$¢PhytoKeys PhytoKeys 259: 131-144 (2025) DOI: 10.3897/phytokeys.259.145861 Research Article Paphiopedilum motuoense (Orchidaceae, Cypripedioideae), a new species from Motuo, Xizang, China Feng-Xia Tang’®, Wen-Hui Rao™®, Ze Zhang, Xin-Yi Wu'®, Meng Wang™, Jian Li'®, Jie-Shan Chen"™®, Xiao-Juan Duan"™®, Mei-Na Wang'® 1 Shenzhen Key Laboratory for Orchid Conservation and Utilization, Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization, The Orchid Conservation & Research Center of Shenzhen and the National Orchid Conservation Center of China, Shenzhen, 518114, Guangdong, China 2 Yunnan Yelantang Biotechnology Co., Ltd, Kunming, 650000, Yunnan, China Corresponding author: Mei-Na Wang (snow-wmn2005@163.com) OPEN Qrceess Academic editor: Murielle Simo-Droissart Received: 4 January 2025 Accepted: 29 April 2025 Published: 30 June 2025 Citation: Tang F-X, Rao W-H, Zhang Z, Wu X-Y, Wang M, Li J, Chen J-S, Duan X-J, Wang M-N (2025) Paphiopedilum motuoense (Orchidaceae, Cypripedioideae), a new species from Motuo, Xizang, China. PhytoKeys 259: 131-144. https://doi.org/10.3897/ phytokeys.259.145861 Copyright: © Feng-Xia Tang et al. This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International - CC BY 4.0). Abstract Paphiopedilum motuoense, a newly discovered orchid from Motuo County, Xizang, China, is described and illustrated. Paphiopedilum motuoense can be distinguished from the related Paphiopedilum species P gingyongii and P venustum by several distinctive features: a significantly smaller staminode above the column, a distinct purplish-red lip with purplish-brown veins, pouched with erect and triangular auriculas on both sides of its mouth, and leaves with pale green and pale yellowish-green tessellations. The novelty is also well supported as a new species by molecular phylogenetic analysis. Additionally, the complete chloroplast genome of P motuoense, 157,218 bp in length, was assembled and annotated. It contains an LSC region of 86,275 bp, SSC region of 949 bp and two IRs of 34,997 bp, with 120 genes, including 76 PCGs, 36 tRNA genes and 8 rRNA genes. Key words: China, new orchid species, Paphiopedilum motuoense, plastid genome Introduction Orchidaceae, one of the two largest angiosperm families with more than 29,000 species, is divided into five subfamilies (Chase et al. 2015; Govaerts et al. 2021; Zhang et al. 2023; Pérez-Escobar et al. 2024). The subfamily Cypripedioideae Lindl. (slipper orchids) is characterized by a pouchlike lip, two fertile stamens, a shield-like staminode, and a synsepal composed of the fused lateral sepals (Lindley 1840; Dressler 1982; Guo et al. 2012). Slipper orchids have a wide dis- tribution, ranging from temperate to tropical regions of Eurasia and America, and the conduplicate-leaved species native to tropical Asia were considered a distinct lineage, which was established the genus Paphiopedilum by Ernst Hugo Heinrich Pfitzer in 1886 (Pfitzer 1886; Cox et al. 1997; Guo et al. 2012). Paphiopedilum is the largest genus of the subfamily Cypripedioideae, com- prising 109 accepted species and 35 natural hybrids (Liu et al. 2009; IPNI 2023; POWO 2024). Based on morphological characteristics, Pfitzer (1903) * The authors contributed equally to this work. 131 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China summarized the taxonomic studies of Paphiopedilum and divided the genus into three subgenera, namely Anotopedilum, Brachypetalum and Otopedilum. Brieger (1971) maintained the delimitation of Brachypetalum subgenus, and divided the genus into four subgenera (Barbata, Brachypetalum, Paphiopedi- lum and Polyantha). Later, Karasawa and Saito (1982) combined the data of chromosome number, divided the genus into six subgenera (Brachypetalum, Cochlopetalum, Paphiopedilum, Parvisepalum, Polyantha and Sigmatipetalum), and all the sections of Barbata subgenus were put into the Sigmatipetalum subgenus; but this delimitation result was not widely accepted. Atwood (1984) merged the delimitation of Karasawa and Saito (1982), and divided the genus into only two subgenera (Brachypetalum and Paphiopedilum) based on the lip morphology. Then, Cox et al. (1997) performed molecular phylogenetic anal- yses based on nuclear rDNA ITS sequences and Cribb (1997), based on mor- phological characteristics of flower inflorescence, leaf type, floral morphology and molecular phylogenetic results, and divided the genus into three subgenera (Brachypetalum, Paphiopedilum and Parvisepalum). The subgenus Paphiopedi- lum, characterized by having a helmet-shaped or slipper-shaped pouch, a rather long claw at the base, and lacking the incurved or involute margins at the apex, comprises 93 accepted species and is divided into five sections (Barbata, Co- chlopetalum, Coryopedilum, Paphiopedilum and Pardalopetalum) (Cribb 1997; Tsai et al. 2020; Guo et al. 2021; POWO 2024). Widespread reticulate evolution within Paphiopedilum, coupled with the discovery of new species like P. canhii (Averyanov 2010) and P. rungsuriyanum (Gruss 2014), results in intricate in- terspecific relationships. Additional data and further research are needed to understand these relationships (Guo et al. 2012, 2021; Tsai et al. 2020). Paphiopedilum is mainly distributed in tropical and subtropical southeast Asia (Cribb 1987; Cribb 1997; Liu et al. 2009; Pemberton 2013). With 33 accept- ed species, China is an important species diversity center of Paphiopedilum (Liu et al. 2009; Liu et al. 2020; POWO 2024; COL-China 2024; IPNI 2025). In China, Paphiopedilum species are predominantly distributed across the south- western to southern regions, with the highest species diversity recorded in Yun- nan Province (Liu et al. 2009; POWO 2024; COL-China 2024). Recent taxonomic studies have identified several novel species within Yunnan Province, such as P notatisepalum Z.J.Liu, Meina Wang & S.R.Lan (Wang et al. 2017). Xizang, con- stituting the principal region of the Qinghai-Tibet Plateau, harbors the world's highest-latitude tropical rainforest and represents a plant diversity hotspot with the greatest elevational range globally. To date, only five Paphiopedilum spe- cies have been documented in Xizang (Chen et al. 2023), including P. fairriea- num (Lindl.) Stein (Stein 1892), P micranthum Tang & F.T.Wang (Tang and Wang 1951), P gingyongii Z.J.Liu & L.J.Chen (Liu and Chen 2010), P venustum (Wall. ex Sims) Pfitzer (Pfitzer 1888) and P wardii Summerh. (Summerhayes 1932). Paphiopedilum species are extremely rare and highly prized for their ornamental value, notably their exceptionally attractive flowers (Cribb 1987; Cribb 1997; Liu et al. 2009; Pemberton 2013). All Paphiopedilum species have been included in Appendix | of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES), the specimens of which are prohibited in international trade except for non-commercial purposes (https://cites.org/eng). In China, all Pa- phiopedilum species have also been listed on the List of Wild Plants under State Priority Conservation, and their illegal collections have been forbidden since 2021. PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 132 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China During a botanical investigation in Motuo County, Xizang, China, in 2014, we found a distinct subpopulation of Paphiopedi/lum under the broad-leaved forest. Some of these plants were transferred to the Yunnan Yelantang Biotechnolo- gy Co., Ltd. (Kunming, China) at that time. Based on the monitoring of those plants and the detailed morphological and molecular comparisons with similar species, they were considered to belong to a new species of Paphiopedilum which is described below. Materials and methods Morphological analysis Morphological characteristics of the new species, including the number, length and width of leaf, the length and width of scape, the diameter and color of flower, the shape, length and width of sepal, petal, and staminode, were observed, mea- sured and photographed based on three flowering living individuals of this species in Motuo, Xizang, China. The terminologies we used follow the terms of Chen et al. (2009) and Liu et al. (2009). The voucher specimen is deposited in the National Or- chid Conservation Center of China (NOCC). We compared its morphological char- acteristics with 87 species descriptions in the literature of Paphiopedilum genus, and with 9 herbarium specimens belonging to P. gingyongii (including holotype) and P. venustum. These herbarium specimens of the two species are housed in NOCC, National Plant Specimen Resource Bank Main Library (PE), Herbarium of Kunming Institute of Botany, Chinese Academy of Sciences (KUN), and Tibetan Plateau Biological Herbarium of the Chinese Academy of Sciences (HNWP). Molecular analysis Total genomic DNA was extracted from fresh leaf (voucher specimen J.B.Chen 01800) using a modified procedure of CTAB (cetyltrimethylammonium bro- mide) (Doyle and Doyle 1987; Li et al. 2013). After purifying the extracted DNA, we prepared paired-end libraries by fragmenting the genomic DNA into short fragments of approximately 350 bp. Subsequently, the libraries were sequenced in paired-end (150 bp) using the Illumina NovaSeq 6000 platform (Illumina, Inc., San Diego, CA, USA) at Novogene Co., Ltd. (Beijing, China). A total of 10 GB reads of genome skimming data were generated. The plastid genome was assembled employing GetOrganelle with appropri- ate parameters (Jin et al. 2020), using the chloroplast genome of P venustum (Wall. ex Sims) Pfitze as the reference sequence (OM066335). The Bandage software was employed to visualize the final assembly graphs of the plastid genomes, assessing their completeness and accuracy (Wick et al. 2015). Then, the obtained scaffolds and contigs were annotated by Geneious Prime (Bio- matters Ltd., Auckland, New Zealand) (Kearse et al. 2012) and Plastid Genome Annotator (Qu et al. 2019). The annotated complete chloroplast genome was deposited in GenBank with accession number OR392426. The circular plastid genome map of P motuoense was drawn and visualized in Chloroplot online software (https://irscope.shinyapps.io/chloroplot/) (Zheng et al. 2020). A total of 62 species from Paphiopedilum were included in the molecular phylogenetic analyses, with Phragmipedium longifolium (Warsz. & Rchb.f.) Rolft (NC_028149) PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 133 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China used as the outgroup (Li et al. 2016; Guo et al. 2021) (Table 1). We used the Py- thon script get_annotated_regions_from_gb.py (https://github.com/Kinggerm/ PersonalUtilities) to extract protein-coding gene sequences from the plastid genome. Each protein-coding gene was individually aligned by MAFFT 7.3 (ffT- NS-i x 1000 strategy) (Katoh and Standley 2013) and removed by trimAl 1.2 with default settings (Capella-Gutiérrez et al. 2009). Finally, the 67 protein-coding genes were rapidly concatenated using the Python script concatenate_fasta.py (https://github.com/Kinggerm/PersonalUtilities) before phylogenetic analyses. Phylogenetic analyses were performed using Maximum Likelihood (ML) and Bayesian inference (BI). The ML analysis was conducted using IQ-TREE v1.6 with SH-aLRT test and ultrafast bootstrap (UFBoot) feature (—alrt 1000 —bb 1000 —nt AUTO) (Nguyen et al. 2015; Hoang et al. 2018). The substitution model for the concatenated alignment was automatically selected using ModelFinder in 1Q- TREE, with the Bayesian Information Criterion (BIC) identifying TVM+F+R2 as the best-fit model. BI analyses of phylogeny were performed in MrBayes v.3.2.6 under partition model (2 parallel runs, 2000000 generations), in which the initial 25% of sampled data were discarded as burn-in. The produced phylogenetic trees were rendered using Figtree v1.4.4 (http://tree.bio.ed.ac.uk/software/fig- tree/) and enhanced with Adobe Illustrator software for improved visualization. Results and discussion Morphological evidence The leaf morphology, petal and lip color, pouch mouth trait, and staminode size that the novelty shows are distinguishing characteristics of Paphiopedilum, which indicates the novelty is placed within the genus. Moreover, the novelty exhibits the closest similarities with P venustum and P gingyongii, but differs in several key characteristics, including triangular auriculas on both sides of its pouch mouth, a significantly smaller staminode above the column, a distinct purplish-red lip with purplish-brown veins, pale yellow-white petals flushed with purple-red spots, and leaves tessellated with pale green and pale yellowish-green spots. These com- pared morphological characters of the novelty, P venustum, and P. gingyongii, are clearly presented in Table 1 and Fig. 1, according to which we speculate that this novelty could be a new species, and we name it P motuoense. Molecular evidence The complete plastid genome of P motuoense is 157,218 bp in length and exhib- its a typical quadripartite structure, including one large single-copy region (LSC) of 86,275 bp, one small single-copy region (SSC) of 949 bp, and two inverted repeat regions (IRs) of 34,997 bp (Fig. 2). The chloroplast genome contains 120 genes, including 76 protein-coding genes (PCGs), 36 tRNA genes and 8 rRNA genes (Table 2, Fig. 2). The total GC content of the whole plastome is 35.7%. The phylogenetic analyses based on 67 shared protein-coding genes from 62 Pa- phiopedilum whole plastomes indicate that P motuoense is close to P gingyongii with high support (SH-aLRT 99.4%, UfBoot 100%) and then sister to P venus- tum also with strong support (SH-aLRT 100%, UfBoot 100%) (Fig. 3). The BI tree (Suppl. material 1) exhibits identical topological relationships to the ML analysis. PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 134 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China Table 1. Morphological comparisons of P motuoense and its two related species. Characters Paphiopedilum motuoense P. venustum P. qingyongii Leaf pattern and leaves adaxially tessellated with leaves adaxially tessellated with | leaves adaxially without tessellation color pale green and pale yellowish dark green and gray- or yellow- or scarcely tessellated green brownish green Petal color petals pale yellowish white, flushed _ petals yellow-whitish with green | petals downward curvy, white-green with purplish red veins, flushed with dark red in densely spotted with purple-red apical half Lip color lip purplish red with purplish brown | lip pale yellow, distinctly veined with | lip pale yellow, distinctly veined with veins, pouch margin pale yellowish | green and slightly flushed dark red, | deep green, pouch margin yellow white pouch margin pale yellow Pouch mouth of lip | pouch triangular auriculate on both | pouch auriculate on both sides of pouch auriculate on both sides of sides of its mouth its mouth its mouth Staminode shape staminode smaller, reniform- staminode larger, reniform- staminode larger, broadly oblong, and size obcordate, 2.9-3.2 x 5-5.3 mm obcordate, 6-7 x 9-13 mm 6.9-7.1 x 8.8-9 mm hee Se : go ov “4 ie 4 r [ Dark Purplish Grey A GREYED-GREEN 193 GROUP. a Pale Yellow Green A GREYED-PURPLE GROUP. Nite Dark Greyish Purple B Ew GREEN 77 GROUP. “) 3 | Dark Yellowish Green A | ‘Yellowish Green _ 4 . i Figure 1. Paphiopedilum motuoense and its allies al-d1 P motuoense a1 whole plant b1 leaf c1 flower d1 staminode. a2-d2 P venustum a2 whole plant b2 leaf c2 flower d2 staminode. a3—d3 P. gingyongii a3 whole plant b3 leaf c3 flower d3 staminode. PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 135 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China no} a o > Paphiopedilum motuoense 76 genes 157,218 bp GC: 35.7% Gv fey (0) ao’ photosystem | photosystem II cytochrome b/f complex ATP synthesis RubisCO larg subunit RNA polymerase small ribosomal protein large ribosomal protein clpP, matK, infA hypothetical reading frame transfer RNA ribosomal RNA other Q 8 > Figure 2. Chloroplast genome map of Paphiopedilum motuoense. Our morphological and molecular evidences indicate that P motuoense is distinctive for identification as a Separate species from the other Paphiopedilum species. The following is a description of the P motuoense. Taxonomy treatment Paphiopedilum motuoense M. N. Wang, F. X. Tang & W. H. Rao, sp. nov. urn:lsid:ipni.org:names:77364576-1 Figs 4,5 Chinese name. ae} 924 Type. CHINA + Xizang, Linzhi City, Motuo County, alt. 800 m, 23 Apr 2021, J.B.Chen 01800 (holotype: NOCC). Diagnosis. Paphiopedilum motuoense is similar to P venustum and P gin- gyongii in morphology, but differs from them by having a smaller staminode, petals flushed with purplish red, a purplish red lip with purplish brown veins, pouched with erect and triangular auriculas on both sides of mouth, and leaves adaxially tessellated with pale green and pale yellowish green (Table 1, Fig. 1). PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 136 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China 99,5/100 | Paphiopedilum dayvanum | * Paphiopedilum sugiyamanum 783/90 100/100 | — Paphiopedilum mastersianum 100/100 — Paphiopedilum parnatanum Paphiopedilum hennisianum 94.6/87 | ost '— Paphiopedilum violascens S900; Paphiopedilum purpuratum |— Paphiopedilum appletonianum 94.999 F388 "—— Paphiopedilum barbatum Paphiopedilum bullenianum Lf 100/100 Paphiopedilum callosum 100/100 _ -—— Paphiopedilum motuoense 99.4/100 Too00 Paphiopedilum qingyongii — Paphiopedilum venustum reat soon00 -— Paphiopedilum sukhakulii 92.6/100 '— Paphiopedilum wardii Paphiopedilum rungsurtvanum Paphiopedilum insigne 91.4/90 Paphiopedilum tigri aT aphiopedilum tigrinum 953/66 | Paphiopedilum charlesworthii O57 Paphiopedilum druryi 100/100 Paphiopedilum rhizomatosum r} 96.4/97 100/100 ~ Paphiopedilum henryanum Paphiopedilum vejvarutianum (PYphiopedilun barbigerum 715/19 Paphiopedilum notatisepalum 0/59 Paphiopedilum helenae 95.196 Paphiopedilum tranlienianum 71.7/65 993/100 Paphiopedilum gratrixianum 100/100 Paphiopedilum spicerianum 100/100 ne Ee Paphiopedilum guangdongense 1; ee Paphiopedilum villosum Paphiopedilum hirsutissimum Paphiopedilum fairrieanum (— Paphiopedilum kolopakingii 90.1/99 — Paphiopedilum platyphyllum doa shir platyphy Paphiopedilum gigantifolium 100/100 1OH100 Paphiopedilum philippinense 100/100 ‘i ag Cpitapedt um rothschildianum 99.7; 100/100 Paphi , , aphiopedilum sanderianum 100/100 Paphiopedilum parishii 100/100 ha cise Paphiopedilum dianthum Paphiopedilum lowii 84.8/77 Paphiopedilum glaucophyllum 99.8/100 100/100 Paphiopedilum prinulinum 100/100. is R — Paphiopedilum lienianum 91.2/95 |4e1/80 Paphiopedilum moquetteanum 100/93 — Paphiopedilum victoria-mariae Paphiopedilum canhii -—— Paphiopedilum bellatulum 100/100 Paphiopedilum wenshanense 59.4/84 | Paphiopedilum godefroyae 100/100 Paphiopedilum leucochilum 100/100 1 /} Paphiopedilum concolor 100/100 Paphiopedilum niveum -—— Paphiopedilum emersonii 99.8/100 1850. Paphiopedilum hangianum 100/100 _|___—____—._Paphiopedilum malipoense ry 87.4/94 "—— Paphiopedilum micranthum 100/100 Paphiopedilum vietnamense 100/100 Paphiopedilum delenatii Paphiopedilum armeniacum Phragmipedium longifolium 0.004 Figure 3. Phylogram of Paphiopedilum spp., based on 67 shared protein-coding genes from 62 Paphiopedilum whole plastomes. The numbers near the nodes are the values of SH-aLRT test (left) and the ultrafast bootstrap (right). PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 137 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China Table 2. Genes presented in the chloroplast genome of P motuoense. Group of genes Photosystem | Photosystem II Cytochrome b/f complex ATP synthase NADH dehydrogenase Rubis CO large subunit gene RNA polymerase Small ribosomal proteins Large ribosomal proteins rRNA tRNA Gene psaA, psaB, psaC*, psal, psaJ psbA, psbB, psbC, psbD, psbE, psbF, psbH, psbl, psbJ, psbK, psbL, psbM, psbN, psbT, psbZ petA, petB, petD, petG, petL, petN atpA, atpB, atpE, atpF, atpH, atpl rbcL rpOA, rpoB, rpoC7, rpoC2 rps2, rps3, rps4, rps7*, rps8, rps11, rps12, rps14, rps15*, rps16, rps718, rps19* rpl2*, rp!14, rp!16, rpl20, rpl22, rpl23*, rp!32, rp!33, rpl36 mn4.5*, rrnd*, rrn16*, rrn23* trnA-UGC%, trnC-GCA, trnD-GUC, trnE-UUC, trnF-GAA, trnfM-CAU, trnG-GCC, trnG-UCC, trnH-GUG*, trnI-CAU*, trnI-GAU*, trnL-CAA*, trnL-UAA, trnL-UAG, trnM-CAU, trnN-GUU%, trnP-UGG, trnQ-UUG, trnR- ACG*, trnR-UCU, trnS-GCU, trnS-GGA, trnS-UGA, trnT-GGU, trnT-UGU, trnV-GAC%, trnV-UAC, trnY-GUA Translational initiation factor intA Maturase matk C-type cytochrome synthesis ccsA* Subunits of Acetyl-CoA-carboxylase accD Protease clpP Conserved open reading frames ycf1*, ycf2*, ycf3, ycf4 Note: *means duplicated gene in IRs. Description. Plants terrestrial. Leaves 5-6, basal, oblong, 5-8 x 1.6-1.9 cm, apex acute and slightly 2-lobed, adaxially tessellated with pale green and pale yellowish green, abaxially densely spotted with purple, base contracted into pet- iole shape. Scape erect, up to 5.3-7.1 cm long, purple, densely shortly hirsute, 1-flowered, terminal; bract broadly ovate, 2.1-2.3 x 1.6-1.8 cm, margin ciliate; pedicel and ovary 2.6-3.1 cm long, pubescent. Flower 5.7-6.2 cm in diam.; dorsal sepal and synsepal white with green veins; petals pale yellowish white, flushed with purplish red, with a few large maroon warts mainly in basal half; lio purplish red with purplish brown veins, margin pale yellowish white; stami- node pale yellow with green veins and flushed with pale purplish red. Dorsal sepal obcordate, 3.2—3.5 x 2.6-2.7 cm, apex acuminate, abaxially pubescent, margin ciliate; synsepal ovate, 3.2—3.4 x 1.8-2 cm, apex acute, abaxially pubes- cent, margin ciliate. Petals obovate-oblong, 4.4—4.7 x 1.6-1.8 cm, apex obtuse or subacute, margins long-ciliate; lip helmet-shaped, 2.4-2.7 cm long; lateral lobes without verruca or slightly verrucose; pouch 1.4-1.6 x 1—1.1 cm, slightly narrowed toward base, with triangular auriculas on both sides of its mouth, margins erect at mouth apex, villose at inner bottom, minutely papillate-puberu- lent outside. Staminode reniform-obcordate, 2.9-3.2 x 5-5.3 mm, with a sinus at apex, sinus with mucro at middle, adaxially finely papillate. Etymology. The species epithet “motuoense’” refers to the locality name of type specimen of this new species, Motuo County, Linzhi City, Xizang, China. Distribution and habitat. Paphiopedilum motuoense is currently known only from the type locality Motuo County, Xizang, China. It is terrestrial in humus-rich places and semi-epiphytic in the humus of rocky places in forest margins at elevations of 800 m. Phenology. Flowering in March and April. PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 138 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China Figure 4. Images of living plants of Paphiopedilum motuoense A whole plant B, C flower (side view and back view) D, E dorsal sepal (front view and back view) F, G synsepal (front view and back view) H, | petal (front view and back view) J-L lip (front view, side view and back view) M lip (vertical section) N ovary and column (with bract, synsepal and staminode) O-Q column (back view and side view) R, S staminode (front view and back view). PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China Figure 5. Paphiopedilum motuoense A whole plant B, C flower (side view and back view) D, E dorsal sepal (front view and back view) F, G synsepal (front view and back view) H, I petal (front view and back view) J-L lip (front view, side view and back view) M lip (vertical section) N ovary and column (with bract, synsepal and staminode) O-Q column (back view and side view) R, S staminode (front view and back view). PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 140 Feng-Xia Tang et al.: Paphiopedilum motuoense, a newly discovered orchid from China Acknowledgements We appreciate the valuable comments from the reviewers and the editors. We would like to thank Prof. Fuwu Xing and Dr. Cheng Zhang for their suggestions that improved the 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 financially supported by the Science, Technology and Innovation Commission of Shenzhen Municipality (Grant No. JCYJ20210324123013037), the National Natural Science Foundation of China (Grant No. 32001245) and the Science, Technology and Innovation Commission of Shenzhen Municipality (Grant No. KCX- FZ20211020164200001). Author contributions Data curation: MNW, XYW. Funding acquisition: MNW. Investigation: MNW, ZZ, FXT, WHR. Methodology: MNW, XYW. Project administration: MNW, JL. Software: XYW. Su- pervision: MNW. Visualization: MNW, JSC, XJD. Writing-original draft: MNW, FXT, WHR, MW. Writing-review and editing: MNW, FXT, WHR, MW, JL. Author ORCIDs Feng-Xia Tang © https://orcid.org/0000-0003-2575-9665 Wen-Hui Rao © https://orcid.org/0000-0002-2177-6700 Ze Zhang © https://orcid.org/0000-0002-8774-4867 Xin-Yi Wu © https://orcid.org/0000-0002-8623-1867 Meng Wang ® hittps://orcid.org/0000-0002-1665-3642 Jian Li © https://orcid.org/0000-0002-0096-6257 Jie-Shan Chen ® https://orcid.org/0009-0000-8859-6053 Xiao-Juan Duan © https://orcid.org/0009-0000-0429-8594 Mei-Na Wang ® https://orcid.org/0000-0001-5998-6028 Data availability All of the data that support the findings of this study are available in the main text. References Atwood JT (1984) The relationships of the slipper orchids (subfamily Cypripedioideae, Orchidaceae). Selbyana 7: 129-147. Averyanov L (2010) The orchids of Vietnam illustrated survey part 2 subfamily Orchidoideae. Turczaninowia 13(2): 5-98. Brieger FG (1971) Unterfamilie: Cypripedioideae. In: Schlechter R (Ed.) Die Orchideen. Paul Parey, Berlin, 161-198. 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Link: https://doi.org/10.3897/phytokeys.259.145861.suppl1 PhytoKeys 259: 131-144 (2025), DOI: 10.3897/phytokeys.259.145861 144