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Phylogeny of the Pleurothallidinae. The single most-parsimonious, successively weighted, gap-coded matK/trnL/ ITS DNA data set of Pridgeon et al. (2001) but with revised epithets. Number above branches are Fitch lengths, and those below branches are equally weighted bootstrap percentages >50%. Letters indicate the main clades according to Pridgeon et al. (2001); * indicates the clade included in the recent circumscription of the subtribe that previously belonged to Laeliinae. Outgroups are represented by Arpophyllum giganteum and Isochilus amparoanus. Reproduced from Pridgeon & Chase (2001) with permission.
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Desarrollamos un proyecto dirigido a determinar, a traves de polinizacion experimental, la auto- incompatibilidad de los generos representativos de los linajes mas importantes de Pleurothallidinae, buscando el grupo donde posiblemente aparecio por primera vez y cuantas veces ha ocurrido desde entonces. Adicionalmente estudiamos la biologia floral d...
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... by Diptera. The species are characterized morphologically by the absence of pseudobulbs and the presence of an articulation between the ovary and the pedicel. An exception is seen in the clade composed by three small genera from Central America (Dilomilis, Neocogniauxia, and Tomzanonia), currently included in the subtribe Pridgeon et al., , 2005; (Fig. 1). In recent molecular phylogenetic analyses, in spite of being a monophyletic and easily recognizable group, its original circumscription was enlarged to include that small clade (a total of only eight species) of bird-pollinated, self-compatible species that previously were placed in Laeliinae (Pridgeon et al., 2001). In the combined ...
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... were able to maintain high levels of genetic variability. This hypothetical evolutionary scenario could be weakened if we found basal mellitophilous species (before the advent of myophily) with self-incompatibility or myophilous taBLE 1. List of taxa examined of representative genera of the main clades of Pleurothallidinae (according to ; see Fig. 1). All sampled species in this work are native to Brazil. n= sample ...
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... systems -We sampled 22 species in eight genera belonging to seven of the main clades of Pleurothallidinae as defined by ; Fig. 1, Table 1): Acianthera (six species), Anathallis (four spp.), Masdevallia (one), Myoxanthus (one), Octomeria (four), Specklinia (two), Stelis (three), and Zootrophion (one). It was not possible to sample clade G, because we could not obtain individuals of Phloeophila species, the only genus of this lineage. ...
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... FAA. Approximately 300 seeds from each fruit were examined by optical microscopy and classified as viable or non-viable on a morphological basis only, according to the morphology and relative size of the embryo: seeds with well-developed embryos were considered viable, and seeds with no embryo or a rudimentary embryo were considered inviable ( Fig. 3A; Borba et al., 2001a). Here we present only a summary description of the pollination of the species with the identity of pollinators, because a detailed description of the reproductive biology of the species is being published elsewhere ( Barbosa et al., ...
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... biology of Octomeria species-Octomeria crassifolia was pollinated exclusively by both males and females in similar proportions of four species of Bradysia (Diptera, Sciaridae; Fig. 1G). During 63 hours of observation, a total of 92 visits were recorded and 31 pollinarium removals and six pollinarium depositions were observed. The visits occurred more frequently early in the morning, between 06:00 and 08:00. Pollinators of O. grandiflora were rarely observed in flowers of O. crassifolia. However, pollination did not ...
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... in some species, as observed in a previous study in Acianthera (Borba et al., 2001a), fruit set in self-pollination was about the half that observed in cross-pollination experiments. As emphasized by Borba et al. (2001a), strict self-incompatibility and selfcompatibility are extremes of a continuum between which there is often no clear-cut difference, and relatively few species fit exactly in these extremes. ...
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... diaphana, O. wawrae, Myoxanthus exasperatus, M. punctatus, and additional Acianthera and Specklinia species. However, the results were not included here because of the still low sample size. Occurrence of self-incompatibility in Myoxanthus species is particularly important to our study, since they represent a clade not sampled in this study (see Fig. ...
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... And even though, a recent review gathered new evidence of honest signaling in orchids across subfamilies and tribes that were previously assumed to be outright deceitful (which suggests that the rewardlessness prevalence among these plants might be overestimated: i.e. Shrestha et al. 2020, and information from nearly 2900 orchid species arround the world showed that pollinator attraction is mainly based on rewards i.e. 54%: Ackerman et al. 2023) the idea of rewardlessness as the most common pollination strategy in orchids is widespread (Ackerman 1986;Borba et al. 2011;Johnson and Schiestl 2016;Schiestl 2005). However, it is necessary to increase our knowledge of orchids' pollination systems to understand which strategies may be involved in attracting and keeping visitors. ...
Pollination by deception is assumed as the general rule among pleurothallid orchids. However, considering the exceptional diversity of these orchids (44 genera and over 5100 species) and the relatively limited number of available studies (pollination ecology has been assesed in only 17 genera), generalized trends about their pollination systems might disregard a wide variety of specific life-history traits and inconspicuous honest signals/rewards for pollinators. Known associations of pleurothallid orchids with a large assortment of fly taxa further support this assumption. We investigated the natural pollination system of Masdevallia hortensis, a strictly endemic species of cloud forests in the Western Andes of Colombia. Masdevallia hortensis exhibited a sophisticated and customized pollination mechanism, producing sugary secretions in the lateral sepals along purple dotted patches, fed upon by different visiting species of fruit flies (Drosophilidae). The sucrose concentration in these secretions varied throughout the day and was significantly lower after removing the pollinaria. Visiting fruit flies appeared to be guided towards a chamber between the mobile lip and the column by the dotted lines in the lateral sepals. During visitations, individuals of the most abundant species in our observations (Drosophilidae sp. 1 [AO]) were singly entrapped in the chamber until eventually freeing themselves with the pollinaria attached to their bodies. We also demonstrated that M. hortensis is strictly self-incompatible, which makes fly pollination an essential process for the maintenance of natural populations of the species. The flowers of M. hortensis offer rewards for visiting insects, an aspect that should also be evaluated among congenerics. In this way, we urge integrative ecological studies to understand the evolutionary patterns of this group of orchids.
... Self-incompatibility is a process leading to recognition and rejection of own or related pollen and it is rare in orchids (De Nettancourt, 1977;Van der Pijl and Dodson 1966). Although self-incompatibility has been reported in some orchids groups, mainly in the subfamily Epidendroideae such as Dendrobiinae (i.e., Dendrobium), Pleurothallidinae, Oncidiinae (Johansen 1990;Borba et al. 2011;Castro et al. 2019), it is more often present in species that are pollinated by Diptera, insects which pay long visits to a flower, in addition to visiting many flowers of the same plant Semir 1998b, 2001;Borba et al. 2001aBorba et al. , 2011. This pollinator behavior may promote self-pollination, leading to autogamy in the absence of mechanical or Handling Editor: Ferhat Celep. ...
... Self-incompatibility is a process leading to recognition and rejection of own or related pollen and it is rare in orchids (De Nettancourt, 1977;Van der Pijl and Dodson 1966). Although self-incompatibility has been reported in some orchids groups, mainly in the subfamily Epidendroideae such as Dendrobiinae (i.e., Dendrobium), Pleurothallidinae, Oncidiinae (Johansen 1990;Borba et al. 2011;Castro et al. 2019), it is more often present in species that are pollinated by Diptera, insects which pay long visits to a flower, in addition to visiting many flowers of the same plant Semir 1998b, 2001;Borba et al. 2001aBorba et al. , 2011. This pollinator behavior may promote self-pollination, leading to autogamy in the absence of mechanical or Handling Editor: Ferhat Celep. ...
... genetic barriers Borba et al. 2001b). This phenomenon has been observed in Neotropical subtribe Pleurothallidinae (nearly 4100 species), in which selfincompatibility is a main trait of the group (Borba et al. 2011). Bulbophyllum and Pleurothallidinae are the two largest plant groups pollinated by Diptera ( Van der Pijl and Dodson 1966). ...
A vast majority of Orchidaceae species are self-compatible, and many present mechanical or temporal barriers that prevent self-pollination. Self-incompatibility is present in some orchid groups with pollinated by insects that pay long visits to a flower, in addition to visiting many flowers of the same plant, as observed in Diptera. The genus Bulbophyllum, despite being pollinated by flies, predominantly comprises self-compatible species, and some of them present pre-pollination selfing barriers. However, there are some reports of Bulbophyllum sect. Micranthae presenting self-incompatibility. This section comprises 12 species, some of which show complex taxonomy possibly due by introgression. Here, we analyzed the morphology and development of pollen tube following intra- and interspecific experimental pollinations in eight species of Bulbophyllum sect. Micranthae to understand their reproductive system and isolation. All species in the section are self-incompatible, except for the species sister to the remaining section, Bulbophyllum mentosum, and need vectors for pollination. Epifluorescence microscopy revealed that the major site of incompatibility is present at the beginning of the column while in some others the pollen grains did not germinate. However, in some species with partial self-incompatibility, pollen tubes turned tortuous and anomalous upon reaching the ovary. We believe that this self-incompatibility reaction in the ovary is an extended gametophytic self-incompatibility reaction, as reported in other myophilous orchids, but diallelic crossing experiments are necessary to confirm our hypotheses. In the studied species of B. sect. Micranthae, reproductive isolation is mainly due to pre-pollination barriers, and fruit set is possible in most interspecific crossings, except for those involving the basal most species B. mentosum.
... The diversity of existing species in Orchidaceae is a product of their ability to exploit a wide range of ecological niches (Ricklefs and Renner, 1994), their habitat specialization (Gravendeel et al., 2004) and the ecological interactions that occur in this group. Several studies have linked orchid diversity to specialized interactions with mycorrhizal fungi and to a greater extent with specific pollinators (van der Pijl and Dodson, 1966;Dressler, 1981;Tremblay, 1992;Cozzolino andWidmer, 2005 Jersáková et al., 2006;Borba et al., 2011;Zhang et al., 2017). ...
... The flowers that present this pollination syndrome are very small and with different colors ranging from purple-pink and yellow with spots and lines. Furthermore, they do not produce nectar to attract pollinators (Borba et al., 2011). In this case, the flowers produce fetid odors, which in many cases are not perceptible to humans, but are easy for flies to smell since they resemble rotten meat, decomposing fruits, etc. ...
Objective: To describe the pollination syndromes of the orchids of Megamexico and the importance of the interactions between the orchids and their respective pollinators for the conservation of both groups. Design/Methodology/Approach: An exhaustive search was carried out on the pollinators of each of the orchids that grow in Megamexico. With the information sources available, a data matrix was prepared that includes orchids and all their pollinators. Subsequently, it was quantified which group of pollinators the orchids interact with the most. Finally, it is described what physiological adaptations and morphologies orchids have developed to attract specific pollinators. Results: Orchids from Megamexico maintain close relationships with specific pollinators. Said interaction is mediated by the shape, size, production of aromas, nectar, and the color of the flower. Thus, four large groups of pollinators are those that interact with the orchids of Megamexico, with the Hymenoptera being the group of pollinators that pollinates the most orchid species in Megamexico and birds to a lesser extent. Study Limitations/Implications: This study describes the importance of pollinators and their interactions with orchids for orchid prevalence. Findings/Conclusions: It is of vital importance to include orchid pollinators in conservation programs to ensure that interactions between orchids and pollinators continue to be effective and thus guarantee the permanence of the two groups.
... Within Orchidaceae, some groups are expressive such as the Pleurothallidinae subtribe, which comprises about 5100 Neotropical species and represents about 18 % of all orchid species (Karremans, 2016). In this subtribe, most species are myophilous and self-incompatible (Borba et al., 2011), and there is a great diversity of pollination systems (Karremans & Díaz-Morales, 2019). For instance, there are rewarding species that offer nectar (Barbosa et al., 2009;Borba et al., 2011) and shelter or rendezvous sites (Endara et al., 2010), and rewardless species that may be pollinated by food deception (Karremans et al., 2015;Bogarín et al., 2018), brood site imitation Pansarin et al., 2016), and sexual mimicries (Blanco & Barboza, 2005;Duque-Buitrago et al., 2014). ...
... In this subtribe, most species are myophilous and self-incompatible (Borba et al., 2011), and there is a great diversity of pollination systems (Karremans & Díaz-Morales, 2019). For instance, there are rewarding species that offer nectar (Barbosa et al., 2009;Borba et al., 2011) and shelter or rendezvous sites (Endara et al., 2010), and rewardless species that may be pollinated by food deception (Karremans et al., 2015;Bogarín et al., 2018), brood site imitation Pansarin et al., 2016), and sexual mimicries (Blanco & Barboza, 2005;Duque-Buitrago et al., 2014). ...
... Little is known about the pollination ecology and reproductive biology of Masdevallia. To date, only one study with a population from a Brazilian Masdevallia species was conducted (Borba et al., 2011), without any information from a species from the center of diversity of the genus. Also, information on distribution, phenology, reproductive systems, and pollinators are still incipient. ...
Knowledge of how natural and anthropogenic factors can limit the distribution of rare species is key to develop conservation strategies. We show data on how both factors potentially affect the distribution of a rare high Andean orchid. We chose Masdevallia coccinea as a model because it is endangered and the most commercialized species of the entire genus. Using ecological niche modeling, we confirm its rarity, with a restricted potential occurrence equivalent to only 0.00005 % of South America, 0.0003 % of the Andes, and 0.0009 % of Colombia. We projected the future potential occurrence of M. coccinea and found that it can have a reduction of 43.49 % in the best-case scenario and 89.06 % in the worst-case scenario in 2100. We describe the flowering and fruiting phenologies and demonstrate that the precipitation two months before may determine the first one. Masdevallia coccinea is a rewardless species and, unlike most Pleurothallidinae species, we show that it is self-compatible but non-autonomous selfer. Thus, it is dependent on biotic pollination, performed only by Leucophenga sp. (Drosophilidae: Diptera) which has low efficiency (8.6 % of fruit set). This is the first pollinator report for the genus. While self-compatibility may be related to reproductive assurance, apparently inbreeding depression may affect populations since selfed fruits had less viable seeds. Nevertheless, self-pollination can be avoided by the retention of the anther cap on pollinia, giving more time until the pollinator flies away to another flower. We consider that the rarity of M. coccinea may be a result of both anthropogenic (habitat loss, climate change, and overcollection) and natural factors (high elevation distribution, self-compatibility, rewardless strategy, pollinator specificity, and low fruit set). The several new facets investigated here can be used for future conservation strategies of this emblematic species as well as other high Andean threatened species.
... Pollen tube growth, fruit set, and seed abortion rate are often used as the main indicators of SI, especially the growth of the pollen tube, as it is one of the most direct and important indicators of SI [16,17]. Although SI has been reported in various orchid groups, it is most common in a few groups, such as Dendrobiinae (i.e., Dendrobium) [18], Pleurothallidinae [19], Oncidiinae [14,20], Malaxidinae [21][22][23], Laeliinae [14,24], Aeridinae [25], Angraecinae [25], and Neottieae (i.e., Epipactis) [14], which all belong to the subfamily Epidendroideae ( Figure 1). Here, we mainly review the pollen tube growth in orchid SI species. ...
... SI analysis of Restrepia, belonging to clade B [19] of the Pleurothallidinae, revealed that 45% of the species are self-incompatible, and seed abortion is lower for cross-pollinated plants (both intraspecific and interspecific cross-pollination) than for self-pollinated plants. ...
... In addition to pollen tubes, some Pleurothallidinae individuals can produce fruits with no seeds after self-pollination [19,34]. Seed abortion of A. johannensis after self-pollination may be caused by SI or by inbreeding depression [37]. ...
Self-incompatibility affects not only the formation of seeds, but also the evolution of species diversity. A robust understanding of the molecular mechanisms of self-incompatibility is essential for breeding efforts, as well as conservation biology research. In recent years, phenotypic and multiple omics studies have revealed that self-incompatibility in Orchidaceae is mainly concentrated in the subfamily Epidendroideae, and the self-incompatibility phenotypes are diverse, even in the same genus, and hormones (auxin and ethylene), and new male and female determinants might be involved in SI response. This work provides a good foundation for future studies of the evolution and molecular mechanisms of self-incompatibility. We review recent research progress on self-incompatibility in orchids at the morphological, physiological, and molecular levels, provide a general overview of self-incompatibility in orchids, and propose future research directions.
... The subtribe Pleurothallidinae is the largest monophyletic orchid group pollinated by Diptera (Pridgeon et al. 2001;Pridgeon et al. 2010;Borba et al. 2011). The subtribe's taxonomy is troublesome and has been under multidisciplinary investigations aiming to properly circumscribe the group (Pridgeon et al. 2001(Pridgeon et al. , 2010Borba et al. 2011;Chiron et al. 2012;Karremans 2016;Cardoso-Gustavson et al. 2017). ...
... The subtribe Pleurothallidinae is the largest monophyletic orchid group pollinated by Diptera (Pridgeon et al. 2001;Pridgeon et al. 2010;Borba et al. 2011). The subtribe's taxonomy is troublesome and has been under multidisciplinary investigations aiming to properly circumscribe the group (Pridgeon et al. 2001(Pridgeon et al. , 2010Borba et al. 2011;Chiron et al. 2012;Karremans 2016;Cardoso-Gustavson et al. 2017). According to these studies, all synapomorphies proposed are related to floral characters (Borba et al. 2011;Cardoso-Gustavson et al. 2017) that undoubtedly reflect their conspicuous pollination syndromes. ...
... The subtribe's taxonomy is troublesome and has been under multidisciplinary investigations aiming to properly circumscribe the group (Pridgeon et al. 2001(Pridgeon et al. , 2010Borba et al. 2011;Chiron et al. 2012;Karremans 2016;Cardoso-Gustavson et al. 2017). According to these studies, all synapomorphies proposed are related to floral characters (Borba et al. 2011;Cardoso-Gustavson et al. 2017) that undoubtedly reflect their conspicuous pollination syndromes. ...
Pleurothallidinae orchids have been the focus of many multidisciplinary studies due to their challenging systematics and taxonomy. The synapomorphies already recognized in the group are mostly related to floral characters, the last proposed being the occurrence of alkanes in the floral fragrance. The composition of the floral bouquet varied significantly among the studied species, leading us to hypothesize that the variations in volatiles emitted could be linked to the structure of osmophores, especially when comparing the myophilous and sapromyophilous pollination syndromes. Sepals and labellum at different developmental stages of seven Brazilian Pleurothallidinae species were examined using light, scanning, and transmission electron microscopy. Nectar reabsorption was assessed by Lucifer Yellow CH tracer and imaged under confocal microscopy. Nectaries were restricted to the labellum of the myophilous species, whereas osmophores occurred in the dorsal and/or lateral sepals, varying according to species. In the sapromyophilous species, floral nectaries were not detected and osmophores were restricted to the labellum. Osmophore structure was correlated with the volatiles emitted, being the trichome osmophores notably present on the sepals of both myophilous species that possess nectaries. For the first time, we demonstrated reabsorption of the released nectar in Pleurothallidinae and the occurrence of a unique gland named sticky-exudate glands, which occurred in the lateral sepals and labellum of Echinosepala aspasicensis, a sapromyophilous species, that released a heterogeneous exudate composed of polysaccharides and lipids. Similar glands have been reported in Bulbophyllum, highlighting the convergence between both groups.
... The Neotropical Pleurothallidinae are the most diverse subtribe in the family [44 genera, 5114 species, divided into nine genera 'affinities' proposed by Karremans (2016)] and are pollinated by Diptera (Borba et al., 2011;Karremans et al., 2016;Karremans section Pleurothallis section Fractiflexae (invalidly published). ...
... Nevertheless, the placement of this group within the subtribe, as stated by Karremans (2016), is merely interpretative. The same DNA evidence permits the placement of Dilomilis affinity as a sister subtribe, which in fact does not share the general morphology of Pleurothallidinae and is birdpollinated (Borba et al., 2011). ...
Pabstiella consists of c. 130 epiphytic species in the Neotropics. We present a phylogenetic analysis based on nrITS, matK and trnH-psbA sequences from 59 species of the genus and 40 Pleurothallidinae and two Laeliinae and one Bletiinae as an outgroup, using maximum likelihood, Bayesian inference and maximum parsimony. We also performed molecular dating, biogeographical analyses and ancestral morphological character reconstruction. Our results confirm the monophyly of Pabstiella with strong support. Ten clades are inferred and are herein proposed as sections. Pabstiella originated in the Andes and the Atlantic Rainforest in the Late Miocene (c. 7.93 Mya) in an epoch when these biomes were probably connected. A main vicariance event divided an early-diverging lineage that inhabited the Andes from an Atlantic Rainforest lineage that diversified in this region during the Pliocene and Pleistocene, mainly in the Serra do Mar in south-eastern Brazil. Our findings also suggest that the Atlantic Rainforest may have played an important role in the origin of subtribe Pleurothallidinae. The morphological character reconstruction showed high levels of homoplasy, with few recognized synapomorphies associated with stems and petals. Other characters related to the habit and stems were identified as important in the evolutionary history of the genus.
... 25,000 species and ~ 800 genera 1,2 are one of two of the most diverse and widely distributed flowering plant families on Earth and have captivated the attention of scientists for centuries 3 . The family has a striking morphological and ecological diversity and evolved complicated interactions with fungi, animals and other plants 4,5 and a diverse array of sexual systems [6][7][8] . Numerous efforts have been made to understand the natural history, evolution and phylogenetic relationships within the family 2,7,[9][10][11][12][13] . ...
Recent phylogenomic analyses based on the maternally inherited plastid organelle have enlightened evolutionary relationships between the subfamilies of Orchidaceae and most of the tribes. However, uncertainty remains within several subtribes and genera for which phylogenetic relationships have not ever been tested in a phylogenomic context. To address these knowledge-gaps, we here provide the most extensively sampled analysis of the orchid family to date, based on 78 plastid coding genes representing 264 species, 117 genera, 18 tribes and 28 subtribes. Divergence times are also provided as inferred from strict and relaxed molecular clocks and birth-death tree models. Our taxon sampling includes 51 newly sequenced plastid genomes produced by a genome skimming approach. We focus our sampling efforts on previously unplaced clades within tribes Cymbidieae and Epidendreae. Our results confirmed phylogenetic relationships in Orchidaceae as recovered in previous studies, most of which were recovered with maximum support (209 of the 262 tree branches). We provide for the first time a clear phylogenetic placement for Codonorchideae within subfamily Orchidoideae, and Podochilieae and Collabieae within subfamily Epidendroideae. We also identify relationships that have been persistently problematic across multiple studies, regardless of the different details of sampling and genomic datasets used for phylogenetic reconstructions. Our study provides an expanded, robust temporal phylogenomic framework of the Orchidaceae that paves the way for biogeographical and macroevolutionary studies. OPEN
... For example, Johnson et al. (1998) studied the patterns of adaptive radiation in Disa P.J.Bergius, suggesting that floral diversity in this orchid genus is the result of adaptation to pollinators. This leads to unexpected adaptations of orchids and contributes to pollinator attraction (Borba et al. 2011). Most orchid species require an external pollinating agent (Dressler 1981), among the variety of pollination systems only abiotic and mammal pollination are absent (Tremblay et al. 2005). ...
... Similar results have been observed in Liparis makinoana Schltr. (Soo et al. 2001 (Borba et al. 2011). Brassavola cebolleta Rchb.f. also did not present fruit production in spontaneous self-pollination treatments or in agamospermia, indicating that it too requires pollinators for sexual reproduction, and apomixis is not observed, as it is in R. cervantesii (Rech et al. 2010). ...
... For the years 2014 and 2015, greater fruit production was observed in the R. cervantesii cross-pollination treatments, while a lower percentage was recorded in fruit production from self-pollination. Our results are similar to those obtained for Laelia speciosa (Ávila-Díaz & Oyama 2007), Cattleya luteola and Prosthechea vespa (Quiroga et al. 2010), and representatives of Pleurothallidinae (21 of the 22 species studied) (Borba et al. 2011), presenting high values of fruit production from cross-pollination treatments that were significantly greater than those produced by selfpollination. The ability to obtain fruits with both types of pollination (self-pollination and cross-pollination) in the same species can be an advantage to ensure the production of progeny under adverse environmental conditions or in colonization events, when availability is variable, of pollinators and/or reproductive couples (Vogler & Kalisz 2001;Kalisz et al. 2004;Haleigh & Wagner 2018). ...
Background and aims – The Orchidaceae family is vulnerable, because of the destruction of their habitat, as well as the extraction of individuals from natural populations. This is the case of the genus Rhynchostele Rchb.f.; among the actions considered important for appropriate conservation strategies for this genus is the generation of fundamental knowledge, such as on its reproductive biology. The objective of this work is to understand the mating system and reproductive success of Rhynchostele cervantesii, an endangered epiphytic orchid endemic to Mexico.Material and methods – Manual and open-pollination treatments were conducted during 2014 and 2015 in a cloud forest in Michoacan, Mexico. In each period, 30 to 40 randomly selected inflorescences were subjected to the following treatments: a) spontaneous-self-pollination, b) emasculation, c) self-pollination, d) cross-pollination, and e) open-pollination. The developed fruits were counted and harvested, the viability of the seeds was determined, through the observation and evaluation of embryos using microscopy.Key results – Significant differences were recorded between the treatments in both 2014 and 2015, with higher fruit production in cross-pollination than in self-pollination and natural-pollination. There were significant differences in seed viability, with higher values for seeds from open-pollination and cross-pollination and lower values for seeds from self-pollination.Conclusions – Rhynchostele cervantesii is a species that requires pollinators for sexual reproduction because there is no fruit production with spontaneous-self-pollination. Under pollen limitation, the fruit set of natural pollination was a lot lower than in cross-pollination although fruits were the same quality. R. cervantesii had a mixed mating system with a tendency to exogamy, presenting high values of female reproductive success compared to other tropical epiphytic orchid species reported in the literature.
... The Neotropical Pleurothallidinae are the most diverse subtribe in the family [44 genera, 5114 species, divided into nine genera 'affinities' proposed by Karremans (2016)] and are pollinated by Diptera (Borba et al., 2011;Karremans et al., 2016;Karremans section Pleurothallis section Fractiflexae (invalidly published). ...
... Nevertheless, the placement of this group within the subtribe, as stated by Karremans (2016), is merely interpretative. The same DNA evidence permits the placement of Dilomilis affinity as a sister subtribe, which in fact does not share the general morphology of Pleurothallidinae and is birdpollinated (Borba et al., 2011). ...
Pabstiella consists of c. 130 epiphytic species in the Neotropics. We present a phylogenetic analysis based on nrITS, matK and trnH-psbA sequences from 59 species of the genus and 40 Pleurothallidinae and two Laeliinae and one Bletiinae as an outgroup, using maximum likelihood, Bayesian inference and maximum parsimony. We also performed molecular dating, biogeographical analyses and ancestral morphological character reconstruction. Our results confirm the monophyly of Pabstiella with strong support. Ten clades are inferred and are herein proposed as sections. Pabstiella originated in the Andes and the Atlantic Rainforest in the Late Miocene (c. 7.93 Mya) in an epoch when these biomes were probably connected. A main vicariance event divided an early-diverging lineage that inhabited the Andes from an Atlantic Rainforest lineage that diversified in this region during the Pliocene and Pleistocene, mainly in the Serra do Mar in southeastern Brazil. Our findings also suggest that the Atlantic Rainforest may have played an important role in the origin of subtribe Pleurothallidinae. The morphological character reconstruction showed high levels of homoplasy, with few recognized synapomorphies associated with stems and petals. Other characters related to the habit and stems were identified as important in the evolutionary history of the genus.
