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28 February 2024 Phylogenetic and biogeographical analyses of Thismia (Thismiaceae) support T. malipoensis as the eighth species in China
Ji-Dong Ya, Hai-Yao Chen, Wei Zhang, Ren-Bin Zhu, Jie Cai, Wen-Bin Yu
Author Affiliations +
Abstract

Thismia Griff. (Thismiaceae) is a holo-mycoheterotrophic genus with more than 100 species. In this study, phylogenetic analyses supported that T. malipoensis from Yunnan is a new species in T. sect. Glaziocharis. Morphologically, this new species can be distinguished from its phylogenetic sister species T. abei by having the annulus of the flower expanded and modified into a cucullate (hood-like) structure with zygomorphic symmetry with one opening on one side. Biogeographical inference showed that SW China to Indo-Burma and the Sunda Shelf region was suggested as the ancestral distribution region of Thismia s.s., then eastward to SE China and Japan, and southward to New Guinea to Australia, respectively. The Chinese species should have originated from at least two different ancestral sources, and geographical isolation caused the divergence between T. malipoensis and T. abei at 17.47 Mya.

Citation: Ya J.-D. Chen H.-Y. Zhang W. Zhu R.-B. Cai J. & Yu W.-B. 2024: Phylogenetic and biogeographical analyses of Thismia (Thismiaceae) support T. malipoensis as the eighth species in China. – Willdenowia 54: 47–63.

Version of record first published online on 28 February 2024 ahead of inclusion in April 2024 issue.

Introduction

Heterotrophic plants exhibit distinct mechanisms for nutrient acquisition when compared to autotrophic plants. To date, more than 880 mycoheterotrophic plants that rely upon associations with fungi for their supply of organic nutrients have been documented (Merckx & al. 2013b). Due to their highly reduced leaves and lack of chlorophyll (Cameron & Leake 2007; Leake & Cameron 2010), holo-mycoheterotrophic are only visible during brief periods of flowering and fruiting. As large-scale expeditions are conducted across different seasons and remote areas, numerous leafless holo-mycoheterotrophic species have been discovered and classified as new to the field of science. Currently, most of these species are only found in small populations within one or a few locations, making them susceptible to being categorized as Critically Endangered (CR). Therefore, efforts to conserve these species are crucial (Merckx & al. 2013a).

Thismia Griff. is a holo-mycoheterotrophic genus with remarkable flowers in its peculiar appearance and exceptionally complicated and quite diverse floral morphology (Kumar & al. 2017; Shepeleva & al. 2020; Nuraliev & al. 2021). Traditionally, Thismia was treated as a member of the tribe Thismieae in the family Burmanniaceae (Maas-van de Kamer 1998; Merckx 2008; Li & al. 2020), with four other holo-mycoheterotrophic genera, i.e. Afrothismia Schltr., Haplothismia Airy Shaw, Oxygyne Schltr. and Tiputinia P. E. Berry & C. L. Woodw. Recent phylogenetic studies have supported the tribe Thismieae as a separate family Thismiaceae, because Thismiaceae is sister to the family Taccaceae and rather distantly related to the family Burmanniaceae s.s. (Merckx & al. 2006; Lam & al. 2016; Shepeleva & al. 2020). The most recent phylogenetic study showed that Thismia species are polyphyletic in that the Old World species formed five well-supported groups, and the neotropical species are clustered with Tiputinia, which probably represents a separate genus (Shepeleva & al. 2020), i.e. Ophiomeris Miers. In this study, we tentatively followed the traditional delimitation of Thismia s.l., which is characterized by having a prominent hypanthium (also called a flower tube or flower chamber) that bears six tepals and six stamens. To date, Thismia includes about 105 species and one variety (IPNI 2023; POWO 2023) ( Supplementary appendix S1 (wi.54.54102_Supplementary_appendix_S1.xlsx)), which are distributed mainly in tropical and subtropical Asia to temperate Australia and tropical America (Shepeleva & al. 2020).

Thismia taiwanensis Sheng Z. Yang & al. was the first species to be described from China until 2002 (Yang & al. 2002). To date, seven species have been recognized in China (Yang & al. 2002; Ho & al. 2009; Chiang & Hsieh 2011; Li & Bi 2013; Mar & Saunders 2015; Xu & al. 2020; Li & al. 2023), and T. guangdongensis X. J. Li & al. was most recently described in 2023. Of these, Guangdong, Hong Kong and Taiwan have recorded two species, and Hainan and Yunnan have one species each, with T. hongkongensis Mar & R. M. K. Saunders found in Guangdong and Hong Kong. During botanical surveys in SE Yunnan in 2019, a unique flowering species of Thismia was found and collected (Fig. 14). After investigations on morphological characters and herbarium specimens from China and adjacent regions, as well as a literature review, we confirmed that this species was new to science, which was also supported by phylogenetic analyses using two nuclear ribosomal DNA (nrDNA) and two mitochondrial DNA (mtDNA) sequence datasets. Herein, this new species is named as T. malipoensis J. D. Ya & W. B. Yu by providing a morphological description, type specimen, colour photographs (Fig. 13) and line drawings (Fig. 4). Moreover, the phylogenetic and biogeographic significance of this new species is inferred and discussed, and conservation suggestions and recommendations are proposed.

Material and methods

Morphological observation

Voucher specimens of Thismia malipoensis were collected from the Laoshan Provincial Nature Reserve of Malipo County during a field expedition in 2019. The morphological characters of this new species were observed, measured and photographed based on living individuals. Fresh flowering specimens were identified by checking herbarium specimens and reviewing the relevant literature (Yang & al. 2002; Larsen & Averyanov 2007; Ho & al. 2009; Chiang & Hsieh 2011; Li & Bi 2013; Mar & Saunders 2015; Kumar & al. 2017; Dančák & al. 2020a; Nuraliev & al. 2020; Xu & al. 2020; Siti-Munirah & al. 2021; Li & al. 2023). DNA tissues of T. malipoensis were collected from flowering shoots, then dried using silica-gel. The type specimen was deposited at the Herbarium of the Kunming Institute of Botany, Chinese Academy of Sciences (KUN; herbarium code following Thiers 2023+).

Phylogenetic analyses

In order to elucidate the phylogenetic association between Thismia malipoensis and other species within the Thismiaceae, phylogenetic analyses were conducted using sequences of nrDNA 18S and ITS, as well as mtDNA atpA and matR. A comprehensive analysis was performed on a total of 48 taxa, encompassing four genera of Thismiaceae, with Tacca palmatifida Baker (Taccaceae) as the outgroup. Appendix 1 provides voucher information for the GenBank accession numbers associated with the sequences generated or obtained from the studies of Dančák & al. (2020b, 2020c), Merckx & al. (2017), Shepeleva & al. (2020) and Sochor & al. (2018) for this study.

Genomic DNA of the new species was isolated from silica-gel dried shoot tissues of the type collection using the CTAB method (Doyle & Doyle 1987). Library preparation for the Next Generation Sequencing followed the procedure of Zeng & al. (2018). Around 500 pg purified genomic DNA was fragmented to approximately 350 bp in size by sonication, then built into a blunt-end DNA library using the NEBNext Ultra II DNA library prep kit for Illumina (New England BioLabs) following the standard protocol. The 150 bp pair-end reads were generated using the Illumina Sequencing Platforms. Raw data were assembled de novo for the mitochondrial and nuclear ribosome DNA contigs/sequences using the GetOrganelle toolkit (Jin & al. 2020). Assembled mitochondrial and nuclear ribosome DNA sequences of the new species were annotated in Geneious (Kearse & al. 2012) using nrDNA 18S and nrITS, and mtDNA atpA and matR sequences of Thismia species from NCBI as references, then four targeted DNA regions were extracted from the annotated sequences, respectively.

Four DNA regions were aligned separately using MAFFT 7.505 (Katoh & Standley 2013). Then the four regions were concatenated into a supermatrix ( Supplementary appendix S2 (wi.54.54102_Supplementary_appendix_S2.nex)) using SequenceMatrix package of TaxonDNA 1.78 (Meier & al. 2006). Bayesian inference (BI) and maximum likelihood (ML) methods were used to reconstruct phylogenies. The BI analysis was performed using MrBayes 3.26 (Ronquist & al. 2012). The total dataset was partitioned, and the DNA substitution model of the Bayesian information criterion (BIC) for two DNA regions was estimated using jModeltest 2 (Darriba & al. 2012; Darriba & al. 2019). Markov chain Monte Carlo (MCMC) analysis was performed using MrBayes for 10,000,000 generations for the total dataset, with two simultaneous runs, each run comprising four incrementally heated chains. The BI analysis was started with a random tree and sampled every 1000 generations. The ML analysis was conducted with RAxML 8.2.10 (Stamatakis 2014) using the GTR substitution model with gamma (Γ) distribution rate heterogeneity among sites and the proportion of invariable sites estimated from the data. Support values for nodes/clades were estimated from 1000 bootstrap replicates.

Fig. 1.

Thismia malipoensis – A: plant; B, C: plants in habitat. – Source of material: J.-D. Ya & W. Zhang 19CS18569. – Photographed by J.-D. Ya.

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Divergence time estimation and ancestral area reconstruction

The concatenated dataset of 18S rDNA, ITS, atpA and matR from 49 taxa was used to estimate the divergence time among Thismia species. According to the previous studies (Merckx & Smets 2014), we chose the secondary calibration time for two nodes: (1) 63–96 Ma for the root node between Thismiaceae s.s. and Tacca J. R. Forst. & G. Forst.; and (2) 54–86 Ma for the crown node of Thismiaceae age (Merckx 2008). The divergence time was estimated using the MCMCTree program in PAML 4.10.5 (Yang 2007). Using the independent rate model (clock = 2) and the GTR + Γ model (model = 7). We ran two separate MCMC iterations for 400,000 generations with sampling every 10 iterations, the first 160,000 iterations were discarded as burn-in, and the mcmcfile was used to check ESS values and convergence in Tracer 1.7.2 (Rambaut & al. 2018).

Distribution information of 44 taxa of Thismiaceae and the outgroup Tacca palmatifida was collected from POWO (2023), and that of four accessions without name just using source information of the voucher (Shepeleva & al. 2020). Ten biogeographical areas of distribution range were classified according to previous biogeographical studies (Thomas & al. 2012; Sirichamorn & al. 2014; Liu & al. 2021): (A) Japan; (B) SE China; (C) SW China to Indo-Burma (excluding the Malay Peninsula); (D) India and Sri Lanka; (E) the Sunda Shelf region; (F) Philippines; (G) Wallacea; (H) New Guinea to Australia; (I) South America; and (J) Africa (Fig. 6). The dispersal-extinction cladogenesis (DEC) model (Ree & Smith 2008), dispersal-vicariance analyses (DIVA) model (Ronquist 1997), and BAYAREALIKE model (Landis & al. 2013), as well as the other three model with “+J” parameter (founder event speciation) in the R package BioGeoBEARS (Matzke 2018) that was used to reconstruct ancestral areas of Thismia. The maximum number of regions was set to 2 and the lowest corrected Akaike information criterion (AICc) was considered the “best” model.

Results and Discussion

Phylogenetic analyses

The nrDNA 18S matrix was 1639 bp in length including 384 variable sites and 247 parsimony-informative sites. The nrDNA ITS matrix was 1012 bp in length including 721 variable sites and 613 parsimony-informative sites. The mtDNA atpA matrix was 1112 bp in length including 146 variable sites and 79 parsimony-informative sites. The mtDNA matR matrix was 1065 bp in length including 419 variable sites and 190 parsimony-informative sites. The best-fit BIC model of 18S, mrITS, atpA and matR datasets was GTR+G11, TPM1uf+G, TPM1+G11, and GTR+I+G, respectively. The major-rule consensus tree of ML analysis with support values of both ML and BI analyses is shown in Fig. 5, which is consistent with the topology in the study by Shepeleva & al. (2020). Five well-supported clades were recovered in Thismia s.s., and each clade can correspond to four revised sections and one subsection in the subgenus division of Thismia proposed by Kumar & al. (2017). In addition, six species were not placed into any clades of the five recognized clades, with two Chinese species, T. hongkongensis and T. tentaculata K. Larsen & Aver., which are well supported as a monophyletic clade (MLBS/BIPP=98/1.00).

According to the study by Shepeleva & al. (2020), the modified Thismia sect. Glaziocharis (Taub. ex Warm.) Hatus. is a monophyletic group corresponding to Clade 1, which consists of five species from E Asia (China, Japan) and mainland SE Asia (Thailand, Laos). Herein, our phylogenetic analyses supported T. malipoensis as a member of T. sect. Glaziocharis, and allied to T. abei (Akasawa) Hatus., which are both putatively as the sister to T. gongshanensis Hong Qing Li & Y. K. Bi with weak support by ML analysis (MLBS = 53). Of the six sampled Chinese species, four species were fell in this clade, with the exception of T. hongkongensis and T. tentaculata.

Morphological comparisons and species delimitation

Morphologically, Thismia malipoensis stands out significantly from the other six species in T. sect. Glaziocharis of the Clade 1 (Fig. 5), as well as other four recognized Chinese species by having a cucullate (hood-like) and zygomorphic annulus structure of the flower that covering the apical floral tube with an opening on one side. As the phylogenetic sister species of T. malipoensis, T. abei is characterized by having connected inner tepals as a loose dome, a ring-like and actinomorphic annulus structure, short tepal appendages (5–6 mm) and glabrous stigma. To date, the cucullate annulus of has been reported only in T. belumensis Siti-Munirah & Suhaimi-Miloko and T. labiata J. J. Sm., which were supposed to be members of T. sect. Glaziocharis (Siti-Munirah & al. 2021). Thismia malipoensis is distinguished from T. belumensis in characters of the tepal appendages: both outer and inner tepal appendages are spreading; inner tepal appendages are shorter than 2 cm and equal to the outer ones; and inner tepal appendages have a fimbriate base and a subulate tip. Geographical information is also important for species delimitation between T. malipoensis and T. belumensis. Thismia malipoensis is distributed in Malipo, SE Yunnan, China, and grows under evergreen broad-leaved forests in limestone mountains at an elevation of 1100 m, whereas T. belumensis is endemic to Perak, Peninsular Malaysia, under the shade of lowland dipterocarp forests at an elevation of 260–290 m (Siti-Munirah & al. 2021). Due to the lack of molecular data for T. belumensis, it is not clear whether T. malipoensis and T. belumensis are sister species or are sharing similar floral forms and structures by convergent evolution.

Fig. 2.

Thismia malipoensis, different views of flower – A, D: bottom view; B: lateral view; C: top view; E, F: front view. – Source of material: J.-D. Ya & W. Zhang 19CS18569. – Photographed by J.-D. Ya.

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Divergence time estimation and ancestral area reconstruction

The crown age of Thismiaceae was estimated to be 75.84 Mya (95 % HDP: 42.85–94.53 Mya), and the diversification time of Thismia s.s. started at 62.06 Mya (95 % HDP: 34.75–79.37 Mya) in tropical and subtropical Asia (Fig. 6). For the five major clades or sections, the diversification times varied from 14.26 Mya (95 % HDP: 6.15–26.92 Mya) in the Clade 2 to 38.91 Mya (95 % HDP: 20.16–58.05 Mya) in the Clade 1. Rapid speciation started at c. 23.80 Mya from the late Oligocene to the present. In the Clade 1, T. thaithongiana Chantanaorr. & Suddee diverged from the remaining six species at 38.91 Mya (95 % HDP: 20.16–58.05 Mya). Then, the two subclades, T. malipoensisT. gongshanensis and T. nigricoronataT. taiwanensis, separated at 31.49 Mya (95 % HDP: 16.09–48.46 Mya), and T. malipoensis diverged from T. abei at 17.47 Mya (95 % HDP: 7.23–31.34 Mya).

Ancestral area reconstruction indicated that the ancestral distribution region of Thismiaceae was not clear, because the earliest divergent Oxygyne species are found in Africa and Japan of E Asia, and the following clade Haplothismia exannulataThismia panamensis is mainly distributed in Central to South America, as well as the only species Haplothismia exannulata Airy Shaw is in India of S Asia (Fig. 6). Thismia s.s. is restricted in E Asia to S and SE Asia. The Sunda Shelf and SW China to Indo-Burma regions were suggested as the ancestral distribution region of Thismia s.s., and its common ancestor might be originated from the Sunda Shelf region at 64.95 Mya. The back-and-forth migration and dispersal between SW China to Indo-Burma (excluding the Malay Peninsula) and the Sunda Shelf region have happened at least five to six times in Clades 1, 4 and 5, which are highly associated with land-bridge connection between the SE Asia mainland and the Sunda Shelf region before 5.0 Mya (Hall 2009, 2012, 2013; Tan & al. 2020). The common ancestor of T. sect. Glaziocharis and T. sect. Sarcosiphon (Blume) Jonker should have originated in the SW China to Indo-Burma region at 38.91 Mya and 16.87 Mya, respectively, then spread eastward to SE China at 22.45 Mya and Japan at 17.47 Mya in the section Glaziocharis (Clade 1) and in situ diversification in T. sect. Sarcosiphon (Clade 4) started at 16.87 Mya. Thismia sect. Rodwaya (Schltr.) Jonker from New Guinea to Australia might have originated from the Sunda Shelf region at 43.50 Mya, then in situ diversification started at 14.26 Mya as New Guinea to Australia isolated from the Sunda Shelf region (Hall 2012; Zhang & al. 2023). The common ancestor of the clade T. gardneriana + T. hongkongensis + T. tentaculata should be from the Sunda Shelf region, but the ancestral region of T. gardneriana + T. hongkongensis + T. tentaculata was ambiguous, which might be caused by incomplete sampling and an unresolved relationship between T. gardneriana and T. hongkongensis + T. tentaculata. In addition, long-distance dispersal cannot be excluded for T. gardneriana spreading to India.

Biodiversity and conservation of extremely rare Thismia species

Thismia species almost exclusively distributed in tropical and subtropical areas of Asia and America, with the exception that T. americana N. Pfeiff. in the C United States, which has been supposed to be extinct. Since the genus was established in 1845, species discovery progressed very slowly with only 40 species discovered until 1999 ( Supplementary appendix S1 (wi.54.54102_Supplementary_appendix_S1.xlsx)). After 2011, at least one new species per year was described, and seven and 15 new species were described in 2017 and 2018, respectively. To date, there are 106 species and one variety recognized ( Supplementary appendix S1 (wi.54.54102_Supplementary_appendix_S1.xlsx)). However, most Thismia species are found or collected only once or a few times with small population sizes (Dančák & al. 2020a; Nuraliev & al. 2020). As holo-mycoheterotrophic herbs, Thismia species are highly reliant on the specialized fungal host, so the distribution range might be restricted by the availability of host fungi (Merckx & Bidartondo 2008; Gomes & al. 2017; Merckx & al. 2017). Meanwhile, Thismia plants are usually very small, inconspicuous and of ephemeral emergence in the flowering and fruiting seasons. To date, only a few Thismia species have been observed in more than three locations; c. 65 % of Thismia species are known only from the type localities and adjacent areas; c. 45 % of them were found only by their discoverers; and some species had just a single individual (Dančák & al. 2020a; Nuraliev & al. 2020). Therefore, most Thismia species could be locally endemic, rare and critically endangered (CR) in accordance with the IUCN Red List categories and criteria (IUCN Standards and Petitions Committee 2022).

It is concerning that in the current geographic distribution range of eight Thismia species in China, six species are known only from the type locality, and only two species, T. hongkongensis and T. tentaculata, have been recorded in two or more populations. In China, T. hongkongensis and T. tentaculata were originally found in Hong Kong, the range of T. hongkongensis extended to Guangdong and the type locality of T. tentaculata was Quang Tri Province, Vietnam, but the total number of individuals was few. Moreover, T. jianfenglingensis Han Shu & al. is endemic to the Jianfengling National Nature Reserve, Ledong County, Hainan Island, and had only six flowering individuals when found for the first time in 2017, whereas no flowering individuals were found in 2018. Herein, T. malipoensis is known only from Malipo County, SW Yunnan, where only ten flowering individuals were found in 2019. New populations of some species will be found in further investigations. However, most Thismia species have extremely small population sizes, which may be more sensitive to environmental changes and more vulnerable to extinction. As mentioned above, holo-mycoheterotrophic Thismia species are highly reliant on the specialized fungal host for seed germination and seeding development, and those fungi should be associated with the forest and soil ecosystem. Therefore, the most effective strategies are in situ conservation.

Fig. 3.

Thismia malipoensis – A: flower, longitudinal section; B: tepals, sparsely papillate outer surface; C–E: floral tube, lateral view (C), top view (D), front view (E); F: stamens, bottom view; G: stamens, lateral view; H: stigma, lateral view; I: stigma, top view; J: ovary, cross-section; K: ovary, longitudinal section. – Source of material: J.-D. Ya & W. Zhang 19CS18569. – Photographed by J.-D. Ya. – All scale bars = 5 mm.

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Fig. 4.

Thismia malipoensis – A: plant; B: flower, top view; C: flower, longitudinal section; D: stamens, lateral view; E: stamens, bottom view; F: stigma, lateral view; G: ovary, longitudinal section. – Source of material: J.-D. Ya & W. Zhang 19CS18569. – Drawn by Z.-D. Han.

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Taxonomic treatment

Thismia malipoensis J. D. Ya & W. B. Yu, sp. nov.Fig. 14.

Holotype: China, Yunnan Province, Wenshan Prefecture, Malipo County, Tianbao Town, 1191 m a.s.l., forest on limestone, 13 Aug 2019, J.-D. Ya & W. Zhang 19CS18569 (KUN 1584483!).

DiagnosisThismia malipoensis belongs to T. sect. Glaziocharis and can be distinguished from its phylogenetic sister species T. abei by having the annulus of the flower expanded and modified into a cucullate (hood-like) structure with zygomorphic symmetry with one opening on one side. Moreover, T. malipoensis can be distinguished from the morphologically similar T. belumensis, in that the outer and inner tepal appendages are spreading (vs erect) and equal in length by less than 2 cm (vs outer tepal appendage 1.5–2.5 cm and inner tepal appendage c. 3 cm), the inner tepal base is fimbriate at the margin (vs entire) and the inner tepal tip is subulate (vs clavate).

DescriptionHerbs terrestrial, achlorophyllous, c. 7 cm tall. Roots vermiform, creeping, branched, fleshy, terete, pale brownish. Stems erect, unbranched, 2–4 cm long, densely deflexed papillate hairy. Leaves scale-like, white, translucent, lanceolate, c. 8 × 2 mm, base appressed to stem, apex acuminate. Involucral bracts 3, pale brownish, lanceolate, c. 1 cm long, densely deflexed papillate hairy, base appressed, margin entire, apex acuminate. Pedicel c. 3 mm long (post anthesis). Flowers terminal, solitary, zygomorphic, c. 4.5 × 4 cm (including appendages); hypanthium pyriform, dark green, translucent, c. 8 mm long, c. 7 mm wide near apex, narrowed to c. 4 mm wide just above ovary, outer surface densely deflexed papillate hairy, with 12 raised dark brown longitudinal veins, inner surface smooth; outer tepals 3, pale brown, broadly ovate, c. 2.5 × 3 mm, 2 on opposite side of annulus opening and 1 below annulus opening, outer surface sparsely papillate, apex acute, each outer tepal with a tentacle-like spreading appendage, pale brown, 0.8–1.3 cm × c. 1 mm; inner tepals 3, pale brown, translucent, glabrous, broadly obovate, c. 3.5 × 4 mm, apically broadly fused and tightly adpressed and completely overlapping cucullate part of annulus, each inner tepal with a tentacle-like spreading appendage c. 1.7 cm long, pale brown, fimbriate at basal margin, subulate at apex. Annulus expanded and modified into a cucullate (hood-like) structure, covering apical part of floral tube and forming a lateral floral aperture; cucullate outer surface white with 3 brown lines, glabrous; cucullate inner surface light green, covered with numerous white trichomes. Stamens 6, light yellow, pendent from apical part of floral tube; each connective c. 3.5 mm long, outer surface with 2 linear to filiform thecae, each c. 2 mm long, inner surface smooth, outer surface and apex covered with transparent trichomes; supraconnective apex rounded and curved outward; filaments short, connected to floral tube and annulus. Ovary obovoid, c. 3.5 × 4.8 mm, 1-loculed, with 3 parietal placentae; style c. 0.6 mm long; stigma 3-lobed, lobes triangular-pyramidal, c. 1.2 mm long, apex truncate and densely papillate. Fruit cup-shaped; seeds brown.

Phenology — Observed flowering in August, fruiting in September.

Distribution and habitatThismia malipoensis is currently known only from the type locality in Laoshan Provincial Nature Reserve of Malipo County, SE Yunnan. The population consists of about ten individuals under evergreen broad-leaved forest on limestone mountains at an elevation of 1191 m a.s.l. The forest includes the Lithocarpus tephrocarpus (Drake) A. Camus (Fagaceae), Alseodaphnopsis andersonii (King ex Hook. f.) H. W. Li & J. Li and Litsea yunnanensis Y. C. Yang & P. H. Huang (both Lauraceae) and the bamboo Melocalamus arrectus T. P. Yi.

Etymology — The specific epithet “malipoensis” refers to the county name Malipo of Wenshan Prefecture, the type locality of the new species.

Chinese name — 麻栗坡水玉杯 (má lì pō shuǐ yù bēi).

Conservation status — We evaluated the conservation status of Thismia malipoensis according to the IUCN Red List categories and criteria (IUCN Standards and Petitions Committee 2022). The new species is known only from the type locality, our field surveys identified only one population with about ten individuals. The conservation status of the species is therefore considered to be Critically Endangered (CR A2acd; B1ab(i,iii,v)+2ab(i,iii,v); C2a(i); D).

Key to species of Thismia in China

1. Corolla tube apex covered by a cucullate and zygomorphic annulus, with an opening on one side T. malipoensis

– Corolla tube apex with a ring-like and actinomorphic annulus 2

2. Tepals free and mitre absent 3

– Tepals connected into a loose dome or mitre 4

3. Inner tepal appendage c. 17 mm long, longer than perianth tube T. tentaculata

– Inner tepal appendage 1–4 mm long, shorter than perianth tube T. guangdongensis

4. Inner tepals fused into a mitre; stigma 2-lobed T. gongshanensis

– Inner tepals forming a loose dome; stigma 3-lobed 5

5. Inner tepal appendage c. 30 mm long T. taiwanensis

– Inner tepal appendage < 5 mm long 6

6. Inner tepal appendage c. 1 mm long; stamen connective apex with glandular hairs T. huangii

– Inner tepal appendage 3–4 mm long; stamen connective apex without glandular hairs 7

7. Perianth tube pinkish white; outer lobes separated from dome T. hongkongensis

– Perianth tube deep orange-red; outer lobes embedded in dome T. jianfenglingensis

Fig. 5.

Phylogeny of Thismiaceae inferred from the concatenated two nrDNA and two mitochondrial DNA sequences dataset. Maximum likelihood bootstrap (MLBS) and Bayesian inference posterior probability (BIPP) values are presented above the branches. The bottom scale bar represents the number of substitutions per site.

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Author contributions

Ji-Dong Ya, Wen-Bin Yu and Jie Cai conceived the study; Ji-Dong Ya and Wei Zhang contributed to field surveys; Ji-Dong Ya and Wen-Bin Yu provided the species description and collected the data; Wen-Bin Yu and Hai-Yao Chen analysed the data and interpreted the results; Ji-Dong Ya, Hai-Yao Chen, Wen-Bin Yu and Jie Cai wrote and revised the manuscript; Ren-Bin Zhu and Wen-Bin Yu checked and generated the checklist of Thismia; all authors approved the final version of the manuscript.

Fig. 6.

Divergent time estimation and ancestral area reconstruction of Thismiaceae. The map shows the ten biogeographical areas. The lineages through time plot shows the diversification of Thismiaceae.

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Acknowledgements

We are grateful to Zhou-Dong Han for his outstanding illustration; to Ming-Feng Long for his kind assistance in the field; to the physical support of the National Wild Plant Germplasm Resource Center of Kunming Institute of Botany, Chinese Academy of Sciences; and to the HPC Platform of the Information Center, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences. This study was financially supported by the Science and Technology Basic Resources Investigation Program of China (2021FY100200), the Large-scale Scientific Facilities of the Chinese Academy of Sciences (2017-LSFGBOWS-02), the Key Basic Research Program of Yunnan Province, China (202101BC070003), the Yunnan Revitalization Talent Support Program “Young Talent” and “Innovation Team” Projects, the 14th Five-Year Plan of Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences (XTBG-1450303), and Ecological and Environmental Conservation Program from the Department of Ecology and Environment of Yunnan Province. We also thank Martin Dančák (Palacký University, Olomouc, Czech Republic) and an anonymous reviewer for their comments on an earlier version of this paper.

© 2024 The Authors ·

This open-access article is distributed under the  CC BY 4.0 licence

References

1.

Cameron D. D. & Leake J. R. 2007: A different kind of parasitic plant: a brief history of mycoheterotrophy and parasitism. – Haustorium 50: 4–6. Google Scholar

2.

Chiang P. Y. & Hsieh T. H. 2011: Thismia huangii (Thismiaceae), a new species from Taiwan. – Taiwania 56: 138–142. Google Scholar

3.

Dančák M., Hroneš M. & Sochor M. 2020a: Thismia: the rarest of the rare? Ranges of some Bornean species are much larger than previously believed. – Phytotaxa 455: 245–261.  https://doi.org/10.11646/phytotaxa.455.4.2 Google Scholar

4.

Dančák M., Hroneš M. & Sochor M. 2020b: Thismia minutissima (Thismiaceae), a remarkable new mycoheterotrophic species from Sarawak, Borneo. – Kew Bull. 75(29).  https://doi.org/10.1007/S12225-020-09886-4 Google Scholar

5.

Dančák M., Hroneš M. & Sochor M. 2020c: Thismia ornata and T. coronata (Thismiaceae), two new species from Sarawak, Borneo. – Willdenowia 50: 65–76.  https://doi.org/10.3372/wi.50.50106 Google Scholar

6.

Darriba D., Posada D., Kozlov A. M., Stamatakis A., Morel B. & Flouri T. 2019: ModelTest-NG: a new and scalable tool for the selection of DNA and protein evolutionary models. – Molec. Biol. Evol. 37: 291–294.  https://doi.org/10.1093/molbev/msz189 Google Scholar

7.

Darriba D., Taboada G. L., Doallo R. & Posada D. 2012: jModelTest 2: more models, new heuristics and parallel computing. – Nature, Meth. 9: 772.  https://doi.org/10.1038/nmeth.2109 Google Scholar

8.

Doyle J. J. & Doyle J. L. 1987: A rapid DNA isolation procedure for small quantities of fresh leaf tissue. – Phytochemistry 19: 11–15. Google Scholar

9.

Gomes S. I. F., Aguirre-Gutiérrez J., Bidartondo M. I. & Merckx V. S. F. T. 2017: Arbuscular mycorrhizal interactions of mycoheterotrophic Thismia are more specialized than in autotrophic plants. – New Phytol. 213: 1418–1427.  https://doi.org/10.1111/nph.14249 Google Scholar

10.

Hall R. 2009: Southeast Asia's changing palaeogeography. – Blumea 54: 148–161.  https://doi.org/10.3767/000651909X475941 Google Scholar

11.

Hall R. 2012: Late Jurassic–Cenozoic reconstructions of the Indonesian region and the Indian Ocean. – Tectonophysics 570–571: 1–41.  https://doi.org/10.1016/j.tecto.2012.04.021 Google Scholar

12.

Hall R. 2013: The palaeogeography of Sundaland and Wallacea since the Late Jurassic. – J. Limnol. 72: 1–17.  https://doi.org/10.4081/jlimnol.2013.s2.e1 Google Scholar

13.

Ho G. W. C., Mar S. S. & Saunders R. M. K. 2009: Thismia tentaculata (Burmanniaceae tribe Thismieae) from Hong Kong: first record of the genus and tribe from continental China. – J. Syst. Evol. 47: 605–607.  https://doi.org/10.1111/j.1759-6831.2009.00037.x Google Scholar

14.

IPNI 2023: International Plant Names Index. – Published at https://www.ipni.org/[accessed 5 Jun 2023]. Google Scholar

15.

IUCN Standards and Petitions Committee 2022: Guidelines for using the IUCN Red List categories and criteria. Version 15.1 (July 2022). Prepared by the Standards and Petitions Committee of the IUCN Species Survival Commission. – Published at https://www.iucnredlist.org/documents/RedListGuidelines.pdf  Google Scholar

16.

Jin J.-J., Yu W.-B., Yang J.-B., Song Y., dePamphilis C. W., Yi T.-S. & Li D.-Z. 2020: GetOrganelle: a fast and versatile toolkit for accurate de novo assembly of organelle genomes. – Genome Biol. 21(241).  https://doi.org/10.1186/s13059-020-02154-5 Google Scholar

17.

Katoh K. & Standley D. M. 2013: MAFFT multiple sequence alignment software version 7: improvements in performance and usability. – Molec. Biol. Evol. 30: 772–780.  https://doi.org/10.1093/molbev/mst010 Google Scholar

18.

Kearse M., Moir R., Wilson A., Stones-Havas S., Cheung M., Sturrock S., Buxton S., Cooper A., Markowitz S., Duran C., Thierer T., Ashton B., Meintjes P. & Drummond A. 2012: Geneious basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. – Bioinformatics 28: 1647–1649.  https://doi.org/10.1093/bioinformatics/bts199 Google Scholar

19.

Kumar P., Gale S. W., Li J. H., Bouamanivong S. & Fischer G. A. 2017: Thismia nigricoronata, a new species of Burmanniaceae (Thismieae, Dioscoreales) from Vang Vieng, Vientiane Province, Laos, and a key to subgeneric classification. – Phytotaxa 319: 225–240.  https://doi.org/10.11646/phytotaxa.319.3.2 Google Scholar

20.

Lam V. K. Y., Merckx V. S. F. T. & Graham S. W. 2016: A few-gene plastid phylogenetic framework for mycoheterotrophic monocots. – Amer. J. Bot. 103: 692–708.  https://doi.org/10.3732/ajb.1500412 Google Scholar

21.

Landis M. J., Matzke N. J., Moore B. R. & Huelsenbeck J. P. 2013: Bayesian analysis of biogeography when the number of areas is large. – Syst. Biol. 62: 789–804.  https://doi.org/10.1093/sysbio/syt040 Google Scholar

22.

Larsen K. & Averyanov L. V. 2007: Thismia annamensis and Thismia tentaculata, two new species of Thismiaceae from central Vietnam. – Rheedea 17: 13–19. Google Scholar

23.

Leake J. R. & Cameron D. D. 2010: Physiological ecology of mycoheterotrophy. – New Phytol. 185: 601–605.  https://doi.org/10.1111/j.1469-8137.2009.03153.x Google Scholar

24.

Li D.-Z., Chen Z.-D., Wang H., Lu A.-M., Luo Y. & Yu W.-B. (ed.). 2020: The families and genera of Chinese vascular plants. – Beijing: Science Press. Google Scholar

25.

Li H.-Q. & Bi Y.-K. 2013: A new species of Thismia (Thismiaceae) from Yunnan, China. – Phytotaxa 105: 25–28.  https://doi.org/10.11646/phytotaxa.105.1.4 Google Scholar

26.

Li X.-J., Liu A. & Zhang D.-X. 2023: Thismia guangdongensis (Thismiaceae), a new mycoheterotrophic species from China. – Nordic J. Bot. 2023(e03819).  https://doi.org/10.1111/njb.03819 Google Scholar

27.

Liu J., Lindstrom A. J., Nagalingum N. S., Wiens J. J. & Gong X. 2021: Testing the causes of richness patterns in the paleotropics: time and diversification in cycads (Cycadaceae). – Ecography 11: 1606–1618.  https://doi.org/10.1111/ecog.05910 Google Scholar

28.

Maas-van de Kamer H. 1998: Burmanniaceae. – Pp. 154–164 in: Kubitzki K. (ed.), The families and genera of vascular plants III. Flowering plants: Monocotyledons: Lilianae (except Orchidaceae). – Berlin: Springer.  https://doi.org/10.1007/978-3-662-03533-7_21 Google Scholar

29.

Mar S. S. & Saunders R. M. K. 2015: Thismia hongkongensis (Thismiaceae): a new mycoheterotrophic species from Hong Kong, China, with observations on floral visitors and seed dispersal. – PhytoKeys 46: 21–33.  https://doi.org/10.3897/phytokeys.46.8963 Google Scholar

30.

Matzke N. J. 2018: BioGeoBEARS: biogeography with Bayesian (and likelihood) evolutionary analysis with R scripts. Version 1.1.1. – Published on GitHub on 6 Nov 2018.  https://rdrr.io/github/nmatzke/BioGeoBEARS/  Google Scholar

31.

Meier R., Shiyang K., Vaidya G. & Ng P. K. L. 2006: DNA barcoding and taxonomy in diptera: a tale of high intraspecific variability and low identification success. – Syst. Biol. 55: 715–728.  https://doi.org/10.1080/10635150600969864 Google Scholar

32.

Merckx V. 2008: Myco-heterotrophy in Dioscoreales. Systematics and evolution. – Leuven: Katholieke Universiteit Leuven. Google Scholar

33.

Merckx V. & Bidartondo M. I. 2008: Breakdown and delayed cospeciation in the arbuscular mycorrhizal mutualism. – Proc. Roy. Soc. Biol. Sci. Ser. B 275: 1029–1035.  https://doi.org/10.1098/rspb.2007.1622 Google Scholar

34.

Merckx V. S. F. T., Freudenstein J. V., Kissling J., Christenhusz M. J. M., Stotler R. E., Crandall-Stotler B., Wickett N., Rudall P. J., Maas-van de Kamer H. & Maas P. J. M. 2013b: Taxonomy and classification. – Pp. 19–101 in: Merckx V. (ed.), Mycoheterotrophy: the biology of plants living on fungi. – New York: Springer.  https://doi.org/10.1007/978-1-4614-5209-6_2 Google Scholar

35.

Merckx V. S. F. T., Gomes S. I. F., Wapstra M., Hunt C., Steenbeeke G., Mennes C. B., Walsh N., Smissen R., Hsieh T.-H., Smets E. F. & Bidartondo M. I. 2017: The biogeographical history of the interaction between mycoheterotrophic Thismia (Thismiaceae) plants and mycorrhizal Rhizophagus (Glomeraceae) fungi. – J. Biogeogr. 44: 1869–1879.  https://doi.org/10.1111/jbi.12994 Google Scholar

36.

Merckx V., Schols P., Maas-van de Kamer H., Maas P., Huysmans S. & Smets E. 2006: Phylogeny and evolution of Burmanniaceae (Dioscoreales) based on nuclear and mitochondrial data. – Amer. J. Bot. 93: 1684–1698.  https://doi.org/10.3732/ajb.93.11.1684 Google Scholar

37.

Merckx V. S. F. T. & Smets E. F. 2014: Thismia americana, the 101st anniversary of a botanical mystery. – Int. J. Pl. Sci. 175: 165–175.  https://doi.org/10.1086/674315 Google Scholar

38.

Merckx V. S. F. T., Smets E. F. & Specht C. D. 2013a: Biogeography and conservation. – Pp. 103–156 in: Merckx V. (ed.), Mycoheterotrophy: the biology of plants living on fungi. – New York: Springer.  https://doi.org/10.1007/978-1-4614-5209-6_3 Google Scholar

39.

Nuraliev M. S., Yudina S. V., Shepeleva E. A., Truong B. V., Do T. X., Beer A. S. & Remizowa M. V. 2021: Floral structure in Thismia (Thismiaceae: Dioscoreales): new insights from anatomy, vasculature and development. – Bot. J. Linn. Soc. 195: 501–531.  https://doi.org/10.1093/botlinnean/boaa066 Google Scholar

40.

Nuraliev M. S., Yudina S. V., Truong B. V., Do T. X., Luu H. T., Kuznetsov A. N. & Kuznetsova S. P. 2020: A revision of the family Thismiaceae (Dioscoreales) in Cambodia, Laos and Vietnam. – Phytotaxa 441: 229–250.  https://doi.org/10.11646/phytotaxa.441.3.1 Google Scholar

41.

POWO 2023: Plants of the World Online. – Published at https://powo.science.kew.org/[accessed 6 Jun 2023]. Google Scholar

42.

Rambaut A., Drummond A. J., Xie D., Baele G. & Suchard M. A. 2018: Posterior summarization in Bayesian phylogenetics using Tracer 1.7. – Syst. Biol. 67: 901–904.  https://doi.org/10.1093/sysbio/syy032 Google Scholar

43.

Ree R. H. & Smith S. A. 2008: Maximum likelihood inference of geographic range evolution by dispersal, local extinction, and cladogenesis. – Syst. Biol. 57: 4–14.  https://doi.org/10.1080/10635150701883881 Google Scholar

44.

Ronquist F. 1997: Dispersal-vicariance analysis: a new approach to the quantification of historical biogeography. – Syst. Biol. 46: 195–203.  https://doi.org/10.1093/sysbio/46.1.195 Google Scholar

45.

Ronquist F., Teslenko M., van der Mark P., Ayres D. L., Darling A., Höhna S., Larget B., Liu L., Suchard M. A. & Huelsenbeck J. P. 2012: MrBayes 3.2: efficient Google Scholar

46.

Bayesian phylogenetic inference and model choice across a large model space. – Syst. Biol. 61: 539–542.  https://doi.org/10.1093/sysbio/sys029 Google Scholar

47.

Shepeleva E. A., Schelkunov M. I., Hroneš M., Sochor M., Dančák M., Merckx V. S. F. T., Kikuchi I. A. B. S., Chantanaorrapint S., Suetsugu K., Tsukaya H., Mar S. S., Luu H. T., Li H. Q., Logacheva M. D. & Nuraliev M. S. 2020: Phylogenetics of the mycoheterotrophic genus Thismia (Thismiaceae: Dioscoreales) with a focus on the Old World taxa: delineation of novel natural groups and insights into the evolution of morphological traits. – Bot. J. Linn. Soc. 193: 287–315.  https://doi.org/10.1093/botlinnean/boaa017 Google Scholar

48.

Sirichamorn Y., Thomas D. C., Adema F. A. C. B. & van Welzen P. C. 2014: Historical biogeography of Aganope, Brachypterum and Derris (Fabaceae, tribe Millettieae): insights into the origins of palaeotropical intercontinental disjunctions and general biogeographical patterns in Southeast Asia. – J. Biogeogr. 41: 882–893.  https://doi.org/10.1111/jbi.12262 Google Scholar

49.

Siti-Munirah M. Y., Suhaimi-Miloko Z. & Ahmad M. I. Z. 2021: Thismia belumensis (Thismiaceae), a remarkable new species from The Royal Belum State Park, Gerik, Perak, Peninsular Malaysia. – PhytoKeys: 121–134.  https://doi.org/10.3897/phytokeys.172.59336 Google Scholar

50.

Sochor M., Hroneš M. & Dančák M. 2018: New insights into variation, evolution and taxonomy of fairy lanterns (Thismia, Thismiaceae) with four new species from Borneo. – Pl. Syst. Evol. 304: 699–721.  https://doi.org/10.1007/s00606-018-1504-5 Google Scholar

51.

Stamatakis A. 2014: RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. – Bioinformatics 30: 1312–1313.  https://doi.org/10.1093/bioinformatics/btu033 Google Scholar

52.

Tan K., Malabrigo P. L. & Ren M.-X. 2020: Origin and evolution of biodiversity hotspots in Southeast Asia. – Acta Ecol. Sin. 40: 3866–3877. Google Scholar

53.

Thiers B. M. 2023+ [continuously updated]: Index herbariorum: a global directory of public herbaria and associated staff. New York Botanical Garden's virtual herbarium. – Published at https://sweetgum.nybg.org/science/ih/[accessed 25 Dec 2023]. Google Scholar

54.

Thomas D. C., Hughes M., Phutthai T., Ardi W. H., Rajbhandary S., Rubite R., Twyford A. D. & Richardson J. E. 2012: West to east dispersal and subsequent rapid diversification of the mega-diverse genus Begonia (Begoniaceae) in the Malesian archipelago. – J. Biogeogr. 39: 98–113.  https://doi.org/10.1111/j.1365-2699.2011.02596.x Google Scholar

55.

Xu H., Yang H.-J., Lin M.-X., Corrales A., Hogan J.-A., Li Y.-D. & Fang S.-Q. 2020: Thismia jianfenglingensis (Thismiaceae), a new species of fairy lantern from Hainan Island, China. – Phytotaxa 429: 179–185.  https://doi.org/10.11646/phytotaxa.429.2.9 Google Scholar

56.

Yang S. Z., Saunders R. M. K. & Hsu C. J. 2002: Thismia taiwanensis sp. nov. (Burmanniaceae tribe Thismieae): first record of the tribe in China. – Syst. Bot. 27: 485–488. Google Scholar

57.

Yang Z. 2007: PAML 4: phylogenetic analysis by maximum likelihood. – Molec. Biol. Evol. 24: 1586–1591.  https://doi.org/10.1093/molbev/msm088 Google Scholar

58.

Zeng C.-X., Hollingsworth P. M., Yang J., He Z.-S., Zhang Z.-R., Li D.-Z. & Yang J.-B. 2018: Genome skimming herbarium specimens for DNA barcoding and phylogenomics. – Pl. Methods 14(43).  https://doi.org/10.1186/s13007-018-0300-0 Google Scholar

59.

Zhang L. G., Li X. Q., Jin W. T., Liu Y. J., Zhao Y., Rong J. & Xiang X. G. 2023: Asymmetric migration dynamics of the tropical Asian and Australasian floras. – Pl. Diversity 45: 20–26.  https://doi.org/10.1016/j.pld.2022.05.006 Google Scholar

Appendices

Supplemental content online

See  https://doi.org/10.3372/wi.54.54102

 Supplementary appendix S1 (wi.54.54102_Supplementary_appendix_S1.xlsx). Checklist of accepted species of Thismia s.l. (Thismiaceae), including authorship(s), distribution range and publication information.

 Supplementary appendix S2 (wi.54.54102_Supplementary_appendix_S2.nex). Combined matrix of the four DNA markers (nrDNA 18S and ITS, mtDNA atpA and matR) in Nexus format. The matrix is partitioned by regions.

Appendix 1

Summary of taxon sampling for this study. Taxon names are followed by geographic origin of the sample, voucher information and GenBank accession numbers for the genes sampled: 18S, ITS, atpA and matR. Voucher information is from Dančák & al. (2020b, 2020c), Merckx & al. (2017), Shepeleva & al. (2020) and Sochor & al. (2018). Herbarium codes follow Thiers (2023+).

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img-z17-1_47.gif
Ji-Dong Ya, Hai-Yao Chen, Wei Zhang, Ren-Bin Zhu, Jie Cai, and Wen-Bin Yu "Phylogenetic and biogeographical analyses of Thismia (Thismiaceae) support T. malipoensis as the eighth species in China," Willdenowia 54(1), 47-63, (28 February 2024). https://doi.org/10.3372/wi.54.54102
Received: 9 August 2023; Accepted: 20 December 2023; Published: 28 February 2024
KEYWORDS
biogeography
China
conservation
new species
phylogeny
taxonomy
Thismia
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