–24 cm, adaxially green with sparse simple trichomes, abaxially light green, with sparse simple trichomes, denser and longer on the main vein, base cordate, margin serrate or crenate, apex acute to acuminate. Verticillasters axillary, 8–20-flowered; floral leaves with petioles 5–35 mm long, lanceolate, base rounded to cuneate, 1–6 × 0.5–4 cm, gradually reduced upwards; bracts subulate, 6–10 mm long, with sparse long simple trichomes, ca. 2 mm long. Calyx tubular, 10–11 × 4–5 mm, pubescent outside with sparse long simple trichomes on veins, conspicuously 10-veined; teeth 5, truncate, ca. 1.5 mm long, apical spines 3–4 mm long, with sparse long simple trichomes. Corolla light purple to pink, ca. 2.1 cm long, 2-lipped; posterior lip ca. 7 mm long, galeate, densely stellate tomentose outside, margin denticulate, bearded inside; anterior lip 3-lobed, ca. 7 × 8 mm, middle lobe largest, oblong, ca. 5 × 3 mm, lateral lobes ovate; tube glabrous outside, ca. 1.5 cm, annulate pilose inside. Stamens 4, included, with cobwebby indumentum, posterior filaments with reflexed appendages at base. Style unequally 2-lobed. Nutlets oblong-globose, glabrous.
The new species is named after the collector Augustine Henry (1857–1930), who collected more than 15,000 dry specimens and seeds from China.
Flowering from July to September and fruiting from October to December.
Based on present collections, P. henryi is only known from its type locality, i.e. Muyang Mountain in Jianshui County, Yunnan Province, China. It is restricted to the edge of the forest at an elevation near 1280 m.
jiàn shuǐ cǎo cāo sū (建水草糙苏).
Phlomoides henryi: Paratypes. China, Yunnan Province, Honghe Hani and Yi Autonomous Prefecture, Jianshui County, 6 September 2019, Jianshui Exped. 2164 (KUN); Yunnan Province, Mengtze, 1898, A. Henry 10216 (K000928267, K without barcode, LE without barcode); Yunnan Province, “Lao Kouy [Chan], 11 August 1907, F. Ducloux 369 (E).
Phlomoides bracteosa: India. Choor & Kidarlonta, 1832, J.F. Royle 633 (Type: K, K000894384); State of Punjab: Kangra, Lahul, Bhaga Valley, alt. 3000 m, 26 July 1933, T.R. Chand 74A (MICH, 1519061); State of Uttarakhand: Bhyundar Valley – Valley of flowers (N.E. road Josimath to Badrinath), alt. 3430 m, 16 August 1975, Anonymous 6583 (PE, 1290791).
Phlomoides breviflora: China. Xizang Province: Yadong County, on the way from Yadong to Dingga, alt. 2850 m, 11 June 1975, Qinghai-Tibet Exped. 750283 (Holotype: KUN, 1218974!; Isotype: PE, 00835569!, 00835570!); Nielamu County, Qu Town, alt. 3240 m, 12 September 2019, Y.P. Chen, Y. Zhao & B.Y. Zhang EM1139 (KUN). India. Sikkim: Nayathang-Phalut, alt. 3500 m, 2 June 1960, H. Hara et al. 5551 (TI); West Bengal: Darjeeling, Phalut, alt. 3500 m, 5 June 1960, H. Hara et al. 5555 (TI); Darjeeling, Garibans-Tanglu, alt. 2700 m, 7 June 1969, H. Hara et al. 5557 (TI). Nepal. Bagmati Province: Rasuwa District, Gossain than (Gosainkund), Benth in Wall. Cat. Herb. Ind. n. 2066 (Type: K, K001115039!).
Phlomoides macrophylla: Nepal. Gandaki Province: Mustang, Annapurna Conservation area, Trekking route Jomosom-Nayapul, Near Ghorepani village (way to Tikhedhunga). China. Xizang Province: Yadong County, Shang Yadong Village, alt. 3448 m, 30 August 2023, Y. Zhao et al. XCL2703 (KUN); Yadong County, on the way from Yadong County to the Pass of Naiduila Mountain, 9 September 2019, Y.P. Chen, Y. Zhao & B.Y. Zhang EM1145 (KUN).
Phlomoides nyalamensis: China. Xizang Province: Nyalam County, Zhangmu Town, on the way from Lixin to Xuebugang, open spaces in forests, alt. 2700–2800 m, 29 June 1975, Qinghai-Xizang Comp. Exped. 6622 (Holotype, KUN 1218985! Isotype, KUN 1218984!); Nyalam County, Zhangmu Town, Lixin Village, damp hillside, alt. 2800 m, 24 August 1972, Tibetan herbal medicine Exped. 1461 (PE, 00835826); Nyalam County, Zhangmu Town, on the way from Lixin to Xuebugang, edge of forests, alt. 2896 m, 13 September 2019, Y.P. Chen, Y. Zhao & B.Y. Zhang EM1145 (KUN).
Phlomoides tibetica: China. Xizang Province: in open alpine pastures of turf in well drained situations, alt. 4200–4500 m, 6 July 1924, F. K. Ward 5901 (Holotype: K, K000894378!; isotype: BM, BM000950520, E, E00301982!); Linzhi City, Bomi County, Northern Galongla Mountain, 3800–3900 m, 16 August 1982, S.Z. Cheng & B.S. Li 00160 (PE, 00923472); Linzhi City, on the way from Lulang Town to the Pass of Sejila Mountain, 4108 m, 15 September 2016, C.L. Xiang et al. 1456 (KUN). Bhutan. Trashigang District, Shingbe Town, 3800 m, 27 May 1949, F. Ludlow, G. Sherriff & J.H. Hicks 20673 (BM).
Phlomoides milingensis: China. Xizang Province: Mainling County, Zedanggang, alt. 4400 m, 26 July 1972, Tibetan herbal medicine Exped. 3883 (holotype: PE, 00835583; isotype: PE, 00923457); Mainling County, Nanyi Mountain, alt. 3400–3500 m, 28 July 1972, Tibetan herbal medicine Exped. 4228 (PE, 00835584; PE, 00923459); Gongbo’gyamda County, Xueka, 10 August 1974, Anonymous 2129 (PE, 00832244).
Phlomoides rotata: China: Xizang, Shannan City, Cona County, near radar station, alt. 4280 m, 17 July 1975, C.Y. Wu et al. 75-893 (KUN, 0216402); Naqu City, Lhari County, meteorological station, alt. 4500 m, 8 June 1976, Qinghai-Xizang Comp. Exped. 10491 (KUN, 0216409); Nyingchi City, Gongbo’gyamda County, Mira pass, alt. 4920 m, 30 August 1974, Qinghai-Xizang Comp. Exped. 74-2027 (KUN, 0216384); Sichuan, Liangshan Yi Autonomous Prefecture, Muli County, on the way from Mogalaji to Nonsa pasture, alt. 4312 m, 30 August 2015, C.L. Xiang 1219 (KUN, 1264607); Yunnan, Diqing Tibetan Autonomous Prefecture, Deqin County, Baimaxueshan, alt. 4300 m, 4 June 2000, Z.K. Zhou et al. 159 (KUN, 0699320); Qinghai, Yushu Tibetan Autonomous Prefecture, Yushu County, Hongtu Mountain, alt. 4900 m, 3 September 2013, J.W. Zhang et al. Zh678 (KUN, 1260768).
The authors are very grateful to the curators of the following herbaria for their kind assistance in research facilities: K, CDBI, E, GXMI, HIB, IBK, IBSC, K, KUN, KYO, MW, NAS, PE, SZ, TI. We also thank Mr. Zhi-Jia Gu for his technical assistance with SEM.
The authors have declared that no competing interests exist.
No ethical statement was reported.
This study was supported by the National Natural Science Foundation of China (Grant Nos. 32161143015, 31872648), International Partnership Program of Chinese Academy of Sciences (Grant No. 070GJHZ202211FN), the Yunnan Fundamental Research Projects (Grant No. 202001AS070016) awarded to CLX, the Postdoctoral Research Funding Projects of Yunnan Province and the Postdoctoral Directional Training Foundation of Yunnan Province awarded to FZ.
YZ and CLX conceived this research. YZ, FZ, JFX, YPC and CLX collected materials. YZ and FZ performed the experiments. YZ, AP, YPC and CLX analysed the data. All co-authors contributed to the manuscript and revised it critically.
Yue Zhao https://orcid.org/0000-0001-6850-5070
Fei Zhao https://orcid.org/0000-0001-7328-2262
Alan J. Paton http://orcid.org/ 0000-0002-6052-6675
Jin-Fei Xiao https://orcid.org/0000-0001-7729-2548
Ya-Ping Chen http://orcid.org/0000-0002-7502-1848
Chun-Lei Xiang https://orcid.org/0000-0001-8775-6967
DNA sequences associated with this work are available from GenBank (https://www.ncbi.nlm.nih.gov/genbank/) or from the corresponding author upon reasonable request.
Specimen information (taxon, voucher, herbarium, country) for samples newly-sequenced in the present study with GenBank accession numbers for atpB-rbcL, psbA-trnH, rpl16, rpl32-trnL, rps16, trnK, trnL-trnF, trnS-trnG and trnT-L, respectively. Herbarium abbreviations are listed after the vouchers. The accession numbers marked with an asterisk represent sequences newly generated. Only GenBank accession numbers are listed for sequences downloaded from NCBI.
Phlomis composita Pau, ON820555, ON820616, ON835580, ON835627, ON835674, ON835721, ON843184, OR632134, ON843231; Phlomis fruticosa L., ON820556, ON820615, ON835581, ON835628, ON835675, ON835722, ON843185, OR632135, ON843232; Phlomis herba-venti subsp. pungens (Willd.) Maire ex DeFilipps, ON820557, ON820614, ON835582, ON835629, ON835676, ON835723, ON843186, ON815620, ON843233; Phlomoides alpina (Pall.) Adylov, Kamelin & Makhm., OR631925, OR631967, OR642305, OR632009, OR632051, OR642347, OR642389, OR632137, OR632093; Phlomoides atropurpurea (Dunn) Kamelin & Makhm., OQ672946, OQ672946, OQ672946, OQ672946, OQ672946, OQ672946, OQ672946, OQ672946, OQ672946; Phlomoides betonicoides (Diels) Kamelin & Makhm., MN617020, MN617020, MN617020, MN617020, MN617020, MN617020, MN617020, MN617020, MN617020; Phlomoides bracteosa (Royle ex Benth.) Kamelin & Makhm., C.R. Lancaster 72 (BM), India, Srinagar, Pahalgam, Gully, alt. 2286 m, OR674852*, OR674855*, OR674856*, OR674858*, OR674860*, OR674863*, OR674864*, OR674867*, OR674869*; Phlomoides breviflora (Benth.) Kamelin & Makhm., OQ672923, OQ672923, OQ672923, OQ672923, OQ672923, OQ672923, OQ672923, OQ672923, OQ672923; Phlomoides burmanica (Mukerjee) Kamelin & Makhm., ON820563, ON820630, ON835588, ON835635, ON835682, ON835729, ON843192, OR632142, ON843239; Phlomoides chinghoensis (C.Y. Wu) Kamelin & Makhm., ON820580, ON820611, ON835605, ON835652, ON835699, ON835746, ON843209, ON815622, ON843256; Phlomoides congesta (C.Y. Wu) Kamelin & Makhm., ON820567, ON820608, ON835592, ON835639, ON835686, ON835733, ON843196, OR632145, ON843243; Phlomoides dentosa (Franch.) Kamelin & Makhm., OR631929, OR631973, OR642309, OR632015, OR632057, OR642351, OR642395, OR632149, OR632099; Phlomoides deserticola Sennikov, OQ672935, OQ672935, OQ672935, OQ672935, OQ672935, OQ672935, OQ672935, OQ672935, OQ672935; Phlomoides forrestii (Diels) Kamelin & Makhm., OR631934, OR631976, OR642313, OR632018, OR632060, OR642355, OR642398, OR632153, OR632102; Phlomoides franchetiana (Diels) Kamelin & Makhm., ON820561, ON820621, ON835586, ON835633, ON835680, ON835727, ON843190, OR632155, ON843237; Phlomoides hamosa (Benth.) Mathiesen, ON820558, ON820604, ON835583, ON835630, ON835677, ON835724, ON843187, OQ672937, ON843234; Phlomoides henryi Y.Zhao & C.L.Xiang, F. Zhao, Y. Zhao & C.L. Xiang XCL2222 (KUN), China, Yunnan Province, Jianshui County, Limin Town, Muyang Mountain, alt. 2179 m, OR674853*, OR674854*, OR674857*, OR674859*, OR674861*, OR674862*, OR674865*, OR674866*, OR674868*; Phlomoides inaequalisepala (C.Y. Wu) Kamelin & Makhm., OR631937, OR631979, OR642317, OR632021, OR632063, OR642359, OR642401, OR632160, OR632105; Phlomoides jeholensis (Nakai & Kitag.) Kamelin & Makhm., OR631938, OR631980, OR642318, OR632022, OR632064, OR642360, OR642402, OR632162, OR632106; Phlomoides koraiensis (Nakai) Kamelin & Makhm., OR631939, OR631981, OR642319, OR632023, OR632065, OR642361, OR642403, OR632163, OR632107; Phlomoides liangwangshanensis Y. Zhao, H.L. Zheng & C.L. Xiang, OR631940, OR631982, OR642320, OR632024, OR632066, OR642362, OR642404, OR632165, OR632108; Phlomoides likiangensis (C.Y. Wu) Kamelin & Makhm., OR631942, OR631984, OR642322, OR632026, OR632068, OR642364, OR642406, OR632167, OR632110; Phlomoides longiaristata (C.Y. Wu & H.W. Li) Salmaki, ON820559, ON820603, ON835584, ON835631, ON835678, ON835725, ON843188, ON815625, ON843235; Phlomoides longicalyx (C.Y. Wu) Kamelin & Makhm., OR631943, OR631985, OR642323, OR632027, OR632069, OR642365, OR642407, OR632168, OR632111; Phlomoides macrophylla (Benth.) Kamelin & Makhm., OR631944, OR631986, OR642324, OR632028, OR632070, OR642366, OR642408, OR632169, OR632112; Phlomoides maximowiczii (Regel) Kamelin & Makhm., ON820565, ON820622, ON835590, ON835637, ON835684, ON835731, ON843194, OR632170, ON843241; Phlomoides mazzettii Lazkov, OR631945, OR631987, OR642325, OR632029, OR632071, OR642367, OR642409, OR632171, OR632113; Phlomoides medicinalis (Diels) Kamelin & Makhm., OR631946, OR631988, OR642326, OR632030, OR632072, OR642368, OR642410, OR632172, OR632114; Phlomoides megalantha (Diels) Kamelin & Makhm., OR631947, OR631989, OR642327, OR632031, OR632073, OR642369, OR642411, OR632173, OR632115; Phlomoides melanantha (Diels) Kamelin & Makhm., OR631948, OR631990, OR642328, OR632032, OR632074, OR642370, OR642412, OR632174, OR632116; Phlomoides milingensis (C.Y. Wu & H.W. Li) Kamelin & Makhm., OR631949, OR631991, OR642329, OR632033, OR632075, OR642371, OR642413, OR632175, OR632117; Phlomoides moluccelloides (Bunge) Salmaki, OQ672938, OQ672938, OQ672938, OQ672938, OQ672938, OQ672938, OQ672938, OQ672938, OQ672938; Phlomoides mongolica (Turcz.) Kamelin & A.L. Budantzev, ON820576, ON820617, ON835601, ON835648, ON835695, ON835742, ON843205, OR632176, ON843252; Phlomoides muliensis (C.Y. Wu) Kamelin & Makhm., OR631950, OR631992, OR642330, OR632034, OR632076, OR642372, OR642414, OR632177, OR632118; Phlomoides nyalamensis (H.W. Li) Y. Zhao & C.L. Xiang, OR631952, OR631994, OR642332, OR632036, OR632078, OR642374, OR642416, OR632179, OR632120; Phlomoides oreophila (Kar. & Kir.) Adylov, Kamelin & Makhm., OR631953, OR631995, OR642333, OR632037, OR632079, OR642375, OR642417, OR632180, OR632121; Phlomoides ornata (C.Y. Wu) Kamelin & Makhm., ON820570, ON820618, ON835595, ON835642, ON835689, ON835736, ON843199, OR632181, ON843246; Phlomoides paohsingensis (C.Y. Wu) Kamelin & Makhm., OR631954, OR631996, OR642334, OR632038, OR632080, OR642376, OR642418, OR632182, OR632122; Phlomoides pedunculata (Y.Z.Sun) Kamelin & Makhm., OQ672936, OQ672936, OQ672936, OQ672936, OQ672936, OQ672936, OQ672936, OQ672936, OQ672936; Phlomoides pratensis (Kar. & Kir.) Adylov, Kamelin & Makhm., ON820579, ON820612, ON835604, ON835651, ON835698, ON835745, ON843208, ON815626, ON843255; Phlomoides rotata (Benth. ex Hook.f.) Mathiesen, ON820564, ON820602, ON835589, ON835636, ON835683, ON835730, ON843193, ON815627, ON843240; Phlomoides ruptilis (C.Y. Wu) Kamelin & Makhm., OR631955, OR631997, OR642335, OR632039, OR632081, OR642377, OR642419, OR632183, OR632123; Phlomoides sagittata (Regel) C.L.Xiang & Y.Zhao, ON820578, ON820620, ON835603, ON835650, ON835697, ON835744, ON843207, ON815617, ON843254; Phlomoides setifera (Bureau & Franch.) Kamelin & Makhm., OR631956, OR631998, OR642336, OR632040, OR632082, OR642378, OR642420, OR632186, OR632124; Phlomoides speciosa (Rupr.) Adylov, Kamelin & Makhm., ON820592, ON820631, ON835617, ON835664, ON835711, ON835758, ON843221, ON815629, ON843268; Phlomoides strigosa (C.Y. Wu) Kamelin & Makhm., OR631957, OR631999, OR642337, OR632041, OR632083, OR642379, OR642421, OR632187, OR632125; Phlomoides tatsienensis (Bureau & Franch.) Kamelin & Makhm., OR631960, OR632002, OR642340, OR632044, OR632086, OR642382, OR642424, OR632190, OR632128; Phlomoides tibetica (C. Marquand & Airy Shaw) Kamelin & Makhm., OR631961, OR632003, OR642341, OR632045, OR632087, OR642383, OR642425, OR632193, OR632129; Phlomoides tuberosa (L.) Moench, ON820575, ON820624, ON835600, ON835647, ON835694, ON835741, ON843204, ON815631, ON843251; Phlomoides umbrosa (Turcz.) Kamelin & Makhm., ON820571, ON820605, ON835596, ON835643, ON835690, ON835737, ON843200, OR632198, ON843247; Phlomoides younghusbandii (Mukerjee) Kamelin & Makhm., MW405448, MW405448, MW405448, MW405448, MW405448, MW405448, MW405448, MW405448, MW405448; Phlomoides zenaidae (Popov) Adylov, Kamelin & Makhm., OQ672945, OQ672945, OQ672945, OQ672945, OQ672945, OQ672945, OQ672945, OQ672945, OQ672945.