Checklist and molecular phylogenetics reveal three taxonomic novelties in Habenaria (Orchidaceae, Orchidoideae) from Chapada dos Veadeiros, Goiás, Brazil

. A comprehensive checklist of Habenaria from Chapada dos Veadeiros, State of Goiás, was performed alongside morphologic and molecular phylogenetic studies, revealing three new taxa endemic to this region. A total of 61 taxa (59 species and two varieties) of Habenaria are recorded for Chapada dos Veadeiros, representing a two-fold increase compared to previous lists and comprising one of the greatest diversities of the genus in Brazil. Of this total, four taxa are locally endemic. Habenaria cultellifolia , until recently known only from the type collection, was rediscovered in the region after 127 years without records and represents this species’ only known extant population. Three proposed new taxa of Habenaria ( H. minuticalcar J.A.N. Bat. & Bianch. sp. nov., H. proiteana J.A.N. Bat., A.A. Vale


Introduction
Habenaria Willd.(Orchidaceae, Habenariinae) is a genus of terrestrial orchids distributed throughout tropical and subtropical regions of the world (Cribb 2001).Currently, around 910 species of Habenaria are accepted (Govaerts et al. 2021), making the genus one of the largest in the family.Brazil, with 164 to 177 taxa (Flora e Funga do Brasil 2022; Govaerts et al. 2021) has the largest number of Habenaria species in the New World and possibly also worldwide and this genus is the largest of the family Orchidaceae in the country.Despite the recent progress made in the taxonomic knowledge of this genus in Brazil, there are still large gaps in almost all other aspects, such as the distribution, the regional and the local diversity, the ecology, the pollination and the reproductive biology of the species.Higher regions, with altitudes above 1,000 m a.s.l., are the ones with the greatest diversity (Brade 1951;Toscano de Brito 1995;Batista & Bianchetti 2003;Carvalho et al. 2013;Vieira & Barros 2017), but surveys are few and a systematic analysis of the distribution of the genus diversity in Brazil is lacking.
A particular area with great potential for terrestrial orchids is the Chapada dos Veadeiros region, State of Goiás, due to the large extension of savannic grasslands and the mild climate.The floristic knowledge of this region is still incomplete, with the first floristic inventory being the study of Munhoz & Proença (1998) for the municipality of Alto Paraíso de Goiás.This study listed 1,310 species and 120 families of vascular plants, including 47 species and 18 genera of Orchidaceae.In a more recent checklist for Chapada dos Veadeiros, Mendonça et al. (2007) listed 2,661 species distributed in 150 families.Orchidaceae, with 84 species, appears as the eighth family with the highest number of species.Both in the checklist by Munhoz & Proença (1998) and by Mendonça et al. (2007), Habenaria corresponds to the Orchidaceae genus with the largest number of species in this region, with 17 (35% of the family total) and 28 (33%) species, respectively.Besides checklists, some studies described new species of Orchidaceae for Chapada dos Veadeiros, including Encyclia chapadensis L.C.Menezes (Menezes 1992), Cleistes pusilla Pansarin (Pansarin 2004), Sacoila cerradicola Meneguzzo (Meneguzzo 2013), Habenaria irwiniana J.A.N.Bat.& Bianch., and H. minuta J.A.N.Bat.& Bianch.(Batista et al. 2017).Additionally, taxonomic treatments for Bulbophyllum Thouars (Santos & Silva 2019) and Epidendrum L. (Santos & Silva 2020) are also available for the Chapada dos Veadeiros National Park (PNCV).
Since 1991, two of the authors of the present study (JANB and LBB) have been exploring the diversity of Orchidaceae from the Chapada dos Veadeiros region within the scope of a larger Orchidaceae study from the Cerrado biome.Among the specimens found and collected throughout these years, several specimens of Habenaria did not correspond to any of the currently recognised Neotropical species and could not be assigned to any species rank.Subsequent morphological and molecular studies of these specimens revealed that some correspond to new species.In this study, we present the results of the Habenaria checklist from Chapada dos Veadeiros alongside morphological and molecular studies revealing three new taxa of Habenaria endemic to this region.

Taxon sampling for the phylogenetic analyses
To investigate the phylogenetic relationships of the putative new taxa we performed an initial analysis using the same dataset as Batista et al. (2013), which comprises ca 51% of the Neotropical species of Habenaria and 1,389 bp of the complete nuclear internal transcribed spacer (ITS) region and part of the plastid matK gene.Based on this analysis, we selected some terminals for a second analysis using the dataset of Pedron et al. (2014), including some species addressed in later studies (Batista et al. 2014(Batista et al. , 2016(Batista et al. , 2017;;Cruz-Lustre et al. 2022;Lau et al. 2021) and 37 new sequences from 11 species (12 terminals) that were generated for this study.For the outgroup, encompassing old world taxa, we used the same dataset as Batista et al. (2022a), which consists of a selected sampling based on the studies of Jin et al. (2017) and Ngugi et al. (2020), comprising representatives of the main clades of Asian and African Habenaria and related genera.The final data set consisted of the combined ITS, ETS, matK-trnK and rps16-trnK DNA sequences of 105 terminals of 103 taxa, including 85 terminals of 83 Neotropical Habenaria taxa containing most subclades of Neotropical Habenaria (Batista et al. 2013).We used Stenoglottis longifolia Hook.f., from subtribe Orchidinae, to root all trees based on the results of Ngugi et al. (2020).Voucher information, geographic origins, and GenBank accession numbers are provided in Supplementary file 1.

Phylogenetic analyses
DNA sequence alignments were analysed using parsimony and Bayesian inference.To assess incongruences between the datasets we performed searches using both individual and combined matrices.In most cases, the concatenation was done with the same individuals sequenced for the different markers.We performed maximum parsimony (MP) phylogenetic analyses using PAUP* version 4 (Swofford 2002)  consisted of 2,000 replicates of random taxon additions, with branch swapping using the tree-bisection and reconnection (TBR) algorithm, saving ≤ 10 trees per replicate to avoid extensive swapping on suboptimal islands.We evaluated internal support by character bootstrapping (Felsenstein 1985) using 2,000 replicates, simple addition, and TBR branch swapping, saving ≤ 15 trees per replicate.For bootstrap support levels, we considered bootstrap percentages (BP) of 50-70% as weak, 71-85% as moderate, and > 85% as strong (Kress et al. 2002).
We performed Bayesian analyses using MrBayes 3.2.7a(Ronquist et al. 2012) as implemented in the Cyberinfrastructure for Phylogenetic Research (CIPRES) Portal 2.0 (Miller et al. 2010), treating each DNA region as a separate partition.We selected an evolutionary model for each DNA region in MrModeltest 2 (Nylander 2004) using the Hierarchical Likelihood Ratio Tests (hLRTs).The unlink command was used to estimate model parameters separately for each partition.Each analysis consisted of two independent runs with four chains for 5,000,000 generations, sampling one tree every 1,000 generations and a temperature parameter of 0.2.Convergence between the runs was evaluated using the average standard deviation of split frequencies (< 0.01) and the Potential Scale Reduction Factor -PRSF (= 1.0) and was achieved after 705,000 generations.After discarding the first 1,500 trees (30%) as the burn-in, the remaining trees were used to assess topology and posterior probabilities (PP) in a majority-rule consensus.Because PP in Bayesian analysis are not equivalent to BP but are generally much higher (Erixon et al. 2003), we used criteria similar to a standard statistical test, considering groups with PP > 0.95 as strongly supported, groups with PP ranging from 0.90-0.95as moderately supported, and groups with PP < 0.90 as weakly supported.

Morphological analyses
Characters regarding habit, perianth, and gynostemium were examined from herbarium specimens, dissected flowers, and alcohol-fixed flowers.For the analyses of the gynostemium, we used a stereoscopic microscope and measured it using a digital calliper.Data concerning flowering times, habitats, and geographic distributions were based on herbarium labels and field observations.Morphological terminology is based on Stearn (1992) and Simpson (2006).

Conservation assessment
The distribution map was elaborated with ArcMap 10.5 (ESRI 2016).Occurrence records were obtained from herbaria and all records included had their identification revised according to the species protologues and the taxonomic circumscriptions of Batista et al. (2011 a, b).For specimens without coordinates, the latitude and longitude were inferred using Google Earth.We estimated the extent of occurrence (EOO) and area of occupancy (AOO) using a beta version of GeoCAT (Bachman et al. 2011) as implemented in the Royal Botanic Gardens, Kew Web site, with the default setting of 2 km cell width for AOO.Conservation status was inferred using the World Conservation Union Red List Categories and Criteria (IUCN 2012) and the guidelines for using the IUCN red list categories and criteria (IUCN 2016).To estimate the number of populations of each species, we considered each known record, if geographically separated, as a population.

Survey of Habenaria in Chapada dos Veadeiros
A total of 61 taxa (59 species and two varieties) of Habenaria were recorded for Chapada dos Veadeiros (Table 1) in the municipalities of Alto Paraíso de Goiás, Cavalcante, Colinas do Sul, São João d'Aliança, and Teresina de Goiás (Fig. 1).The species with the highest number of records and the most common were: H. obtusa Lindl.( 50 1).27 taxa in this study are known from three or fewer records and can be considered rare or uncommon in the region.Among these stands out, H. minuta, endemic to Chapada dos Veadeiros and known so far from only three collections.Another important result was the rediscovery of H. cultellifolia (Fig. 5G-I).This species was described in 1877 from material collected in Uberaba, located 600 km to the south, in the State of Minas Gerais, and never collected again, and until its collection in Chapada dos Veadeiros was known only from the type collection.
Regarding   25).The number of collections of the genus in the region is high (542 herbarium collections; 9 collections/taxa), which reflects both the specific sampling effort for Orchidaceae carried out by the authors and the high frequency of the genus in the region.

Phylogenetic analyses
The concatenated ITS, ETS, matK-trnK, and rps16-trnK DNA matrix consisted of 4,372 aligned characters, of which 841 (19%) were parsimony-informative.Table 2 presents the general features of the datasets and parsimony statistics along with a summary of the models implemented for each partition.Missing data accounted for 8.4% of the combined matrix, most of which were from the complete matK-trnK, rps16-trnK and ETS sequences that were missing from Old World taxa.Missing data for the Neotropical sequences accounted for just 2.4% of the combined matrix.Comparable to previous molecular phylogenetic analyses with similar datasets (Pedron et al. 2014;Lau et al. 2021), no significant incongruences were detected between nuclear and plastid datasets, therefore only the results of the combined matrix are presented and discussed.The parsimony analyses retained a total of 17,551 most parsimonious trees, with a tree length of 3,013 steps, a consistency index (CI) of 0.63, and a retention index (RI) of 0.78.The strict consensus tree from the parsimony analysis was, for the most part, congruent with the Bayesian majority-rule consensus tree.Still, as the latter was more fully resolved and had stronger overall support, it was chosen for presentation and discussion (Fig. 2).For comparison, bootstrap percentages from the bootstrap analysis are shown in the Bayesian tree.
The relationships recovered were similar to those of earlier molecular phylogenetic studies (Batista et al. 2013(Batista et al. , 2014(Batista et al. , 2016(Batista et al. , 2017(Batista et al. , 2022a;;Pedron et al. 2014;Lau et al. 2021), with the New World Habenaria species forming a well-supported monophyletic group (1.00 PP, 83% BP) and recovering several wellsupported subclades within the New World clade that are numbered accordingly (subclades 1 to 21) (Fig. 2).The two new species addressed in this study (see Taxonomic treatment) were recovered within the Cerrado clades (1.00 PP, 96% BP), which comprises clades and species distributed mainly in the Cerrado Phytogeographic Domain of Central Brazil (Batista et al. 2013)

Etymology
From the Latin 'minutus' (very small) and 'calcar' (spur), referring to the very small spur, an unusual feature in the genus and one of the smallest among New World Habenaria.

Distribution, Habitat and Phenology
Habenaria minuticalcar sp.nov. is restricted to Chapada dos Veadeiros, State of Goiás, central Brazil.The new species grows in seasonally humid grassland (Fig. 3A), associated with 'murundus' or 'campos rupestres' (rocky fields), with dark, shallow, rocky, sandy-clay soils, at 1,100 to 1,600 m a.s.l.The soil can be humid during some periods in the rainy season (Fig. 3B) but usually dries out completely during the dry season.The plants are difficult to find in their habitat as they are small and slender, grow among grasses and can only be found when they are in flower.Flowering occurs during the peak of the rainy season, from December to January.Like other Habenaria and terrestrial grassland orchids, the flowering of the new species is enhanced by seasonal brush fires.All our collections of the species were made in areas burned from the previous dry season.

Conservation Assessment
Habenaria minuticalcar sp.nov. is currently known from only four populations, all from Chapada dos Veadeiros, and shows an EOO estimated at 194 km 2 and a small AOO of 16 km 2 .Based on the IUCN Red List Categories and Criteria and its guidelines (IUCN 2012(IUCN , 2016)), the species can be tentatively classified as Endangered EN B1ab(iii)+2ab(iii); C2a(i).

Taxonomic notes
Habenaria minuticalcar sp.nov.resembles H. guilleminii in the size and morphology of the spur.However, it is distinguished by the linear, narrow, 2-3 mm wide leaves (Fig. 4D-F) (vs lanceolate or elliptical, 5-23 mm wide in H. guilleminii), narrow, 0.9-1.2mm wide lateral sepals (vs 2-3 mm wide), petal anterior segment 1.5-2.2mm long, reaching half the size of the petal posterior segment (vs reduced to a tooth-like projection) and lip lateral segments at most half the length of the median segment (Fig. 4G-I) (vs lateral segments longer than the median segment).Habenaria minuticalcar sp.nov. is also similar to H. pansarinii var.minuscula J.A.N.Bat.& Bianch. in general morphology and size of the spur.However, it differs from this taxon by the floral bract measuring 5-17 mm long, approximately the same length as the pedicellate ovary (Fig. 4J-K) (vs 2.2-3.5 mm long, shorter than the pedicellate ovary), the slightly larger flowers (e.g., dorsal sepal 3-5 × 3-3.5 mm vs 2.4-3 × 1.8-2.4mm in H. pansarinii var.minuscula), the petal posterior segment 2.5-4.2mm long, oblong with an obtuse apex (vs 2.1-2.7 mm long, lanceolate, apex acute), and the geographical distribution restricted to Chapada dos Veadeiros (vs restricted to the southern and central regions of the Espinhaço Range in the State of Minas Gerais).

Etymology
From the Greek 'xanthos' (yellow) and 'dactylos' (finger), referring to the yellow segments of the petals and lip.

Distribution, Habitat and Phenology
Habenaria lavrensis var.xanthodactyla var.nov. is so far only known from Chapada dos Veadeiros, Goiás, in the municipalities of Alto Paraíso de Goiás and São João da Aliança.In the northern part of Chapada dos Veadeiros, in the Distrito Federal and the state of Minas Gerais, where we have collected or examined specimens of this species, only H. lavrensis var.lavrensis is known.Habenaria lavrensis var.xanthodactyla var.nov.grows in dry, hillside or campo rupestre, in shallow, dry, gravelly soil.Flowering occurs during the peak of the rainy season, from January to February.In common with other grassland species of the genus, bushfires favoured flowering during the dry period.All populations we observed in flowering grew in areas that burned in the previous dry period.The habitat and the flowering period are about the same between the two varieties.

Conservation Assessment
Habenaria lavrensis var.xanthodactyla var.nov. is currently known from only seven populations from six localities and shows an EOO estimated at 1,647 km 2 and an AOO of 32 km 2 .Based on the IUCN Red List Categories and Criteria and its guidelines (IUCN 2012(IUCN , 2016)), the species can be tentatively classified as Vulnerable VU B1ab(iii)+2ab(iii).

Taxonomic notes
Habenaria lavrensis var.xanthodactyla var.nov. is identical in vegetative and flower morphology to H. lavrensis var.lavrensis but distinct by the yellow lateral segments of petals and lip (Fig. 3G-H).In contrast, the entire flower is completely white or, at most segments, slightly creamy-white in H. lavrensis var.lavrensis (Fig. 3F).Both H. lavrensis var.lavrensis and H. lavrensis var.xanthodactyla var.nov.grow at the Chapada dos Veadeiros as homogeneous populations, spatially separated, and we have never found mixed populations or intermediate specimens.In defining the taxonomic rank of this taxon, we followed Stace (1989), who defined a variety as "a population of one or several biotypes, forming more or less distinct local races of a species."

Etymology
We named this species after Karina Proite for her support, companionship, help in the field and contribution to numerous collections, and for finding the population used to characterise, describe, and typify the newly proposed species.
Similar to other regions with surveys for Habenaria (Brade 1951;Batista & Bianchetti 2003;Batista et al. 2004Batista et al. , 2008;;Carvalho et al. 2013), the number of endemic species is small, reinforcing that cases of restricted endemism are rare for the genus in Brazil (Table 3).This result contrasts with other groups that present high diversity in mountainous complexes of Southeastern and Central Western Brazil, such as Eriocaulaceae and the genus Mimosa (Simon & Proença 2000), which present a high number of endemic species.For the subfamily Paepalanthoideae (Eriocaulaceae), for example, more than 50% of the 42 species that occur in Chapada dos Veadeiros are endemic to the area (Trovó & Echternacht 2022).
Our results suggest that a significant portion of the diversity of the genus in the region does not occur or is underrepresented within the PNCV (Table 1).However, this data must be considered with reservations since many records reported in this study do not show coordinates or precise geographical locations, making it impossible to know exactly where they were collected.Collections in the region are centred along access roads, mainly on the GO-118 highway between Alto Paraíso de Goiás and Teresina de Goiás and GO-239 between Alto Paraíso and the village of São Jorge (Fig. 1).Large extensions of the region have not yet been sufficiently botanically collected or have no collections at all, including the central and north-western parts of the park as well as the extensive area added to the park in 2016 (Fig. 1), which includes some localities with the highest altitudes in the region and for which there are currently no herbarium collections available.More comprehensive and systematic explorations are expected to reveal new records and eventual taxonomic novelties within the Habenaria from this region, considering the exceptional richness of the genus in the Chapada dos Veadeiros.In this work, for example, we describe three new taxa and record the only known extant population of H. cultellifolia, a species known only from type material collected at least 127 years ago.
These discoveries indicate that although some parts of the region have been extensively explored, new novelties are still to be expected, particularly among the terrestrial, small and ephemerous species.
Efforts to resolve the taxonomy of Brazilian species of Habenaria have intensified in the last years with the study of species complexes and the description of several new species (Batista et al. 2017(Batista et al. , 2022a, b;, b;Cruz-Lustre et al. 2020, 2022;Lau et al. 2021).Nevertheless, for the Chapada dos Veadeiros at least three morphotypes of the Habenaria nuda complex and some taxa associated with H. montis-European Journal of Taxonomy 891: 51-86 (2023) wilhelminae Renz, H. rodeiensis Barb.Rodr., and H.pseudoculicina (Table 1) are still unresolved and additional studies are needed to uncover the identity and taxonomy of these taxa.

Phylogenetic analyses
In terms of overall results of the phylogenetic analyses, the main differences between our results and some of the earlier molecular phylogenetic studies (Batista et al. 2013(Batista et al. , 2014(Batista et al. , 2016(Batista et al. , 2018;;Cruz-Lustre et al. 2022;Lau et al. 2021) were the position of H. monorrhiza (Sw.)Rchb.f. and subclade 1 (H.sect.Pratenses Kraenzl.), which were recovered in a polytomy at the base of the Neotropical clade, while in our results, they were internally recovered in the Neotropical clade (Fig. 2).Nevertheless, in both cases, the results had low support, showing that the phylogenetic relationships of these taxa within the Neotropical clade are not yet resolved.In another difference, species from subclades 16, 17 and 18 were intermingled into a single subclade, while in the topology of Batista et al. (2013) these subclades were recovered separately with high support.Although our dataset included a greater number of markers and characters (five markers and 4,372 characters vs two markers and 1,389 characters in the study of Batista et al. 2013), our sampling of taxa from these subclades was smaller (6 taxa vs 15 taxa), which may have reduced the resolution of the relationships between them.
As for the new species described here, both were recovered with high support as sister to species with different floral morphology, but with similar vegetative morphology, habitat and ecology (8.0-11.7 mm long) is about the same length as the pedicellate ovary (12-15 mm long).On the other hand, the morphology of the column is similar between the two species.In particular, the rostellum is very similar, is completely held between the anther loculi, and the pollinaria are joined by the viscidia in both species.However, these features are not unique and are shared with other non-directly related species groups, such as the species in subclade 20 (H.sect.Microdactylae Kraenzl.) and subclade 15, among others.
For the pair H. proiteana sp.nov./ H. psammophila the differences are greater than the similarities.Vegetatively, H. psammophila has sub-patent, papery, somewhat rigid leaves with the limb mostly detached from the stem.In floral morphology, the flowers of H. psammophila are smaller, the spur (3.0-4.5 mm long) is shorter than the pedicellate ovary (7-11 mm long), and the morphology of the column is quite different between the two species.The only obvious similarities between H. proiteana sp.nov./ H. psammophila are the anterior segment of the petals longer than the posterior segment and the lateral segments of the lip longer than the median segment.However, these features are also found in all species of the H. nuda complex and in several other Neotropical groups and species of the genus.An additional and perhaps more striking, but less obvious similarity, is the spur of H. psammophila.
Although much smaller, it is also clavate and occasionally also more or less genuflexus, which are distinctive features of H. proiteana sp.nov.. Our results show that H. proiteana sp.nov. is not directly related to the species of the H. nuda complex (subclade 21), with which it shares a similar overall morphology in both vegetative and floral terms.This result suggests that both this general morphology as also the anterior petal segment inserted above the posterior segment's base, a characteristic shared with H. rodriguesii, and H. australis, are probably a result of convergence.On the other hand, H. rodriguesii and H. australis form a highly supported species pair and this morphological feature probably constitutes a synapomorphy for this pair within subclade 21.
Most subclades of Neotropical Habenaria, such as subclades 1, 2, 3 (Cruz-Lustre et al. 2022), 4, 8, 15, 20 (Batista et al. 2018) (Batista et al. 2022a).In addition to these differences within the subclades, some species pairs from the Cerrado Phytogeographic Domain recovered as sisters with high support, such as H. caldensis Kraenzl./ H. magniscutata and now H. minuticalcar sp.nov./ H. pseudoculicina and H. proiteana sp.nov./ H. psammophila (Fig. 2) differ greatly in floral morphology.Since habitat, ecology and vegetative morphology are similar between these species pairs, and the pairs are often sympatrical, these results suggest speciation driven by different pollinators (Harder & Johnson 2009).In line with this interpretation, Pedron et al. (2012) showed that H. montevidensis is pollinated by butterflies, while other species of H. sect.Pentadactylae, are pollinated by moths or present floral morphology associated with moth pollination, suggesting that the morphological differences of H. montevidensis are a result of a shift to diurnal pollinators.

Fig. 1 .
Fig. 1.Distribution map of Habenaria records from Chapada dos Veadeiros.A. Map with all records.B. Map with records of new taxa plus H. lavrensis Hoehne var.lavrensis.The area incorporated into the park in 2016 is indicated by a dashed line.Habenaria lavrensis denotes specimens with unknown intraspecific identification.

Fig. 2 .
Fig. 2. Bayesian 50% majority-rule consensus tree of the combined ITS, ETS, matK-trnK and rps16-trnK datasets.Numbers next to the nodes represent the posterior probabilities (PP) from the Bayesian analyses and bootstrap percentages (BP) from the parsimony analyses.Only the values of the main clades are shown.Neotropical subclades are numbered according to Batista et al. (2013).The new species described here are highlighted in bold and indicated by an arrow.Species with the petal anterior segment inserted above the base of the posterior segment are highlighted by an asterisk.Subclades in which it was possible to associate with one of the sections of Kränzlin's sectional treatment are identified accordingly.

Table 1
(continued on the next page).Habenaria Willd.taxa recorded for Chapada dos Veadeiros, Goiás.Endemic taxa are indicated by an asterisk.Vouchers correspond to the first record of the taxon for the region.PNCV = Parque Nacional da Chapada dos Veadeiros (Chapada dos Veadeiros National Park).

Table 1
Batista et al. 2022b) taxa recorded for Chapada dos Veadeiros, Goiás.Endemic taxa are indicated by an asterisk.Vouchers correspond to the first record of the taxon for the region.PNCV = Parque Nacional da Chapada dos Veadeiros (Chapada dos Veadeiros National Park).BATISTA J.A.N. et al., Habenaria from Chapada dos Veadeiros do Brasil 2022), while six taxa are shared with other montane regions of the States of Minas Gerais and Bahia: H. ciliatisepala J.A.N.Bat.&Bianch., H. lavrensis Hoehne, H. magniscutata Catling, H. nuda Lindl.var.nuda,H.psammophila, and H. subrepens J.A.N.Bat., B.L.Lau & I.G.C.Machado-Costa (Flora e Funga do Brasil 2022;Batista et al. 2022b).Regarding distribution of records and taxa within the PNCV, of the total records, 175 (33%) were made in the PNCV, and of the total taxa recorded for the region, 22 (36%) have no record for the PNCV, while another 17 (28%) taxa have only 1-2 records for this park (Table1).From a historical perspective, the first records for the region were made by A.F.M. Glaziou, who collected four species in 'Chapadão dos Viadeiros' between 1894 and 1895.The following records were only made in the 60s, when H.S. Irwin and colleagues intensively collected in the region between 1966 and 1971, resulting in 19 new records of Habenaria for Chapada dos Veadeiros (Table1).In the 70s, the main collectors were W.R. Anderson, G. Hatschbach, and Gates & Estabrook, whose collections resulted in 13 new records for the region.From the 1990s onwards, the collections in the region were made mainly by researchers and postgraduate students from regional institutions such as CENARGEN, Universidade de Brasília and Jardim Botânico de Brasilia, contributing with five new records.And from 1991 onwards, field collections within this study began and lasted until 2018, contributing to 17 new records of Habenaria for this region.According to our data, of a total of 63 collectors, the ones with the highest number of collections are J.

Table 2 .
. Both H. minuticalcar sp.nov./ H. pseudoculicina and H. proiteana sp.nov./ H. psammophila, occur in 'campos rupestres', in seasonally humid, sandy soils in isolated montane areas of central Brazil.In Characterization of the markers used in the phylogenetic analyses.Ter = number of terminals; Char = number of aligned characters in base pairs; MD = percentage of missing data; VU = number of variable uninformative characters; VI = number of variable informative characters; CI = consistency index; RI = retention index.hLRTs = hierarchical likelihood ratio tests.BATISTA J.A.N. et al., Habenaria from Chapada dos Veadeiros the pair H. minuticalcar sp.nov./H.pseudoculicina both plants are slender with linear and reduced leaves, while in H. pseudoculicina the anterior segment of the petals (5.0-6.5 mm long) is about 1.7 times longer than the posterior segment(2.8-3.8 mm long), the lateral segments of the lip (5.0-7.5 mm long) are 1.3 to 1.4 times longer than the median segment (3.5-5.0 mm long) and the spur