EVALUACIÓN DE UN SISTEMA DE REGENERACIÓN POR EMBRIOGÉNESIS SOMÁTICA DE NEEM (Azadirachta indica) / Evaluation of a Somatic Embryogenesis Regenetation System for Neem (Azadirachta indica)

Azadirachta indica, también conocida como neem, es una especie arbórea leñosa perteneciente a la familia Meliaceae, de gran importancia en diversas disciplinas científicas, tales como la forestal y la médico-farmacéutica. Se estableció un método para la propagación in vitro de esta planta, evaluándose como explantes, secciones foliares de vitro-plantas, cotiledones y esquejes. Se emplearon medios semisólidos con combinaciones variables de la citocinina 6-benzylaminopurina (BAP) y las auxinas ácido 2,4-diclorofenoxiacético (2,4-D) y ácido indolacético (AIA). Se observó la formación de callo regenerativo, a partir del cual se generó embriogénesis somática primaria y secundaria, mediante los reguladores de crecimiento BAP (1,0 mg.L-1) y 2,4-D (0,2 mg.L-1), mientras que la formación de callo no regenerativo fue promovida por concentraciones mayores a 0,3 mg.L-1 de 2,4-D. De los explantes evaluados, la mayor frecuencia de regeneración de plantas (~67 %) se presentó con secciones cotiledonares.

Azadirachta indica, also known as neem, is a woody tree species belonging to the Meliaceae family, of great importance in various scientific disciplines, such as forestry, medicinal and pharmacist. It was established a method for in vitro propagation of this plant, evaluating explants as leaf sections of vitro-plants, cotyledons and stem cuttings, using semi-solid medium with various combinations of cytokinin 6-benzylaminopurine (BAP) and auxins 2,4-dicholorophenoxyacetic acid (2,4-D) and indol-3-acetic acid (IAA). Regenerative callus formation was observed, from which primary and secondary somatic embryogenesis was generated by growth regulators BAP (1.0 mg.L-1) and 2,4-D (0.2 mg.L-1), whereas non-regenerative callus formation was promoted by concentrations higher than 0,3 mg.L-1 of 2,4-D. Between the used explants, the highest frequency of plant regeneration (~ 67%) presented from cotyledon sections.

Leaf morphological strategies of seedlings and saplings of Rhizophora mangle (Rhizophoraceae), Laguncularia racemosa (Combretaceae) and Avicennia schaueriana (Acanthaceae) from Southern Brazil

AbstractThe initial phase of a plant life cycle is a short and critical period, when individuals are more vulnerable to environmental factors. The morphological and anatomical study of seedlings and saplings leaf type enables the understanding of species strategies of fundamental importance in their establishment and survival. The objective of this study was to analyze the structure of seedlings and saplings leaf types of three mangrove species, Avicennia schaueriana, Laguncularia racemosa, Rhizophora mangle, to understand their early life adaptive strategies to the environment. A total of 30 fully expanded cotyledons (A. schaueriana and L. racemosa), 30 leaves of seedlings, and 30 leaves of saplings of each species were collected from a mangrove area in Guaratuba Bay, Paraná State, Brazil. Following standard methods, samples were prepared for morphological (leaf dry mass, density, thickness) and anatomical analysis (epidermis and sub-epidermal layers, stomata types, density of salt secretion glands, palisade and spongy parenchyma thickness). To compare leaf types among species one-way ANOVA and Principal Component Analysis were used, while Cluster Analysis evaluated differences between the species. We observed significant structural differences among species leaf types. A. schaueriana showed the thickest cotyledons, while L. racemosa presented a dorsiventral structure. Higher values of the specific leaf area were observed for seedlings leaves of A. schaueriana, cotyledons of L. racemosa and saplings leaves of A. schaueriana and R. mangle. Leaf density was similar to cotyledons and seedlings leaves in A. schaueriana and L. racemosa, while R. mangle had seedlings leaves denser than saplings. A. schaueriana and R. mangle showed hypostomatic leaves, while L. racemosa amphistomatic; besides, A. chaueriana showed diacytic stomata, while L. racemosa anomocytic, and R. mangle ciclocytic. Seedling leaves were thicker in R. mangle (535 µm) and L.racemosa (520 µm) than in A. schaueriana (470.3 µm); while saplings leaves were thicker in L. racemosa (568.3 µm) than in A. schaueriana seedlings (512.4 µm) and R. mangle (514.6 µm). Besides, seedlings leaves palisade parenchyma showed increasing thickness in L. racemosa (119.2 µm) <A. schaueriana (155.5 µm) <R. mangle (175.4 µm); while in saplings leaves as follows R. mangle (128.4 µm) <A. schaueriana (183.4 µm) <L. racemosa (193.9 µm). Similarly, seedlings leaves spongy parenchyma thickness values were as follows A. schaueriana (182.6 µm) = R. mangle (192.8 µm) <L. racemosa (354.4 µm); while in saplings were A. schaueriana (182.6 µm) = R. mangle (187.3 µm) <L. racemosa (331.3 µm). The analyzed traits, in different combinations, represent morphological adjustments of leaf types to reduce water loss, eliminate salt excess, increase the absorption of light, allowing a higher efficiency on the maintenance of physiological processes in this initial growth stage. Rev. Biol. Trop. 64 (1): 305-317. Epub 2016 March 01.

ResumenLa fase inicial del ciclo de vida de una planta es un período corto y crítico, cuando los individuos son más vulnerables a factores ambientales. El estudio morfológico y anatómico del tipo de hojas de las plántulas y árboles pequeños, permite la comprensión de las estrategias de las especies, que es de importancia fundamental para su establecimiento y supervivencia. El objetivo de este estudio fue analizar la estructura de los tipos de hojas de las plántulas y árboles pequeños de tres especies de mangle: Avicennia schaueriana, Laguncularia racemosa y Rhizophora mangle, para entender sus estrategias de vida tempranas de adaptación al ambiente. Un total de 30 cotiledones completamente abiertos (A. schaueriana y L. racemosa), 30 hojas de plántulas, y 30 hojas de árboles pequeños de cada especie se recolectaron en una área de manglar en Bahía Guaratuba, Estado de Paraná, Brasil. Siguiendo los métodos estándares, se prepararon muestras para análisis morfológicos (biomasa de hoja seca, densidad y espesor) y anatómicos (epidermis y capas sub-epidérmicas, tipos de estomas, densidad de glándulas secretoras de sal y grosor del parénquima empalizado y del esponjoso). Para comparar los tipos de hojas entre las especies se utilizaron un modelo lineal y Análisis de Componentes Principales, mientras que un análisis de conglomerados evaluó las diferencias entre las especies. Observamos diferencias estructurales significativas entre tipos de hoja en las especies. A.schaueriana mostró cotiledones más gruesos, mientras que L. racemosa presenta una estructura dorsiventral. Se observaron valores más altos del área foliar específica para las hojas de las plántulas de A. schaueriana, cotiledones de L. racemosa y hojas de árboles pequeños de A. schaueriana y R. mangle. La densidad de la hoja fue similar a la de los cotiledones y hojas de plántulas de A. schaueriana y L. racemosa, mientras que R. mangle tenía las hojas de las plántulas más gruesas que los árboles pequeños. A. schaueriana y R. mangle mostraron hojas hipostomáticas; L. racemosa anfiestomáticas; por otro lado A. chaueriana mostró estomas diacíticos, L. racemosa anomocíticos y R. mangle ciclocíticos. Las hojas de las plántulas eran más gruesas en R. mangle (535 micras) y L. racemosa (520 micras) que en A. schaueriana (470.3 m); mientras que las hojas de las plántulas eran más gruesas en L. racemosa (568.3 m) que en A. schaueriana (512.4 micras) y R. mangle (514.6 m). Además el parénquima empalizado de las plántulas mostró un aumento de espesor en L. racemosa (119.2 m) <A. schaueriana (155.5 m) <R. mangle (175.4 m); mientras que en las hojas de los árboles pequeños fue de siguiente manera: R. mangle (128.4 m) <A. schaueriana (183.4 m) <L. racemosa (193.9 m). Del mismo modo, en las hojas de las plántulas los valores del espesor del parénquima esponjoso fueron: A. schaueriana (182.6 m) = R. mangle (192.8 m) <L. racemosa (354.4 m); mientras que en los árboles pequeños: A. schaueriana (182.6 m) = R. mangle (187.3 m) <L. racemosa (331.3 m). Los rasgos analizados, en diferentes combinaciones, representan ajustes morfológicos de tipos de hojas para reducir la pérdida de agua, eliminar el exceso de sal, aumentar la absorción de la luz, lo que permite una mayor eficiencia en el mantenimiento de los procesos fisiológicos en esta etapa de crecimiento inicial.

Cambios microestructurales en los granos de Phaseolus vulgaris endurecidos / Microstructural changes in hardened beans (Phaseolus vulgaris)

El endurecimiento de los granos de Phaseolus vulgaris almacenados a alta temperatura y alta humedad relativa es una de las principales limitantes para su consumo. El objetivo de esta investigación fue evaluar por microscopia de barrido electrónico los cambios estructurales ocurridos en los cotiledones y en la testa de los granos endurecidos. Los granos recién cosechados se almacenaron durante doce meses bajo dos condiciones: 5°C-34% HR y 37°C-75% HR. Esta última con el fin de propiciar el endurecimiento. Los granos almacenados crudos y cocidos se liofilizaron y se fracturaron. Las secciones de testa y cotiledones se observaron en un microscopio electrónico JSM-6390. Al cabo de doce meses se constató que los granos almacenados a 37°C-75% HR aumentaron su dureza en un 503%, mientras que los granos almacenados a 5°C-34% HR no incrementaron su dureza significativamente. A nivel microestructural, en los cotiledones de los granos crudos se notaron claras diferencias en el aspecto de la pared celular, en el tamaño del espacio intercelular y en la textura de la matriz proteica. Mientras que en la testa de los granos crudos se evidenciaron diferencias en la compactación de la empalizada y de la capa sub-epidérmica. En los granos cocidos se observó una total separación entre las células de los cotiledones de los granos blandos y una muy limitada separación en los granos duros. Se concluye que las diferencias observadas en los granos duros y blandos, demostraron una participación importante de ambas estructuras, cotiledones y testa, en el endurecimiento de los granos.

The hardening of Phaseolus vulgaris beans stored at high temperature and high relative humidity is one of the main constraints for consumption. The objective of this research was to evaluate by scanning electron microscopy, structural changes in cotyledons and testa of the hardened beans. The freshly harvested grains were stored for twelve months under two conditions: 5 ° C-34% RH and 37 ° C-75% RH, in order to promote hardening. The stored raw and cooked grains were lyophilized and fractured. The sections of testa and cotyledons were observed in an electron microscope JSM-6390. After twelve months, grains stored at 37 ° C-75% RH increased their hardness by 503%, whereas there were no significant changes in grains stored at 5 ° C-34% RH. At the microstructural level, the cotyledons of the raw grains show clear differences in appearance of the cell wall, into the intercellular space size and texture matrix protein. There were also differences in compaction of palisade and sub-epidermal layer in the testa of raw grains. After cooking, cotyledon cells of the soft grains were well separated while these of hard grains were seldom separated. In conclusion, the found differences in hard and soft grains showed a significant participation of both structures, cotyledons and testa, in the grains´ hardening.

Seedling structure of Euphorbia L. and Chamaesyce Gray species / Estrutura da plântula de espécies de Euphorbia L. e Chamaesyce Gray

Chamaesyce prostrata (Aiton) Small, Euphorbia heterophylla L. and Euphorbia graminea Jacq. are weedy species that occur with relative frequency in the region of Maringá, Paraná State, Brazil. Data about the morphology and anatomy of seedlings of these species were obtained, with emphasis on significant structural differences of seedlings between species of Euphorbia and Chamaesyce. Seedlings are phanerocotylar and epigeal. Root is axial and triarch. Cotyledons are leaf-like and dorsiventral. Eophylls and prophylls are simple and dorsiventral. Morphological variations occur in the cotyledon shape of Euphorbia seedlings. It is remarkable the Kranz structure of leaves that occurs only in Chamaesyce.

Chamaesyce prostrata (Aiton) Small, Euphorbia heterophylla L. e E. graminea Jacq. (Euphorbiaceae) são espécies invasoras frequentes na região de Maringá, Paraná. Foram obtidos dados sobre a morfologia e anatomia das plântulas dessas espécies, com ênfase se há caracteres estruturais significativos das plântulas entre as espécies de Euphorbia e Chamaesyce. As plântulas são fanerocotiledonares e epígeas. A raiz é axial e triarca. Os cotilédones são foliáceos e dorsiventrais. Os eofilos e profilos são folhas simples e dorsiventrais. As variações mofológicas ocorrem no formato dos cotilédones das plântulas de Euphorbia. Sob o aspecto anatômico é especialmente notável a estrutura Kranz das folhas que ocorrem somente em Chamaesyce.

Descrição morfológica para identificação das plântulas de nove espécies lenhosas de uma floresta de restinga / Morphological description to identify the seedlings of woody species

O estágio de plântula é uma importante fase do ciclo vegetal, pois fornece informações relevantes que podem ser utilizadas em estudos taxonômicos e ecológicos. Estudos que enfocam a descrição morfológica revelam caracteres que, muitas vezes tendem a desaparecer ao longo do desenvolvimento do vegetal, mas que podem ser relevantes para a diferenciação dos táxons. Além disso, através da caracterização dos distintos tipos de plântulas presentes nos diferentes ecossistemas é possível inferir sobre a historia evolutiva dos grupos vegetais, servindo também como subsídio para estudos de regeneração. Frente ao exposto, o presente estudo objetiva caracterizar a morfologia das plântulas de nove espécies lenhosas, com cotilédones CHR, com vistas à identificação das espécies, fornecendo subsídios para trabalhos taxonômicos, filogenéticos e ecológicos, além de montar uma chave de identificação para estas espécies. A área de estudo foi uma floresta de restinga localizada no litoral sul de Pernambuco, sementes foram obtidas de frutos maduros e semeadas em bandejas contendo substrato arenoso, sendo mantidas em casa de vegetação onde seu desenvolvimento foi acompanhado diariamente. Como resultado, foi possível verificar que algumas espécies, tais como Andira fraxinifolia, Casearia javitensis, Eugenia hirta, Maytenus obtusifolia, Protium heptaphyllum e Ximenia americana, na fase de plântula apresentaram características morfológicas diferentes da fase adulta como a quantidade de folíolos, textura e forma dos eofilos. Estes dados foram similares aos encontrados em outros estudos para florestas tropicais. A partir dos resultados, pode-se concluir que os diferentes caracteres morfológicos entre as plântulas estudadas são importantes para serem utilizados na identificação e reconhecimento das mesmas em campo, bem como na caracterização de suas espécies.

The seedling stage is an important stage of the cycle plant because it provides relevant information that can be used in taxonomic and ecological studies. Studies that focus on morphological description reveal characters who often tend to disappear during the development of the plant, but that may be relevant to the differentiation of taxa. Furthermore, through the characterization of the different types of seedlings present in different ecosystems is possible to infer the evolutionary history of plant groups, serving also as subsidy for regeneration studies. Based on these, this study aims to characterize the morphology of seedlings of nine woody species with cotyledons CHR, with a view to identifying the species, providing information for taxonomic, phylogenetic and ecological, besides setting up a dichotomous key. The study area was a forest restinga located on the southern coast of Pernambuco, seeds were collected from mature fruits and sown in trays containing sandy soil, and kept in a greenhouse where his development was monitored daily. As a result, we found that some species, such as Andira fraxinifolia, Casearia javitensis, Eugenia hirta, Maytenus obtusifolia, Protium heptaphyllum and Ximenia american at seedling stage showed different characteristics of young plant as quantity of leaflets, texture and shape of eophylls. These data were similar to those found in other studies for tropical forests. From the results it can be concluded that the different morphological features between the seedling analyzed are important for use in the identification and recognition of the same field, as well as the characterization of its species.