Terrestrial Orchid Research Articles

Diversity of orchid types in the Oeluan Tourism Forest, Bijeli Village, Noemuti District, North Central Timor Regency

Indonesia as a mega biodiversity country has a variety of fauna and flora. One of the floras that is often found in Indonesia is orchids. Orchids have high economic value and are able to balance the ecosystem in their habitat. This study was aimed at determining the diversity of orchid species in the Oeluan tourism forest. The current study employed a quantitative descriptive study by using the transect and the purposive sampling method. The research was conducted in February at the Oeluan Tourism Forest. The results of this study indicated that there were four terrestrial orchid genera and three epiphytic orchid genera. The analysis of the diversity index of orchid species in the Oeluan tourism forest resulted in the diversity index of (H’) 1.0438 in the medium category. The measured environmental factors at the study site were an average soil pH of 7, an average humidity of 76.33% and an average air temperature of 27.400C. The diversity of orchid species in the Oeluan tourism forest, bijeli village, Noemuti district, North Central Timor regency, East Nusa Tenggara Province, was included in the medium category.

Didymoplexiella siamensis (Gastrodieae, Orchidaceae), a new generic record for the flora of India

<i>Didymoplexiella siamensis</i> (Gastrodieae, Orchidaceae), a new generic record for the flora of India

Marsh-orchids of Canada: long-standing mysteries partially solved

SummaryBetween 1959 and 1988, three populations of purple-flowered terrestrial orchids attributable to Dactylorhiza subgenus Dactylorhiza were discovered in Canada. The populations at Timmins, Ontario, and St John's, Newfoundland were strongly marked on both flowers and leaves, in contrast with the anthocyanin-deficient population at Tilt Cove, Newfoundland. All three populations have since experienced a wide range of taxonomic assignments; debates are also ongoing regarding their origin and most appropriate conservation status. Here, we address these questions by combining detailed in situ morphometric analyses based on 52 characters with allozyme profiles and data from nrITS, 15 plastid microsatellites and seven nuclear microsatellites. The allozyme data alone are sufficient to both confirm allopolyploidy and categorically refute past assignments of these populations to D. incarnata, D. maculata, D. fuchsii, D. majalis or D. purpurella. Several morphometric characters, nuclear microsatellites and nrITS all reliably distinguish each of the three study populations, whereas the two sampled subpopulations from St John's proved near-identical morphologically. In contrast, morphological variation within each of the three populations is strikingly low, particularly in characters other than those influenced by plant vigour. Similarly, compared with 14 European populations, the three Canadian populations proved genetically impoverished (two were near-invariant) and likely experienced recent, extreme genetic bottlenecks during establishment. The three populations differ substantially, both morphologically and molecularly, therefore probably representing independent immigration events. Although clearly attributable to D. praetermissa, all three populations deviate significantly in morphology and DNA data from comparable populations sampled across Europe, preventing identification of their precise geographic origins. Any attempt to determine their mode or origin — through natural long-distance transport, or accidental or deliberate introduction by humans – is challenged to explain why three lineages of a single European Marsh-orchid species, each in different ways atypical of that species, arrived independently in North America whereas no other European dactylorchid species has become established there.

Three helmet-orchid species share abundant fungi of Serendipitaceae regardless of altitude

The tiny terrestrial orchids (plant height less than 8 cm) in the genus Corybas dependent on mycorrhizal fungal (OMF) partners for seed germination and seedling development. The OMF community of the Corybas remains poorly understood, although the relevant knowledge is very important for in situ and ex situ conservation of these orchids. In this study, we characterized OMF richness and compositions of three helmet-orchid species, i.e., Corybas geminigibbus, C. himalaicus, and C. shanlinshiensis, from their natural habitats by using Illumina sequencing of the internal transcribed spacer 2 region. We found that fungal colonization was restricted in the rhizomes of the helmet-orchids instead of their tuberoids, and serendipitoid fungi were predominant, while tulasnelloid were absent in the three investigated Corybas species regardless of their altitude. The three Corybas species shared 27 serendipitoid operational taxonomic units that are different to those of their related orchids, the genera of Cyrtostylis and Stigmatodactylus. Corybas shanlinshiensis alone had a range of ectomycorrhizal fungi (mainly russuloid and thelephoroid) broader than C. himalaicus and C. geminigibbus. Our study provides new information about terrestrial orchid–fungi associations and may further contribute to orchid conservation.

Reproductive biology in Gomphichis valida Rchb. f. (Orchidaceae: Orchidoideae, Cranichidinae): Generalist pollination in a high‐Andean terrestrial orchid with long‐lived flowers

AbstractFloral features, the pollination process, the breeding system, and fruiting success are documented in the Andean terrestrial orchid Gomphichis valida. Studies were performed at the Department of Ancash, Peru, at ca. 4250 m a.s.l., from June to November 2022. The breeding system was studied through controlled pollinations in plants excluded from pollinators (bagged). Natural pollination was studied in the field and through photos and videos. The flowers are nectariferous and non‐protandrous and present the longest lifespan reported so far for a neotropical terrestrial orchid (53–80 days). This species is pollinator dependent (unable to set fruit and seed without the agency of pollinators) and self‐compatible. The observed pollinators belong to three functional groups that appeared successively (in order of appearance): hummingbirds (Chalcostigma stanleyi), bumblebees (Bombus coccineus and Bombus rubicundus), and solitary Halictidae bees (Augochlorella sp.). All recorded pollinators probe the flowers for nectar. The pollinaria adhere to the bee mouthparts and onto the lateral side of the hummingbird bill. Bumblebees were the most frequent and efficient functional group (up to 62.07% of the observed pollination events), followed by Halictidae (31.03%) and hummingbirds (6.90%). Natural fruiting success is high (66.66%–94.49%). Such high natural fruiting success is unexpected at high altitudes, but it is likely explained by a combination of factors, including the long floral lifespan, nectar secretion, and self‐compatibility. This is the first report of hummingbirds as pollinators of Cranichidinae orchids, and G. valida is, to the best of our knowledge, a neotropical orchidoid orchid with the most generalist pollination biology recorded so far.

List of Orchid Species along the Northern Trekking Route of Mount Binaiya, Seram Island, Maluku - Indonesia

Orchidaceae is widely distributed on Earth with major distribution in the tropical regions, including Indonesia. Seram Island located in Maluku Province Indonesia has a high potential to become a habitat for various species of Australasian orchids. Orchid diversity tends to be related to altitude variations in the area that indirectly contribute to creating microclimate variations. One area on Seram Island with various variations in altitude is Mount Binaiya within the Manusela National Park area. This study aims to identify orchid species found on Mount Binaiya. The exploration method was used on the northern trekking route of Mount Binaiya, divided into Waisamata, Kanikeh, Waiansela, and Waihuhu areas. Based on the result, 47 species of orchids were obtained, which belong to the subfamilies Epidendroideae (35 species), Orchidoideae (11 species), and Vanilloideae (1 species). Based on the life form, 25 species were epiphytes, while 22 species were terrestrial orchids. Waisamata had the highest number of species (23 species), followed by Waiansela (14 species), Waihuhu (14 species), and Kanikeh (8 species). Of all these species, some that need further research are Corybas spp., Cyrtosia nana, Pterostylis papuana, Glomera papuana, and Mediocalcar pygmaeum. Some species are new records in their distribution or rediscoveries of existing records.

Genome-Wide Identification and Drought Stress Response Pattern of the NF-Y Gene Family in Cymbidium sinense.

Cymbidium sinense, a type of orchid plant, is more drought-resistant and ornamental than other terrestrial orchids. Research has shown that many members of the NUCLEAR FACTOR Y (NF-Y) transcription factor family are responsive to plant growth, development, and abiotic stress. However, the mechanism of the NF-Y gene family's response to abiotic stress in orchids has not yet been reported. In this study, phylogenetic analysis allowed for 27 CsNF-Y genes to be identified (5 CsNF-YAs, 9 CsNF-YBs, and 13 CsNF-YC subunits), and the CsNF-Ys were homologous to those in Arabidopsis and Oryza. Protein structure analysis revealed that different subfamilies contained different motifs, but all of them contained Motif 2. Secondary and tertiary protein structure analysis indicated that the CsNF-YB and CsNF-YC subfamilies had a high content of alpha helix structures. Cis-element analysis showed that elements related to drought stress were mainly concentrated in the CsNF-YB and CsNF-YC subfamilies, with CsNF-YB3 and CsNF-YC12 having the highest content. The results of a transcriptome analysis showed that there was a trend of downregulation of almost all CsNF-Ys in leaves under drought stress, while in roots, most members of the CsNF-YB subfamily showed a trend of upregulation. Additionally, seven genes were selected for real-time reverse transcription quantitative PCR (qRT-PCR) experiments. The results were generally consistent with those of the transcriptome analysis. The regulatory roles of CsNF-YB 1, 2, and 4 were particularly evident in the roots. The findings of our study may make a great contribution to the understanding of the role of CsNF-Ys in stress-related metabolic processes.

The complete chloroplast genome sequence of Calanthe sieboldii (orchidaceae)

Calanthe sieboldii Decne. ex Regel is a terrestrial orchid with high ornamental and commercial value. In the present study, the chloroplast genome of C. sieboldii was characterized using Illumina technology. The chloroplast genome is 158,345 bp in length with a total AT content of 63.28%. There are 127 genes, comprising 37 tRNA genes, 82 protein-coding genes, and 8 rRNA genes. Phylogenetic relationship analysis was performed using common protein-coding genes extracted from 13 chloroplast genomes of Orchidaceae. It was revealed that C. sieboldi was sister to C. hancockii and closely clustered with C. aristulifera and C. henryi. These findings provide valuable genomic resources that are helpful for further phylogenetic and evolutionary studies of Calanthe.

Middle Pleistocene climate change influenced northward divergence and cold adaptation in an Asian lady’s slipper orchid, Cypripedium macranthos Sw. (Orchidaceae)

Abstract The Pleistocene was characterized by climate fluctuations with rapid transitions between glacial and interglacial periods. East Asia’s land bridge dynamics and monsoon system during the Middle Pleistocene transition (MPT) further constrained species distributions and dispersals, which promoted local adaptation and species diversification, making East Asia an excellent location to study species biogeography. Cypripedium macranthos Sw., a terrestrial orchid, diverged in the Pleistocene and became widespread in Russia, China, Korea, and Japan. Many populations are now at risk due to illegal harvesting or habitat loss. With comprehensive sampling including 171 individuals among 22 populations across the entire distribution range, together with plastid DNA sequencing, we have explored the origin and impact of palaeoclimate on its spatial genetic structure and historical biogeography. Regression-based analyses commonly used in landscape genetics were also applied to investigate the effects of precipitation, temperature, and wind on gene flow. The results revealed northeastern China as the probable cradle of C. macranthos, with northeastward divergence and range expansion during the MPT, aided by monsoon shifts and wind-driven seed dispersal. Isolation by environment was detected with winter temperature as a driver for north–south divergence controlling for an effect of isolation by distance, revealing cold adaptation of the northern populations or temperature-driven genetic differentiation. We also emphasize the founder speciation of an unique variety on Rebun Island of Japan. The widespread but genetically and ecologically divergent C. macranthos populations highlight the need for more conservation attention on northern populations under global warming.

Inventory of Orchid Species in Customary Forest Area of Cipta Karya Village, Sungai Betung District, Bengkayang Regency

Orchids are one of the flowering plants that have many species and are widely distributed, as well as having different attractions and characteristics in each species and habitat. The aim of this research to inventory the types of orchids in Customary Forest Area of Cipta Karya Village, Bengkayang Regency. The research was conducted using the cruise method. The characteristics observed were the morphology of orchids such as roots, pseudobulbs, stems, leaves, and flowers. The inventory results found 13 species of orchids. Epiphytic orchids were found more commonly in this area, with a total of 9 species, while terrestrial orchids were found with 4 species. The epiphytic orchid species foundware composed of Appendicula sp., Bulbophyllum sp., Coelogyne speciosa, Cymbidium sp., Dendrobium crumenatum, Dendrobium anosmum, Luisia sp., Micropera pallida and Oberonia sp. The terrestrial orchid species found ware composed of Acanthephippium javanicum, Malaxis latifolia, Nephelaphyllum pulchrum, and Spathoglottis plicata.

KEANEKARAGAMAN JENIS ANGGREK DI JALUR PENDAKIAN WIRAYANA GUNUNG CAKRABUANA KABUPATEN MAJALENGKA

The existence of orchids in nature continues to decline due to habitat destruction and overexploitation. Conservation efforts that have been carried out so far have often experienced difficulties due to insufficient data and information as a basis for management, including conservation areas. The more widespread forest destruction will result in these species being threatened with extinction. This study aims to determine the diversity of orchid species, on the Wirayana Mount Cakrabuana Climbing Path. The information is expected to provide an initial contribution to the area management and orchid conservation efforts on the Wirayana Mount Cakrabuana Climbing Path. This research was conducted by means of a vegetation survey, sampling and documentation by exploring the existence of orchids. The sampling technique used the path transect method. The number of transects used is one line transect with a line length of 1500 m with a total of 22 sample plots. Data analysis was carried out by calculating the Important Value Index (INP), Species Dominance Index (Di) and Species Diversity Index (H'). The results showed that in the natural forest area of ​​Mount Cakrabuana there were 22 species of individuals which included 7 species of terrestrial orchids and 15 species of epiphytic orchids with a total of 243 individuals found. From the results of the calculation of species diversity, it is stated that the diversity of orchid species in the natural forest area of ​​Mount Cakrabuana is classified as moderate (2,973). This information is very important, especially for area managers in preserving the biodiversity of Mount Cakrabuana, especially orchids in the future.

Identification of Orchid Plants Species in the East Tomohon Forest Area

Orchids which belong to the Orchidaceae family, are popular ornamental plants that are loved by many people. It is estimated that there are around 5,000 species of orchids in Indonesia. Orchids live in various habitats including epiphytic, terrestrial, lithophytic, and saprophytic. Environmental factors such as temperature, air humidity, soil pH, and light intensity can affect the growth, development, and spread of orchids. Tomohon City is known as one of Indonesia's largest flower producers, one of that is orchids. In the East Tomohon forest area, various types of orchids were found, but the types of orchids that grow have not been identified. The purpose of this study was to identify the types of orchids that grow in the East Tomohon forest area based on their morphology. The identification of orchid plants is done by observing the morphological characteristics of the shape of leaves, roots, stems, colors, and flower shapes in detail so that their taxonomy can be known. The research was conducted on August 5-19, 2022 using the cruising method (exploratory survey) the acquisition of data was analyzed descriptively. The results of research on Orchid Plant Species in the East Tomohon Forest Area found 192 individuals consisting of 10 orchid species, 4 species are terrestrial orchids, they are Calanthe tripilcata (18), Nervilia aragoana (13), Phaius thankervilleae (21), Vanda sp. (25), and 6 species are epiphytic orchids, they are Coelogyne asperata (16), Coelogyne pandurata (17), Cattleya sp. (21), Dendrobium crumenatum, Oncidium ornithorhynchum (18), Phalaenopsis amabilis (27).

Invitro germination and optimization of basal media for protocorm-like bodies proliferation in Dieniaophrydis(J. Koenig) Seidenf

Dienia ophrydis is an endangered terrestrial orchid used for its ornamental and therapeutic purposes. This orchid possesses an anti-malarial alkaloid known as Malaxin. There is ongoing exploitation in the wild due to illegitimate herbal collection and climate change. To control the potential risk of extinction, there are no conservational measures or in vitro protocols established in this genus. The study focuses on addressing a germination protocol and enhancement of protocorm-like bodies (PLB) proliferation by optimizing the basal media and vitamins. It also aims to improve the misinterpretation of tetrazolium viability results in orchid testa along with a permeability test. Further, the seeds were subjected to 1% TTC staining to perform embryo viability test and 0.4% trypan blue (TB) dye for testa permeability. Ten basal media such as Gamborg's medium, Mitra medium, Thomale GD medium, Vacin and Went medium, Knudson C medium, Knop's medium, MS medium and ½ MS medium, ¼ MS medium, and modified MS medium (MMS) were tested for seed germination. The Half-strength MS medium responded with 92% of the highest germination and T-GD medium is not suitable for seed germination. The seed viability and permeability result showed 93% of permeability, 72% viability, and 9% dead seeds. Protocorms were transferred to the modified mMS medium containing half-strength macronutrients, full micronutrients, and 10 ppm of thiamine in vitamin formulation. PLB proliferation was observed and 12 shoots emerged from a single protocorm explant. Subsequently, the seedlings were acclimatized with 8-0-24 ratios of nitrogen, phosphorus, and potassium (NPK) as determined by soil testing. The study contributes in experimenting with seed germination of terrestrial orchid seeds asymbiotically and to establishing a protocol for conservation programs, especially in this genus.

Cheirostylis Blume (Orchidaceae): a new generic addition to the orchid flora of Manipur state, Northeast India

The genus Cheirostylis, belonging to the Orchidaceae family, is represented by numerous species distributed worldwide. In India, the genus is represented by 10 species, with two species reported from Western Ghats and Eastern Ghats, and the remaining eight species from the Northwestern Himalayas and Northeastern regions. This study reports the occurrence of Cheirostylis griffithii Lindl. in Manipur, Northeast India, which is a new generic record for the state. The taxon was discovered in Pudunamei village and identified based on herbarium specimens and available literature. Detailed morphological descriptions and measurements were made using living plant specimens. The taxonomic treatment includes information on plant characteristics, distribution, and a comprehensive list of Indian Cheirostylis species. The species is a terrestrial orchid with fleshy stems, lanceolate bracts, pubescent flowers, connate sepals forming a tube, and a 3-lobed lip with deeply laciniate lobes. The occurrence of Cheirostylis in Manipur expands its distributional range within India. The study contributes to the documentation of the orchid flora of Manipur, which encompasses a diverse range of orchid taxa.

Terrestrial orchid diversity in Hosanagara taluk of Shimogga district Central Western Ghats region of KarnatakaTerrestrial orchid diversity in Hosanagara taluk of Shimogga district Central Western Ghats region of Karnataka.

The current article evaluates the study of terrestrial orchids in the Hosanagara taluk of Shimogga district, reports a total of 30 terrestrial orchid species in the Hosanagara taluk region Shimogga district Karnataka from the period of January 2019-January 2022, and provides information on their distribution, phenology aspects, and biological status in the Hosanagara taluk of Shimogga district. Extensive field surveys were carried out periodically in different parts of the taluk, particularly in grasslands and hilly regions, to study the plant's diversity and phenological character. It was noticed that, when compared to other species, the Habenaria species is richly distributed in the Hosanagara taluk region. During field trips, some anthropogenic activities that have a significant impact on the diversity of terrestrial orchids were also observed. To learn more about the variety and current state of terrestrial orchid species in the Hosanagara taluk in the Shimogga District of Karnataka, further research must be conducted.

Study of morphology and orchid mycorrhizal associations in Malaxis rheedei

The mycorrhizal association in orchids is a striking example of mutualism, where the orchid and the associated mycorrhizal fungi exchange essential nutrients and support each other’s growth and reproduction. The present study focused on a selected terrestrial orchid which is noteworthy for its medicinal value. We aimed to investigate the specific mycorrhizal associations to understand how these interactions influence the morphology of their roots. The morphological characteristics of roots were studied using light microscopy. Transmission electron microscopy studies were conducted to analyze and measure fungal hyphae and peloton sizes as well as to know about fungal colonization distribution. Phylogenetic analysis was conducted to identify the mycorrhizal fungi. Colonization was seen in the 91% root cortex proving high mycorrhizal association. Hyphae appeared in the form of peloton patches at the cortex, occupying up to 62% of the cortical space. The root length with an intact peloton was noted 30% more than the degraded peloton. A single fungus, Tulasnella calospora was isolated from the roots. It was identified using phylogenetic analysis of ITS sequences. Identification of this orchid-mycorrhizal association has the potential to address ecological challenges, contribute to its conservation, and unlock the phytochemical potential of plant-fungus interactions for drug development.

Conservation of Gayo’s Endemic Orchid (Paphiopedilum primulinum) Through In Vitro Seed Germination and Development with Coconut Water

Paphiopedilum primulinum is one of endemic orchids in Gayo, Aceh, Indonesia. This terrestrial orchid is registered as endangered species due to IUCN. The low availability of this orchids in nature is due to habitat lost, over-collection for commercial purpose, and low seed germination rate which affected by unidentified factors, yet the study of propagating this orchid is rarely discussed. This experimental research is aim to see the effect of coconut water (CW) concentration to P. primulinum seed germination and development through in vitro culture. The sample of the study used the seed of P. primulinum which age 14 weeks after pollination. The research use ½ Murashige and Skoog media (MS) with four different coconut water (CW) concentration (0,10%, 20%, 30%) and incubated for 90 days. The seed germination rate is calculated due to the percentage of its total germinate seed and seed development indicator divided into 5 stages (0-5). The result shows that the seed of Paphiopedilum primulinum germinate best at 30% CW concentration and only in this concentration the seedling stage reach stage 5 (4.8%). This result implies that 30% CW concentration in ½ MS medium is the best medium to germinate this seed.

Ancient tea gardens play important role on in situ conservation of epiphytic orchids in southwest Yunnan, China

Southwest Yunnan of China is the homeland of famous Pu’er tea and preserved the largest area of ancient tea gardens in the world. The ancient tea gardens keeping a large number of shade trees have prevailed for hundreds of years persist and formed typical agroforestry systems. To assess the potential of the ancient tea gardens for in situ orchid conservation, in this study, three typical ancient tea gardens located at different elevations, JiNuo Mountain ancient tea garden (JNM, 1150 m asl.), JingMai Mountain ancient tea garden (JMM, 1345 m asl.), and LaoBanZhang ancient tea garden (LBZ, 1787 m asl.), were selected to investigate species diversity and composition, and conservation status of all orchids. Floral visitors, reproductive success, and seedling recruitment of selected epiphytic orchids were also investigated from 2019 to 2021. We identified 88 orchid species (85 epiphytic and 3 terrestrial orchids) in 33 genera in the three ancient tea gardens, including one critically endangered species (CR), four endangered species (EN) and 16 vulnerable species (VU). Although the orchid species diversity was not significantly different, the species composition varied greatly among the three ancient tea gardens. For the randomly selected 40 epiphytic orchid species, all species could set fruits naturally, and the natural fruit-sets ranged from 1.11 ± 6.0% to 62.07 ± 15.4% with great variations among species and years. For 60 epiphytic orchid species observed, seedling recruitment for 26 species (43.3%) were observed and confirmed. Among 42 insect visitors observed, 21 species including bees, flies and ants, were confirmed as effective pollinators to 16 epiphytic orchids. These results revealed that the ancient tea gardens maintained high species diversity of epiphytic orchids and provided pollinator services for reproductive success and suitable micro-environment for seed germination and seedling recruitment. We strongly suggested that the ancient tea gardens in the area could serve as important supplements for in situ conservation of epiphytic orchids.

In vitro asymbiotic propagation of the vulnerable slipper orchid Cypripedium cordigerum D. Don

This paper presents an attempt to establish a protocol for the conservation of Cypripedium cordigerum D. Don a vulnerable species of orchid using in vitro symbiotic seed germination. The suitability of four orchids seed germination media [Terrestrial orchid medium (BM), modified terrestrial orchid seed germination medium (BM-1), Malmgren modified terrestrial orchid medium (MM), Knudson C medium (KC)] was tested using different photoperiods i.e. 24-h dark or 12-h light per day. The seed capsules were harvested at two different stages of development in order to determine the effect of capsule maturity on seed germination. The maximum percentage germination of seed from intact capsules in the dark was 65.00 ± 0.12% on the BM-1 medium. Release of brownish exudates was stopped by the addition of activated charcoal to the cultures. Seedlings developed in 26.17 ± 0.17 weeks. The current study is the first to report the ex-situ conservation of C. cordigerum.

Prioritizing conservation of terrestrial orchids: A gap analysis for Italy

Protected areas (PAs) are a strategic tool for biodiversity conservation, and conservation planning approaches are used to optimize PAs capacity to preserve specific target groups. Orchidaceae is one of the most threatened plant families, as most species are vulnerable to habitat changes because of their strong ecological specialization. Italy plays a key role in biogeography as a result of its geographical position and hosts one of the most diverse orchid floras in the Mediterranean Basin. The aim of this work is to depict the degree of protection granted to orchids by the current network of PAs across the entire Italian country, testing whether distributional and ecological features affect species' conservation representativeness, and identifying a priority list of species to be further protected. We compiled a dataset comprising 71,693 occurrence records, the spatial conservation representativeness was calculated as the percent of occurrences falling within the borders of PAs. Generalized Linear Models were run to assess differences in the spatial conservation representativeness among species according to the preferred habitat, endemicity, chorology, and protection by the Habitats Directive. We produced a list of species to be used for prioritizing conservation planning. Our findings show that emphasis is needed on adopting orchid species linked to primary or well mature habitats to select additional PAs with high conservation significance. Our findings reiterate the urgency of targeted conservation actions that can protect orchids and prevent their decline.