The communal breeding system of the smooth-billed ani (Crotophaga ani) was studied at two wildlife refuges in southwest Puerto Rico. Groups ranged in size from 2 to 9 adults, and communal clutches ranged in size from 11 to more than 25 eggs. Microsatellite DNA loci were isolated and used to examine the mating system and the extent of reproductive skew in individual groups. Loci were also screened in guira cuckoos (Guira guira) to examine the relationships suggested by Quinn et al. (1994). In the four focal smooth-billed ani groups, I found suggestions of monogamy, polygamy, and incidents of intraspecific brood parasitism. In all groups, incubated clutch size and microsatellite data indicate that subordinate females are gaining some reproductive success. At one nest, evidence suggested a decrease in the intensity of reproductive skew between successive nests. Although the data are limited, they suggest that alternative strategies to monogamy may be more prevalent than previously believed, and that reproductive skew may be affected by the availability of limiting resources within the breeding season. / Thesis / Master of Science (MS)

Aciphylla species (wild spaniard/speargrass) are an iconic component of the Australasian high country flora, but their reproductive system is enigmatic. They are insect-pollinated dioecious mast seeders (synchronous highly variable seed production), which seems maladaptive. The resource supply to pollinators is highly variable, yet dioecious plants are dependent on pollinators, and dioecious masting requires male and female plants to flower synchronously. Floral display in Aciphylla is relatively large, with tall inflorescences bearing thousands of flowers, suggesting that plants would not have the resources to produce such large stalks every year. But why do they have such huge inflorescences in the first place? I tested whether pollinator attraction is providing an economy of scale which favours intermittent production of very large inflorescences, by manipulating floral display size during a high-flowering year and measuring insect visitation rates and seed set (female reproductive success). Using space-for-time substitution and selective removal of male inflorescences, I also tested whether female seed set was affected by distance to flowering male plants (i.e. changes in local pollen availability) to see if flowering asynchrony would reduce pollination success. Bags were used to exclude pollination by insects and test for wind pollination, and hand pollination was done to test for pollen limitation. Insect surveys suggest that Aciphylla has a generalist pollination system (to avoid satiating a specialist pollinator during 'mast' years'). Male inflorescences received significantly more visits than females, and some seeds were set inside bags (although only 20-30%), suggesting wind pollination may occur at low levels. Seed set rate was higher for taller inflorescences with greater flowering length in A. aurea but tall inflorescences with excess flowers led to a decrease in seed set rates in A. scott-thomsonii. Hand pollination significantly increased seed set rates although these effects were not as large as expected (e.g. 10% increases from natural to hand-pollinated inflorescences were typical). There was no evidence for resource limitation in any species. Female plants in dense flowering populations had higher seed set rates, and individual floral display size in females was particularly important when females were 'isolated' from males. Insect visitation rates were generally higher on inflorescences with a larger floral display, suggesting that display size is important for pollinator attraction. Overall, these results suggest that the pollinator-attraction benefits of such a large floral display (at both the plant and population level) are possibly providing an economy of scale, although the relative effects are small.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A cactÃcea Cereus jamacaru DC à uma espÃcie emblemÃtica para o nordeste brasileiro. Tem se desenvolvido muitos estudos e usos da planta, mas nÃo se tem estabelecido qual o polinizador em Ãreas de distribuiÃÃo natural. O objetivo do trabalho foi identificar qual o polinizador mais efetivo para forma e funÃÃo da flor de C. jamacaru. A coleta de dados foi feita em um fragmento de Caatinga onde foram coletados dados sobre fenologia, morfologia e biologia floral alÃm de busca por visitantes. Foram contados 695 botÃes, 175 flores e 21 frutos. O mandacaru apresenta floraÃÃo do tipo multiple bang. As flores sÃo hermafroditas, protÃndricas, de simetria dorsoventral esternotrÃbica. O hipanto à cÃnico 6,5 cm de comprimento, 3 cm diÃmetro na abertura e 0,6 cm na base. A cÃmara nectarÃfera à formada pela base dos estames e possui a abertura com uma espÃcie de fechadura pelo estilete. De acordo com indÃcios morfolÃgicos ao comparar a polinizaÃÃo feita por morcegos e mariposas observou-se a grande maioria dos argumentos a favor de quiropterofilia. Simulamos visitas por Glossophaga soricina e obtivemos perfeita compatibilidade, porÃm nÃo observamos visita de nenhum animal com encaixe perfeito. O visitante que mais se aproximou do comportamento polinizador foi Xylocopa sp, uma abelha de grande porte. Provavelmente esta abelha foi a responsÃvel pela formaÃÃo dos poucos frutos. / The Cereus jamacaru DC cactus is an important species for the Brazilian Northeast. Has developed many studies and uses of the plant, but has not been established which pollinator in areas of natural distribution. The aim was to identify the most effective pollinator for form and function C. jamacaru flower. Data collection was done in a fragment of Caatinga where we collected data on phenology, morphology and floral biology beyond search visitors. Fruits were numbered 695young flowers, 175 flowers and 21 fruits. The cactus flowering displays multiple bang type. The flowers are hermaphrodite. The hypanthium is tapered 6.5 cm length, 3 cm in diameter and 0.6 cm opening in the base. The nectariferous chamber is formed by the base of the stamens and has the opening with some kind of lock by the stylus. According to the morphological evidence comparing pollination by bats and moths observed most of the arguments in favor of chiropterophily. We simulate visits per Glossophaga soricina and got perfect compatibility, however we did not observe any animals visit with optimal fit. The visitor who came closest pollinator behavior was Xylocopa sp, a large bee. Probably this bee was responsible for the formation of the few fruits.

The dynamics of a plant-pathogen interaction vary both within and among species. Both spatial structuring and specific genetic and life-history characteristics will affect the interaction and the outcome of a potential co-evolution between the two organisms. In this thesis I have studied the interaction between the wild perennial herb Silene dioica and its automictic, obligate anther smut Microbotryum violaceum MvSd. From the plant perspective, I have examined different aspects of biochemical resistance in S. dioica to M. violaceum MvSd. From the pathogen perspective, I have focused on the breeding system of M. violaceum MvSd and its connection to fitness and distribution of genetic diversity. I have used varying methods; glasshouse trails involving inoculation of plants with the pathogen, classical Mendelian analysis involving controlled crosses between plants, microscopic studies of spores and molecular DNA-analysis. With the results I demonstrate that resistance to M. violaceum MvSd in S. dioica can be specific to the attacking pathogen strain and also spatially highly diverse both within and among populations within a metapopulation. Together, these factors are likely to delay the establishment of the disease within host populations and reduce the spread and amount of disease, once it has been established. The results also suggest that the specific resistance expressed against two different M. violaceum MvSd strains were determined by separate gene systems and that, in both cases, the resistance was simply inherited. This implies a potential for relatively rapid response to M. violaceum-induced selection in S. dioica populations variable for resistance. My results also show that automixis clearly is the predominating breeding system of M. violaceum MvSd, similarly to what earlier has been shown for M. violaceum MvSl. Furthermore, I found lower levels of neutral genetic diversity in M. violaceum MvSd in the northern parts of Sweden, compared to what has been found in populations in more southern Europe. This result is consistent with predictions that populations in the outer regions of a species distribution have lower levels of genetic variation. Moreover, populations were highly differentiated in northern Sweden, which could have been generated by high selfing rates, genetic drift and high population turnover rates, all factors that coincide with life-history and ecology of M. violaceum MvSd. However, despite the general low variability in neutral genetic markers, I did find variation in fitness related traits, both within and among populations, as well as differences in infection ability between strains, suggesting there is a potential for co-evolution between S. dioica and M. violaceum MvSd in the area. To summarize, this thesis reflect a plant-pathogen system that is highly influenced by constant colonisation-extinction dynamics, which is likely to have influenced both the genetics of resistance in the plant and the breeding system of the pathogen and thus also the interaction between the two organisms.

Aciphylla species (wild spaniard/speargrass) are an iconic component of the Australasian high country flora, but their reproductive system is enigmatic. They are insect-pollinated dioecious mast seeders (synchronous highly variable seed production), which seems maladaptive. The resource supply to pollinators is highly variable, yet dioecious plants are dependent on pollinators, and dioecious masting requires male and female plants to flower synchronously. Floral display in Aciphylla is relatively large, with tall inflorescences bearing thousands of flowers, suggesting that plants would not have the resources to produce such large stalks every year. But why do they have such huge inflorescences in the first place? I tested whether pollinator attraction is providing an economy of scale which favours intermittent production of very large inflorescences, by manipulating floral display size during a high-flowering year and measuring insect visitation rates and seed set (female reproductive success). Using space-for-time substitution and selective removal of male inflorescences, I also tested whether female seed set was affected by distance to flowering male plants (i.e. changes in local pollen availability) to see if flowering asynchrony would reduce pollination success. Bags were used to exclude pollination by insects and test for wind pollination, and hand pollination was done to test for pollen limitation. Insect surveys suggest that Aciphylla has a generalist pollination system (to avoid satiating a specialist pollinator during 'mast' years'). Male inflorescences received significantly more visits than females, and some seeds were set inside bags (although only 20-30%), suggesting wind pollination may occur at low levels. Seed set rate was higher for taller inflorescences with greater flowering length in A. aurea but tall inflorescences with excess flowers led to a decrease in seed set rates in A. scott-thomsonii. Hand pollination significantly increased seed set rates although these effects were not as large as expected (e.g. 10% increases from natural to hand-pollinated inflorescences were typical). There was no evidence for resource limitation in any species. Female plants in dense flowering populations had higher seed set rates, and individual floral display size in females was particularly important when females were 'isolated' from males. Insect visitation rates were generally higher on inflorescences with a larger floral display, suggesting that display size is important for pollinator attraction. Overall, these results suggest that the pollinator-attraction benefits of such a large floral display (at both the plant and population level) are possibly providing an economy of scale, although the relative effects are small.

Made available in DSpace on 2016-08-17T15:00:42Z (GMT). No. of bitstreams: 1 FERNANDA NOGUEIRA MENDES.pdf: 1972021 bytes, checksum: d44e6b10ae8938f3d3e7315d887558ba (MD5) Previous issue date: 2007-02-26 / FUNDAÇÃO DE AMPARO À PESQUISA E AO DESENVOLVIMENTO CIENTIFICO E TECNOLÓGICO DO MARANHÃO / (Pollination and Reproduction System of Byrsonima umbellata Mart. (Malpighiaceae) in Cerrado at Maranhão, Brazil). The reproductive biology of Byrsonima umbellata was studied in a Cerrado area close to Bom Jesus (03°12'28 '' S, 43°24'12 '' W) far from 46 km of the municipal district of Urbano Santos, northeast of Maranhão, Brazil, from Mar to Oct/2005 and Mar to Oct/2006. Flowering phenology, morphology and floral biology were studied and bees visitors were observed. The breeding system was verified through experimental pollinations in the field. Byrsonima umbellata is a specie with hermaphrodite flowers, of diurnal anthesis and high pollen viability. The flowering pattern is annual, extensive and irregular, having production of buds and flowers more or less constant along the months. The controlled pollinations showed that the species is partially self-incompatible. Forty two species of bee visitors were observed. The tribe Meliponini presented larger abundance, while the wealth was larger in the Centridini tribe. / (Sistema de Polinização e Reprodução de Byrsonima umbellata Mart. (Malpighiaceae) em Área de Cerrado no Maranhão, Brasil). A biologia reprodutiva de Byrsonima umbellata foi estudada em uma área de cerrado próxima ao povoado de Bom Jesus (03°12 28 S, 43°24 12 W), distante 46 km da sede do município de Urbano Santos, nordeste do Maranhão, Brasil, durante o período de mar a out/2005 e mar a out/2006. Foram realizados estudos sobre fenologia, morfologia e biologia floral e observadas as abelhas visitantes. O sistema reprodutivo foi determinado através de experimentos de polinização feitos em campo. Byrsonima umbellata é uma espécie que possui flores hermafroditas, de antese diurna e alta viabilidade polínica. O padrão de floração é anual, extensivo e irregular, havendo produção de botões e flores mais ou menos constante ao longo dos meses. Os resultados dos experimentos de polinização indicam que a espécie é parcialmente autoincompatível. Foram observadas 42 espécies de abelhas visitantes. A tribo Meliponini apresentou maior abundância, enquanto a riqueza foi maior na tribo Centridini.

A key feature of social Hymenoptera is the division of labor in reproduction between one or a few fertile individuals – the queen(s) – and many sterile nestmates that function as helpers – the workers. The reproductive altruism of workers has long been considered as one of the most important paradox of Evolution. Today, kin selection (Hamilton 1964a,b) is recognized as a prime selective force for the evolution reproductive altruism in Hymenoptera. Hamilton’s kin selection theory states that workers may benefit helping relatives reproduce as long as the relatives they aid share a higher than average proportion of their genes with the workers and effectively pass on copies of the workers' genes to the next generation. Relatedness between colony members is therefore pivotal in kin selection theory, because it directly influences the benefits from indirect fitness. In social Hymenoptera, within-colony relatedness is usually high, because of the haplodiploid sex determinism system. However, several factors of the breeding system are known to affect the colony genetic structure and, hence, the workers’ indirect inclusive fitness: the number of breeders, their genetic relationships and their relative contribution to the reproduction. On the other hand, dispersal strategies influence the population genetic structure, which in turn may result in different interaction patterns between members from neighboring colonies. Despite its central role in the evolution of cooperation and reproductive altruism in animals, kin selection also predicts conflicts between colony members. Because the individuals from a colony are not genetically identical, their reproductive interests may be different (Trivers 1974). These conflicts are diversified, both regarding their actors and their causes (Trivers & Hare 1976; Bourke & Franks 1995; Keller 1995; Chapuisat & Keller 1999b; Keller & Reeve 1999; Sundström & Boomsma 2001). The first part of this work deals with the population genetic structure, mating system and dispersal strategies of two Pheidole ants, the Mediterranean ant Pheidole pallidula and the Mojave Desert ant Pheidole tucsonica. Chapter 1 investigates the population genetic structure, the breeding system, the colony kin composition and the colony genetic structure of the Mediterranean ant P. pallidula. This study was performed by using highly polymorphic DNA microsatellite marker loci. The results show that a single, unrelated male inseminates each queen. Colonies are monogynous (i.e. headed by one reproductive queen) or polygynous (i.e. headed by 2 to 4 functional queens). Moreover, they are genetically differentiated and form a population exhibiting significant isolation-by-distance, suggesting that some colonies originate through budding. Chapter 2 reports cross-species amplifications of microsatellite markers developed for the ant P. pallidula on 13 ant species belonging to the sub-family Myrmicinae. Moreover, levels of genetic diversity within a colony, as well as relationship among colonies are studied for the black ant Pheidole tucsonica. Chapter 3 characterizes the level of inter-nest aggression, the spatial distribution and the genetic structure of a P. tucsonica population. The results show that inter-colony aggression varies from none to “all out” fights and that it is largely non-transitive. No effect of geographical distance or genetic structure on inter-nest aggression levels is detected. Moreover, genetic data reveal high rates of polygyny and/or polyandry. Overall, these results do not support the idea of a simple mechanism of nestmate recognition through queen or worker-produced pheromones or environmental cues. The second part of this work is devoted to the queen-queen conflict over reproduction, and the queen-workers conflict over sex allocation in P. pallidula. Chapter 4 is a detailed analysis on the partitioning of reproduction among queens in polygynous colonies of the species. Our results show a significant departure from equal contribution of queens to reproductive female, male and worker production. Reproductive skew is greater for male production than for queen and worker production. There is no relationship between the magnitude of the reproductive skew and (i) the number of reproductive queens per colony, (ii) their relatedness and (iii) the overall colony productivity, some of the factors predicted to influence the extent of reproductive skew. Finally, this study reveals a trade-off in the relative contribution of nestmate queens to reproductive female and worker production. The queens contributing more to reproductive female production contribute significantly less to worker production. To our knowledge, such a trade-off is shown for the first time in the Formicidae. Chapter 5 focuses on queen-workers conflict over sex allocation. Colonies of the Mediterranean ant P. pallidula show a strong split sex ratio, with 85% colonies producing more than 80% sexuals of one sex. Genetic analyses reveal that this species has an unusual breeding system, with colonies being headed by a single or a few unrelated queens. As expected in such a breeding system, our results show no variation in relatedness asymmetry between monogynous (single queen per colony) and polygynous colonies. Nevertheless, sex allocation is tightly associated with the breeding structure, with monogynous colonies producing a male-biased brood and polygynous colonies almost only females. In addition, sex allocation is closely correlated with colony total sexual productivity. Overall, our data show that when colonies become more productive (and presumably larger) they shift from monogyny to polygyny and from male production to female production, a pattern that has never been reported in social insects so far. A new explanation based on the concept of the “tragedy of the commons” is proposed to explain the strong sex ratio specialization observed in P. pallidula and in other species characterized by facultative polygyny. Chapter 6 investigates the relationship between the breeding system (monogynous vs. polygynous colonies) and the biosynthesis rate of juvenile hormone. Previous works in P. pallidula showed (i) that maternal effects induced by hormones and/or other compounds transferred to the eggs could influence the caste fate of female eggs and (ii) that sex specialization is tightly associated with the breeding structure (monogynous colonies produce a male-biased brood and polygynous colonies almost only females - Chapter 5). This study reveals a strong relationship between the biosynthetic rate of juvenile hormone (JH) production and the breeding system. Because in this species the breeding structure is closely associated with colony sex ratio, we propose that the rate of JH in queens could be a critical parameter in colony sex ratio determination. Queens of P. pallidula would exert partial control over sex ratio by laying different proportions of worker-destined eggs and queen-destined eggs according to the structure monogynous or polygynous of their colony. To conclude, some perspectives for future research on the different topics presented in this work are suggested. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

<p>In this thesis, I examined variation in sex expression and mating patterns in the sexually polymorphic, wind-pollinated herb <i>Plantago maritima</i>. With a combination of field studies, greenhouse experiments, and genetic analyses, I (a) examined factors influencing sex ratio variation in gynodioecious plants (in which hermaphrodites and females coexist), (b) discovered variation in breeding system, (c) investigated density-dependence of seed production, and (d) documented genetic variation within and among populations close to the northern range margin in Europe. </p><p>In a survey of 104 <i>P. maritima</i> populations, I documented considerable variation in sex ratio (range 0-70% females, median 6.3% females). As predicted, females were more frequently missing from small than from large populations, and the variance in sex ratio increased with decreasing population size. Among twelve populations sampled for seed production, the frequency of females was positively related to relative fecundity of females and negatively related to population size. The results suggest that the local sex ratio is influenced both by the relative fecundity of females and hermaphrodites, and by stochastic processes in small populations.</p><p>A comparative field study showed that plant fecundity decreased with increasing distance to nearest pollen donor both within and among populations in an archipelago in southern Sweden, where self-incompatibility was confirmed in controlled crosses. In contrast, plant fecundity was overall higher and was not density-dependent in the Skeppsvik archipelago in northern Sweden, where controlled crosses showed that plants are self-compatible. The results were consistent with the prediction that evolution of self-fertility should reduce density-dependence of pollination success.</p><p>I quantified the genetic structure within and among populations from eastern Sweden and western Finland based on variation at four polymorphic microsatellite loci. The genetic diversity was low in northern Sweden, which may be the result of a history of small population sizes and periods of frequent self-fertilization.</p>

In this thesis, I examined variation in sex expression and mating patterns in the sexually polymorphic, wind-pollinated herb Plantago maritima. With a combination of field studies, greenhouse experiments, and genetic analyses, I (a) examined factors influencing sex ratio variation in gynodioecious plants (in which hermaphrodites and females coexist), (b) discovered variation in breeding system, (c) investigated density-dependence of seed production, and (d) documented genetic variation within and among populations close to the northern range margin in Europe. In a survey of 104 P. maritima populations, I documented considerable variation in sex ratio (range 0-70% females, median 6.3% females). As predicted, females were more frequently missing from small than from large populations, and the variance in sex ratio increased with decreasing population size. Among twelve populations sampled for seed production, the frequency of females was positively related to relative fecundity of females and negatively related to population size. The results suggest that the local sex ratio is influenced both by the relative fecundity of females and hermaphrodites, and by stochastic processes in small populations. A comparative field study showed that plant fecundity decreased with increasing distance to nearest pollen donor both within and among populations in an archipelago in southern Sweden, where self-incompatibility was confirmed in controlled crosses. In contrast, plant fecundity was overall higher and was not density-dependent in the Skeppsvik archipelago in northern Sweden, where controlled crosses showed that plants are self-compatible. The results were consistent with the prediction that evolution of self-fertility should reduce density-dependence of pollination success. I quantified the genetic structure within and among populations from eastern Sweden and western Finland based on variation at four polymorphic microsatellite loci. The genetic diversity was low in northern Sweden, which may be the result of a history of small population sizes and periods of frequent self-fertilization.

Dentre as orquídeas terrícolas brasileiras, dois grupos são particularmente importantes do ponto de vista taxonômico, sistemático e biogeográfico: as orquídeas das subtribo Chloraeinae e Spiranthinae. Na sua atual delimitação, Chloraeinae compreende aproximadamente 70 espécies em três gêneros: Chloraea, Bipinnula e Gavilea. Por outro lado, Spiranthinae compreende cerca de 470 espécies exclusivamente neotropicais, distribuídas em 42 gêneros. As análises filogenéticas mais recentes suportam a existência de quatro grupos: os clados Stenorhynchos, Pelexia, Eurystyles+Lankesterella e Spiranthes. Assim, os objetivos deste estudo são: 1) realizar um estudo taxonômico e nomenclatural das orquídeas brasileiras do clado Pelexia; 2) documentar a morfologia floral e vegetativa, com ênfase nos caracteres diagnósticos; 3) realizar um estudo taxonômico e nomenclatural das espécies brasileiras de Chloraeinae; 4) construir uma filogenia molecular mais completa e representativa do clado Pelexia; 5) estabelecer se os gêneros do clado Pelexia são monofiléticos; 6) construir uma filogenia molecular mais completa e representativa de Chloraeinae, com a inclusão de táxons brasileiros; 7) estabelecer um conjunto de caracteres diagnósticos para os gêneros envolvidos neste estudo e, se necessário, propor uma nova delimitação genérica; e 8) propor cenários para a provável evolução de caracteres de importância ecológica. Os resultados de um estudo detalhado, bem como de análises morfológicas e moleculares são: 1) propostas de 48 tipificações, novas sinonímias e notas taxonômicas para as espécies de Goodyerinae e Spiranthinae descritas por José Velloso e Barbosa Rodrigues; 2) novos registros de Pteroglossa e Lyroglossa para o Rio Grande do Sul; 3) uma correção nomenclatural de Spiranthes bicolor e 4) uma sinopse taxonômica das Chloraeinae brasileiras; 5) uma análise filogenética molecular para testar a monofilia dos gêneros atualmente circunscritos no clado Pelexia; 6) uma atualização das relações filogenéticas de Chloraeinae, incluindo espécies Pampeanas de Bipinnula e a espécie tipo do gênero, B. biplumata; 7) um panorama sobre a biologia reprodutiva de duas espécies de Pachygenium e dados adicionais sobre Brachystele e Cyclopogon s.l.; e 8) um estudo detalhado da morfologia floral, biologia da polinização e sistema reprodutivo de Chloraea membranacea (Chloraeinae). / Among the Brazilian terrestrial orchids, two groups are particularly important from a taxonomic, systematic and biogeographical aspects: orchids of subtribe Chloraeinae and Spiranthinae. In its current delimitation, Chloraeinae comprises approximately 70 species distributed in three genera: Chloraea, Bipinnula and Gavilea. On the other hand, Spiranthinae comprises about 470 exclusively neotropical species distributed in ca. 42 genera. The most recent phylogenetic analyses support the existence of four clades: Stenorrhynchos, Pelexia, Eurystyles+Lankesterella and Spiranthes clade, respectively. Thus, the aims of this study are: 1) to make a taxonomic and nomenclatural study of Brazilian orchids of Pelexia clade; 2) to document the floral and vegetative morphology, with emphasis on diagnostic features; 3) to make a taxonomic and nomenclatural study of the Brazilian species of Chloraeinae; 4) to assemble a more complete and representative molecular phylogeny of the Pelexia clade; 5) to establish if the genera of the Pelexia clade are monophyletic; 6) to construct a more complete and representative molecular phylogeny of the Chloraeinae, with the inclusion of Brazilian taxa; 7) to establish sets of diagnostic characters for the genera involved in this study and, if necessary, propose a new generic delimitation; and 8) to propose scenarios for the evolution of characters of ecological importance. The main results are: 1) 48 typifications, new synonymies and taxonomic notes for Spiranthinae and Goodyerinae species described by José Velloso and Rodrigues Barbosa; 2) new records of Pteroglossa and Lyroglossa to Rio Grande do Sul; 3) a nomenclatural correction of Spiranthes bicolor and 4) a taxonomic synopsis of Brazilian Chloraeinae; 5) molecular phylogenetic analyses to test the monophyly of the currently circumscribed genera in the Pelexia clade; 6) an update of the phylogenetic relationships of the Chloraeinae, including Pampean species of Bipinnula, and the type species of the genus, B. biplumata; 7) an overview on the reproductive biology of two Pachygenium species and additional data on Brachystele and Cyclopogon s.l.; and 8) a detailed study of the floral morphology, pollination biology and reproductive system of Chloraea membranacea (Chloraeinae).