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Research Article
Molecular and morphological evidence for a new species of Stachys (Lamiaceae) from Hunan, China
expand article infoLing Xue‡, Jia-Hua Cai‡, Min Zhan‡, Xiao-Ping Li§, Lei Wu‡, Ya-Ping Chen|
‡ Central South University of Forestry and Technology, Changsha, China
§ Yingzuijie National Nature Reserve of Hunan Province, Huaihua, China
| Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China

Corresponding author: Lei Wu ( wuleiibk@163.com )

Corresponding author: Ya-Ping Chen ( chenyaping@mail.kib.ac.cn )

Academic editor: Chun-Lei Xiang
© 2023 Ling Xue, Jia-Hua Cai, Min Zhan, Xiao-Ping Li, Lei Wu, Ya-Ping Chen.
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: Xue L, Cai J-H, Zhan M, Li X-P, Wu L, Chen Y-P (2023) Molecular and morphological evidence for a new species of Stachys (Lamiaceae) from Hunan, China. PhytoKeys 236: 121-134. https://doi.org/10.3897/phytokeys.236.112741
Open Access

Abstract

Stachys yingzuijieensis, a new species from western Hunan, China, is described and illustrated. Molecular phylogenetic analyses based on three nuclear ribosomal DNA loci (ETS, ITS and 5S-NTS) recovered S. yingzuijieensis within the Stachys clade and as a sister group of S. arrecta. The two species can be easily distinguished by the morphology of lamina, corolla and nutlet. A key to all species of Stachydeae from China is also provided.

Key words

Eurystachys clade, Lamioideae, micromorphology, Stachydeae, taxonomy

Introduction

As one of the largest genera in Lamiaceae, Stachys L. comprises over 365 species distributed worldwide (Bhattacharjee 1980; Harley et al. 2004; POWO 2023). Together with other 11 genera, Stachys belongs to the largest tribe in subfamily Lamioideae, i.e. Stachydeae (Zhao et al. 2021). However, the intergeneric relationship within the tribe is taxonomically challenging and Stachys has been continuously shown to be non-monophyletic in previous molecular phylogenetic studies. While exploring the phylogenetic position of the Hawaiian endemic mints with respect to Stachys, Lindqvist and Albert (2002) showed that three genera endemic to Hawaii (Haplostachys (A. Gray) Hillebr., Phyllostegia Benth. and Stenogyne Benth.), as well as Prasium L., Phlomidoschema (Benth.) Vved. and Sideritis L., were embedded within Stachys. Scheen et al. (2010) and Bendiksby et al. (2011) further added the Asian genera Chamaesphacos Schrenk ex Fisch. & C.A. Mey., Hypogomphia Bunge, Suzukia Kudô and Thuspeinanta T. Durand to the list of taxa nested within Stachys in their lamioid-wide studies. The most comprehensive phylogenetic analyses of Stachydeae were performed by Salmaki et al. (2013, 2019), based on multiple nuclear ribosomal and plastid DNA loci. Salmaki et al. (2019) recognised 12 well-supported clades within the Eurystachys clade, a name suggested by Salmaki et al. (2013) to include all genera of Stachydeae, except the monotypic genus Melittis L. Though the synapomorphies for Stachydeae remain unclear, members of the tribe usually share campanulate or weakly 2-lipped calyx with spiny lobes and hairy throat, strongly 2-lipped corolla and apically rounded nutlets (Scheen et al. 2010).

A total of 18 species of Stachys are recorded from China and eight of them are endemic (Li and Hedge 1994). Except for Stachys, China also accommodates another three genera of Stachydeae, i.e. Chamaesphacos (1 sp.), Sideritis (2 spp.) and Suzukia (2 spp.) (Li and Hedge 1994; Liu and Zhang 2004). Recently, a potential new species of Stachys was discovered during our field investigation in western Hunan Province, China. By carrying out comprehensive molecular phylogenetic and morphological studies, we confirmed its status as a species new to science. It was, thus, named Stachys yingzuijieensis L. Wu & Y.P. Chen and described below.

Materials and methods

Molecular phylogenetic analyses

The phylogenetic placement of the new species within Stachydeae was evaluated based on the framework of Salmaki et al. (2019). A total of 90 accessions representing 88 taxa from all 12 clades and 11 genera of Eurystachys, as well as Melittis melissophyllum L., were sampled as the ingroups. Two genera that are closely related to Stachydeae – Betonica L. and Galeopsis L. – were selected as the outgroups. Except for one accession of the new species and one accession for each of eight species of Stachys from China that were newly sequenced here, all remaining sequences were downloaded from GenBank. Voucher information for newly-sequenced samples and GenBank accession numbers for all sequences are listed in Appendix 1.

Total genomic DNA was extracted from silica-gel-dried leaf material using the modified CTAB method (Doyle and Doyle 1987). According to Salmaki et al. (2019), three nuclear ribosomal DNA loci, i.e. the internal and external transcribed spacers (ITS and ETS) and the 5S non-transcribed spacer (5S-NTS), were used to reconstruct the phylogenetic relationships. Polymerase chain reaction primers and protocols of ITS and ETS followed those used by Chen et al. (2019) and that of 5S-NTS followed Roy et al. (2013).

Raw sequences were assembled and edited using Geneious v.11.0.3 (Kearse et al. 2012). Data matrices were aligned using MUSCLE (Edgar 2004) and then manually adjusted in Geneious. After removing the ambiguously aligned regions in the ITS dataset, the three DNA loci were concatenated for phylogenetic reconstruction. Partitioned Bayesian Inference (BI) and partitioned Maximum Likelihood (ML) analyses were performed on the web server Cyberinfrastructure for Phylogenetic Research Science (CIPRES) Gateway (http://www.phylo.org/; Miller et al. 2010), using RAxML-HPC2 (Stamatakis 2014) and MrBayes v.3.2.2 (Ronquist et al. 2012), respectively. Detailed settings for the two analyses followed those described in Chen et al. (2019). The resulting trees were visualised in TreeGraph 2 (Stover and Müller 2010).

Morphological studies

Morphological similarities and differences between the new species and other taxa of Stachydeae were compared, based on our previous field investigations and specimen examination. Images of specimens (including type specimens) and living plants of Stachydeae from JSTOR (https://www.jstor.org/), Global Biodiversity Information Facility (GBIF, https://www.gbif.org/), Chinese Virtual Herbarium (CVH, https://www.cvh.ac.cn/) and Plant Photo Bank of China (PPBC, http://ppbc.iplant.cn/) were examined. Protologues and other taxonomic and floristic literature related to Stachydeae (Knorring 1954; Ball 1972; Nelson 1981; Bhattacharjee 1982; Codd 1985; Li and Hedge 1994; Turner 1994; Paton et al. 2009; Salmaki et al. 2012) was also reviewed.

Trichomes on the lamina and calyx, as well as the nutlet and pollen morphology of the new species, were investigated using scanning electron microscopy (SEM). All materials were directly mounted on to stubs and sputter-coated with gold for 90 s at 20 mA. Micromorphological observations were conducted using a Zeiss EVO LS10 scanning electron microscope (Carl Zeiss NTS, Oberkochen, Germany) at 10 kV. Terminologies used for trichome, nutlet and pollen description followed those of Salmaki et al. (2008a, 2008b, 2009), Karaismailoğlu and Güner (2019, 2021) and Totmaj and Salmaki (2022).

Results

Phylogenetic results

The aligned length of the combined nuclear dataset was 1,381 bp (589 bp for ITS, 456 bp for ETS and 336 bp for 5S-NTS). The topologies of the BI and ML trees were largely consistent with each other, but the BI tree provided higher resolution. Thus, only the Bayesian 50% majority-rule consensus tree was presented (Fig. 1), the posterior probabilities (PP) and Bootstrap support (BS) values being superimposed on the nodes.

Figure 1. 

Bayesian 50% majority-rule consensus tree of Stachydeae based on combined nuclear (ITS, ETS and 5S-NTS) dataset. Support values ≥ 0.50 PP or 50% BS are displayed above the branches (an “*” indicates a support value = 1.00 PP or 100% BS and a “-” indicates a conflicting node in the BI and ML trees). Species marked in bold represent samples newly sequenced in the present study. Multiple accessions of the same species are numbered according to Appendix 1.

Our molecular phylogenetic result (Fig. 1) revealed that Melittis was sister to the remaining Stachydeae, i.e. the Eurystachys clade (PP = 0.99, BS = 58%). Two large clades were resolved within the Eurystachys clade: the first one (PP = 0.91, BS = 71%) mainly included temperate North American, Hawaiian and several Old World taxa and the second one (PP = 0.99, BS = 61%) only comprised Old World (mostly Mediterranean) taxa. Twelve robustly supported small clades (PP = 1.00, BS > 90%) can be further recognised, with two clades (Eriostomum clade and Stachys clade) in the first Eurystachys clade and the remaining (Burgsdorfia clade, Distantes clade, Empedoclia clade, Hesiodia clade, Marrubiastrum clade, Olisia clade, Prasium clade, Setifolia clade, Sideritis clade and Swainsoniana clade) in the second Eurystachys clade. Species distributed in China were mostly recovered in the Stachys clade, including the new species. Although relationships within the Stachys clade were poorly resolved, Stachys yingzuijieensis was strongly supported as sister to Stachys arrecta L.H. Bailey (PP = 0.99, BS = 81%).

Morphological results

Stalked glandular and simple non-glandular trichomes were found on both surfaces of the lamina as well as the calyx of the new species (Fig. 2A–C). The abaxial surface of lamina and the outside surface of the calyx were more densely covered with longer trichomes. Pollen grains of Stachys yingzuijieensis were tricolpate with reticulate exine sculpturing (Fig. 2D–F), while nutlets were ovate with glabrous and reticulate surface (Fig. 2G–I).

Figure 2. 

Trichome, pollen, and nutlet micromorphology of Stachys yingzuijieensis A trichomes on the adaxial surface of lamina B trichomes on the abaxial surface of lamina C trichomes on the outside surface of calyx D polar view of pollen E equatorial view of pollen F surface sculpturing of pollen G dorsal view of nutlet H ventral view of nutlet I surface sculpturing of nutlet.

Discussion

The backbone of Stachydeae in the present study and the 12 clades recovered in the Eurystachys clade (Fig. 1) were consistent with that of Salmaki et al. (2019). The Stachys clade, which was referred to as the “Stachys core clade” in Salmaki et al. (2013) and the Stachys s.s. clade in Lindqvist and Albert (2002), is one of the largest monophyletic groups in the Eurystachys clade and comprises five genera (Haplostachys, Phyllostegia, Stachys, Stenogyne and Suzukia) and over 100 species. No synapomorphy has been found for this clade due to large morphological and geographical diversity (Salmaki et al. 2019). Next-generation sequencing data and comprehensive morphological studies are needed to further clarify the synapomorphies and relationships within this taxonomically problematic and important group.

Only several representatives of Stachydeae from China had been included in previous molecular phylogenetic studies and no morphological study had been carried out for Chinese Stachys. In this study, nine species of Stachys from China were newly sequenced and included in the phylogenetic analyses. Our results showed that most species that were collected from or reported to be occurring in China were recovered within the Stachys clade, including the new species (Fig. 1). Stachys yingzuijieensis was further revealed to be sister to Stachys arrecta, a species distributed in the evergreen broad-leaved forests at altitudes of 1500–2000 m in central China.

Stachys yingzuijieensis differs from all other Chinese Stachydeae in its densely villosus and glandular pubescent plants, as well as the white corollas with tube included in the calyces (Figs 3, 4). For example, the corollas of Stachys arrecta are pink with purple spots and the corolla tubes are exerted from the calyces (Li and Hedge 1994). Except for above differences, Stachys yingzuijieensis also has oblong to oblong-lanceolate laminae with crenulate margin, whereas the laminae of Stachys arrecta are cordate with coarsely serrate margin. Moreover, they can be distinguished in the nutlet surface, which is smooth in the new species (Fig. 2), but verrucate in Stachys arrecta. More detailed differences between the two species are listed in Table 1. Here, we also provided a key to all species of Stachydeae from China below.

Table 1.
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Morphological comparisons between Stachys yingzuijieensis and S. arrecta.

Characters S. yingzuijieensis S. arrecta
Lamina Oblong to oblong-lanceolate, 10–16 × 4–6 cm, margin crenulate Cordate, 2.5–6.5 × 1.5–3 cm, margin coarsely serrate
Calyx Approximately 7 mm long, teeth ca. 3 mm long, ovate-lanceolate Approximately 5 mm long, teeth 2–2.5 mm long, narrowly triangular
Pedicel Absent Approximately 1 mm long
Corolla White without spots, ca. 1 cm long, tube included in calyx Pink with purple spots, ca. 1.2 cm long, tube exerted from calyx
Nutlet Surface smooth Surface verrucate
Figure 3. 

Morphology of Stachys yingzuijieensis from the type locality A habitat B habit C–D inflorescence E adaxial view of lamina F abaxial view of lamina G roots and rhizomes H frontal view of corolla I lateral view of calyces J dissected corolla K nutlets (A–J photographed by Lei Wu K photographed by Ya-Ping Chen).

Figure 4. 

Holotype specimen of Stachys yingzuijieensis.

Key to the species of Stachydeae from China

1 Creeping herbs 2
– Erect herbs 3
2 Middle lobe of lower corolla lip entire Suzukia shikikunensis
– Middle lobe of lower corolla lip irregularly incised Suzukia luchuensis
3 Lamina spinescent-aristate Chamaesphacos ilicifolius
– Lamina not spinescent-aristate 4
4 Calyx tubular-campanulate; corolla included in calyx 5
– Calyx campanulate; corolla exserted from calyx 6
5 Corolla yellow, middle lobe of lower lip incised Sideritis montana
– Corolla purple, middle lobe of lower lip entire Sideritis balansae
6 Annual herbs Stachys arvensis
– Perennial herbs 7
7 Bracteoles over half as long as calyx 8
– Bracteoles less than half as long as calyx, early deciduous 9
8 Plants densely sericeous-lanate; verticillasters in compact spikes Stachys lanata
– Plants pilose; verticillasters in widely spaced spikes Stachys melissifolia
9 Lamina oblong, lanceolate to oblong-lanceolate 10
– Lamina ovate, ovate-oblong, or cordate 16
10 Corolla white Stachys yingzuijieensis
– Corolla pink, purple to red-purple 11
11 Lamina densely villous-tomentose abaxially Stachys oblongifolia
– Lamina hispid, puberulent, or glabrous abaxially 12
12 Stems densely retrorse villous Stachys palustris
– Stems spreading hispid, glabrous, or subglabrous 13
13 Calyx densely villous-hispid outside Stachys baicalensis
– Calyx sparsely villous-hispid or glandular puberulent outside 14
14 Calyx teeth obtuse at apex; corolla tube long exserted from calyx Stachys adulterina
– Calyx teeth spinescent at apex; corolla tube included in calyx 15
15 Lamina sparsely minutely hispid or subglabrous adaxially; calyx sparsely villous-hispid along veins outside Stachys chinensis
– Lamina glabrous adaxially; calyx glandular puberulent outside Stachys japonica
16 Corolla white or yellow 17
– Corolla pink or purple 18
17 Corolla white; calyx teeth triangular, less than 2 mm long Stachys taliensis
– Corolla yellow; calyx teeth ovate-triangular, over 2 mm long Stachys xanthantha
18 Rhizomes not enlarged or succulent 19
– Rhizomes enlarged, succulent 21
19 Lamina over 8 cm long Stachys sylvatica
– Lamina less than 5 cm long 20
20 Calyx teeth ovate-lanceolate Stachys strictiflora
– Calyx teeth triangular Stachys kouyangensis
21 Calyx teeth linear-lanceolate, reflexed Stachys pseudophlomis
– Calyx teeth narrowly triangular to triangular, straight 22
22 Lamina ovate-oblong; nutlet smooth Stachys geobombycis
– Lamina ovate to cordate; nutlet tuberculate 23
23 Lamina cordate; calyx ca. 5 mm long Stachys arrecta
– Lamina ovate to oblong-ovate; calyx ca. 9 mm long Stachys sieboldii

Taxonomic treatment

Stachys yingzuijieensis L.Wu & Y.P.Chen, sp. nov.

urn:lsid:ipni.org:names:77332759-1
Figs 3, 4

Type

China, Hunan, Huitong County, Yingzuijie National Nature Reserve, alt. 300–800 m, 26°56′N, 109°54′E, 3 Aug 2022, L. Wu et al. YZJ0145 (holotype: CSFI079941!; isotype: CSFI!).

Diagnosis

Stachys yingzuijieensis is most closely related to S. arrecta, but differs in its lamina oblong to elliptic-lanceolate (vs. cordate) with margin crenulate (vs. coarsely serrate), corolla white (vs. pink with purple spots) with tube included in calyx (vs. exerted from calyx) and nutlet surface smooth (vs. verrucate).

Herbs perennial. Rhizomes white, densely glandular pubescent. Stems erect, simple, 50–75 cm long, quadrangular, densely villous and glandular pubescent. Leaves opposite; petioles 2–4 cm long, densely villous and glandular pubescent; lamina oblong to oblong-lanceolate, papery, 10–16 × 4–6 cm, apex acute, margin crenulate, base cordate, adaxially green, sparsely villous and glandular pubescent, abaxially light green, densely villous and glandular pubescent, lateral veins 4–5-paired, conspicuously elevated abaxially. Verticillasters 6-flowered, flowers sessile; bracts leaf-like, upper ones sessile, lanceolate, densely villous and glandular pubescent on both surfaces, longer than verticillasters; bracteoles linear, 1–2 mm long. Calyx campanulate, ca. 7 mm long, 10-veined, densely villous and glandular pubescent outside, glandular pubescent inside, fruiting calyx dilated, ca. 9 mm long; teeth 5, subequal, ovate-lanceolate, ca. 3 mm long, apex spinescent. Corolla white, ca. 1 cm long, tube ca. 7 mm long, ca. 1.5 mm wide, pubescent annulate inside at 1/3 distance from base; 2-lipped, upper lip erect, concave, subcircular, ca. 3 mm in diam., densely pubescent and glandular pubescent outside, glabrous inside, lower lip spreading, sparsely pubescent and glandular pubescent to glabrescent outside, glabrous inside, ca. 6 mm long, 3-lobed, medium lob largest, trapeziform, ca. 3 mm long, ca. 4 mm wide, apex entire or emarginate, lateral lobs oblong, ca. 2 mm long, ca. 1 mm wide. Stamens 4, straight, included, filaments pubescent and glandular pubescent, anther cells 2, divergent. Style included, glabrous, apex subequally 2-lobed, lobes subulate. Ovary rounded at apex, glabrous. Nutlets 4, dark brown, ovoid, ca. 1.5 mm in diam., smooth and glabrous.

Phenology

Flowering from July to September, fruiting from August to October.

Distribution and habitat

Currently, S. yingzuijieensis is only known from the Yingzhuijie National Nature Reserve and a total of 50 mature plants were found during our field investigation. The new species usually grows in shady and moist places in evergreen broad-leaved forests at an altitude of 300–800 m.

Etymology

The specific epithet is derived from the type locality of the new species, i.e. the Yingzuijie National Nature Reserve in western Hunan Province, China.

Chinese name (assigned here)

yīng zhuǐ jiè shuǐ sū (鹰嘴界水苏).

Additional specimen examined

China. Hunan: Huitong County, Yingzuijie National Nature Reserve, 8 Aug 2022, L. Wu et al. YZJ0654 (CSFI!).

Acknowledgements

We would like to thank Ms. Jia-Yi Zeng for her help with data collection and Mr. Zhi-Jia Gu for his technical assistance in SEM. We are also grateful to the anonymous referees for their valuable suggestions that greatly improved our manuscript.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This work was supported by the Natural Science Foundation of Hunan (2021JJ31152) and the Investigation Foundation of Yingzuijie National Nature Reserve (90102/68228166) to LW and the Yunnan Fundamental Research Projects (202301AT070303) to Y-PC.

Author contributions

LX, LW and Y-PC conceptualized the study. LW, J-HC and X-PL collected the samples. LX, MZ and Y-PC conducted the analyses. LX drafted the manuscript. Y-PC and LW revised the manuscript. All authors read and approved the final manuscript.

Author ORCIDs

Ya-Ping Chen https://orcid.org/0000-0002-7502-1848

Data availability

All of the data that support the findings of this study are available in the main text.

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Appendix 1

Specimen information (taxon, voucher, herbarium, country) for samples newly sequenced in the present study with GenBank accession numbers for ITS, ETS and 5S-NTS, respectively. A “-” indicates a missing sequence. 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.

Betonica officinalis L., KF529533, MK909580, MK909684; Betonica scardica Griseb., KF529534, MK909581, -; Chamaesphacos ilicifolius Schrenk ex Fisch. & C.A. Mey., KF529540, MK909585, MK909689; Galeopsis angustifolia Ehrh. ex Hoffm, KF529535, -, MK909685; Galeopsis pubescens Besser, KF529536, -, MK909686; Haplostachys haplostachya (A. Gray) H.St. John, KF529541, -, MK909690; Hypogomphia bucharica Vved., KF529542, MK909587, MK909691; Hypogomphia turkestana Bunge, KF529543, MK909588, MK909692; Melittis melissophyllum L., KF529544, KF235787, MK909693; Phyllostegia velutina (Sherff) H.St. John, KF549547, KF235809, AF308212; Phyllostegia waimeae Wawra, KF529548, KF235811, KF235752; Prasium majus L. 1, KF529549, MK909590, MK909694; Prasium majus L. 2, KF529550, MK909591, AF501919; Sideritis canariensis L., AF335605, MK909593, MK909695; Sideritis clandestina (Bory & Chaub.) Hayek, AF335616, MK909595, -; Sideritis endressii subsp. emporitana Willk, AF335627, MK909598, MK909698; Sideritis incana L., AF335634, MK909601, -; Sideritis leucantha Cav., AF335636, MK909603, MK909700; Sideritis macrostachys Poir., AF335609, -, AF501920; Sideritis montana L. 1, AF335612, -, MK909701; Sideritis montana L. 2, KF529551, -, MK909702; Sideritis nutans Svent., DQ900767, MK909604, MK909703; Sideritis perfoliata L., AF335618, -, MK909705; Sideritis romana L. 1, AF335614, -, MK909706; Sideritis romana L. 2, KF529552, -, AF501922; Sideritis scardica Griseb., AF335619, MK909606, MK909707; Sideritis syriaca L., AF335620, -, MK909708; Sideritis tragoriganum Lag., AF335639, MK909608, MK909710; Stachys aculeolata Hook. f., KF529556, KF235814, AF501924; Stachys aethiopica L., KF529559, KF235815, KF235753; Stachys affinis Bunge, MH703287, KF235816, AF501925; Stachys albens A. Gray, KF529560, MK909613, AF501928; Stachys alpigena T.C.E. Fr., KF529561, KF235822, KF235755; Stachys arabica Hornem., KF529564, MK909616, MK909711; Stachys arenaria Vahl, KF529566, MK909618, -; Stachys arrecta L.H. Bailey, H.J. Dong et al. HGNU-0485 (KUN), Hubei, China, OR878465*, OR887616*, OR887626*; Stachys arvensis (L.) L., KF529567, MK909619, MK909712; Stachys baicalensis Fisch. ex Benth., Y.P. Chen & Y. Zhao EM1459 (KUN), Hebei, China, OR878468*, OR887619*, OR887629*; Stachys bullata Benth., KF529576, KF235831, KF235759; Stachys burchelliana Launert, KF529574, MK909624, MK909713; Stachys byzantina K. Koch, KF529577, KF235832, AF501938; Stachys chinensis Bunge ex Benth., B. Liu et al. 7252 (PE), Beijing Botanical Garden (cultivated), China, OR878467*, OR887618*, OR887628*; Stachys chrysantha Boiss. & Heldr., KF529580, MK909628, AF501939; Stachys circinata L’Hér., KF529581, MK909629, -; Stachys corsica Pers., KF529582, KF235836, KF235762; Stachys cretica L., KF529583, KF235838, AF501948; Stachys debilis Kunth, KF529584, KF235839, KF235763; Stachys distans Benth., KF529585, MK909630, MK909716; Stachys dregeana Benth., KF529586, MK909631, MK909717; Stachys durandiana Coss., KF529587, MK909632, MK909718; Stachys eriantha Benth., KF529589, KF235842, AF501951; Stachys floridana Shuttlew. ex Benth, KF529590, KF235843, AF501952; Stachys graeca Boiss. & Heldr., KF529595, MK909636, MK909719; Stachys grandidentata Lindl., KF529596, KF235845, KF235766; Stachys grandifolia E. Mey., KF529597, MK909637, -; Stachys heraclea All., KF529598, MK909638, MK909720; Stachys hildebrandtii Vatke, KF529599, MK909639, MK909721; Stachys hyssopoides Burch. ex Benth, KF529600, MK909640, MK909722; Stachys inflata Benth., KF529601, MK909641, MK909723; Stachys ionica Halácsy, KF529602, MK909642, MK909724; Stachys kouyangensis (Vaniot) Dunn, Y.P. Chen et al. EM482 (KUN), Yunnan, China, OR878463*, OR887614*, OR887624*; Stachys lamioides Benth., KF529607, KF235852, KF235773; Stachys latidens Small, KF529608, KF235854, AF501956; Stachys lavandulifolia Vahl, KF529609, MK909646, MK909725; Stachys macraei Benth., KF529611, KF235857, KF235774; Stachys maritima Gouan, KF529612, MK909647, MK909726; Stachys melissifolia Benth., Y.P. Chen et al. EM1206 (KUN), Xizang, China, OR878466*, OR887617*, OR887627*; Stachys natalensis Hochst., KF529619, MK909654, -; Stachys nigricans Benth., KF529622, MK909657, -; Stachys oblongifolia Wall. ex Benth., H. Peng et al. FJ881 (KUN), Guizhou, China, OR878472*, OR887623*, OR887632*; Stachys ocymastrum (L.) Briq., KF529623, MK909658, MK909727; Stachys palustris L., KF529624, MK909659, MK909728; Stachys pilosa Nutt., KF529628, KF235861, MK909730; Stachys pubescens Ten., KF529629, MK909663, MK909731; Stachys reptans Hedge, KF529632, MK909664, MK909732; Stachys rupestris Montbret & Aucher ex Benth., KF529633, MK909665, MK909733; Stachys saxicola Coss. & Balansa, KF529634, MK909666, MK909734; Stachys schtschegleevii Sosn. ex Grossh., KF529637, MK909667, MK909736; Stachys setifera subsp. iranica (Rech. f.) Rech. f., KF529636, -, MK909735; Stachys setifera subsp. setifera C.A. Mey., KF529635, -, AF501976; Stachys sieboldii Miq., Y.P. Chen & Y. Zhao EM1492 (KUN), Ningxia, China, OR878469*, OR887620*, OR887630*; Stachys subaphylla Rech.f., KF529641, MK909671, MK909737; Stachys swainsonii Benth., KF529642, KF235871, AF501977; Stachys sylvatica L., KF529643, MK909672, MK909738; Stachys tenuifolia Willd., OR392565, -, AF501981; Stachys tetragona Boiss. & Heldr., KF529646, MK909673, MK909739; Stachys trinervis Aitch. & Hemsl., KF529647, MK909674, MK909740; Stachys turcomanica Trautv., KF529649, MK909676, MK909742; Stachys xanthantha C.Y. Wu, Y.P. Chen & H.M. Li EM620 (KUN), Chongqing, China, OR878464*, OR887615*, OR887625*; Stachys yingzuijieensis L. Wu & Y.P. Chen, L. Wu et al. YZJ0654 (CSFI), Hunnan, China, OR878471*, OR887622*, OR887613*; Stenogyne bifida Hillebr., KF529652, KF235876, AF308221; Suzukia luchuensis Kudô, KF529653, MK909678, MK909744; Suzukia shikikunensis Kudô, KF529655, KF235889, KF235782; Thuspeinanta brahuica (Boiss.) Briq., KF529656, MK909679, MK909745; Thuspeinanta persica (Boiss.) Briq., KF529657, MK909681, MK909746.

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