Optimization of callus cultures at Echinacea purpurea L. for the amount of caffeic acid derivatives

BACKGROUND: In order to produce an effective callus in Echinacea purpurea L.; determination of the explant type and growth regulators that best respond to callus induction and the optimization of the culture conditions to increase the amount of caffeic acid derivatives (CADs) in the obtained callus. CADs contents of callus cultures of E. purpurea were evaluated by establishing an effective callus induction system in vitro. RESULTS: Various medium containing different growth regulators were tested using leaf, petiole, cotyledon and root as the explants. The best callus development was achieved in MS medium with 1.0 mg l 1 2,4- D + 2.0 mg l 1 BAP in leaf, 1.0 mg l 1 NAA + 0.5 mg l 1 TDZ in petiole, 2.0 mg l 1 NAA + 1.0 mg l 1 TDZ in cotyledon and 0.5 mg l 1 NAA + 0.5 mg l 1 BAP in roots. Upon optimisation of callus growth, each type of explant was cultured for 4, 6, 8 and 10 weeks in medium for the analyses of caftaric acid, chlorogenic acid, caffeic acid and chicoric acid contents. The highest amounts of caftaric acid (4.11 mg/g) and chicoric acid (57.89 mg/g) were found from petiole explants and chlorogenic acid (8.83 mg/g) from root explants at the end of the 10-week culture time. CONCLUSIONS: As a result of the present study, the production of caffeic acid derivatives was performed by providing the optimization of E. purpurea L. callus cultures. Effective and repeatable protocols established in this study may offer help for further studies investigating the production of caffeic acid derivatives in vitro.

Defoliation heights for palisade grass cv. Marandu in silvopastoral system / [Alturas de desfolhação para capim-marandu em sistema silvipastoril]

The objective was to test the response of Urochloa brizantha cv. Marandu in monoculture and in silvopastoral system (SPS), at two distances from the trees, and define the best defoliation height for SPS. Four intermittent defoliation heights (25, 35, 45 and 55cm) and two distances from tree lines (2.5 and 5.0m) were evaluated in the SPS with a control defoliated with 25cm in full sun. The experiment was performed in a randomized block design with 3 replicates in a 4 × 2 + 1 split plot scheme. The control had higher forage accumulation (46.9kg/ha. day) than the SPS (31.1kg/ha. day). The bulk density was also higher in the control (0.89mg/cm³) than in SPS (0.48mg/cm³). The percentage of leaves (78.06%) and leaf/stem ratio (6.04) did not differ among the treatments. In the SPS, there was an increase of 31.07% in forage accumulation from 25 to 55cm. The forage accumulation and bulk density of Urochloa brizantha cv. Marandu in monoculture is greater than in the SPS regardless of the management goal and the distance from trees. The goal of 55cm in the SPS presented greater forage accumulation.(AU)

O objetivo foi testar a resposta de pastos de Urochloa brizantha cv. Marandu em monocultivo e em sistema silvipastoril (SSP), a diferentes distâncias das árvores, bem como definir qual altura de desfolhação é a mais indicada para manejo em SPS. Foram avaliadas quatro alturas de desfolhação intermitente (25, 35, 45 e 55cm) e duas distâncias das linhas de árvores (2,5 e 5,0m), no SPS, e uma testemunha desfolhada com 25cm, em pleno sol. O delineamento foi em blocos ao acaso, com três repetições, em esquema de parcelas subdivididas 4 × 2 + 1. O controle apresentou maior acúmulo de forragem (46,9kg/ha.dia) que a média do SSP (31,1kg/ha.dia). A densidade volumétrica da forragem do controle (0,89mg/cm³) foi maior que a do SSP (0,48mg/cm³). A porcentagem de folhas (78,06%) e a relação folha/colmo (6,04) não diferiram entre os tratamentos. O aumento das alturas de 25 para 55cm no SSP resultou em aumento de 31,07% no acúmulo de forragem. O acúmulo e a densidade volumétrica da forragem são maiores no monocultivo, independentemente da meta de manejo e da distância das árvores no sistema silvipastoril. A meta de 55cm no silvipastoril apresenta maior acúmulo de forragem.(AU)

Trichospilus diatraeae (Hymenoptera: Eulophidae): development and reproduction in Lepidoptera palm oil pests / Trichospilus diatraeae (Hymenoptera: Eulophidae): desenvolvimento e reprodução em lepidópteros-praga da palma de óleo

Abstract Brazil is a leading palm oil producer, but the defoliating caterpillars Opsiphanes invirae Hübner Brassolis sophorae L. (Lepidoptera: Nymphalidae) can reduce the productivity of this crop. The aim of this study was to evaluate the development and reproduction of the parasitoid Trichospilus diatraeae Cherian & Margabandhu (Hymenoptera: Eulophidae) in pupae of these oil palm defoliators. Ten O. invirae or B. sophorae pupae with up to two days old were exposed each to 30 T. diatraeae females for 48 hours. Parasitism and emergence of the progeny of T. diatraeae were similar in pupae of both Lepidoptera defoliators. The life cycle of this parasitoid was shorter in O. invirae (21.50 ± 0.42 days) pupae than with those of B. sophorae (27.60 ± 1.80 days). The number of the progeny (669.00 ± 89.62) and dead immature (217.13 ± 58.18) of T. diatraeae were higher in B. sophorae pupae than in those of O. invirae with 447.83 ± 51.52 and 13.50 ± 5.23, respectively. The sex ratio and female and male longevity of T. diatraeae emerged from these hosts were similar. The reproductive traits, especially the number of individuals (offspring) of T. diatraeae were better with B. sophorae pupae than with those of O. invirae.

Resumo O Brasil é um dos principais produtores de óleo de palma, porém as lagartas desfolhadoras Opsiphanes invirae Hübner e Brassolis sophorae L. (Lepidoptera: Nymphalidae) podem comprometer a produtividade dessa cultura. O objetivo deste trabalho foi avaliar o desenvolvimento e aspectos reprodutivos do parasitoide Trichospilus diatraeae Cherian & Margabandhu (Hymenoptera: Eulophidae) em pupas desses desfolhadores da palma de óleo. Dez pupas de O. invirae ou de B. sophorae com até dois dias de idade, foram expostas, cada uma, ao parasitismo por 30 fêmeas de T. diatraeae , por 48 horas. O parasitismo e a emergência da progênie de T. diatraeae foram semelhantes em pupas de ambas as espécies de lepidópteros desfolhadores da palma de óleo. O ciclo de vida desse parasitoide foi mais curto em pupas de O. invirae (21,50 ± 0,42 dias) que com as de B. sophorae (27,60 ± 1,80 dias). O número de progênie (669,00 ± 89,62) e de imaturos mortos (217,13 ± 58,18) de T. diatraeae foram maiores em pupas de B. sophorae que naquelas de O. invirae com 447,83 ± 51,52 e 13,50 ± 5,23, respectivamente. A razão sexual e a longevidade de fêmeas e machos de T. diatraeae emergidos desses hospedeiros foram semelhantes. As características reprodutivas, especialmente, o número de indivíduos (progênie) de T. diatraeae foram melhores com pupas de B. sophorae que com as de O. invirae .

Physiological response of Cucurbita pepo var. pepo mycorrhized by Sonoran desert native arbuscular fungi to drought and salinity stresses

ABSTRACT Plants response to symbiosis with arbuscular mycorrhizal fungi (AMF) under water stress is important to agriculture. Under abiotic stress conditions native fungi are more effective than exotics in improving plant growth and water status. Mycorrhization efficiency is related to soil fungi development and energy cost-benefit ratio. In this study, we assessed the effect on growth, water status and energy metabolism of Cucurbita pepo var. pepo when inoculated with native AMF from the Sonoran desert Mexico (mixed isolate and field consortium), and compared with an exotic species from a temperate region, under drought, low and high salinity conditions. Dry weights, leaf water content, water and osmotic potentials, construction costs, photochemistry and mycorrhization features were quantified. Under drought and low salinity conditions, the mixed isolate increased plant growth and leaf water content. Leaf water potential was increased only by the field consortium under drought conditions (0.5-0.9 MPa). Under high salinity, the field consortium increased aerial dry weight (more than 1 g) and osmotic potential (0.54 MPa), as compared to non-mycorrhized controls. Plants inoculated with native AMF, which supposedly diminish the effects of stress, exhibited low construction costs, increased photochemical capacity, and grew larger external mycelia in comparison to the exotic inoculum.

Leaf area estimation from linear measurements in different ages of Crotalaria juncea plants

ABSTRACT The goal of this study was to estimate the leaf area of Crotalaria juncea according to the linear dimensions of leaves from different ages. Two experiments were conducted with C. juncea cultivar IAC-KR1, in the 2014/2015 sowing seasons. At 59, 82, 102, 129 days after sowing (DAS) of the first and 61, 80, 92, 104 DAS of the second experiment, 500 leaves were collected, totaling 4,000 leaves. In each leaf, the linear dimensions were measured (length, width, length/width ratio and length × width product) and the specific leaf area was determined through Digimizer and Sigma Scan Pro software, after scanning images. Then, 3,200 leaves were randomly separated to generate mathematical models of leaf area (Y) in function of linear dimension (x), and 800 leaves for the models validation. In C. juncea, the leaf areas determined by Digimizer and Sigma Scan Pro software are identical. The estimation models of leaf area as a function of length × width product showed superior adjustments to those obtained based on the evaluation of only one linear dimension. The linear model Ŷ=0.7390x (R2=0.9849) of the real leaf area (Y) as a function of length × width product (x) is adequate to estimate the C. juncea leaf area.

Growth, photosynthetic pigments and production of essential oil of long-pepper under different light conditions

ABSTRACT Piper hispidinervum C. DC. is popularly known as long-pepper and it owns a commercial value due to the essential oil it produces. Long-pepper oil is rich in safrole and eugenoln components that have insecticidal, fungicidal and bactericidal activity. It has been establish that to medicinal plants light influences not only growth but also essential oil production. The growth, the content of photosynthetic pigments and the essential oil production of Piper hispidinervum at greenhouses with different light conditions was evaluated. The treatments were characterized by cultivation of plants for 180 days under different light conditions, produced by shading greenhouses with 50% and 30% of natural incident irradiance, two colored shading nets red (RN) and blue (BN) both blocking 50% of the incident radiation and one treatment at full-sun (0% of shade). The results showed that the treatments of 50% shade and RN and BN were the ones which stimulated the greater growth. Blue and red light also had the best production of photosynthetic pigments. Essential oil yielded more under full sun therefore this is the most indicated condition to produce seedlings for the chemical and pharmaceutical industry.

Anatomical traits related to stress in high density populations of Typha angustifolia L. (Typhaceae) / Características anatômicas relacionadas ao estresse em altas densidades populacionais de Typha angustifolia L. (Typhaceae)

Abstract Some macrophytes species show a high growth potential, colonizing large areas on aquatic environments. Cattail (Typha angustifolia L.) uncontrolled growth causes several problems to human activities and local biodiversity, but this also may lead to competition and further problems for this species itself. Thus, the objective of this study was to investigate anatomical modifications on T. angustifolia plants from different population densities, once it can help to understand its biology. Roots and leaves were collected from natural populations growing under high and low densities. These plant materials were fixed and submitted to usual plant microtechnique procedures. Slides were observed and photographed under light microscopy and images were analyzed in the UTHSCSA-Imagetool software. The experimental design was completely randomized with two treatments and ten replicates, data were submitted to one-way ANOVA and Scott-Knott test at p<0.05. Leaves from low density populations showed higher stomatal density and index. These modifications on stomatal characteristics were more evident on the leaf abaxial surface. Plants from low density populations showed thicker mesophyll and higher proportion of aerenchymal area. Roots from low density populations showed a higher proportion of the vascular cylinder. Whereas, plants from higher density populations showed greater thickness of the endodermis, exodermis, phloem and root cortex. Higher density populations showed a higher proportion of aerenchymal gaps in the root cortex. Therefore, cattail plants from populations growing under high density population show anatomical traits typical of plants under stress, which promotes the development of less functional anatomical modifications to aquatic environments.

Resumo Algumas espécies de macrófitas podem apresentar um elevado potencial de crescimento, colonizando extensas áreas de ambientes aquáticos. O crescimento descontrolado da taboa (Typha angustifolia L.) causa problemas para a biodiversidade local e para atividades antrópicas, isso também pode levar à competição e problemas para a própria espécie. Dessa forma, o objetivo desse trabalho foi verificar se ocorrem modificações anatômicas em plantas de Typha angustifolia L. de diferentes densidades populacionais, uma vez que essas podem auxiliar na compreensão da biologia dessa espécie. Raízes e folhas de T. angustifolia foram coletadas de populações naturais com alto e baixo adensamento populacional. Esse material vegetal foi fixado e submetido a procedimentos usuais de microtécnica vegetal. As lâminas foram observadas e fotografadas em microscopia de luz e as imagens analisadas no software UTHSCSA-Imagetool. O delineamento experimental foi inteiramente casualizado com dois tratamentos e dez repetições, os dados foram submetidos à análise de variância e ao teste de Scott-Knott para p<0,05. As folhas das populações com baixa densidade demonstraram maior densidade e índice estomáticos. Essas características foram mais evidentes na superfície abaxial em comparação com a superfície adaxial. As plantas de populações com baixo adensamento demonstraram maior espessura do mesofilo e área de aerênquima foliar. As raízes das populações com baixo adensamento demonstraram maior proporção do cilindro vascular. Por outro lado, as plantas de populações com alto adensamento demonstraram maior espessura da endoderme, exoderme, floema e do córtex nas raízes. Populações de alto adensamento demonstraram maior proporção de aerênquima no córtex radicular. Portanto, plantas de taboa sob alto adensamento populacional estão sob estresse o qual promove o desenvolvimento de modificações anatômicas menos funcionais para os ambientes aquáticos.

Phenological characterization and temperature requirements of Annona squamosa L. in the Brazilian semiarid region

ABSTRACT The phenological growth stages of various species of monocotyledonous and dicotyledonous plants can be uniformly coded using the Biologische Bundesanstalt, Bundessortenamt, und Chemische Industrie (BBCH) scale. The aim of the present study was to determine the duration of different phenological stages and the temperature requirements of the sugar-apple, Annona squamosa, during two crop cycles in the semiarid region of Brazil. Phenological stages were divided into eight of ten possible principal stages: (0) bud development, (1) leaf development, (3) shoot/branch development, (5) inflorescence emergence, (6) flowering, (7) fruit development, (8) fruit maturity and (9) senescence and the beginning of dormancy. The phenological cycle of the sugar-apple from having closed leaf buds to the fruit ripening stage lasted 149 and 164 days with temperature requirements of 1684.5 and 1786.7 degree days (DD) for the first and second crop cycles, respectively. The results provided important information that will inform the correct timing for crop management practices.

Seedling growth and survival of five tree species in secondary forests and adjacent pastures in the montane rain forests of Southern Costa Rica / Crecimiento y supervivencia de plántulas de cinco especies de árboles en bosques secundarios y pastizales adyacentes en un bosque lluvioso montano del sur de Costa Rica

Abstract:Successful forest restoration in tropical environments is limited by the paucity of studies on the initial establishment and early survival requirements of seedlings of most native tropical tree species under disturbed conditions. Here, we evaluated the initial growth responses and the regeneration potential of seedlings of five tree species native to Costa Rica (Hasseltia floribunda, Inga densiflora, Persea americana, Tapirira mexicana and Trichilia pittieri). Seedlings were grown in secondary forests and adjacent open pastures under contrasting conditions of light availability. We quantified seedling growth, survival and herbivory from August 2010 to August 2011 on a monthly basis, and measured differences in leaf mass per area (LMA) at the end of the experiment. We found significant variation in growth responses between the understory of secondary forests and pastures. Growth in height was highest in pastures across all species, with I. densiflora, P. americana and T. mexicana showing the most striking differences. In contrast, H. floribunda and T. pittieri did not show differences in diameter growth between environments. Except for T. mexicana, herbivory increased throughout the experiment in all the species. Herbivory increased faster in pastures for H. floribunda and T. pittieri and showed higher rates in the forest understory for I. densiflora and P. americana. Seedling survival showed significant differences for all species across environments. Survival of H. floribunda and I. densiflora was higher in secondary forests, whereas the other species showed higher survival in pastures. LMA showed higher values in the pastures across all species, showing that individuals exposed to high light conditions had great photosynthetic rate and great leaf construction capacity. Due to their rapid growth and high survival, I. densiflora and T. mexicana showed great potential to restore abandoned pastures and secondary forests. Increasing our knowledge on the response of seedlings under disturbed conditions in tropical ecosystems is critical for improving the restoration of altered environments by matching the ecological amplitude of native species with specific environmental conditions. Rev. Biol. Trop. 64 (4): 1565-1583. Epub 2016 December 01.

Resumen:La restauración exitosa del bosque en ambientes tropicales está limitada por la carencia de estudios sobre los requisitos de hábitat, el establecimiento inicial, y la sobrevivencia de las plántulas de la mayoría de las especies de árboles tropicales nativos. En este estudio, evaluamos las respuestas en el crecimiento inicial y el potencial de regeneración de las plántulas de cinco especies de árboles nativos de Costa Rica (Hasseltia floribunda, Inga densiflora, Persea americana, Tapirira mexicana y Trichilia pittieri). Sembramos las plántulas en bosques secundarios y en pastizales abiertos adyacentes con condiciones de luz contrastantes. Cuantificamos el crecimiento, la supervivencia y el daño por herbivoría de las plántulas de forma mensual entre agosto 2010 y agosto 2011, y medimos las diferencias en el la masa foliar por área de la hoja al final del experimento. Existió una variación significativa en el crecimiento de las plántulas expuestas a la sombra en los bosques secundarios y aquellas que crecieron en pastizales. El crecimiento en altura fue mayor en pastizales en todas las especies; I. densiflora, P. americana y T. mexicana mostraron la mayor diferencia entre ambientes. En con- traste, H. floribunda y T. pittieri no mostraron diferencias en el crecimiento del diámetro entre ambientes. Con la excepción de T. mexicana, todas las especies mostraron un aumento en herbivoría durante el estudio. En los pastizales, el incremento de la herbivoría fue más rápido en H. floribunda y T. pittieri, mientras que para I. densiflora y P. americana fue mayor en el bosque. La sobrevivencia de plántulas fue diferente entre ambientes para todas las especies. La sobrevivencia de H. floribunda e I. densiflora fue mayor en el bosque secundario, mientras que en las demás especies hubo mayor sobrevivencia de plántulas en los pastizales. La masa foliar por área fue mayor en las plántulas que crecieron en los pastizales que en bosque secundario para todas las especies, lo que sugiere que los individuos que crecieron en condiciones de alta incidencia de luz tuvieron mayor tasa fotosintética y mayor capacidad de construcción de la hoja. Debido al rápido crecimiento y alta supervivencia I. densiflora y T. mexicana tienen gran potencial para la restauración de bosques secundarios y pastizales abandonados. Generar información sobre la respuesta de plántulas creciendo en bosques secundarios y pastizales en regeneración es crítico para mejorar los procesos de restauración de ambientes alterados. La restauración de estos bosques depende de la correspondencia entre las condiciones específicas de hábitat y la plasticidad ecológica de las especies nativas.

Regression models for estimating leaf area of seedlings and adult individuals of Neotropical rainforest tree species / Modelos de regressão para estimativa da área foliar de indivíduos jovens e adultos de espécies arbóreas de floresta neotropical

Abstract Individual leaf area (LA) is a key variable in studies of tree ecophysiology because it directly influences light interception, photosynthesis and evapotranspiration of adult trees and seedlings. We analyzed the leaf dimensions (length – L and width – W) of seedlings and adults of seven Neotropical rainforest tree species (Brosimum rubescens, Manilkara maxima, Pouteria caimito, Pouteria torta, Psidium cattleyanum, Symphonia globulifera and Tabebuia stenocalyx) with the objective to test the feasibility of single regression models to estimate LA of both adults and seedlings. In southern Bahia, Brazil, a first set of data was collected between March and October 2012. From the seven species analyzed, only two (P. cattleyanum and T. stenocalyx) had very similar relationships between LW and LA in both ontogenetic stages. For these two species, a second set of data was collected in August 2014, in order to validate the single models encompassing adult and seedlings. Our results show the possibility of development of models for predicting individual leaf area encompassing different ontogenetic stages for tropical tree species. The development of these models was more dependent on the species than the differences in leaf size between seedlings and adults.

Resumo Área foliar individual (AF) é uma variável chave em estudos sobre a ecofisiologia de arbóreas, porque influencia diretamente a interceptação de luz, a fotossíntese e a evapotranspiração das árvores adultas e das mudas. Foram analisadas as dimensões foliares (comprimento - C e largura - L) de indivíduos adultos e de mudas de sete espécies arbóreas de florestas neotropicais (Brosimum rubescens, Manilkara maxima, Pouteria caimito, Pouteria torta, Psidium cattleyanum, Symphonia globulifera e Tabebuia stenocalyx), com o objetivo de testar a viabilidade de modelos de regressão linear para estimar a AF de indivíduos adultos e mudas. No sul da Bahia, Brasil, um primeiro conjunto de dados foi coletado entre março e outubro de 2012. A partir das sete espécies analisadas, apenas duas (P. cattleyanum e T. stenocalyx) apresentaram relações muito semelhantes entre e AF e CL, em ambos os estádios ontogenéticos. Para estas duas espécies, um segundo conjunto de dados foi coletado em agosto de 2014, a fim de validar os modelos únicos que englobam folhas de indivíduos adultos e mudas. Nossos resultados mostram a possibilidade de desenvolvimento de modelos para a predição da área foliar, abrangendo diferentes estádios ontogenéticos para espécies arbóreas tropicais. O desenvolvimento destes modelos foi mais dependente das espécies do que das diferenças entre o tamanho das folhas de mudas e de indivíduos adultos.

Plant structure predicts leaf litter capture in the tropical montane bromeliad Tillandsia turneri / A estrutura da planta determina a folhagem interceptada pela bromelia Tillandsiaturneri

Abstract Leaves intercepted by bromeliads become an important energy and matter resource for invertebrate communities, bacteria, fungi, and the plant itself. The relationship between bromeliad structure, defined as its size and complexity, and accumulated leaf litter was studied in 55 bromeliads of Tillandsia turneri through multiple regression and the Akaike information criterion. Leaf litter accumulation in bromeliads was best explained by size and complexity variables such as plant cover, sheath length, and leaf number. In conclusion, plant structure determines the amount of litter that enters bromeliads, and changes in its structure could affect important processes within ecosystem functioning or species richness.

Resumo As folhagens interceptadas pelas bromélias é um importante recurso para a comunidade de invertebrados, bactérias, fungos e para a própria planta. Estudou-se a relação entre a estrutura de 55 bromélias de Tillandsia tumeri, definida como o tamanho, a complexidade da planta, a folhagem acumulada por meio de regressão múltipla e o critério de informação de Akaike. Encontrou-se que as variáveis de tamanho, cobertura, comprimento da bainha e a variável de complexidade do número de folhas explicam a acumulação de folhas nas bromélias. Em conclusão, a estrutura do planta determina a quantidade de folhas armazenada na bromélia e os câmbios da estrutura da bromélia poderiam afetar importantes processos de funcionamento do ecossistema ou a riqueza de espécies.

Biomass allocation and nutrients balance related to the concentration of Nitrogen and Phosphorus in Salvinia auriculata (Salviniaceae) / Alocação de biomassa e o Balanço de nutrientes relacionados à concentração de Nitrogênio e Fósforo em Salvinia auriculata (Salviniaceae)

Abstract Aquatic plants can use differential allocation (trade-off) of carbon among their structures depending on the nutrition concentration. Given that N and P are limiting in the growth of plants, our questions were: Are the N and P concentrations in S. auriculata related to the biomass allocation to its structures? Is a differential allocation of N and P between floating and submerged leaves? We evaluated the relation between the nutrients and the biomass allocation, and the trade-off among the leaves using the Spearman correlation. Our results showed that N and P concentrations in S. auriculata are related to the biomass allocation to its structures, and that there is no trade-off of these nutrients between “shoot and root”. Thus, we can see the importance of N and P concentration in the biomass of S. auriculata, and why this plant is capable to development in different environments as a weedy.

Resumo Plantas aquáticas podem realizar alocação diferencial (trade-off) de carbono entre as suas estruturas dependo da disponibilidade de nutrientes. Considerando que N e P são limitantes para o crescimento de plantas, nossas perguntas foram: As concentrações de N e P em S. auriculata estão relacionadas com a alocação de biomassa para suas estruturas? Existe alocação diferencial de N e P entre os folíolos aéreos e a “raiz”? Avaliamos a relação entre os nutrientes e a biomassa, e o trade-off entre as folhas utilizando correlação de Spearman. Os resultados encontrados mostram que as concentrações de N e P em S. auriculata estão relacionadas com a alocação de biomassa para as suas estruturas, e que não há um trade-off de N e P entre os folíolos aéreos e a “raiz”. Dessa maneira, é possível observar a importância de N e P no ganho de biomassa de S. auriculata, e porque a espécie é capaz de se desenvolver em diferentes ambientes como uma planta daninha.

Phytotoxic potential of young leaves from Blepharocalyx salicifolius (Kunth) O. Berg (Myrtaceae) / Potencial fitotóxico de folhas jovens de Blepharocalyx salicifolius (Kunth) O. Berg

Abstract The loss of leaves by plant species found in the Cerrado (Brazilian savanna) is an energetically expensive process due to adverse environmental conditions and predation by herbivory. The mature leaves have adaptations which minimize these events. However, the young individuals lack these structures and produce high leaf concentrations of secondary metabolites as a form of protection. These compounds can be used in bioprospection of natural herbicides. Thus, this study aimed to evaluate the phytotoxicity of hexane, ethyl acetate and aqueous extracts of young leaves from Blepharocalyx salicifolius (Kunth) O. Berg on the elongation of wheat coleoptiles (Triticum aestivum L.) and evaluate the potential phytotoxic of ethyl acetate extract on germination, growth and cell size of metaxylem of sesame (Sesamum indicum L.) seedlings. The hexane and ethyl acetate extracts inhibited the elongation of wheat coleoptiles at all concentrations; however, the most promising results were observed in coleoptile fragments treated with the ethyl acetate extract. This treatment changed the mean germination time and the synchrony of sesame seeds, inhibited the growth of shoots and roots, reduced the dry weight of seedlings, led to abnormalities in the seedlings and reduced the length of the metaxylem cells in the sesame seedlings. These results demonstrated the phytotoxic potential of young leaf extracts of B. salicifolius and the high phytotoxicity of the ethyl acetate extract in the initial development of S. indicum.

Resumo A perda das folhas pelas plantas do Cerrado é um processo energeticamente custoso devido às condições ambientais adversas e intensa predação por herbivoria. As folhas maduras possuem adaptações que minimizam esses eventos. Entretanto, os indivíduos jovens carecem dessas estruturas e produzem elevadas concentrações foliares de metabólitos secundários como forma de proteção. Esses compostos podem ser utilizados na bioprospecção de herbicidas naturais. Dessa forma, objetivamos com este estudo avaliar a fitotoxicidade dos extratos hexânico, acetato etílico e aquoso de folhas jovens de Blepharocalyx salicifolius (Kunth) O. Berg sobre o alongamento de coleóptilos estiolados de Triticum aestivum L. (trigo) e avaliar o potencial fitotóxico do extrato acetato etílico sobre a germinação, crescimento e comprimento das células do metaxilema de Sesamum indicum (gergelim). Os extratos hexânico e acetato etílico inibiram o alongamento dos coleóptilos em todas as concentrações testadas, entretanto os resultados mais promissores foram observados nos fragmentos submetidos ao extrato acetato etílico. Este extrato alterou o tempo médio de germinação e a sincronia das sementes de gergelim, inibiu o crescimento da parte aérea e subterrânea com redução da massa seca das plântulas, levou ao surgimento acentuado de anomalias e causou redução no comprimento médio das células do metaxilema de plântulas de gergelim. Assim, ficou comprovado o potencial fitotóxico dos extratos de folhas jovens de B. salicifolius e a acentuada fitotoxicidade do extrato acetato etílico sobre o desenvolvimento inicial de S. indicum.

Phenology of two Ficus species in seasonal semi-deciduous forest in Southern Brazil / Fenologia de duas espécies de Ficus em floresta estacional semidecidual no sul do Brasil

Abstract We analyzed the phenology of Ficus adhatodifolia Schott ex Spreng. (23 fig tree) and F. eximia Schott (12 fig tree) for 74 months in a remnant of seasonal semi-deciduous forest (23°27’S and 51°15’W), Southern Brazil and discussed their importance to frugivorous. Leaf drop, leaf flush, syconia production and dispersal were recorded. These phenophases occurred year-round, but seasonal peaks were recorded in both leaf phenophases for F. eximia and leaf flushing for F. adhatodifolia. Climatic variables analyzed were positively correlated with reproductive phenophases of F. adhatodifolia and negatively correlated with the vegetative phenophases of F. eximia. In despite of environmental seasonality, little seasonality in the phenology of two species was observed, especially in the reproductive phenology. Both species were important to frugivorous, but F. adhatodifolia can play a relevant role in the remnant.

Resumo Foi analisada a fenologia de Ficus adhatodifolia Schott ex Spreng. (23 indivíduos) e F. eximia Schott (12 indivíduos), por 74 meses em um remanescente de floresta estacional semidecidual (23°27’S e 51°15’W, centro de visitantes) no sul do Brasil e discutido sua importância para os frugívoros. Foram registradas as fenofases: queda e brotamento de folhas, produção e dispersão de siconios. Estas fenofases ocorreram ao longo do ano, porém picos sazonais foram registrados em abscisão e brotamento foliares para F. eximia e brotamento foliar para F. adhatodifolia. As variáveis climáticas analisadas foram positivamente correlacionadas com as fenofases reprodutivas de F. adhatodifolia e negativamente correlacionadas com as fenofases vegetativas de F. eximia. Apesar da sazonalidade ambiental, foi observada baixa variação sazonal na fenologia das duas espécies, especialmente na fenologia reprodutiva. Ambas as espécies foram consideradas importantes para os frugivoros, porém F. adhatodifolia pode desempenhar papel de destaque no fragmento florestal.

Functional leaf traits of understory species: strategies to different disturbance severities / Traços funcionais foliares de espécies de sub-bosque: estratégias às diferentes intensidades de perturbação

The specific leaf area (SLA) has been related to environmental disturbances, showing a positive correlation between the disturbances intensities and SLA in a plant community. These studies, however, assessed the responses of plant community as a whole, neglecting species attributes, such as the position in the vertical stratum of forests. Considering the importance of SLA to understand forest ecological processes, this study aimed to determine the influence of the disturbance regime on the SLA of understory species, considering that, unlike for communities as a whole, an increase in the disturbance intensity implies a decrease in SLA of understory species. This study was conducted in nine understories of seasonal forests in Brazil. The most abundant species were selected and their SLA were evaluated. The variability of SLA among populations in different forests was analyzed by Student’s t-tests. The SLA of the understories (SLAU) was also compared by an adaptation of the Community-weighted mean index. The comparison of species SLA showed significant differences among the populations of understories under different disturbance regime, showing a decrease in SLA with an increase in the disturbance intensity. Similar results were found for the SLA of understories communities (SLAU), corroborating our hypothesis. The correlation between a reduction in species SLA and in SLA of understory with an increase in disturbance intensity, contradicted the trend observed in the literature for the community as a whole. This study highlights the importance of the evaluation of SLA in understories, as an indicator of the successional stage of communities.

A área foliar específica (SLA) tem sido relacionada a distúrbios ambientais, apresentando uma correlação positiva entre a intensidade de perturbação e a SLA da comunidade vegetal. Estes estudos, no entanto, avaliaram as respostas da comunidade vegetal como um todo, negligenciando os atributos por espécies, tais como a posição vertical no estrato florestal. Considerando a importância da SLA para entender os processos ecológicos das florestas, este estudo teve como objetivo determinar a influência do regime de perturbação na SLA de espécies de sub-bosque, cuja hipótese é que, ao contrário de comunidades como um todo, um aumento na intensidade de perturbação implica na diminuição da SLA de espécies de sub-bosque. Este estudo foi realizado em nove sub-bosque de florestas estacionais no Brasil. As espécies mais abundantes foram selecionados e suas SLA foram avaliadas. A variabilidade de SLA entre as populações em diferentes florestas foi analisada pelo teste t de Student. O SLA dos sub-bosque (SLAu) também foi comparado por uma adaptação do índice de média ponderada da comunidade. A comparação de SLA das espécies mostraram diferenças significativas entre as populações de sub-bosque sob um regime de distúrbios diferentes, mostrando um decréscimo na SLA com um aumento na intensidade de perturbação. Resultados semelhantes foram encontrados para o SLA dos sub-bosque (SLAu), corroborando nossa hipótese. A correlação entre a redução no SLA espécies e SLA do sub-bosque com um aumento na intensidade de perturbação contradiz a tendência observada na literatura para a comunidade como um todo. Este estudo destaca a importância da avaliação de SLA em sub-bosque, como um indicador do estágio sucessional das comunidades.

Plant Growth Promotion and Root Colonization by EPS Producing Enterobacter sp. RZS5 under Heavy Metal Contaminated Soil.

The heavy metal resistant bacterium isolated from field soil and identified as Enterobacter sp. RZS5 tolerates a high concentration (100-2000 mM) of various heavy metal ions such as Mn2+, Ni2+, Zn2+, Cu2+, CO2+ and Fe2+ when grown in such environment and produces exopolysaccharides (EPS). Here, we have demonstrated EPS production by Enterobacter sp. RZS5 during 60 h of growth in yeast extract mannitol broth (YEMB). The yield increased by two fold after the addition of 60 M of Ca2+; 50 M of Fe2+ and 60 M of Mg2+ ions in YEMB, and the optimization of physico-chemical parameters. EPS was extracted with 30% (v/v) of isopropanol as against the commonly used 50% (v/v) isopropanol method. EPS-rich broth promoted seed germination, shoot height, root length, number of leaves and chlorophyll content of wheat (Triticum aestivum) and peanut (Arachis hypogaea) seeds. The higher colony-forming unit of Enterobacter sp. in soil inoculated with EPS rich broth of Enterobacter sp. indicated the root colonizing potential and rhizosphere competence of the isolate. The FTIR spectra of the EPS extract confirmed the presence of the functional group characteristics of EPS known to exhibit a high binding affinity towards certain metal ions. This overall growth and vigour in plants along with the effective root colonization, reflected the potential of the isolate as an efficient bio-inoculant in bioremediation.

Genetic variability in G2 and F2 region between biological clones of human respiratory syncytial virus with or without host immune selection pressure

Human respiratory syncytial virus (HRSV) is an important respiratory pathogens among children between zero-five years old. Host immunity and viral genetic variability are important factors that can make vaccine production difficult. In this work, differences between biological clones of HRSV were detected in clinical samples in the absence and presence of serum collected from children in the convalescent phase of the illness and from their biological mothers. Viral clones were selected by plaque assay in the absence and presence of serum and nucleotide sequences of the G2 and F2 genes of HRSV biological clones were compared. One non-synonymous mutation was found in the F gene (Ile5Asn) in one clone of an HRSV-B sample and one non-synonymous mutation was found in the G gene (Ser291Pro) in four clones of the same HRSV-B sample. Only one of these clones was obtained after treatment with the child's serum. In addition, some synonymous mutations were determined in two clones of the HRSV-A samples. In conclusion, it is possible that minor sequences could be selected by host antibodies contributing to the HRSV evolutionary process, hampering the development of an effective vaccine, since we verify the same codon alteration in absence and presence of human sera in individual clones of BR-85 sample.

Reliable screening technique for evaluation of wild crucifers against mustard aphid Lipaphis erysimi (Kalt.).

Wild crucifers namely Arabidopsis thaliana, Brassica fruticulosa, B. rugosa, B. spinescens, B. tournefortii, Camelina sativa, Capsella bursa-pastoris, Crambe abysinnica, Cronopus didymus, Diplotaxis assurgens, D. gomez-campoi, D. muralis, D. siettiana, D. tenuisiliqua, Enatharocarpus lyratus, Lepidium sativum and Sinapis alba along with five cultivated Brassica species including B. rapa (BSH-1), B. juncea (Rohini), B. napus (GSC-6), B. carinata (DLSC-2) and Eruca sativa (T-27) were screened against mustard aphid Lipaphis erysimi (Kalt.) with a standardized technique under definite level of aphid pressure developed using specially designed cages. Observations have revealed that B. fruticulosa, B. spinescens, Camelina sativa, Crambe abysinnica and Lepidium sativum were resistant to mustard aphid L. erysimi with aphid infestation index (AII) ≤1. Capsella bursa-pastoris was highly susceptible to bean aphid, Aphis fabae during its vegetative stage (with 100% mortality). Other genotypes were found in the range of ‘susceptible’ to ‘highly susceptible’ with AII ranging 3-5.

Mannitol-induced drought stress on calli of Trigonella foenum-graecum L. Var. RMt-303.

Different explants of fenugreek, T. foenum-graecum L. (Var. RMt-303), were compared for their callus induction and subsequent shoot regeneration capabilities on Murashige and Skoog media supplemented with different phytohormones in varying concentration. The highest percentage of callus induction frequency was observed in 1ppm benzylaminopurine (BAP). Maximum shoots were induced on media supplemented with 0.5ppm BAP using leaf and stem tissues as explants. However, root tissues showed only callusing with no subsequent shooting. Cotyledonary node responded better than hypocotyls in terms of shoot induction on media supplemented with thidiazuron (0.1ppm). The callus was subjected to drought stress as simulated by reduced water potential of growth media due to addition of mannitol. Calli could withstand -2 MPa water potential till 30 days indicating that the drought stress tolerance mechanisms are functional in this variety. Chlorophyll a and b and total chlorophyll, proline and total phenolic contents, total peroxidase and catalase activities increased under stress conditions suggesting the tolerance of callus to drought stress. However, ascorbate peroxidase, guaiacol peroxidase activities were found to decrease slightly. Malondialdehyde and H2O2 contents were found to decrease while only a slight disturbance was found in membrane stability index. These results underline the mechanisms that are crucial for drought stress tolerance in fenugreek.

In vitro regeneration and ploidy level analysis of Eulophia ochreata Lindl.

Various parameters including explant-type, medium compositions, use of phytohormones and additives were optimized for direct and indirect regeneration of E. ochreata, a medicinal orchid under threat. Protocorm-like-bodies (PLBs) proved to be the best explants for shoot initiation, proliferation and callus induction. Murashige and Skoog’s (MS) medium containing 2.5 mg L-1 6-benzylaminopurine (BAP), 1.0 mg L-1 kinetin (Kin) and additives (adenine sulfate, arginine, citric acid, 30 mg L-1 each and 50 mg L-1 ascorbic acid) was optimal for shoot multiplication (12.1 shoots and 7.1 PLBs per explant with synchronized growth), which also produced callus. Shoot number was further increased with three successive subcultures on same media and ~40 shoots per explant were achieved after 3 cycles of 30 days each. Additives and casein hydrolysate (CH) showed advantageous effects on indirect shoot regeneration via protocorm-derived callus. Optimum indirect regeneration was achieved on MS containing additives, 500 mg L-1 CH, 2.5 mg L-1 BAP and 1.0 mg L-1 Kin with 30 PLBs and 6 shoots per callus mass (~5 mm size). The shoots were rooted (70% frequency) on one by fourth-MS medium containing 2.0 mg L-1 indole-3-butyric acid, 200 mg L-1 activated charcoal and additives. The rooted plantlets were hardened and transferred to greenhouse with 63% survival rate. Flow-cytometry based DNA content analysis revealed that the ploidy levels were maintained in in vitro regenerated plants. This is the first report for in vitro plant regeneration in E. ochreata.