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Pollination of the pantropical Vanilla has been linked to melittophily and food deception. Here we investigated the role of flower traits on the reproduction of Neotropical Vanilla. We also studied the evolution of pollination systems in order to understand the origin of production of flower resources and the diversification of pollinators in this orchid genus. Our study was founded on data of adaptations in flower morphology, production of resources, scent release, pollinators and breeding systems of Vanilla and presenting new data on reproductive biology of V. palmarum. Data on reproductive biology of Vanilla were mapped onto a phylogeny to address our queries on the evolution of pollination systems in this genus. Vanilla palmarum shows a mixed mating system, with its facultative autogamous flowers being pollinated by hummingbirds. Its yellow flowers are scentless and produces nectar. Mapping of the pollination system onto trees resulted in one origin for bird pollination and at least two origins for autogamy in Vanilla. Nectar secretion has a single origin in the Neotropical thick-leafed lineage. Bird pollination of Vanilla is shown for the first time. The origin of ornithophily within a bee-pollinated clade is supported by flower morphology. Floral transitions to ornithophily have been favoured by the occupation of a distinct niche from that of the other thick-leafed Vanilla species. Despite its specialized pollination, V. palmarum is autogamous. A mixed mating system can promote reproductive assurance in the case of a decline in pollinator populations, or in areas where pollinator services are irregular or absent.

A new variety, Lecanorchis japonica var. insularis (Orchidaceae: Vanilloideae), a mycoheterotrophic orchid from Jejudo Island, Korea, is described and illustrated. It usually grows with L. japonica var. japonica, L. japonica var. hokurikuensis and L. kiusiana in evergreen forests. The newly described variety is distinguished from L. japonica vars. japonica, hokurikuensis, and kiiensis by having fully opened, larger flowers and an obtuse labellum apex.

We confirm the occurrence of Vanilla hartii Rolfe in Brazil. This species was collected in FLONA Carajás, Amazonia, at the municipality of Parauapebas, state of Pará. This work contributes to the knowledge of the geographical distribution of this rare vanilla.

To elucidate the taxonomy of the Lecanorchis nigricans Honda, 1931 species complex, the present study investigated the detailed morphology of three L. nigricans varieties in Japan. While L. nigricans var. patipetala Y.Sawa, 1980 and L. nigricans var. yakusimensis T.Hashim., 1990 have often been treated as synonyms of L. nigricans var. nigricans, the present study demonstrates that the three varieties are morphologically distinct. More specifically, L. nigricans var. nigricans only produces complete cleistogamous flowers and is distinct from the plants currently called “L. nigricans”, which are identical to the chasmogamous variety L. nigricans var. patipetala. The other chasmogamous variety L. nigricans var. yakusimensis can be easily distinguished from L. nigricans var. patipetala by its more spatulate tepals and higher cucullate lip. Therefore, the present study provides emended description of the three L. nigricans varieties based on type specimens and specimens collected from type localities. In addition, the isotype specimen of L. nigricans var. patipetala is designated as the lectotype because the holotype has been lost.

WebComparing the whole genome sequence of Apostasia shenzhenica with transcriptome and genome data from five orchid subfamilies permits the reconstruction of an ancestral gene toolkit, providing insight into orchid origins, evolution and diversification. Supplementary information The online version of this article (doi:10.1038/nature23897) contains supplementary material, which is available to authorized users., Orchid origins Around 10 per cent of flowering plant species are orchids, with a broad diversity in both morphology and lifestyle. Apostasia is one of the earliest-diverging genera of Orchidaceae. To study the evolution and diversity of Orchidaceae, Zhong-Jian Liu, Yves Van de Peer and colleagues sequenced the genome of Apostasia shenzhenica, a self-pollinating species found in southeast China. The authors also report improved genomes for two species of Epidendroideae, Phalaenopsis equestris and Dendrobium catenatum, as well as transcriptome analysis of representatives of subfamilies of Orchidaceae. Their analyses provide insights into orchid origins, genome evolution, adaptation and diversification. Supplementary information The online version of this article (doi:10.1038/nature23897) contains supplementary material, which is available to authorized users., Constituting approximately 10% of flowering plant species, orchids (Orchidaceae) display unique flower morphologies, possess an extraordinary diversity in lifestyle, and have successfully colonized almost every habitat on Earth1,2,3. Here we report the draft genome sequence of Apostasia shenzhenica4, a representative of one of two genera that form a sister lineage to the rest of the Orchidaceae, providing a reference for inferring the genome content and structure of the most recent common ancestor of all extant orchids and improving our understanding of their origins and evolution. In addition, we present transcriptome data for representatives of Vanilloideae, Cypripedioideae and Orchidoideae, and novel third-generation genome data for two species of Epidendroideae, covering all five orchid subfamilies. A. shenzhenica shows clear evidence of a whole-genome duplication, which is shared by all orchids and occurred shortly before their divergence. Comparisons between A. shenzhenica and other orchids and angiosperms also permitted the reconstruction of an ancestral orchid gene toolkit. We identify new gene families, gene family expansions and contractions, and changes within MADS-box gene classes, which control a diverse suite of developmental processes, during orchid evolution. This study sheds new light on the genetic mechanisms underpinning key orchid innovations, including the development of the labellum and gynostemium, pollinia, and seeds without endosperm, as well as the evolution of epiphytism; reveals relationships between the Orchidaceae subfamilies; and helps clarify the evolutionary history of orchids within the angiosperms. Supplementary information The online version of this article (doi:10.1038/nature23897) contains supplementary material, which is available to authorized users.

The number of integuments found in angiosperm ovules is variable. In orchids, most species show bitegmic ovules, except for some mycoheterotrophic species that show ovules with only one integument. Analysis of ovules and the development of the seed coat provide important information regarding functional aspects such as dispersal and seed germination. This study aimed to analyze the origin and development of the seed coat of the mycoheterotrophic orchid Pogoniopsis schenckii and to compare this development with that of other photosynthetic species of the family. Flowers and fruits at different stages of development were collected, and the usual methodology for performing anatomical studies, scanning microscopy, and transmission microscopy following established protocols. P. schenckii have ategmic ovules, while the other species are bitegmic. No evidence of integument formation at any stage of development was found through anatomical studies. The reduction of integuments found in the ovules could facilitate fertilization in this species. The seeds of P. schenckii, Vanilla planifolia, and V. palmarum have hard seed coats, while the other species have seed coats formed by the testa alone, making them thin and transparent. P. schenckii, in contrast to the other species analyzed, has a seed coat that originates from the nucellar epidermis, while in other species, the seed coat originates from the outer integument.

Orchidaceae (orchids) is the largest family in the monocots, including about 25,000 species in 880 genera and five subfamilies. Many orchids are highly valued for their beautiful and long-lasting flowers. However, the phylogenetic relationships among the five orchid subfamilies remain unresolved. The major dispute centers on whether the three one-stamened subfamilies, Epidendroideae, Orchidoideae, and Vanilloideae, are monophyletic or paraphyletic. Moreover, structural changes in the plastid genome (plastome) and the effective genetic loci at the species-level phylogenetics of orchids have rarely been documented. In this study, we compared 53 orchid plastomes, including four newly sequenced ones, that represent four remote genera: Dendrobium, Goodyera, Paphiopedilum, and Vanilla. These differ from one another not only in their lengths of inverted repeats and small single copy regions but also in their retention of ndh genes. Comparative analyses of the plastomes revealed that the expansion of inverted repeats in Paphiopedilum and Vanilla is associated with a loss of ndh genes. In orchid plastomes, mutational hotspots are genus specific. After having carefully examined the data, we propose that the three loci 5′trnK-rps16, trnS-trnG, and rps16-trnQ might be powerful markers for genera within Epidendroideae, and clpP-psbB and rps16-trnQ might be markers for genera within Cypripedioideae. After analyses of a partitioned dataset, we found that our plastid phylogenomic trees were congruent in a topology where two one-stamened subfamilies (i.e., Epidendroideae and Orchidoideae) were sisters to a multi-stamened subfamily (i.e., Cypripedioideae) rather than to the other one-stamened subfamily (Vanilloideae), suggesting that the living one-stamened orchids are paraphyletic.

Orchidaceae constitutes one of the largest families of angiosperms. Owing to the significance of orchids in plant biology, market needs and current sustainable technology levels, basic research on the biology of orchids and their applications in the orchid industry is increasing. Although chloroplast (cp) genomes continue to be evolutionarily informative, there is very limited information available on orchid chloroplast genomes in public repositories. Here, we report the complete cp genome sequence of Dendrobium nobile from Northeast India (Orchidaceae, Asparagales), bearing the GenBank accession number KX377961, which will provide valuable information for future research on orchid genomics and evolution, as well as the medicinal value of orchids. Phylogenetic analyses using Bayesian methods recovered a monophyletic grouping of all Dendrobium species (D. nobile, D. huoshanense, D. officinale, D. pendulum, D. strongylanthum and D. chrysotoxum). The relationships recovered among the representative orchid species from the four subfamilies, i.e., Cypripedioideae, Epidendroideae, Orchidoideae and Vanilloideae, were consistent within the family Orchidaceae.

Monophyllorchis idroboi Szlach., S. Nowak & Baranow sp. nova and M. chocoensis Szlach., S. Nowak & Baranow sp. nova (Orchidaceae, Vanilloideae) from Colombia are described and illustrated on the basis of collections deposited at the National Colombian Herbarium in Bogota. Diagnostic characters of the new taxa are compared with their supposedly closest relatives. A key to the Colombian species of Monophyllorchis and a map showing the distribution of the new species and of the whole genus are presented.

A new species of the mycoheterotrophic genus Lecanorchis Blume (Orchidaceae), L. moritae, is described from Amami-Oshima Island, Japan. It is most similar to L. suginoana but differs in having a transversely elliptic (slightly contracted at base) midlobe (vs. semiorbicular, not contracted at base), more triangular with almost entire margins (vs. relatively rounded, denticulate margins) and reddish apices (vs. somewhat whitish apices) and a lip with white and purple hairs (vs. white and lemon yellow hairs) and sparse unicellular papillae scattered on the disc (vs. dense unicellular papillae). An illustration and nuclear ribosomal internal transcribed spacer (nrITS) sequence (i.e., DNA barcode) of the new species are also provided.

Notes on Cleistes tenuis are presented, including a detailed description, distribution, and herbarium records. This species is particularly interesting because flowers at full anthesis and preserved in alcohol differ so dramatically from those drawn based on herbarium material and descriptions in the literature, that “two species” easily could be distinguished. Differential drying and shrinking in the labellum, however, account for differences in flowers and in this case are referable to the same species.

Two new species of Neotropical Orchidaceae, Epistephium epiphyticum Szlach. & Kolan. and E. cardonae Szlach. & Kolan., are described and illustrated based on Colombian material. So far the new entities are known exclusively from the type locations. Both species are placed within a key to Colombian Epistephium Kunth.

Szlachetko, D. L. & P. Baranow (2013). New species of Palmorchis Barb. Rodr. (Orchidaceae) from Colombia. Candollea 68: 105–113. In English, English and French abstract. Five new species of Palmorchis Barb. Rodr. (Orchidaceae, Vanilloideae) from Colombia, Palmorchis carlos-parrae Szlach. & Baranow, Palmorchis fractiflexa Szlach. & Baranow, Palmorchis maculata Szlach. & Baranow, Palmorchis misas-urretae Szlach. & Baranow and Palmorchis valdiviesoana Szlach. & Baranow, are described and illustrated. Diagnostic characters of the new taxa are compared with the closely related species in Colombia and an identification key is proposed. Ecology, habitat and distribution data for these taxa are also given.

The morphological study of the herbarium material representing Epistephium (Orchidaceae, Vanilloi- deae) led to the discovery of two groups of specimens that significantly differ from all known species of the genus. The results of literature data study and of comparative analysis of those and other specimens suggest that these collections represent new taxa that we describe as E. garayi and E. kubiyuense. The distinguishing features of the spe- cies are indicated. As both new species are reported from Colombia (E. garayi, also from Guyana), the key for the determination of all Colombian representatives of the genus is included. Information on the ecology and distri- bution of newly described taxa is presented.

The genus Vanilla is the most diverse in Vanilloideae, with ca 90 species distributed among tropical regions. Despite their economic importance, studies on pollination of Vanilla are very scarce and data on pollinators of species endemic to Brazil are lacking. Based on fieldwork and laboratory investigations, the floral biology of V. edwallii was studied. The pollinators and pollination process were recorded at the Serra do Japi reserve, state of Sao Paulo, southeastern Brazil, and the presence of floral reward was also investigated. Vanilla edwallii blooms in summer. The lateral inflorescences produce up to four pale green flowers. The white labellum is united to the base of the column forming a mentum. In the studied population V. edwallii is pollinated by Epicharis (Hoplepicharis) affinis, where the males exhibit a territorial behavior, defending flowers from other possible flower visitors. The pollen is deposited on the scutellum of bees when they abandon the flower. The mentum region is dry, suggesting no nectar production. The only secretory structures are osmophores dispersed on the inner surface of the lip responsible for production of a sweet fragrance, which together with color and morphology of flowers is related to bee attraction. The labellum is rich in mucilaginous cells, while the mucilaginous substance is retained inside the cells. The histochemical analysis also detected the presence of phenolic compounds and starch concentrated mainly at the adaxial surface of the lip and around the vascular bundles.

Irrespective of the fact that Borneo is one of the richest areas for mycoheterotrophic plants, only two species of the genus Lecanorchis Blume, a mycoheterotrophic genus of Orchidaceae, have been reported from Borneo. On three occasions during recent botanical surveys in Betung Kerihun National Park, we found an undescribed species of Lecanorchis. Here, we provide a detailed morphological account of this new species, Lecanorchis betung-kerihunensis. We also provide a key to the species of Lecanorchis in Borneo.

The tribe Pogonieae of Vanilloideae (Orchidaceae) consists of six genera, including Pogoniopsis , a myco-heterotrophic taxon with morphological characteristics distinct from the remaining of the tribe. A hypothesis about the phylogeny of the tribe was inferred, involving all currently recognized genera, based on isolated and combined sequence data of 5.8S, 18S and 26S (nrDNA) regions using parsimony and Bayesian analyses. Phylogenetic analyses show that inclusion of Pogoniopsis turns the tribe Pogonieae paraphyletic. All analyses reveal that Pogoniopsis is closely related to members of Epidendroideae. The pantropical Vanilla is monophyletic if Dictyophyllaria is assumed as synonym of Vanilla . Members of Pogonieae are pollinated by several groups of solitary and social bees, two pollination systems being recognized: reward-producing and deceptive. The molecular phylogeny suggests that ancestrals related to Pogonieae gave rise to two evolutionary lines: a tropical one with reward production of flowers, and a predominantly temperate regions invading line with deceptive flowers. Reward-producing flowers characterize the South and Central American clade (= Cleistes ), while deceptive pollination is prominent in the clade that includes North American-Asiatic taxa plus the Amazonian genus Duckeella .

A new species, Lecanorchis tabugawaensis Suetsugu & Fukunaga, sp. nov. from Yakushima Island, Kagoshima Prefecture, Japan, is described and illustrated. Lecanorchis tabugawaensis is similar to Lecanorchis taiwaniana, but it is easily distinguished by the straight column, the glabrous status of the base of the column, the almost entire and narrow labellum morphology, the shorter part of the column fused with the labellum and the glabrous status of the apical part of the adaxial labellum surface. The new species appears to be restricted to two locations, each consisting of only dozens of mature individuals, and is assessed as Critically Endangered [CR D1] according to IUCN Red List Categories and Criteria.

Vanilloideae as currently circumscribed comprises nine genera and two tribes: Vanilleae and Pogonieae. The pantropical genus Vanilla has been frequently assumed to be natural on the basis of its climbing habit and lateral inflorescences. However, the inclusion of the rare Dictyophyllaria dietschiana in phylogenetic analyses makes the genus Vanilla paraphyletic. Within Pogonieae, phylogenetic analyses show that inclusion of Pogoniopsis turns the tribe paraphyletic. All analyses reveal that Pogoniopsis is closely related to members of Epidendroideae. Members of Pogonieae are pollinated by several groups of solitary and social bees, two pollination systems being recognized: reward-producing and deceptive. Molecular phylogeny suggests that the common ancestor to Pogonieae gave rise to two evolutionary lineages: one tropical with a condition of reward production; and one predominantly temperate-invading line with deceptive flowers. Reward-producing flowers characterize South and Central American clade (= Cleistes), while deceptive pollination is prominent in the clade including North American-Asiatic taxa plus Amazonian Duckeella. Species of “orchid bees” have been recorded as pollinators of the genus Vanilla (V. planifolia group and V. pompona group) in the Neotropics. In species of the V. pompona group, these bees are attracted by the fragrance of the flowers. Hummingbirds have been reported to pollinate some species of Vanilla. Vanilla insignis, V. odorata and V. planifolia are known to be pollinated through generalized food deception. Some species of Vanilla yield fruits through spontaneous self-pollination. This form of autogamy has been reported for V. griffithii, V. palmarum, V. planifolia, V. savannarum and V. bicolor. In Brazil, data on the pollination biology of Vanilla are scarce, but conclusive data are available for V. edwallii, which is pollinated by Epicharis (Apidae: Centridini). This species is rewardless, but male Epicharis are attracted to its flowers by their fragrance. Additionally, the Brazilian V. dubia and E. sclerophyllum are pollinated by bees. The mentum region of V. dubia and V. edwallii is dry, whereas that of E. sclerophyllum presents a small quantity of nectar. Flowers of E. sclerophyllum are scentless, while those of V. dubia are odoriferous. Vanilla dubia and V. edwallii are self-compatible and need a pollinator to yield fruits. In contrast, Epistephium sclerophyllum sets fruits through spontaneous self-pollination, but biotic pollination also occurs. Both species are primarily adapted to pollination by euglossine bees. Pollination by Euglossini seems to have evolved at least twice along the evolution of Vanilleae. Furthermore, shifts between rewarding and rewardless flowers and between autogamous and allogamous species have been reported among vanillas. Vanilloideae as currently circumscribed comprises nine genera and two tribes: Vanilleae and Pogonieae. The pantropical genus Vanilla has been frequently assumed to be natural on the basis of its climbing habit and lateral inflorescences. However, the inclusion of the rare Dictyophyllaria dietschiana in phylogenetic analyses makes the genus Vanilla paraphyletic. Within Pogonieae, phylogenetic analyses show that inclusion of Pogoniopsis turns the tribe paraphyletic. All analyses reveal that Pogoniopsis is closely related to members of Epidendroideae. Members of Pogonieae are pollinated by several groups of solitary and social bees, two pollination systems being recognized: reward-producing and deceptive. Molecular phylogeny suggests that the common ancestor to Pogonieae gave rise to two evolutionary lineages: one tropical with a condition of reward production; and one predominantly temperate-invading line with deceptive flowers. Reward-producing flowers characterize South and Central American clade (= Cleistes), while deceptive pollination is prominent in the clade including North American-Asiatic taxa plus Amazonian Duckeella. Species of “orchid bees” have been recorded as pollinators of the genus Vanilla (V. planifolia group and V. pompona group) in the Neotropics. In species of the V. pompona group, these bees are attracted by the fragrance of the flowers. Hummingbirds have been reported to pollinate some species of Vanilla. Vanilla insignis, V. odorata and V. planifolia are known to be pollinated through generalized food deception. Some species of Vanilla yield fruits through spontaneous self-pollination. This form of autogamy has been reported for V. griffithii, V. palmarum, V. planifolia, V. savannarum and V. bicolor. In Brazil, data on the pollination biology of Vanilla are scarce, but conclusive data are available for V. edwallii, which is pollinated by Epicharis (Apidae: Centridini). This species is rewardless, but male Epicharis are attracted to its flowers by their fragrance. Additionally, the Brazilian V. dubia and E. sclerophyllum are pollinated by bees. The mentum region of V. dubia and V. edwallii is dry, whereas that of E. sclerophyllum presents a small quantity of nectar. Flowers of E. sclerophyllum are scentless, while those of V. dubia are odoriferous. Vanilla dubia and V. edwallii are self-compatible and need a pollinator to yield fruits. In contrast, Epistephium sclerophyllum sets fruits through spontaneous self-pollination, but biotic pollination also occurs. Both species are primarily adapted to pollination by euglossine bees. Pollination by Euglossini seems to have evolved at least twice along the evolution of Vanilleae. Furthermore, shifts between rewarding and rewardless flowers and between autogamous and allogamous species have been reported among vanillas.&nbsp

Four new Colombian species of Cleistes (Orchidaceae, Vanilloideae) are described and illustrated: C. uribei Szlach. & Baranow, C. risoraldensis Szlach. & Baranow, C. munchiquensis Szlach. & Baranow and C. idroboi Szlach. & Baranow. Notes concerning their morphological distinctions, habitat, and distribution are provided. A key for determination of the Colombian species of Cleistes is included.

A new orchid species, Vanilla paludosa, native to marshy forests close to cerrado vegetation in the state of Sao Paulo, Brazil, is described and illustrated. The relationship of V. paludosa to other species of the genus is discussed.

This paper presents a re-evaluation of the taxonomic identities of Lecanorchis trachycaula and L. betung-kerihunensis. Consequently, L. trachycaula is reduced to a synonym of L. purpurea while L. betung-kerihunensis is treated as a synonym of L. malaccensis. Because no original material of L. purpurea is existent, we designate its neotype to stabilize its taxonomic status.

Cleistes pallida, a new species of Orchidaceae, is described and illustrated. The relationship of this new species to other taxa of the genus are discussed.

Lecanorchis Blume (1856: 188) comprises about 30 species of mycoheterotrophic orchids (Seidenfaden 1978, Hashimoto 1990, Szlachetko & Mytnik 2000, Govaerts et al. 2017) characterized by having numerous, long, thick, horizontal roots produced from a short rhizome, presence of a calyculus (i.e. a cup-like structure located between the base of the perianth and apex of the ovary) and an elongate column with a pair of small wings on each side of the anther (Seidenfaden 1978, Hashimoto 1990). The genus is distributed across a wide area including China, Korea, India, Indonesia, Japan, Laos, Malaysia, New Guinea, Pacific islands, the Philippines, Taiwan, Thailand and Vietnam (Seidenfaden 1978, Hashimoto 1990, Pearce & Cribb 1999, Szlachetko & Mytnik 2000, Averyanov 2011, 2013).

Four new Colombian species of Cleistes (Orchidaceae, Vanilloideae) are described and illustrated: C. uribei Szlach. & Baranow, C. risoraldensis Szlach. & Baranow, C. munchiquensis Szlach. & Baranow and C. idroboi Szlach. & Baranow. Notes concerning their morphological distinctions, habitat, and distribution are provided. A key for determination of the Colombian species of Cleistes is included.

A new genus, Cleistesiopsis, was segregated from Cleistes based on morphological and molecular characteristics, and two new species of Cleistes: C. batistana and C. elongata, both occurring in the Brazilian Central Plateau (Central-Western Brazil), are described and illustrated. Furthermore, a key to genera currently recognised within Pogonieae is presented.

Background and Aims Most fully mycoheterotrophic (MH) orchids investigated to date are mycorrhizal with fungi that simultaneously form ectomycorrhizas with forest trees. Only a few MH orchids are currently known to be mycorrhizal with saprotrophic, mostly wood-decomposing, fungi instead of ectomycorrhizal fungi. This study provides evidence that the importance of associations between MH orchids and saprotrophic non-Rhizoctonia fungi is currently under-estimated. Methods Using microscopic techniques and molecular approaches, mycorrhizal fungi were localized and identified for seven MH orchid species from four genera and two subfamilies, Vanilloideae and Epidendroideae, growing in four humid and warm sub-tropical forests in Taiwan. Carbon and nitrogen stable isotope natural abundances of MH orchids and autotrophic reference plants were used in order to elucidate the nutritional resources utilized by the orchids. Key Results Six out of the seven MH orchid species were mycorrhizal with either wood- or litter-decaying saprotrophic fungi. Only one orchid species was associated with ectomycorrhizal fungi. Stable isotope abundance patterns showed significant distinctions between orchids mycorrhizal with the three groups of fungal hosts. Conclusions Mycoheterotrophic orchids utilizing saprotrophic non-Rhizoctonia fungi as a carbon and nutrient source are clearly more frequent than hitherto assumed. On the basis of this kind of nutrition, orchids can thrive in deeply shaded, light-limiting forest understoreys even without support from ectomycorrhizal fungi. Sub-tropical East Asia appears to be a hotspot for orchids mycorrhizal with saprotrophic non-Rhizoctonia fungi.

Nucleotide sequences of the plastid genes psbB and psbC were obtained for 34 taxa to represent most genera currently classified within Vanilloideae (Orchidaceae). These genes code for two of the subunits that make up the Photosystem II protein P680, and have only rarely been used for reconstructing phylogenetic relationships among plants. We failed to amplify psbB from the achlorophyllous genera Cyrtosia and Lecanorchis, but were able to align full length copies of psbC sequences from two species of Cyrtosia with the other taxa. This was not the case for an anomalous psbC sequence obtained for Lecanorchis. Nucleotide variation within each of these genes is sufficient to resolve the major relationships among Vanilloideae, and the combined two-gene tree is fully resolved at the genus level and highly supported. These gene trees demonstrate with a high degree of confidence (95% jackknife support) that a clade of mostly achlorophyllous tropical vines including Pseudovanilla and Erythrorchis are the sister group to Vanilla. The two New Caledonian endemic genera of Vanilloideae are sister to this pair, and Epistephium is sister to all remaining Vanillieae.

Parsimony analyses of DNA sequences from the plastid genes atpB and rbcL were completed for 173 species of Orchidaceae (representing 150 different genera) and nine genera from outgroup families in Asparagales. The atpB tree topology is similar to the rbcL tree, although the atpB data contain less homoplasy and provide greater jackknife support than rbcL alone. In combination, the two-gene tree recovers five monophyletic clades corresponding to subfamilies within Orchidaceae, and fully resolves them with moderate to high jackknife support as follows: Epidendroideae are sister to Orchidoideae, followed by Cypripedioideae, then Vanilloideae, and with Apostasioideae sister to the entire family. Although this two-gene hypothesis of orchid phylogeny is an improvement over all single-gene studies published to date, there is still no consensus as to how all the tribes of Epidendroideae are related to one another. Nevertheless, these new topologies help to clarify some of the anomalous results recovered when rbcL was previously analyzed alone, and demonstrate the value of continued plastid gene sequencing within Orchidaceae.

The chromosome numbers of forty-one Brazilian species belonging to 11 genera of preferentially terrestrial orchids (subfamilies Cypripedioideae, Spiranthoideae, Orchidoideae, and Vanilloideae) were examined. Previous records for these subfamilies were reviewed in order to identify the ancestral chromosome numbers of terrestrial orchids. The variation observed within the subfamilies Spiranthoideae (2n=28, 36, 46, 48 and 92), and Orchidoideae (2n=42, 44, ca. 48, ca. 80, 84, and ca. 168) was similar to that previously reported in the literature. In the subfamily Spiranthoideae, some species of Prescottia (subtribe Prescottiinae) and some genera of Spiranthinae showed a bimodal karyotype with one distinctively large pair of chromosomes. The analysis of chromosome numbers of the genera in subfamilies revealed the predominance of the polyploid series 7, 14, 21, 28, 42 with a dysploid variation of ±1 in each ploidy level. These results suggest that the basic chromosome number of terrestrial orchids is x1=7 for the subfamilies Spiranthoideae and Orchidoideae, as well as other Epidendroid orchids, and that the majority of the genera are composed of palaeopolyploids.

An expanded plastid DNA phylogeny for Orchidaceae was generated from sequences of rbcL and matK for representatives of all five subfamilies. The data were analyzed using equally weighted parsimony, and branch support was assessed with jackknifing. The analysis supports recognition of five subfamilies with the following relationships: (Apostasioideae (Vanilloideae (Cypripedioideae (Orchidoideae (Epidendroideae))))). Support for many tribal-level groups within Epidendroideae is evident, but relationships among these groups remain uncertain, probably due to a rapid radiation in the subfamily that resulted in short branches along the spine of the tree. A series of experiments examined jackknife parameters and strategies to determine a reasonable balance between computational effort and results. We found that support values plateau rapidly with increased search effort. Tree bisection-reconnection swapping in a single search replicate per jackknife replicate and saving only two trees resulted in values that were close to those obtained in the most extensive searches. Although this approach uses considerably more computational effort than less extensive (or no) swapping, the results were also distinctly better. The effect of saving a maximal number of trees in each jackknife replicate can also be pronounced and is important for representing support accurately.

Many plants take up soluble monosilicic acid from the soil. Some of these plants subsequently deposit it as cell inclusions of characteristic structure. This article describes the distribution and diversity of opaline silica bodies in monocotyledons in a phylogenetic framework, together with a review of techniques used for their examination, and the ecology, function and economic applications of these cell inclusions. There are several different morphological forms of silica in monocot tissues, and the number of silica bodies per cell may also vary. The most common type is the “druse-like” spherical body, of which there is normally a single body per cell, more in some cases. Other forms include the conical type and an amorphous, fragmentary type (silica sand). Silica bodies are most commonly found either in the epidermis (e.g., in grasses, commelinas and sedges) or in the sheath cells of vascular bundles (e.g., in palms, bananas and orchids). Silica-bearing cells are most commonly associated either with subepidermal sclerenchyma or bundle-sheath sclerenchyma. Silica bodies are found only in orchids and commelinids, not in other lilioid or basal monocots. In orchids, silica bodies are entirely absent from subfamilies Vanilloideae and Orchidoideae and most Epidendroideae but present in some Cypripedioideae and in the putatively basal orchid subfamily Apostasioideae. Among commelinid monocots, silica bodies are present in all palms, Dasypogonaceae and Zingiberales but present or absent in different taxa of Poales and Commelinales, with at least four separate losses of silica bodies in Poales.

The anther is the source of several of the principal characters traditionally used for classification in Orchidaceae, especially in Epidendroideae. In particular, anther and pollinium orientation are important, but little-studied characters; it is not known whether various instances of these states are achieved in the same way. We examined variation in anther and pollinium orientation in Epidendroideae and Vanilloideae using anatomical sections of columns at successive ontogenetic stages and related the observed changes to the final anther morphology to reexamine our homology hypotheses for these characters. Anther bending in vanilloids is achieved primarily by massive expansion of the connective. In nonvandoid epidendroids it is the result of column elongation and tipping of the mature anther, while in vandoids it is due to a redirection of growth in very early ontogenetic stages. Superposed pollinia result from a reorientation of developing thecae; those is Sobralia result from incurving of the thecae, while vandoids have thecae that are directed outward (latrorse). Contrasting ontogenetic and phylogenetic patterns suggest heterochronic changes in these characters, especially in vandoids. These observations allow us to further refine character states that are crucial to our understanding of orchid relationships.

This paper presents a re-evaluation of the taxonomic relationships of Lecanorchis flavicans and L. flavicans var. acutiloba. Based on the comparative study of morphology and type materials, L. flavicans is lectotypified and reduced to a synonym of L. triloba, while L. flavicans var. acutiloba is treated as a synonym of L. vietnamica.

Vanilloideae comprises 15 genera distributed worldwide, among which are Vanilla and Epistephium (tribe Vanilleae). Based on field and laboratory investigations, the pollination biology of V. dubia and E. sclerophyllum was analysed. The former was surveyed in a semi-deciduous mesophytic forest at the biological reserve of Serra do Japi and in a marshy forest at the city of Pradópolis, southeastern Brazil. The latter was examined in rocky outcrop vegetation in the Chapada Diamantina, northeastern Brazil. In the studied populations, the tubular flowers of V. dubia and E. sclerophyllum were pollinated by bees. Pollen was deposited on either their scutellum (V. dubia) or scutum (E. sclerophyllum). The mentum region of V. dubia is dry, whereas that of E. sclerophyllum presents a small quantity of dilute nectar. Flowers of E. sclerophyllum are scentless, while those of V. dubia are odoriferous. Although V. dubia is self-compatible, it needs a pollinator to produce fruit. In contrast, E. sclerophyllum sets fruit through spontaneous self-pollination, but biotic pollination also occurs. Both species are primarily adapted to pollination by euglossine bees. Pollination by Euglossina seems to have occurred at least twice during the evolution of Vanilleae. Furthermore, shifts between rewarding and reward-free flowers and between autogamous and allogamous species have been reported among vanillas.

A new orchid species, Vanilla arcuata, native from costal plane (“restinga” vegetation) of the Atlantic rainforests in the State of São Paulo, Brazil is described and illustrated. The relationship of V. arcuata with other related taxa of the genus is discussed.

The genus Lecanorchis Blume (1856: 188) comprises a group of mycoheterotrophic species that grow in multiple clusters with an erect, branched or unbranched stem (Hashimoto 1990, Sawa et al. 2006, Seidenfaden 1978, Szlachetko & Mytnik 2000). The key characteristic of the species of Lecanorchis is the presence of a calyculus, a cup-like structure between the base of the perianth and the apex of the ovary (Cameron 2003, Hashimoto 1990, Sawa et al. 2006). There are about thirty species and/or varieties in the genus Lecanorchis extending across a large geographical area that includes Thailand, Malaysia, Indonesia, Vietnam, the Philippines, Taiwan, Japan, and New Guinea (Hashimoto 1990, Seidenfaden 1978, Su 2000, Szlachetko & Mytnik 2000, Cameron 2003, Averyanov 2011). Precise identification of the Lecanorchis species is often difficult due to close similarities in morphology and the short durations of flowering periods (Hashimoto 1990, Averyanov 2005, Suddee & Pedersen 2011, Tsukaya & Okada 2013). Furthermore, detailed descriptions are lacking for some species, particularly those described in decades past. Given such difficulties in precise identification, adequate taxonomic studies of this genus have not been conducted.

Arethusa bulbosa, Calopogon tuberosus, and Pognia ophioglossoides are self-compatible but adapted to outcrossing. The flowers produce little or no reward and are probably pollinated primarily by deceit. Fruit production is pollen limited. Pollinators are bees apparently attracted to the large, colorful perianths and ultraviolet absorbing, anther-like brushes on the lips. Larger flowers of C. tuberosus discriminate among pollinators and prevent hybridization with three smaller, ­sympatric species. Reproductive isolation among the smaller flowered taxa is maintained primarily by differences in phenology, habitat, and pollinator preference. The flowers of C. oklahomensis are similar in size to those of C. tuberosus, but this species is hexaploid and blooms earlier.

Xylem of the orchids studied provided unusually favorable material to demonstrate how conductive tissue evolves in monocotyledons. In the end walls of tracheary elements of many Orchidaceae, remnants of pit membranes were observed with scanning electron microscopy and minimally destructive methods. The full range from tracheids to vessel elements, featuring many intermediate stages, was illustrated with SEM in hand sections of fixed roots, stems, and inflorescence axes of 13 species from four subfamilies. Pit membranes in end walls of tracheary elements are porose to reticulate in roots of all species, but nonporose in stems of Cypripedioideae and Vanilloideae and porose to reticulate in stems of Orchidoideae and Epidendroideae. The distribution pattern of pit membranes and pit membrane remnants in end walls of tracheary elements of orchids parallels the findings of others. The position of Cypripedioideae and Vanilloideae as outgroups to Orchidoideae and Epidendroideae, claimed by earlier authors, is supported by clades based on molecular studies and by our studies. Little hydrolysis of pit membranes in tracheary element end walls was observed in pseudobulbs or inflorescence axes of epidendroids. The pervasiveness of network-like pit membranes of various extents and patterns in end walls of tracheary elements in Orchidaceae calls into question the traditional definitions of tracheids and vessel elements, not merely in orchids, but in angiosperms at large. These two concepts, based on light microscope studies, are blurred in light of ultrastructural studies. More importantly, the intermediate expressions of pit membranes in tracheary element end walls of Orchidaceae and some other families of angiosperms are important as indicators of steps in evolution of conduction with respect to organs (more rapid flow in roots than in succulent storage structures) and habitat (less obstruction to flow correlated with a shift from terrestrial to epiphytic).

The phylogeny and the biogeographical history of the genus Vanilla was investigated using four chloroplastic genes (psbB, psbC; psaB and rbcL), on 47 accessions of Vanilla chosen from the ex situ CIRAD collection maintained in Reunion Island and additional sequences from GenBank. Bayesian methods provided a fairly well supported reconstruction of the phylogeny of the Vanilloideae sub-family and more particularly of the genus Vanilla. Three major phylogenetic groups in the genus Vanilla were differentiated, which is in disagreement with the actual classification in two sections (Foliosae and Aphyllae) based on morphological traits. Recent Bayesian relaxed molecular clock methods allowed to test the two main hypotheses of the phylogeography of the genus Vanilla. Early radiation of the Vanilla genus and diversification by vicariance consecutive to the break-up of Gondwana, 95 million years ago (Mya), was incompatible with the admitted age of origin of Angiosperm. Based on the Vanilloideae age recently estimated to 71 million years ago (Mya), we conclude that the genus Vanilla would have appeared approximately 34 Mya in South America, when continents were already separated. Nevertheless, whatever the two extreme scenarios tested, at least three long distance migration events are needed to explain the present distribution of Vanilla species in tropical areas. These transoceanic dispersions could have occurred via transoceanic passageway such as the Rio Grande Ridge and the involvement of floating vegetation mats and migratory birds.

The orchid subfamily Apostasioideae consists of two genera, Apostasia and Neuwiedia. To study the position of Apostasioideae within Orchidaceae and their intra- and intergeneric rela- tionships, a molecular phylogenetic analysis has been conducted on the nuclear ITS region and the two plastid DNA regions trnL-F intron and matK. The two genera traditionally ascribed to Aposta- sioideae are each monophyletic. In Apostasia, A. nuda, with two stamens and no staminode, is sister to a clade comprising three species charac- terised by two stamens and one staminode. Within Neuwiedia, maximum parsimony analyses place N. zollingeri as sister to the clade formed by N. borneensis and N. veratrifolia. A family-wide phylogenetic analysis of matK sequences represent- ing all proposed subfamilies of Orchidaceae pro- duced five moderately to well-supported clades. One of these clades, Apostasioideae, is sister to the clade formed by Vanilloideae, Cypripedioideae, Orchidoi- deae and Epidendroideae. High transition-transver- sion ratio and the absence of stop codons in the individual sequences suggest that matK is at the transition from a possibly functional gene to a pseudogene in Apostasioideae, contrary to what is found in some other groups of Orchidaceae.

Introduction The genus Cleistes Rich. ex Lindl. (Orchidaceae: Vanilloideae: Pogoniinae) comprises about 30 species distributed throughout Central and South America. According to Cameron & Chase (1999) and Pansarin et al. (unpubl. res.), Cleistes is paraphyletic. The North American species C. divaricata and C. bifaria are closer to the North American Pogoniinae genera Isotria and Pogonia than to the Central and South American Cleistes. The Central and South American species of Cleistes form a natural group (Cameron & Chase 1999). They are found in several habitats including disturbed areas on road margins, "campo limpo" (open grasslands), "campo sujo" (grassland with shrubs), "cerrado" (wooded savanna), "vereda" vegetation (wet grassland) in the Central Plateau, "restinga" (open seashore vegetation on sandy soil), sandy rocky soils or among rocks in "campos rupestres" (vegetation of rocky outcrops), marshy areas and "campos de altitude" (high mountain grasslands). This new species, Cleistes pusilla, was found while preparing a taxonomic revision of the genus Cleistes.