Morpho-anatomy of in vitro germination and cryopreservation of the orchid Cattleya crispa (Orchidaceae Morfoanatomía de la germinación in vitro y criopreservación de la orquídea Cattleya crispa (Orchidaceae) / Morfoanatomía de la germinación in vitro y criopreservación de la orquídea Cattleya crispa (Orchidaceae)

Introduction: Cattleya crispa is an ornamental epiphytic orchid with geographic distribution restricted to the Brazilian Atlantic Forest. Due to predatory extractivism and human-induced habitat loss, this species appears on the Red List of Brazilian Flora. Objective: To characterize morpho-anatomical aspects regarding germination and post-seminal development from C. crispa seeds; as well as studying the effect of cryopreservation on these seeds. Methods: We used light microscopy and electron microscopy to describe the microstructure of a 100 ripe seeds. We evaluated seed viability, seed germination, survival rate and protocorm weight in cryopreserved and non-cryopreserved material, with four replicas per treatment using 20 mg of plant material. Results: The seeds are fusiform, whitish yellow with a length from 700 to 900 µm and a water content of 5 %. Germination began seven days after sowing, the formation of the globular protocorm at 30 days and the formation of the seedling occurred 150 days. The persistent seed coat can compress the protocorm and cause it to collapse. The cryopreserved seeds presented 87.15 % viability, 78.32 % germination, 8.48 % survival and protocorms with 104.27 mg five months after sowing. Data wasn't different to non-cryopreserved seeds. Conclusions: The cryocapability of the seeds shows that cryopreservation can be used for long-term conservation. The results of this work contribute to the overall biology of C. crispa and to the propagation and storage of genetic material for conservation purposes.

Introducción: Cattleya crispa es una orquídea epífita ornamental con distribución geográfica restringida a la Mata Atlántica brasileña. Debido al extractivismo depredador y a la pérdida de hábitat inducida por el hombre, esta especie aparece en la Lista Roja de la Flora Brasileña. Objetivo: Caracterizar aspectos morfoanatómicos de la germinación y desarrollo inicial de semillas de C. crispa; así como estudiar el efecto de la criopreservación de estas semillas. Métodos: Utilizamos microscopía óptica, microscopía electrónica de barrido y microscopía electrónica de transmisión para describir la microestructura en 100 semillas maduras. Evaluamos la viabilidad de la semilla, la germinación de la semilla, la tasa de supervivencia y el peso de los protocormos en el material criopreservado y no criopreservado, con cuatro réplicas por tratamiento de 20 mg de material vegetal. Resultados: Las semillas son fusiformes, amarillo blanquecinas, con una longitud de 700 a 900 µm y un contenido de agua del 5 %. La germinación comenzó siete días después de la siembra, la formación del protocormo globular a los 30 días y la formación de la plántula a los 150 días. La cubierta de semilla persistente puede comprimir el protocormo y provocar su colapso. Las semillas criopreservadas presentaron 87.15 % de viabilidad, 78.32 % de germinación, 8.48 % de supervivencia y protocormos con 104.27 mg a los cinco meses de la siembra. Los datos no fueron diferentes a las semillas no criopreservadas. Conclusiones: La capacidad criogénica de las semillas muestra que la crioconservación puede utilizarse para la conservación a largo plazo. Los resultados de este trabajo contribuyen a la biología general de C. crispa y a la propagación y almacenamiento de material genético con fines de conservación.

Morphological Developments of Orchis purpurea Huds. Salep Orchids

Abstract This article aims to monitor the development of Orchis purpurea Huds., salep orchids, of different sizes over a period of two years, and to investigate the relationship between the parameters studied. In the first step, the measurements taken at the time of planting and harvesting of tubers divided into eight different groups according to their size were subjected to variance and Duncan's test. In the second step, the relationship between the parameters was investigated by ignoring seedling groups. The relationship between the two variables was determined by correlation analysis. The significance of the relationships between planting and harvest data sets, and variable contributions were determined by canonical correlation analysis. Finally, leaf area prediction modeling was performed by applying multiple regression analysis. In variance analysis all parameters were significant. The canonical correlation between the first pair of canonical variables was 0.988 (p<0.01). The data obtained from the tubers made the greatest contribution to the explanatory power of the canonical variables. The leaf area model was formulized as LA (mm2) = -1237.0204 + 57.7912 × LW + 16.6211 × LL where LA is leaf area, LW is leaf width, LL is leaf length and a, b, and c are coefficients.

Oncidiinae (Orchidaceae) on the great curve of the Xingu River, Pará state, Brazil / Oncidiinae (Orchidaceae) na volta grande do Rio Xingu, Pará, Brasil

Among the studies on Orchidaceae in the Amazon, none comprised the region of the Great Curve of the Xingu River, located in the lower Xingu river. The aim of this study was to inventory and taxonomically study the species of Oncidiinae (Orchidaceae) in the Great Curve of the Xingu River, Pará state. The floristic survey was performed in the area of the Belo Monte hydroelectric plant, in the Vitória do Xingu municipality, centrally inserted in the called Great Curve of the Xingu River. Botanical collections were accomplished between June 2011 and December 2013. A total of 27 species of Oncidiinae, distributed in 15 genera, was inventoried in the study area. Notylia Lindl. and Trichocentrum Poepp. & Endl. were the richest genera, with five and four species, respectively, followed by Erycina Lindl., Ionopsis Kunth, Lockhartia Hook., Macradenia R.Br., and Ornithocephalus Hook., with two species each. The remaining eight genera are represented by a single species each in the study area. Morphological descriptions, a key for taxonomic identification, illustrations, and comments on distribution, ecology, phenology and morphology are provided for all inventoried species.

Entre os estudos com Orchidaceae na Amazônia, nenhum compreende a região da Volta Grande do rio Xingu, localizada no baixo Xingu. O objetivo deste estudo foi inventariar e estudar taxonomicamente as espécies de Oncidiinae (Orchidaceae) na Volta Grande do rio Xingu, estado do Pará, Brasil. O levantamento florístico foi realizado na área da Usina Hidrelétrica de Belo Monte, no município de Vitória do Xingu, inserido centralmente na chamada Volta Grande do Xingu. Foram realizadas coletas botânicas entre junho de 2011 e dezembro de 2013. Na área de estudo, foram inventariadas 27 espécies de Oncidiinae, distribuídas em 15 gêneros. Notylia Lindl. e Trichocentrum Poepp. & Endl. foram os mais ricos, com cinco e quatro espécies respectivamente, seguidos por Erycina Lindl., Ionopsis Kunth, Lockhartia Hook., Macradenia R.Br., e Ornithocephalus Hook., com duas espécies cada. Os oito gêneros restantes estão representados na área de estudo por uma única espécie. São fornecidas descrições morfológicas, chave taxonômica para identificação, ilustrações e comentários sobre distribuição, ecologia, fenologia e morfologia para todas as espécies inventariadas.

Oncidiinae (Orchidaceae) on the great curve of the Xingu River, Pará state, Brazil / Oncidiinae (Orchidaceae) na volta grande do Rio Xingu, Pará, Brasil

Abstract Among the studies on Orchidaceae in the Amazon, none comprised the region of the Great Curve of the Xingu River, located in the lower Xingu river. The aim of this study was to inventory and taxonomically study the species of Oncidiinae (Orchidaceae) in the Great Curve of the Xingu River, Pará state. The floristic survey was performed in the area of the Belo Monte hydroelectric plant, in the Vitória do Xingu municipality, centrally inserted in the called Great Curve of the Xingu River. Botanical collections were accomplished between June 2011 and December 2013. A total of 27 species of Oncidiinae, distributed in 15 genera, was inventoried in the study area. Notylia Lindl. and Trichocentrum Poepp. & Endl. were the richest genera, with five and four species, respectively, followed by Erycina Lindl., Ionopsis Kunth, Lockhartia Hook., Macradenia R.Br., and Ornithocephalus Hook., with two species each. The remaining eight genera are represented by a single species each in the study area. Morphological descriptions, a key for taxonomic identification, illustrations, and comments on distribution, ecology, phenology and morphology are provided for all inventoried species.

Resumo Entre os estudos com Orchidaceae na Amazônia, nenhum compreende a região da Volta Grande do rio Xingu, localizada no baixo Xingu. O objetivo deste estudo foi inventariar e estudar taxonomicamente as espécies de Oncidiinae (Orchidaceae) na Volta Grande do rio Xingu, estado do Pará, Brasil. O levantamento florístico foi realizado na área da Usina Hidrelétrica de Belo Monte, no município de Vitória do Xingu, inserido centralmente na chamada Volta Grande do Xingu. Foram realizadas coletas botânicas entre junho de 2011 e dezembro de 2013. Na área de estudo, foram inventariadas 27 espécies de Oncidiinae, distribuídas em 15 gêneros. Notylia Lindl. e Trichocentrum Poepp. & Endl. foram os mais ricos, com cinco e quatro espécies respectivamente, seguidos por Erycina Lindl., Ionopsis Kunth, Lockhartia Hook., Macradenia R.Br., e Ornithocephalus Hook., com duas espécies cada. Os oito gêneros restantes estão representados na área de estudo por uma única espécie. São fornecidas descrições morfológicas, chave taxonômica para identificação, ilustrações e comentários sobre distribuição, ecologia, fenologia e morfologia para todas as espécies inventariadas.

Evaluación de diferentes medios de cultivo in vitro en el desarrollo de híbridos de Phalaenopsis (Orchidaceae) / Evaluation of different in vitro culture media in the development of Phalaenopsis hybrid (Orchidaceae)

Los híbridos de Phalaenopsis tienen una gran importancia económica a nivel mundial, como flor cortada y planta ornamental, debido a sus flores vistosas y a la capacidad de adaptación a diferentes condiciones ambientales. Las técnicas de cultivo in vitro resultan indispensables para mejorar la eficacia germinativa, el crecimiento y desarrollo de orquídeas con fines comerciales e investigativos. En esta investigación se determinó el medio de cultivo más apropiado para la germinación in vitro de un híbrido de Phalaenopsis. Inicialmente se evaluó la viabilidad de las semillas utilizando la prueba de tetrazolio (TZ). Las semillas se desinfectaron y se cultivaron aplicando el método de la jeringuilla. El porcentaje de viabilidad en promedio fue de 92,2 % (P≤ 0,05: Tukey HSD), con un porcentaje de germinación entre todos los medios de 95,1 % (P≤ 0,05: Tukey HSD). El medio de cultivo más eficiente para la germinación de híbridos de Phalaenopsis a las 18 semanas de cultivo fue el Murashige & Skoog (MS) suplementado con agua de coco, y jugo de piña con diferencias estadísticamente significativas (P≤ 0,05: Tukey HSD), con respecto a los demás medios de cultivo, contribuyendo de esta manera al uso de componentes orgánicos con el fin de mejorar la germinación y desarrollo de Phalaenopsis.

The Phalaenopsis hybrids have a significant economic importance throughout the world, as ornamental flower or plant. It is because of its attractive flowers and its adaptation capacity into different environments. The different culture media in vitro are vital to improve the efficacy of germination, growing and development of the Orchids for commercial and research purposes. In this research, the most appropriated medium for in vitro propagation of Phalaenopsis hybrid was determined. At first, the seeds viability was evaluated by using tetrazolium test (TZ). The seeds were disinfected and cultivated by means of the syringe method. The viability percentage average was 92.2 % (P≤ 0.05: Tukey HSD), with a percentage of germination of 95.1 % (P≤ 0.05: Tukey HSD) in all the environments. The most efficient culture Medium for Phalaenopsis hybrid phenological development, at 16 weeks, was Murashige & Skoog (MS). Coconut water and pineapple juice were used as supplement showing statistically significant differences (P≤ 0,05: Tukey HSD), in comparison with the other culture media, contributing this way to the usage of organic components, which will be employed to improve the germination and development of the Phalaenopsis.

Anatomía foliar comparada y sistemática del clado-Trichocentrum con énfasis en Cohniella Asparagales: Orchidaceae / Comparative foliar anatomy and systematics of the Trichocentrum-clade with emphasis in Cohniella Asparagales: Orchidaceae

The genera Cohniella, Lophiarella, Lophiaris, and Trichocentrum are included in the Trichocentrum-clade. These genera are distributed from Florida and Northern Mexico to Southern Brazil and Northern Argentina, growing in tropical deciduous forests or tropical rain forests and thorn scrub forests to pine-oak forest, from sea level to 1 700m. The leaf anatomical structure of 23 members of the Trichocentrum-clade was explored as a source of taxonomic and phylogenetic characters. A total of 11 species of Cohniella, three species of Lophiarella, seven species of Lophiaris, two species of Trichocentrum, and other four species were included as outgroup. Anatomical characters were studied by cross sections and paradermic observations of the middle portion of fresh leaves. Although anatomical characters were fairly homogeneous throughout the clade, twelve vegetative anatomical, phylogenetically informative characters were selected and coded for an analysis that was performed using an exhaustive search implicit enumeration implemented through TNT. The strict consensus of 2 692 most parsimonious trees resulted in a poorly resolved polytomy, which however recovers the Trichocentrum-clade with a monophyletic, strongly supported Cohniella nested within it with unifacial leaves and the presence of cellular inclusions in the epidermis as synapomorphies. We concluded that the anatomy characters alone are insufficient to assess the relationships amongst the genera of the Trichocentrum-clade. However, the two synapomorphies recovered for Cohniella strongly support its monophyly when these are analyzed in conjunction with other data sources e.g., molecular and morphological characters.

El clado-Trichocentrum incluye los géneros Cohniella, Lophiarella, Lophiaris y Trichocentrum s.s. Estos géneros se distribuyen desde Florida y el Norte de México hasta el Sur de Brasil y Norte de Argentina, creciendo desde bosques caducifolios, bosques húmedos tropicales hasta matorrales espinosos y bosques de pino-encino, desde el nivel del mar hasta los 1 700m. En este estudio se exploró el valor taxonómico y filogenético de la estructura anatómica de las hojas de 23 especies del clado-Trichocentrum, repartidos en 11 especies de Cohniella, tres de Lophiarella, siete de Lophiaris y dos de Trichocentrum s.s., y de otras cuatro especies incluidas como grupo externo. Se realizaron secciones transversales y observaciones paradérmicas de la porción media de hojas frescas para el estudio de los caracteres anatómicos. Doce caracteres anatómico foliares fueron seleccionados y codificados para el análisis filogenético que se realizó mediante el uso de una búsqueda exhaustiva enumeración implícita con el programa TNT. El consenso estricto de 2 692 árboles más parsimoniosos dio lugar a una politomía que recupera dentro del clado-Trichocentrum a Cohniella como un clado monofilético fuertemente apoyado con sinapomorfías de las hojas unifaciales y la presencia de inclusiones celulares en la epidermis.

Anatomía foliar comparada y relaciones filogenéticas de 11 especies de Laeliinae con énfasis en Brassavola (Orchidaceae)

Comparative leaf anatomy and phylogenetic relationships of 11 species of Laeliinae with emphasis on Brassavola (Orchidaceae). Brassavola inhabits a wide altitude range and habitat types from Northern Mexico to Northern Argentina. Classification schemes in plants have normally used vegetative and floral characters, but when species are very similar, as in this genus, conflicts arise in species delimitation, and alternative methods should be applied. In this study we explored the taxonomic and phylogenetic value of the anatomical structure of leaves in Brassavola; as ingroup, seven species of Brassavola were considered, and as an outgroup Guarianthe skinneri, Laelia anceps, Rhyncholaelia digbyana and Rhyncholaelia glauca were evaluated. Leaf anatomical characters were studied in freehand cross sections of the middle portion with a light microscope. Ten vegetative anatomical characters were selected and coded for the phylogenetic analysis. Phylogenetic reconstruction was carried out under maximum parsimony using the program NONA through WinClada. Overall, Brassavola species reveal a wide variety of anatomical characters, many of them associated with xeromorphic plants: thick cuticle, hypodermis and cells of the mesophyll with spiral thickenings in the secondary wall. Moreover, mesophyll is either homogeneous or heterogeneous, often with extravascular bundles of fibers near the epidermis at both terete and flat leaves. All vascular bundles are collateral, arranged in more than one row in the mesophyll. The phylogenetic analysis did not resolve internal relationships of the genus; we obtained a polytomy, indicating that the anatomical characters by themselves have little phylogenetic value in Brassavola. We concluded that few anatomical characters are phylogenetically important; however, they would provide more support to elucidate the phylogenetic relantionships in the Orchidaceae and other plant groups if they are used in conjunction with morphological and/or molecular characters. Rev. Biol. Trop. 59 (3): 1047-1059. Epub 2011 September 01.

Brassavola crece en un amplio rango altitudinal y de tipos de hábitat desde el Norte de México hasta el Norte de Argentina. En los sistemas de clasificación de las plantas se utilizan normalmente caracteres vegetativos y florales, pero cuando las especies son muy similares, como es el caso de este género, los conflictos surgen en la delimitación de las especies, por lo tanto deben ser aplicados métodos alternativos de identificación. En este trabajo se exploró el valor taxonómico y filogenético de la estructura anatómica de las hojas de Brassavola, se consideró como grupo interno a siete especies de este género y como grupo externo a Guarianthe skinneri, Laelia anceps, Rhyncholaelia digbyana y Rhyncholaelia glauca. Entonces se realizaron secciones transversales de hojas frescas para el estudio de los caracteres anatómicos. Diez caracteres anatómicosvegetativos fueron seleccionados y codificados para el análisis filogenético. La reconstrucción filogenética se llevó a cabo bajo el principio de máxima parsimonia utilizando el programa NONA a través de WinClada. Todas las especies son anatómicamente similares, no obstante, difieren en algunos rasgos como presencia o no: de papilas epidérmicas, de hipodermis, de células con engrosamientos espiralados en la pared secundaria de las células del mesofilo, de inclusiones cristalinas; además en el tipo de hoja de acuerdo al arreglo del mesofilo; en la organización de los haces vasculares y de los paquetes de fibras extravasculares. En el árbol de consenso estricto se obtuvo una politomía. Asimismo, fue evidente que los caracteres anatómicos analizados son filogenéticamente poco informativos; sin embargo, en conjunción con caracteres morfológicos y/o moleculares, podrían dilucidar las relaciones filogenéticas.