Phytochemical profile and biological activities of Bromelia antiacantha extracts / Perfil fitoquímico e atividades biológicas de extratos de Bromelia antiacantha

Reports from popular medicine usually act as a basis for the development of new drugs from natural compounds with therapeutic actions for serious diseases and prevalence such as cancer. Bromelia antiacantha Bertol. is a species of the Bromeliaceae family, considered an unconventional food plant, found in the south and midwest regions of Brazil. Despite the high nutritional content and pharmacological potential of its fruits, few scientific studies report its biological actions. Thus, this study evaluates the phytochemical profile of aqueous and ethanol extracts obtained from B. antiacantha fruits, as well as their possible antioxidant, antitumor, and cytotoxic activities. The aqueous extract exhibited phenolic compounds and flavonoids, while ethanol extracts indicated the presence of flavonoids and coumarin in their composition, regardless of the region of collection. The ethanolic extract demonstrated a more promising antioxidant effect than the aqueous extract and also induced a significant inhibition in the viability of human cervical cancer cells of the SiHa strain. In addition, treatment with both extracts did not alter the viability of non-tumor cells of the immortalized human keratinocyte lineage (HaCaT). These results bring new data about extracts obtained from a native plant, edible and traditionally used in popular medicine, opening new perspectives for its possible therapeutic application.

Relatos da medicina popular costumam atuar como referencial para o desenvolvimento de novos fármacos a partir de moléculas naturais com ações terapêuticas para doenças de alta gravidade e prevalência como o câncer. Bromelia antiacantha Bertol. é uma espécie da família Bromeliaceae, considerada uma planta alimentícia não convencional (PANC), encontrada nas regiões sul e centro-oeste do Brasil. Apesar do alto teor nutritivo e potencial farmacológico de seus frutos, poucos estudos científicos relatam suas ações biológicas. Desta forma, este estudo avalia o perfil fitoquímico de extratos aquoso e etanólico obtidos de frutos de B. antiacantha, bem como a sua possível ação antioxidante, antitumoral e citotóxica. O extrato aquoso apresentou compostos fenólicos e flavonoides, enquanto os extratos etanólicos apontam a presença de flavonóides e cumarina em sua composição, independente da região de coleta. O extrato etanólico demonstrou efeito antioxidante mais promissor do que o extrato aquoso e também induziu uma inibição significativa na viabilidade de células humanas de câncer cervical da linhagem SiHa. Além disso, o tratamento com ambos extratos não alterou a viabilidade de células não tumorais da linhagem de queratinócitos humanos imortalizados (HaCaT). Estes dados trazem novas informações sobre extratos obtidos de uma espécie vegetal nativa, comestível e já utilizada tradicionalmente, mas abrindo novas perspectivas quanto a possíveis aplicações terapêuticas.

Seed mass maturity in the terrestrial bromeliad Hechtia perotensis (Bromeliaceae), endemic to Mexico / Madurez de las semillas en la bromelia terrestre Hechtia perotensis (Bromeliaceae), endémica de México

Abstract Introduction: H. perotensis is a plant with a high potential for ecological restoration because it yields thousands of seeds and grows under low levels of rain, poor soils and contrasting temperatures. However, little is known of the seed mass maturity (high seed germination, low seed fresh weight and low seed moisture content) in this species. Objective: Assess seed germination in the laboratory of H. perotensis during seed development and along the floral stalk (infructescence) in two sites one in rocky location and another near a lake. The hypothesis was that there is a time after flowering in which seeds have highest germination and fresh weight and that the apical, centre and base of the infructescence are different in seed germination and fresh weight in both sites. Methods: Capsules were collected in two sites one in rocky land (Frijol Colorado, Perote, Veracruz) and another near one lake (Alchichica, Puebla), in the months of August, September and November 2016 and January 2017. A repeated measure design (RMD) was used to analyze the effects of infructescence section on seed weight, moisture content and seed germination (41, 87, 152 and 215 days after flowering). Each evaluation time comprised five replicates, each one with 15 seeds. Results: Difference in seed germination, seed weight and moisture content between sections of the infructescence was not significant. However, significant differences were found not only between first and last sample dates, but mainly between first and second dates. Eighty-seven days after flower pollination seed moisture content was lower than 20 % and up to 80 % of seed germinated in both sites of sampling. Conclusions: In this study it was found that the moisture content of H. perotensis seed can be used as an indicator of the physiological maturity of the seed and it is also related to germination of the seed.

Resumen Introducción: H. perotensis es una planta con un alto potencial para la restauración ecológica debido a que produce miles de semillas y crece con niveles bajos de lluvia, suelos pobres y temperaturas contrastantes. Sin embargo, el desarrollo de la madurez de las semillas (germinación alta, baja humedad y bajo peso fresco de semilla) en esta especie es muy poco conocido. Objetivo: Evaluar la germinación en el laboratorio de las semillas de H. perotensis, durante el desarrollo de la semilla y a lo largo del tallo floral (infrutescencia) en dos sitios; uno en un área rocosa y otro cerca de un lago. La hipótesis fue que hay un tiempo después de la floración, en el que las semillas presentan la mayor germinación y menor peso fresco, y que las secciones apical, central y base de la infrutescencia, la germinación y el peso fresco de la semilla son diferentes en los dos sitios. Métodos: se recolectaron las cápsulas en dos sitios uno en un área rocosa (Frijol Colorado, Perote, Veracruz) y otro cerca de un lago (Alchichica, Puebla), en los meses de Agosto, Septiembre y Noviembre de 2016 y Enero de 2017. Se usó un diseño de medidas repetidas (DMR) para analizar los efectos de la sección de la infrutescencia sobre el peso fresco de la semilla, el contenido de humedad y la germinación de la semilla (41, 87, 152 y 215 días, después de la floración). Cada tiempo de evaluación comprendió cinco réplicas y 15 semillas. Resultados: La diferencia en la germinación de la semilla, el peso de la semilla y el contenido de humedad entre las secciones de la infrutescencia no fue significativa. Sin embargo, se encontraron diferencias significativas entre las fechas de muestreo inicial y final, pero sobre todo entre la primera y la segunda fecha. Asimismo, ochenta y siete días después de la polinización de las flores, el contenido de humedad de las semillas fue inferior al 20 % y superior al 80 % en la germinación de las semillas en ambos sitios de muestreo. Conclusiones: en este estudio se encontró que el contenido de humedad de la semilla de H. perotensis, puede usarse como un indicador de la madurez fisiológica de la semilla y también está relacionado con la germinación de la semilla.

Heterotrophic flagellates (Amorpha and Diaphoretiches) in phytotelmata bromeliad (Bromeliaceae) / Flagelados heterotróficos (Amorpha e Diaphoretiches) em fitotelmata de bromélia (Bromeliaceae)

Abstract Many plants may accumulate rainwater, forming phytotelmata, aquatic microhabitats inhabited by various organisms. The aim of this study was to conduct an inventory of heterotrophic flagellates associated with phytotelmata of the bromeliad Aechmea distichantha Lem., found in rocky cliffs on the Upper Paraná River. The bromeliads were removed manually from the rocky wall, the water was removed and cultures of organisms of each plant were mounted in Petri dishes. Sixteen species of heterotrophic flagellate were recorded, drawn and described, among them one species belonging to the Amorpha Domain and 15 species to the Diaphoretiches Domain. The groups with most species were Euglenida and Kinetoplastea. The low diversity of heterotrophic flagellates recorded in this study, compared to the plankton of lakes and reservoirs, is probably related to the fact that phytotelmata are habitats with extreme environmental conditions, thus selecting organisms tolerant to these environments.

Resumo Em sua superfície externa, muitas plantas podem acumular água da chuva formando fitotelmata, ou seja, micro-habitat aquáticos colonizados por diversos organismos, entre eles, os flagelados protistas. Fez-se um inventário de flagelados heterotróficos associados ao fitotelmata da bromélia Aechmea distichantha Lem., encontrada em paredões rochosos em um trecho da bacia do alto rio Paraná. As bromélias foram retiradas manualmente de paredão rochoso e, em laboratório, a água foi retirada dos tanques de cada planta, e foram montadas culturas dos organismos em placas de Petri. Os flagelados heterotróficos foram, então, observados in vivo sob microscópio óptico Olympus BX51, com sistema de contraste de interferência diferencial (DIC) acoplado. Dezesseis espécies flagelados heterotróficos foram registradas, desenhadas e descritas, sendo uma espécie pertencente ao Domínio Amorpha e 15 espécies ao Domínio Diaphoretiches. Os grupos mais especiosos foram Euglenida e Kinetoplastea. A diversidade relativamente reduzida de flagelados heterotróficos, quando comparada ao plâncton de lagos e reservatórios, pode estar relacionada ao fato de os fitotelmata serem habitats limitados, com condições ambientais extremas, selecionando, assim, organismos tolerantes a esses ambientes.

Conservation of Vriesea flammea L. B. Sm., an endemic Brazilian bromeliad: effects of nutrients and carbon source on plant development / Conservação de Vriesea flammea L. B. Sm., uma bromélia Brasileira endêmica: efeitos dos nutrientes e fonte de carbono sobre o desenvolvimento das plantas

Abstract Bromeliaceae is restricted to the Neotropical region and has a high degree of endemism, which contributes to increased biodiversity because of the diverse morphological characteristics of individuals. In order to develop an in vitro conservation technology to obtain plants for reintroduction, seeds of Vriesea flammea L.B.Sm. were collected, sterilized and germinated in culture medium. The plants obtained were cultured for 180 days in MS medium with different concentrations of mineral nutrients (25 and 50% of nitrogenous salts and macronutrients), and different concentrations of sucrose (20, 30, 40, 50 and 60 g L-1), and then acclimatized for 150 days on commercial substrate. When seeds were sterilized directly, only 4% of them were contaminated, whereas sterilization of capsules resulted in 43.6% contaminated seeds. Germination rates above 80% were recorded. Low concentrations of nitrogenous salts and macronutrients produced greater than 76% survival and promoted greater in vitro plant development than the complete MS medium. The development of the aerial system, root system, fresh mass and photosynthetic pigments were positively related to sucrose concentration in vitro. The highest sucrose concentration also indirectly promoted greater development of the aerial system and fresh mass of acclimatized plants. We established conditions for in vitro cultivation and acclimatization for efficient propagation of V. flammea with a view towards conservation of the species or reestablishment of natural populations.

Resumo Bromeliaceae é restrita à região neotropical, com alto grau de endemismo, que contribui para o aumento da biodiversidade, devido às características morfológicas dos indivíduos. Objetivando desenvolver uma tecnologia in vitro de conservação para obtenção de plantas com vistas à reintrodução, sementes de Vriesea flammea L.B.Sm. foram coletadas, esterilizadas, germinadas em meio de cultura e as plantas obtidas foram cultivadas por 180 dias em meio MS com diferentes concentrações de nutrientes minerais (25 e 50% dos sais nitrogenados e dos macronutrientes), bem como em diferentes concentrações de sacarose (20, 30, 40, 50 e 60 g L-1). Após, as plantas foram aclimatizadas por 150 dias em substrato comercial. Quando da esterilização das sementes, apenas 4% destas contaminaram. Por sua vez, a esterilização de cápsulas resultou em 43,6% de sementes contaminadas. Taxas de germinação superiores a 80% foram registradas. Baixas concentrações dos sais nitrogenados e de macronutrientes proporcionaram sobrevivência superior a 76% e promoveram maior desenvolvimento das plantas in vitro do que o meio MS completo. O desenvolvimento do sistema aéreo, radicular, a massa fresca e pigmentos fotossintéticos apresentaram relação positiva com a concentração de sacarose in vitro. A maior concentração de sacarose também propiciou indiretamente maior desenvolvimento do sistema aéreo e massa fresca das plantas aclimatizadas. Estabelecemos as condições de cultivo in vitro e aclimatização para a eficiente propagação de V. flammea com vistas à conservação da espécie ou reestabelecimento das populações naturais.

Micropropagation and in vitroconservation of Alcantarea nahoumii (Bromeliaceae), an endemic and endangered species of the Brazilian Atlantic Forest

Alcantarea nahoumii(Leme) J. R. Grant is a species native to theAtlantic Forest that stands out for ornamental purposes. The objective of this work was to evaluate the in vitrogerminationof A. nahoumiiseeds and establish a micropropagation protocol for production of seedlings so as to minimize the effects of predatory extractivism and develop an in vitroconservation system. Mature seeds were disinfested, established in three culture media (MS, MS½ and MS⅓) and incubated at four temperatures (20, 25, 30 and 35ºC)in a germination chamber. In the micropropagation experiment, stem segments were introduced in MS medium supplemented with 0.5 μM of 1-naphthaleneacetic acid (NAA) and 0.0, 2.2, 4.4 and 6.6 μM of 6-benzylaminopurine (BAP). For the in vitroconservation, plantlets were established in MS or MS½ medium supplemented with 15 g L-1or 30 g L-1of sucrose. The plants were acclimated with commercial substrate. The highest seed germination percentages were promoted by temperature conditions of 20 and 25ºC, with MS culture medium. The highest multiplication rate of shoots was obtained from the treatment without addition of the growth regulator or when combined with 2.2 μM of BAP + 0.5 μM of NAA. The acclimation of the plants occurred with high survival rate. The species can be conserved in vitrounder slow growth condition for 24 months when incubated in MS medium supplemented with 30 g L-1of sucrose.

Cosmarium bromelicola sp. nov. (Desmidiaceae, Zygnematophyceae), a new desmid species from Northeast Brazil / Cosmarium bromelicola sp. nov. (Desmidiaceae, Zygnematophyceae), uma nova espécie de desmídia do nordeste do Brazil

Abstract Cosmarium bromelicola sp. nov. is a new desmid species described from samplings carried out in bromeliad tanks (phytotelmata) from an area of rocky outcrops at Serra da Jiboia, Bahia State, Northeast Brazil. Presence of subtrapeziform cells with a deep depression at the apical region and twisted X-shaped cell in side view are the diagnostic features for the species. Relationships with the morphologically closest taxa are discussed.

Resumo Cosmarium bromelicola sp. nov. é uma nova espécie de desmídia descrita a partir de amostragens realizadas em tanques de bromélias (fitotelmata) de uma área de afloramentos rochosos na Serra da Jiboia, Bahia, nordeste do Brasil. A presença de células trapeziformes com uma depressão profunda na região apical e célula em forma de X em vista lateral são as características diagnósticas da espécie. Relações com os táxons morfologicamente próximos são discutidas.

The Tillandsia genus: history, uses, chemistry, and biological activity / El género Tillandsia: historia, usos, química y actividad biológica

Tillandsia L. genus comprises 649 species, with different uses at different times. T. usneoides L. uses are reported since the late- archaic and pre-Columbian cultures. In XIX-XX centuries, T. usneoides was used in some manufactured products, as polish and packing fruit. Tillandsia has a favorable reputation as medicine: for leucorrhea, rheumatism, ulcers, hemorrhoid treatment, as an anti-diabetic remedy, emetic, analgesic, purgative, contraceptive, antispasmodic and diuretic. Tillandsia chemical composition includes cycloartane triterpenes and hydroxy-flavonoids, which are present in at least 24 species. Several extracts and compounds from Tillandsia spp. have been reported with pharmacological actions, as anti-neoplasia, hypolipidemic, antifungal, anti-HSV-1, hypoglycemic and microbicide. This review communicates the economic importance, ethnobotany, chemistry composition and biological activities of the Tillandsia genus, and analyze its biological and economic perspective. Tillandsia genus has cultural, economic and pharmacological relevance, with a high potential in many essential aspects of the modern society.

El género Tillandsia L. comprende 649 especies, con diferentes usos en diferentes épocas. T. usneoides L. se han reportado desde el arcáico tardío hasta las culturas precolombinas. En los siglos XIX-XX, T. usneoides se usó en productos manufacturados: como abrasivo y embalaje de fruta. Como medicina tradicional, el género Tillandsia se reporta para leucorrea, reumatismo, úlceras, hemorroides, remedio antidiabético, emético, analgésico, purgante, anticonceptivo, antiespasmódico y diurético. Su composición química incluye triterpenos de tipo ciclo-artano e hidroxi-flavonoides, presentes en al menos 24 especies. Los extractos y compuestos del género Tillandsia se han reportado con propiedades antineoplásicas, hipolipidémicas, antifúngicas, anti-HSV-1, hipoglucemiantes y microbicidas. Esta revisión comunica la importancia económica, etnobotánica, composición química y las actividades biológicas del género Tillandsia, y analiza su perspectiva biológica y potencial económica. Tillandsia tiene importancia cultural, económica y farmacológica, con gran potencial en muchos aspectos esenciales de la sociedad moderna.

Distribución geográfica de angiospermas epífitas de la región terrestre prioritaria Cerros Negro-Yucaño, Oaxaca, México / Geographical distribution of epiphytic angiosperms of the priority terrestrial region of Cerros Negro-Yucaño, Oaxaca, Mexico

Resumen El conocimiento sobre la riqueza y la distribución de las plantas epifítas vasculares es aún incompleto en muchas áreas de México. Un ejemplo es la región terrestre prioritaria (RTP) Cerros Negro-Yucaño, que se ubica en el noroeste del estado de Oaxaca y pertenece a la región mixteca alta (Ñuu Savi Sukun, Ñuu vixi). Con base en la revisión de material de herbario en 12 colecciones institucionales mexicanas y en la recolección de especímenes en algunas localidades, principalmente cubiertas por bosque de encino, en 17 de los 18 municipios que incluye la RTP, se compilo un listado de las angiospermas epífitas. Se registró la presencia de 40 especies, distribuidas en 13 géneros y cinco familias; 28 taxa son endémicos de México y tres de ellos se conocen solamente de Oaxaca. Los géneros Tillandsia (18 spp.) y Peperomia (4) fueron los mejor representados en el área. El bosque de encino fue el tipo de vegetación en el cual se encontraron más epífitas (32 spp.) y también el intervalo altitudinal de 1 900 a 2 100 m (19 spp.). Plantas en floración de Artorima erubescens, Laelia albida, L. furfuracea y Prosthechea karwinskii, son utilizadas por pobladores de la región como ornamentales, durante las celebraciones religiosas de Día de Muertos y Semana Santa. Este impacto humano podría representar una amenaza para las poblaciones silvestres de estas especies en el futuro.(AU)

Abstract The knowledge on richness and distribution of epiphytic vascular plants is still incomplete in many areas of Mexico. An example is the terrestrial priority region (RTP) Cerros Negro-Yucaño, which is located in the Northwest portion of Oaxaca and belongs to the Mixteca Alta Region (Ñuu Savi Sukun, Ñuu Vixi). Based on herbarium material revision of 12 Mexican institutional collections and the collection of specimens in some localities, mainly covered by oak forest, in 17 of the 18 municipalities included in the RTP, we compiled a list of epiphytic angiosperms. The presence of 40 species, distributed in 13 genera and five families was recorded; 28 taxa are endemic to Mexico and three of them are only known from Oaxaca. The genera Tillandsia (18 spp.) and Peperomia (4) were the best represented genera in the area. Oak forest was the type of vegetation in which the most of the epiphytes were found (32 spp.), and also the altitudinal range between 1 900 and 2 100 m (19 spp.). Flowering plants of Artorima erubescens, Laelia albida, L. furfuracea, and Prosthechea karwinskii are used by the local people as ornamentals during the religious celebrations of Day of the Dead (Mexico) and Holly Week. This human impact might be a threat to the wild populations of these species in the future.(AU)

Responses of antioxidant enzymes, photosynthetic pigments and carbohydrates in micropropagated Pitcairnia encholirioides L.B. Sm. (Bromeliaceae) under ex vitro water deficit and after rehydration / Respostas de enzimas antioxidantes, pigmentos fotossintéticos e carboidratos em Pitcairnia encholirioides L.B. Sm. (Bromeliaceae) micropropagadas, sob déficit hídrico ex vitro e após reidratação

Abstract In this study, the activities of antioxidant enzymes, photosynthetic pigments, proline and carbohydrate contents in Pitcairnia encholirioides under ex vitro conditions of water deficit were evaluated. Results show that plants under progressive water stress, previously in vitro cultured in media supplemented with 30 g L-1 sucrose and GA3, accumulated more proline and increased peroxidase (POD) activity and the contents of photosynthetic pigments and carbohydrates. For plants previously in vitro cultured with 15 g L-1 sucrose and NAA, no differences were found for proline content and there were reductions in activities of peroxidase (POD), catalase (CAT) and poliphenoloxidase (PPO), and in contents of carbohydrates, with progress of ex vitro water deficit. After rehydration, plants showed physiological recovery, with enzymatic activities and contents of metabolites similar to those found in the controls not submitted to dehydration, regardless of the previous in vitro culture conditions. These results show that micropropagated P. encholirioides has high tolerance to dehydration once in ex vitro conditions, which can ensure the survival of plants from tissue culture when transferred to its natural environment, emphasizing the importance of such biotechnology for the propagation of endangered species.

Resumo Neste estudo, foram avaliadas as atividades de enzimas antioxidantes, pigmentos fotossintéticos, conteúdo de prolina e carboidratos em Pitcairnia encholirioides sob déficit hídrico em condições ex vitro. Os resultados mostraram que as plantas sob estresse hídrico progressivo, previamente cultivadas in vitro em meio de cultura suplementado com 30 g L-1 de sacarose e GA3 acumularam mais prolina e aumentaram a atividade da peroxidase (POD) e os teores de pigmentos fotossintéticos e carboidratos. Para plantas previamente cultivadas in vitro com 15 g L-1 de sacarose e ANA, não foram encontradas diferenças nos conteúdos de prolina e houve reduções nas atividades da peroxidase (POD), catalase (CAT) e polifenoloxidase (PPO), e no conteúdo de carboidratos, com o progresso do déficit hídrico ex vitro. Após a reidratação, as plantas apresentaram recuperação fisiológica, com atividades enzimáticas e conteúdo de metabólitos semelhantes aos encontrados nos controles não sujeitos à desidratação, independentemente das condições de cultivo in vitro. Estes resultados mostram que P. encholirioides micropropagada tem alta tolerância à desidratação uma vez em condições ex vitro, o que pode garantir a sobrevivência de plantas provenientes da cultura de tecidos quando transferidas para seu ambiente natural, enfatizando a importância desta biotecnologia para a propagação de espécies ameaçadas.

Influence of N-P-K fertilization at the acclimatization stage on micropropagated seedlings of Tillandsia bulbosa Hook / Influência da adubação com N-P-K na fase de aclimatização em mudas micropropagadas de Tillandsia bulbosa Hook

Bromeliads are known worldwide for their ornamental potential. In Brazil, species of the genus Tillandsia occur in the Atlantic rainforest, Amazon rainforest, and rocky fields. This work aimed to evaluate the influence of nitrogen, phosphorus, and potassium fertilization on micropropagated seedlings of Tillandsia bulbosa, at the acclimatization stage, and their leaf anatomy. The experiment was carried out in a completely randomized design, in a 4x4+1 factorial scheme, using coconut coir: earthworm humus: sand mixture as substrate (2:1:1). Urea, single superphosphate, and potassium chloride were used as nitrogen, phosphorus and potassium sources, respectively, at proportions of 50, 100, 200, and 400% of the dose recommended. The doses were distributed in four applications, testing total application at planting (1); ½ application at planting and ½ at 80 DAP (2); application at planting, at 50 DAP, and at 100 DAP (3); and » application at planting, » at 30 DAP, » at 60 DAP, and » at 120 DAP (4); and a control (without fertilization). Leaves anatomy was analyzed at 180 days after planting. Fertilization did not significantly influence the development of seedlings during acclimatization. The doses of 50, 100, and 200% provided thicker parenchyma of chlorophyll and aquifer and leaf blade. Tillandsia bulbosa can be acclimatized without fertilizer application.

As bromélias são conhecidas mundialmente por seu potencial ornamental. No Brasil, as espécies de Tillandsia podem ser encontradas na Mata Atlântica, Floresta Amazônica e campos rupestres. O objetivo deste trabalho foi verificar a influência de adubação com nitrogênio, fósforo e potássio em plântulas micropropagadas de Tillandsia bulbosa, na fase de aclimatização e na sua anatomia foliar. O experimento foi implantado em delineamento inteiramente casualizado, em esquema fatorial 4x4+1, utilizando como substrato a mistura de pó de coco: húmus de minhoca: areia (2:1:1). Ureia, super fosfato simples e cloreto de potássio foram utilizados como fontes de nitrogênio, fósforo e potássio, nas proporções de 50, 100, 200 e 400% da dose recomendada. As doses foram distribuídas em quatro aplicações, testando aplicação total no plantio (1); ½ no plantio e ½ aos 80 DAP (2); no plantio, aos 50 DAP e aos 100 DAP (3) e » no plantio, » aos 30 DAP, » aos 60 DAP e » aos 120 DAP (4), e uma testemunha (sem adubação). Foi realizado o estudo anatômico das folhas aos 180 dias. A adubação não influenciou significativamente o desenvolvimento das plântulas durante a aclimatização. As doses de 50, 100 e 200% proporcionaram maior espessura de parênquimas clorofiliano e aquífero e do limbo foliar. A aclimatização de Tillandsia bulbosa pode ser realizada sem a necessidade de adubação.

Pollen morphology and viability in Bromeliaceae

ABSTRACT Pollen morphology characterization is important in taxonomy, conservation and plant breeding, and pollen viability studies can support breeding programs. This study investigated pollen morphology and male fertility in 18 species of Bromeliaceae with ornamental potential. For morphological characterization, pollen grains were acetolyzed and characterization of exine was done using scanning and transmission electron microscopy. Pollen viability was investigated by in vitro germination and histochemical tests. Species belonging to Aechmea and Ananas genera presented medium size pollen, except for Ae. fasciata, with large pollen. Al. nahoumii, P. sagenarius and the Vriesea species analyzed showed large pollen, except for V. carinata, with very large pollen. Pollen of Aechmea, Ananas and P. sagenarius presented bilateral symmetry, diporate, exine varying from tectate to semitectate. Al. nahoumii and Vriesea species presented pollen with bilateral symmetry, monocolpate; exine was semitectate, reticulate and heterobrochate. Germination percentage and tube growth were greater in SM and BKM media. Histochemical tests showed pollen viability above 70% for all species, except for Ananas sp. (40%). Pollen morphology is important for the identification of species, especially in this family, which contains a large number of species. High rates of viability favor fertilization and seed production, essential for efficient hybrid production and conservation.

Diversity and distribution of epiphytic bromeliads in a Brazilian subtropical mangrove

ABSTRACT It is not unusual to find epiphytic bromeliads in mangroves, but most studies on mangrove vegetation do not record their presence. This study aimed to evaluate the diversity and distribution of epiphytic bromeliads in a subtropical mangrove. The richness, abundance and life form (atmospheric and tank) of bromeliads were recorded and compared among host tree species and waterline proximity. The effects of diameter and height of host trees on the abundance of bromeliads were also assessed. The mangrove was composed of Avicennia schaueriana, Laguncularia racemosa and Rhizophora mangle. We recorded seven bromeliad species of the genera Tillandsia and Vriesea. The waterline proximity did not affect the abundance or diversity of bromeliads, but atmospheric forms were predominant near the waterline, whereas tank bromeliads were more frequent in the interior of the mangrove. The three mangrove species hosted bromeliads, but L. racemosa was the preferred host. The species composition showed that the distribution of bromeliads is more related to the host species than to the distance from the waterline. Bromeliad abundance increased with tree size. Bromeliads can be biological indicators of ecosystem health; therefore, inventories and host tree preferences are necessary knowledge for an adequate management of sensitive ecosystems as mangroves.

Viabilidad y germinación de semillas de Hechtiaperotensis (Bromeliaceae) / Viability and germination of Hechtia perotensis (Bromeliaceae) seed

ResumenPoblaciones endémicas de Hechtiaperotensis se han descrito en Puebla y Veracruz, México. La recolección de semillas de buena calidad permite su uso en conservación, investigación y restauración ecológica. Algunos compuestos para evaluar la calidad de las semillas silvestres y endémicas, como nitrato de potasio (KNO3) y ácido giberílico (AG3), se utilizan para incrementar la germinación de las semillas y disminuir la latencia. La prueba de cloruro de trifeniltetrazolio (tetrazolio) se correlaciona con la viabilidad de las semillas porque se basa en la actividad de las deshidrogenasas de tejidos vivos que catalizan la respiración mitocondrial. El objetivo de este estudio fue conocer el tamaño y el peso de las cápsulas y las semillas y la germinación y viabilidad de las semillas de H. perotensis recolectadas en Veracruz en 2012 y 2015. Las hipótesis fueron (1) que la germinación y la viabilidad de las semillas son independientes del año de recolecta, (2) que hay una concentración adecuada de tetrazolio para identificar la viabilidad de las semillas y (3) que el pretratamiento con KNO3 o AG3 incrementa la germinación de las semillas. La germinación se evaluó con un diseño completamente al azar con tres tratamientos (testigo y promotores de la germinación KNO3 0.2 % y AG3 500 mg/L), cuatro tratamientos para la prueba de viabilidad (testigo, 0.2, 0.5 y 1.0 % de tetrazolio) y seis repeticiones de cada tratamiento. La germinación se evaluó en 100 semillas y la viabilidad en 25. Los resultados entre y dentro de años se analizaron con ANDEVA y prueba de comparación múltiple de medias de Tukey. La proporción de semillas no germinadas se cuantificó junto con el número de plántulas normales y anormales, semillas con embrión viable o sin él, y con viabilidad baja o sin viabilidad. En promedio la muestra recolectada en 2012 tuvo 36 % de semillas con embrión viable, 7 con viabilidad baja, 24 % no viables y 33 % sin embrión. Este resultado fue significativamente diferente al de 2015, que presentó 87 % de embriones viables, 10 % con viabilidad baja, 0 % no viables y 3 % sin embrión. La germinación también fue significativamente diferente entre los años (22 y 92 %). Los tratamientos pregerminativos no modificaron la germinación. La germinación y la viabilidad de las semillas de H. perotensis varían significativamente entre los años de recolecta.

Abstract:Endemic populations of Hechtiaperotensis have been described in Puebla and Veracruz, Mexico. Good quality seed collections can be used in conservation, research and ecological restoration. To evaluate seed quality of wild and endemic species, some compounds are used as effective promoters of germination, such as potassium nitrate (KNO3) and gibberellic acid (AG3), because they increase seed germination capacity and reduce latency. The triphenyl tetrazolium chloride (tetrazolium) test correlates seed viability because it is based on the activity of dehydrogenases in live tissues that catalyze mitochondrial respiration. The objective of this study was to obtain information on size and weight of capsules and seeds and seed germination and viability of H.perotensis, collected in Veracruz in the year 2012 and 2015. The hypotheses were 1) that seed germination and viability are independent of the year of collection, 2) that there is a tetrazolium concentration that can identify seed viability better than others, and 3) that pretreatment with KNO3 or AG3 improves seed germination. Seed germination was assessed using a completely randomized design with three treatments (control and the germination promoters 0.2 % KNO3 and 500 mg/L AG3), four treatments for the viability test (control, 0.2, 0.5 and 1.0 % of tetrazolium) and six replicates for each treatment. A total of one hundred seeds for germination experiments, and 25 seeds for the viability test were used. The results between and within years were analyzed with ANOVA and multiple comparison with the Tukey test. The proportion of non-germinated seeds was quantified along with the number of normal and abnormal seedlings, seeds with viable embryo, seeds without embryo, and seeds with low or no viability. On average, for the 2012 collected sample, 36 % had viable embryos, 7 % had low viability, 24 % were not viable and 33 % had no embryo. This result was significantly different from the 2015 sample, for which 87 % of seed showed viable embryos, 10 % had low viability, 0 % was not viable and 3 % had no embryo. Seed germination was also significantly different between years (22 and 92 %) Pregerminative treatments did not improve germination. Seed germination and viability of H. perotensis significantly varied between years of seed collection. Rev. Biol. Trop. 65 (1): 153-165. Epub 2017 March 01.

Effects of climate and forest structure on palms, bromeliads and bamboos in Atlantic Forest fragments of Northeastern Brazil / Efeitos do clima e da estrutura da vegetação em palmeiras, bromélias e bambus em fragmentos de Mata Atlântica no nordeste brasileiro

Abstract Palms, bromeliads and bamboos are key elements of tropical forests and understanding the effects of climate, anthropogenic pressure and forest structure on these groups is crucial to forecast structural changes in tropical forests. Therefore, we investigated the effects of these factors on the abundance of these groups in 22 Atlantic forest fragments of Northeastern Brazil. Abundance of bromeliads and bamboos were assessed through indexes. Palms were counted within a radius of 20 m. We also obtained measures of vegetation structure, fragment size, annual precipitation, precipitation seasonality and human population density. We tested the effects of these predictors on plant groups using path analysis. Palm abundance was higher in taller forests with larger trees, closed canopy and sparse understory, which may be a result of the presence of seed dispersers and specific attributes of local palm species. Bromeliads were negatively affected by both annual precipitation and precipitation seasonality, what may reflect adaptations of these plants to use water efficiently, but also the need to capture water in a regular basis. Bamboos were not related to any predictor variable. As climate and forest structure affected the abundance of bromeliads and palms, human-induced climatic changes and disturbances in forest structure may modify the abundance of these groups. In addition, soil properties and direct measurements of human disturbance should be used in future studies in order to improve the predictability of models about plant groups in Northeastern Atlantic Forest.

Resumo Palmeiras, bromélias e bambus são importantes elementos das florestas tropicais e, entender os efeitos do clima, pressão antrópica e estrutura da floresta nesses grupos é crucial para prever alterações estruturais em florestas tropicais. Portanto, nós investigamos os efeitos desses fatores na abundância desses grupos vegetais em 22 fragmentos de Mata Atlântica no nordeste brasileiro. Abundâncias de bromélias e bambus foram registradas através de índices. Palmeiras foram contadas em um raio de 20 m. Nós também obtemos medidas da estrutura da vegetação, tamanho de fragmento, precipitação anual, sazonalidade na precipitação e densidade populacional humana. Nós testamos os efeitos desses preditores nos grupos vegetais através de análises de caminhos. A abundância de palmeiras foi maior em florestas mais altas, com árvores mais grossas, dossel fechado e sub-bosque aberto, o que deve refletir a presença de dispersores de sementes e atributos específicos das espécies de palmeiras locais. Bromélias foram negativamente afetadas pela precipitação anual e pela sazonalidade na precipitação, o que deve refletir adaptações dessas plantas para o uso eficiente da água, mas também a necessidade de captar água regularmente. Bambus não estiveram relacionados com nenhum dos preditores avaliados. Dado que clima e estrutura florestal afetaram a abundância de bromélias e palmeiras, as mudanças climáticas e distúrbios na estrutura das matas causados por ações antrópicas podem alterar a abundância desses grupos. Adicionalmente, propriedades do solo e medidas diretas de distúrbios antrópicos devem ser usadas em estudos futuros para melhorar o poder preditivo dos modelos sobre a abundância de plantas na Mata Atlântica do nordeste brasileiro.

Volatile compounds profile of Bromeliaceae flowers / Perfil de compuestos volátiles de las flores en las Bromeliaceae

Abstract:Volatile compounds play a vital role in the life cycle of plants, possessing antimicrobial and anti-herbivore activities, and with a significant importance in the food, cosmetic, chemical, and pharmaceutical industry. This study aimed to identify the volatile compounds emitted by flowers of thirteen species belonging to four genera of Bromeliaceae, using headspace solid-phase micro-extraction and detection by gas chromatography-mass spectrometry. A total of 71 volatile compounds belonging to nine chemical groups were identified. The compounds identified represented more than 97 % of the major components in Aechmea bicolor, Ae. bromeliifolia, Ae. distichantha, Ae. fasciata, and Vriesea friburgensis. In the Ananas varieties, over 99 % of the components were identified, and around 90 % in V. simplex. V. friburgensis presented the largest diversity of volatiles with 31 compounds, while Alcantarea nahoumii presented only 14. All three Ananas varieties presented the same 28 compounds in relatively similar abundance, which has been confirmed by principal component analysis. Current taxonomy and pollination syndrome studies available can adequately explain the variation in volatile compounds among species. Rev. Biol. Trop. 64 (3): 1101-1116. Epub 2016 September 01.

ResumenLos compuestos volátiles tienen un papel vital en el ciclo de vida de las plantas. Poseen actividad antimicrobiana y anti-herbivoría biológica y una gran importancia en la industria de alimentos, cosméticos, perfumes, productos químicos y farmacéuticos. Este estudio tuvo como objetivo identificar los compuestos volátiles de trece flores de especies, pertenecientes a cuatro géneros de Bromeliaceae utilizando microextracción en fase sólida mediante cromatografía de gases hifenada con espacio de cabeza acoplada a espectrometría de masas. Se han identificado setenta y un diferentes compuestos volátiles pertenecientes a nueve grupos. Los compuestos identificados representaron más del 97 % de los componentes principales en Aechmea bicolor, Ae. bromeliifolia, Ae. distichantha, Ae. fasciata, Vriesea friburgensis, 99 % en las variedades de Ananas y 90 % en V. Simplex. V. friburgensis mostró la mayor diversidad de compuestos volátiles con 31, mientras que en Al. nahoumii se han encontrado sólo 14 compuestos. Las tres variedades de Ananas presentan los mismos 28 compuestos en cantidades relativamente similares, lo que se confirmó por el análisis de componentes principales. Estudios taxonómicos y síndromes de polinización disponibles podrían explicar la variación de los compuestos volátiles entre especies.

Proline levels, oxidative metabolism and photosynthetic pigments during in vitro growth and acclimatization of Pitcairnia encholirioides L.B. Sm. (Bromeliaceae) / Niveis de prolina, do metabolismo oxidativo e dos pigmentos fotossintéticos durante o crescimento in vitro e aclimatização de Pitcairnia encholirioides L.B. Sm. (Bromeliaceae)

Abstract This study aimed to evaluate the variation in the levels of proline, oxidative metabolism and photosynthetic pigments in plants of Pitcairnia encholirioides grown in vitro under different conditions and after acclimatization. The analyses were performed after 150 days of in vitro cultivation in MS media supplemented with 10 µM GA3 or 0.2 µM NAA, sucrose at 15 or 30 g L–1, in test tubes which allowed gas exchange or in a hermetically sealed system, and 180 days after acclimatization. The in vitro maintenance in hermetically sealed flasks, with GA3 and 15 g L–1 sucrose had adverse metabolic effects, which was demonstrated by the lower proline and photosynthetic pigments accumulation and by the increase in antioxidant enzymes activities. After acclimatization, differences for proline and photosynthetic pigments were no longer found and the enzymatic activities ranged unevenly. The results suggest that the in vitro cultivation in media with 0.2 µM NAA and 30 g L–1 sucrose, in test tubes capped with closures which allowed gas exchange, is more suitable for micropropagation of P. encholirioides, providing a prolonged maintenance of in vitro cultures and plantlets with superior quality for ex vitro development.

Resumo Este trabalho objetivou avaliar a contribuição da prolina, do metabolismo oxidativo e dos pigmentos fotossintéticos na propagação in vitro e aclimatização de Pitcairnia encholirioides, uma bromélia criticamente ameaçada de extinção. As análises foram realizadas após 150 dias de cultivo in vitro em meio MS suplementado com 10 µM de GA3 ou 0,2 µM de ANA, 15 ou 30 g L–1 de sacarose, em tubos de ensaio que permitiam trocas gasosas ou em sistema hermeticamente vedado, e também 180 dias após aclimatização. A manutenção in vitro em frascos hermeticamente fechados, com GA3 e 15 g L–1 de sacarose apresentou efeito metabólico adverso, demonstrado pelo menor acúmulo de prolina e pigmentos fotossintéticos e também pelo aumento das atividades de enzimas antioxidantes. Após aclimatização, as diferenças para prolina e pigmentos fotossintéticos não foram mais encontradas e as atividades enzimáticas variaram de maneira desuniforme. Os resultados sugerem que o cultivo in vitro em meio com 0,2 µM de ANA e 30 g L–1 de sacarose, em tubos fechados com tampas que permitem trocas gasosas, é mais adequado para a micropropagação de P. encholirioides, proporcionando uma manutenção prolongada das culturas in vitro e plântulas com qualidade superior para o desenvolvimento ex vitro.

Vascular epiphytes in a remnant of seasonal semideciduous forest in zona da mata, State of Minas Gerais, Brazil / Epífitas vasculares em um remanescente de floresta estacional semidecidual na zona da mata, Minas Gerais, Brasil

This study was performed in a remnant of Seasonal Semideciduous Forest at Fazenda Fortaleza de Sant'Anna, in the municipality of Chácara, Minas Gerais, Brazil (22.0129S, 43.8628W), in an area of ca. 1 ha, via monthly expeditions throughout the year 2012. We found 91 species of vascular epiphytes, distributed in 44 genera and 12 families. The richest families were Orchidaceae (35 spp.), Bromeliaceae (18 spp.), Polypodiaceae (10 spp.), Piperaceae (nine spp.) and Araceae (seven spp.). Two species were recorded for the first time in Minas Gerais: Rodriguezia sticta and Stelis oligantha (Orchidaceae). Rhipsalis oblonga (Cactaceae) was recollected in the state after about 100 years without records. Four species are cited in the list of endangered plants in Minas Gerais: Nematanthus crassifolius (Gesneriaceae) and Cattleya bicolor (Orchidaceae) are in the category "Vulnerable" (VU), Nidularium azureum (Bromeliaceae) and Pleurothallis pectinata (Orchidaceae) are "Critically Endangered" (CR). Nidularium azureum is also cited as CR in the Red Book of Brazilian Flora. Most species were classified as characteristic holoepiphytes and anemochory was the most frequent dispersion syndrome.

O estudo foi realizado em um remanescente de Floresta Estacional Semidecidual, na Fazenda Fortaleza de Sant'Anna, município de Chácara, Minas Gerais, Brasil (22,0129S, 43,8628W), em uma área aproximada de 1 ha, através de expedições mensais durante o ano de 2012. Foram encontradas 91 espécies de epífitas vasculares, distribuídas em 44 gêneros e 12 famílias. As famílias mais ricas foram Orchidaceae (35 spp.), Bromeliaceae (18 spp.), Polypodiaceae (10 spp.), Piperaceae (nove spp.) e Araceae (sete spp.). Duas espécies foram registradas pela primeira vez para Minas Gerais: Rodriguezia sticta e Stelis oligantha (Orchidaceae). Rhipsalis oblonga (Cactaceae) foi recoletada no estado depois de cerca de 100 anos sem registros. Quatro espécies estão presentes na lista de plantas ameaçadas de extinção em Minas Gerais: Nematanthus crassifolius (Gesneriaceae) e Cattleya bicolor (Orchidaceae) na categoria "Vulnerável" (VU), Nidularium azureum (Bromeliaceae) e Pleurothallis pectinata (Orchidaceae) como "Criticamente em Perigo" (CR). Nidularium azureum também é citado como CR no Livro Vermelho da Flora do Brasil. A maioria das espécies foi classificada como holoepífita característica e a síndrome de dispersão mais frequente foi a anemocoria.

In vitro growth of the bromeliad Alcantarea imperialis (Carrière) Harms with different concentrations of nitrogen / Crescimento in vitro da bromélia Alcantarea imperialis (Carrière) Harms com diferentes concentrações de nitrogênio

In various ecosystems, many plants have been reduced in number or even eliminated from natural habitats, including the endangered ornamental bromeliad Alcantarea imperialis (Carrière) Harms. The in vitro culture is a tool that has been used for conservation of endangered species and widely used for the production of ornamental plants. This study aimed at investigating the influence of different nitrogen concentrations on the growth of plantlets of the bromeliad A. imperialis grown in vitro. Seedlings of A. imperialis were cultured on Murashige and Skoog medium, supplemented with different concentrations of nitrogen (0.00, 3.75, 7.50, 15.00, 30.00, 60.00, 90.00, 120.00, and 175.00 mM) at 30 mmol m- 2 s-1, at 12 hour photoperiod and 26±2°C for six months. The results showed that plants grown with 15.00 and 60.00 mM N showed the best growth.

Em vários ecossistemas, o número de plantas está se reduzindo ou até mesmo erradicadas de seus hábitats naturais, e uma dessas espécies que está em perigo é a bromélia ornamental Alcantarea imperialis (Carrière) Harms. O cultivo in vitro é uma ferramenta que tem sido utilizada na conservação de espécies em extinção e tem sido amplamente utilizado para a produção de plantas ornamentais. Este trabalho teve como objetivo investigar a influência de diferentes concentrações de nitrogênio sobre o crescimento da bromélia A. imperialis cultivadas in vitro. Plântulas de A. imperialis foram cultivadas em meio de Murashige e Skoog, suplementado com diferentes concentrações de nitrogênio (0,00; 3,75; 7,50; 15,00; 30,00; 60,00; 90,00; 120,00 e 175,00 mM) em 30 mmol m-2 s-1, em 12h fotoperíodo, a 26±2°C durante seis meses. Os resultados mostraram que as plantas crescidas com 15,00 e 60,00 mM de N proporcionam o melhor crescimento.

Foraging behaviour of the Scale-throated Hermit Phaethornis eurynome Lesson, 1832 (Aves, Trochilidae) in Vriesea incurvata Gaudich (Bromeliaceae) / Comportamento alimentar de Phaethornis eurynome Lesson, 1832 (Aves, Trochilidae) em Vriesea incurvata Gaudich (Bromeliaceae)

In this study we tested for density-dependent relationships between visitation rates of the Scale-throated Hermit (Phaethornis eurynome) and the plant density and flower number of the bromeliad Vriesea incurvata, by comparing plots with varying densities of this bromeliad. Eight 100 m2 plots were established at least 200 m from each other; four plots contained 10-15 individuals of V. incurvata each, whereas the other four contained 4-5 individuals each. The visitors, number of visits, behaviour (nectar thief or potential pollinator) and the height of foraging were recorded during focal observations on the plants. The number of visits of P. eurynome varied according to the local density of V. incurvata, showing that the heterogeneous distribution of this bromeliad species may promote adjustments in the pollinator populations, through resource variation at a local scale.

Neste estudo foram testadas as relações entre taxas de visitação de Phaethornis eurynome e a densidade e o número de flores de Vriesea incurvata, comparando-se parcelas com diferentes densidades desta bromélia. Oito parcelas de 100 m2 foram estabelecidas a pelo menos 200 m uma da outra; quatro parcelas continham de 10 a 15 indivíduos de V. incurvata cada, enquanto as outras quatro continham de 4 a 5 indivíduos cada. Os visitantes, o número de visitas, o comportamento (pilhador ou potencial polinizador) e altura de forrageamento foram registrados em observações focais das plantas. O número de visitas por P. eurynome variou conforme a densidade de V. incurvata na área, demonstrando que a distribuição heterogênea desta bromélia pode promover ajustes populacionais por parte dos polinizadores, através da variação da oferta de recursos em uma escala local.

Efeito da luz e do sistema de ventilação natural em abacaxizeiro (Bromeliacea) micropropagado / Light effect and natural ventilation system on pineapple micropropagated

O emprego de luz natural e do sistema de micropropagação em ventilação natural proporciona maior irradiância e trocas gasosas no cultivo in vitro de plantas, ocasionando alterações na anatomia foliar e possibilitando um cultivo mais próximo ao fotoautotrófico. O presente trabalho teve por objetivo avaliar o crescimento e anatomia foliar de plantas de abacaxi cultivadas in vitro em dois sistemas de micropropagação (convencional e ventilação natural), com as plantas mantidas em ambiente com luz natural e artificial, e em meio de cultura com diferentes concentrações de sacarose. Os tratamentos constaram de diferentes concentrações de sacarose (0; 15; 30 e 45 g.L-1) em combinação com dois sistemas de micropropagação (convencional e ventilação natural), mantidos sob luz natural. Foram realizados dois tratamentos controle adicionais constando de sistemas de micropropagação (convencional e ventilação natural), acrescido de 30 g.L-1 de sacarose, sendo o cultivo realizado em sala de crescimento com luz e temperatura controlada. Maior crescimento in vitro foi observado em sistema de micropropagação com ventilação natural, em meio de cultura contendo 30 g.L-1 de sacarose, e com as plantas mantidas sob luz natural. A micropropagação em ventilação natural, com as plantas sob luz natural, promoveu alterações na anatomia foliar que proporcionaram maior espessura do parênquima clorofiliano durante o cultivo in vitro. A luz natural em combinação com o sistema de micropropagação em ventilação natural proporciona maior acumulo de biomassa e melhores características de anatomia foliar.

The natural light application on natural ventilation micropropagation systems makes a bigger irradiance and gaseous exchanges on in vitro culture, occasioning foliar anatomy alterations and possibiliting the culture more close of phothoautrofic conditions. The present study had the purpose to evaluate growth in vitro and foliar anatomy of pineapple plants (Ananas comosus L.) incubated in natural or artificial light, using two micropropagation systems (conventional and natural ventilation) with different sucrose concentrations. The treatments were composed of different sucrose concentrations (0; 15; 30 e 45 g. L-1) combined with different micropropagation systems (conventional and natural ventilation), incubated under natural light. Two additional treatments were made with conventional and natural ventilation micropropagation system incubated on growth room with control of irradiance and temperature. The biggest in vitro growth were observed on natural ventilation under natural light with 30 g. L-1 of sucrose on culture media, as well as the foliar anatomy alterations that made bigger chlorophyll parenquimata during in vitro culture. Natural ventilation micropropagation system in combination with natural light produces the greatest biomass production and the better foliar anatomy.