Early changes in gene expression induced by acute UV exposure in leaves of Psychotria brachyceras, a bioactive alkaloid accumulating plant.

UV-B radiation can damage biomolecules, such as DNA, RNA, and proteins, halting essential cellular processes; this damage is partly due to ROS generation. Plant secondary metabolites may protect against UV-B. Psychotria brachyceras Müll. Arg. (Rubiaceae), a subtropical shrub, produces brachycerine, a monoterpene indole alkaloid mainly accumulated in leaf tissues, which displays antioxidant and antimutagenic activities. Exposure of P. brachyceras cuttings to UV-B radiation significantly increases leaf brachycerine concentration. It has been suggested that this alkaloid might contribute to protection against UV-B damage both through its quenching activity on ROS and as UV shield. To identify differentially expressed genes of P. brachyceras in response to UV-B and investigate a possible influence of this stimulus on putative brachycerine-related genes, suppressive subtractive hybridization was applied. Complementary DNA from UV-B-treated leaves for 24 h was used as tester, and cDNA from untreated leaves, as driver. After BLASTX alignments, 134 sequences matched plant genes. Using quantitative RT-PCR, selected genes potentially related to brachycerine showed significant increases in transcription after UV-B exposure: tryptophan decarboxylase, ACC oxidase, UDP-glucose glucosyltransferase, lipase, and serine/threonine kinase. Results suggest a possible involvement of brachycerine in acute UV-B responses and show that alkaloid accumulation seems at least partly regulated at transcriptional level.

Psychotria hoffmansegiana (Willd ex Roem. & Schult.) Mull. Arg. and Palicourea marcagravii st. Hil. (Rubiaceae): potential for forming soil seed banks in a brazilian Cerrado.

The germinability of artificially buried Psychotria hoffmansegiana and Palicourea marcagravii seeds in Cerrado soil was tested, with the aim of evaluating whether dispersed seeds may be able to form a soil seed bank. The assays were carried out at a Cerrado Reserve in São Paulo State, Brazil. Seed samples were placed in nylon bags and buried at two different depths and in two different sites. Samples were periodically exhumed and germination tests were performed with both exhumed and dry stored seeds. In general, soil storage favoured seed survival and germination when compared to dry stored seeds. The seed germination was little affected by soil depth and by burial environment. Seeds of both species remained viable for at least 13 months, considering the time lapse between the collection and the end of the germination tests. It was suggested that both species can potentially form a persistent soil seed bank in Cerrado.

Variation in pollinator assemblages in a fragmented landscape and its effects on reproductive stages of a self-incompatible treelet, Psychotria suterella (Rubiaceae).

Few studies of plant-pollinator interactions in fragmented landscapes evaluate the consequences of floral visitor variation on multiple stages of plant reproduction. Given that fragmentation potentially has positive or negative effects on different organisms, and that self-incompatible plant species depend on pollinators for sexual reproduction, differences in floral visitor assemblages may affect certain plant reproductive stages. We evaluated how pollinator assemblage, availability of floral resources, pollination, reproductive output, and seed and seedling performance of Psychotria suterella Muell. Arg. varied among three fragmentation categories: non-fragmented habitats, fragments connected by corridors, and isolated fragments. Richness and frequency of floral visitors were greater in fragments than in non-fragmented sites, resulting mainly from the addition of species typically found in disturbed areas. Although 24 species visited Psychotria suterella flowers, bumblebees were considered the most important pollinators, because they showed the highest frequency of visits and were present in eight out of ten sites. Additionally, the number of pollen tubes per flower per visit was lower in areas without bumblebees. The increased visitation in fragments seemed to enhance pollination slightly. However, fruit and seed output, germination, and seed and seedling mass were similar in non-fragmented sites, connected sites, and isolated fragments. Our results suggested that, even for a self-incompatible species, responses to habitat fragmentation at different stages of plant reproduction might be decoupled from the responses observed in floral visitors, if fruit set is not pollen limited. If all reproductive stages were considered, variation on the small scale was more important than the variation explained by fragmentation category. In spite of its self-incompatible breeding system, this plant-pollinator system showed resilience to habitat fragmentation, mainly as a result of high availability of potential mates to P. suterella individuals, absence of pollen limitation, and the presence of bumblebees (Bombus spp.) throughout this highly connected landscape.

Psychotria hoffmansegiana (Willd ex Roem. & Schult.) Mull. Arg. and Palicourea marcagravii st. Hil. (Rubiaceae): potential for forming soil seed banks in a brazilian Cerrado

The germinability of artificially buried Psychotria hoffmansegiana and Palicourea marcagravii seeds in Cerrado soil was tested, with the aim of evaluating whether dispersed seeds may be able to form a soil seed bank. The assays were carried out at a Cerrado Reserve in São Paulo State, Brazil. Seed samples were placed in nylon bags and buried at two different depths and in two different sites. Samples were periodically exhumed and germination tests were performed with both exhumed and dry stored seeds. In general, soil storage favoured seed survival and germination when compared to dry stored seeds. The seed germination was little affected by soil depth and by burial environment. Seeds of both species remained viable for at least 13 months, considering the time lapse between the collection and the end of the germination tests. It was suggested that both species can potentially form a persistent soil seed bank in Cerrado.

Observou-se a germinabilidade de sementes de Psychotria hoffmansegiana e Palicourea marcagravii enterradas em solo de Cerrado, buscando-se avaliar o potencial de formação de banco de sementes no solo. Ensaios foram realizados em uma Reserva de Cerrado, São Paulo, Brasil. Amostras de sementes foram colocadas em sacos de tela de nylon e enterradas em duas profundidades, e em dois ambientes. Amostras eram periodicamente desenterradas e submetidas a teste de germinação, juntamente com sementes armazenadas em laboratório. Em geral, o armazenamento no solo favoreceu a sobrevivência e a capacidade de germinação das sementes, em comparação com sementes armazenadas em laboratório. A germinação foi pouco afetada pela profundidade de enterramento e pelo ambiente. Sementes de ambas as espécies conservaram a viabilidade por 13 meses, considerando-se o período entre a coleta e o final dos testes de germinação. É sugerido que ambas as espécies são potencialmente capazes de formar banco de sementes persistente no solo do Cerrado.

Quality of seeds produced by Psychotria tenuinervis (Rubiaceae): distance from anthropogenic and natural edges of Atlantic forest fragment.

The aim of this study was to investigate whether there were differences in the genetic variability and rate and velocity of the seed germination produced by Psychotria tenuinervis located at anthropogenic edges, natural edges, and in the forest interior. The populations of P. tenuinervis showed no differences in genetic variability or structure among the three habitats. There was, however, an indication of inbreeding, which was significantly higher in natural edges than in anthropogenic edges and the forest interior. Within-habitat variation was considerable, but there were no differences in seed mass or rate and velocity of germination among the three habitats. These results suggest that seed characteristics were not influenced by the genetic pattern of P. tenuinervis and that other characteristics of the forest fragment, such as gaps, edge age, and type of matrix exert more influence on seed mass and germination than the distance from the edges.

Storage in cerrado soil and germination of Psychotria vellosiana (Rubiaceae) seeds.

The regeneration of plant communities from seed depends, to a large extent, on the capacity of the seed remaining viable in the soil. The viability and germination of artificially buried Psychotria vellosiana seeds in cerrado soil were studied, with the purpose of discovering some physio-ecological aspects of dispersed seeds and evaluating their potential to constitute a soil seed bank. Seed samples were placed in nylon envelopes and buried in the soil of a Cerrado reserve at two different depths and sites. Buried seeds were retrieved periodically and tested for germination along with dry-stored seeds. In general, there was a reduction in seed germination with storage time, both in soil and dry stored conditions, and in some assays exhumed seeds germinated faster than dry stored ones. In general the soil storage favoured seed viability of ungerminated seeds as compared to dry stored ones, with the seeds remaining partially viable after 10 months of storage. The lack of germination of viable seeds suggests that seeds showed true dormancy and/or required an extended time to germinate. It was observed that some seeds had germinated while buried and such in situ germination tended to increase with rainfall. The water availability in the soil might be a limiting factor for successful germination of P. vellosiana in the field, and the seeds may constitute a persistent soil seed bank in the cerrado as dispersed seeds remain viable in the soil until the following period of seed dispersal.

Integrated analysis of tropical trees growth: a multivariate approach.

BACKGROUND AND AIMS: One of the problems analysing cause-effect relationships of growth and environmental factors is that a single factor could be correlated with other ones directly influencing growth. One attempt to understand tropical trees' growth cause-effect relationships is integrating research about anatomical, physiological and environmental factors that influence growth in order to develop mathematical models. The relevance is to understand the nature of the process of growth and to model this as a function of the environment. METHODS: The relationships of Aphananthe monoica, Pleuranthodendron lindenii and Psychotria costivenia radial growth and phenology with environmental factors (local climate, vertical strata microclimate and physical and chemical soil variables) were evaluated from April 2000 to September 2001. The association among these groups of variables was determined by generalized canonical correlation analysis (GCCA), which considers the probable associations of three or more data groups and the selection of the most important variables for each data group. KEY RESULTS: The GCCA allowed determination of a general model of relationships among tree phenology and radial growth with climate, microclimate and soil factors. A strong influence of climate in phenology and radial growth existed. Leaf initiation and cambial activity periods were associated with maximum temperature and day length, and vascular tissue differentiation with soil moisture and rainfall. The analyses of individual species detected different relationships for the three species. CONCLUSIONS: The analyses of the individual species suggest that each one takes advantage in a different way of the environment in which they are growing, allowing them to coexist.

Storage in cerrado soil and germination of Psychotria vellosiana (Rubiaceae) seeds

A regeneração de comunidades vegetais depende em grande parte da capacidade das sementes de se conservarem viáveis no solo. Estudou-se a viabilidade e germinação de sementes de Psychotria vellosiana manualmente enterradas no cerrado, com o objetivo de se conhecer alguns aspectos fisio-ecológicos de sementes dispersas, bem como seu potencial em formar banco de sementes no solo. Amostras de sementes foram colocadas em invólucros de tela de nylon e enterradas no solo do cerrado, em duas profundidades e em dois ambientes diferentes. As sementes enterradas eram periodicamente recuperadas e testadas quanto à sua germinação, juntamente com sementes armazenadas a seco. Em geral houve uma redução da germinação em função do tempo de armazenamento, seja no solo, seja na estocagem a seco, e em alguns ensaios as sementes exumadas germinaram mais rapidamente do que as armazenadas a seco. Em geral o armazenamento no solo favoreceu a viabilidade de sementes não germinadas, em comparação com as armazenadas a seco, e as sementes conservaram-se viáveis por mais de dez meses. A não germinação de sementes viáveis sugere a ocorrência de dormência e/ou a necessidade de um tempo maior de germinação. Constatou-se que parte das sementes germinou enquanto estavam enterradas, sendo maior a ocorrência no período mais chuvoso. A disponibilidade de água no solo pode ser um fator limitante para a germinação no campo de P. vellosiana, e as sementes podem constituir um banco de sementes persistente, já que sementes dispersas conservaram-se viáveis até o próximo período de dispersão.

Regeneration of Psychotria umbellata and production of the analgesic indole alkaloid umbellatine.

Psychotria umbellata Vell. (Rubiaceae), a Brazilian coastal woody species, produces umbellatine (also known as psychollatine), an analgesic indole alkaloid. An in vitro embryogenic regeneration protocol capable of yielding alkaloid-accumulating plants was developed. Rhizogenic calli, which were obtained from stem segments derived from rooted apical cuttings, were cultured on Murashige and Skoog's (MS) medium containing either 1 mg l(-1) NAA (naphthalene acetic acid) and no kinetin, or 5 mg l(-1) NAA + 1 mg l(-1) kinetin. Calli did not accumulate umbellatine. Segments of rhizogenic callus were cultured on complete MS medium with various concentrations of kinetin and sucrose. Plant regeneration was best in the light with 0.25 mg l(-1) of kinetin and 1.5% sucrose. After 3 months of acclimatization in soil mixture, plant survival was 81%. Leaves of 10-month-old regenerated plants yielded umbellatine concentrations equivalent to those of adult forest-grown plants.

Photochemical efficiency of adult and young leaves of the neotropical understory shrub Psychotria limonensis (Rubiaceae)in response to changes in the light environment

We explored the short-term adjustment in photochemical efficiency (Fv /Fm )in adult and young leaves of the understory neotropical shrub Psychotria limonensis Krause (Rubiaceae)in response to rapid changes in the light environment.Leaves were collected from 20 individual plants growing under sun and shade conditions on Gigante Peninsula,Barro Colorado Natural Monument (Republic of Panama),during the wet season of 1996. Leaves were distributed in four sequences of light treatments (AB leaves were expanded under sun and were transferred to shade,BA leaves experienced the opposite transfer,and the controls AA and BB leaves that were expanded and maintained under sun or shade conditions).Adult and young leaves did not differ in overall photochemical efficiency.Instead,differences were found among light environments,for which leaves transferred from shade to sun showed the lowest F v /F m ratios.There was no relationship between photochemical efficiency and leaf temperature.In P.limonensis,understory plants are susceptible of photoinhibition independently of the leaf ontogenetic stage.The approach utilized in this experiment allowed the rapid exploration of this capacity, and could be applied to poorly studied understory species

Se exploró el ajuste a corto plazo en la eficiencia fotosintética (Fv /Fm )en hojas jovenes y adultas del arbusto del sotobosque neotropical Psychotria limonensis Krause (Rubiaceae)en respuesta a cambios rápidos de luz ambiental. Las hojas fueron recolectadas de 20 plantas individuales bajo condiciones de sol y sombra en Peninsula Gigante, Monumento Natural Barro Colorado (Panamá),durante la estación lluviosa de 1996.Las hojas fueron distribuidas en una secuencia cuatro tratamientos de luz (AB las hojas fueron expandidas bajo el sol y fueron transferidas a la sombra,BA las hojas experimentaron la transferencia contraria,y las hojas controles AA y BB que fueron expandidas y mantenidas bajo condiciones de sol o de sombra).Las hojas adultas o jóvenes no difieren en la eficiencia fotoquímica general.Por el contrario,se encontró diferencias entre los ambientes de luz (iluminados), para los cuales las hojas transferidas de la sombra al sol mostraron las menores tasas Fv /Fm .No hubo relación entre la eficiencia fotoquímica y la temperatura de las hojas. En P.limonensis ,las plantas son suceptibles a la foto -inhibición independientemente del estado ontogenético de la hoja. El enfoque utilizado en este experimento permitió la rápida exploración de esta capacidad y demostró que puede ser utilizado en otras especies poco estudiadas del sotobosque

N,beta-D-Glucopyranosyl vincosamide, a light regulated indole alkaloid from the shoots of Psychotria leiocarpa.

From leaves of Psychotria leiocarpa, an indole alkaloid was isolated to which the structure N,beta-D-glucopyranosyl vincosamide (1) was assigned. This represents the first report of an N-glycosylated monoterpenoid indole alkaloid. In field-grown plants highest amounts of 1 were found in the leaves (2.5% of dry wt) and fruit pulp (1.5% dry wt). Lower amounts were found in the stems (0.2% dry wt) and the seeds (0.1% of dry wt), whereas the alkaloid was not detected in the roots. The accumulation of 1 in aseptic seedlings was also restricted to the shoots and increased with plant age and light exposure, independent of the supply of sucrose in the culture medium.

A functional analysis of the crown architecture of tropical forest Psychotria species: do species vary in light capture efficiency and consequently in carbon gain and growth?

The crown architectures of 11 Psychotria species native to Barro Colorado Island, Panama were reconstructed from field measurements of leaf and branch geometry with the three-dimensional simulation model Y-plant. The objective was to assess the role of species differences in architecture in light capture and carbon gain in their natural understory environment. When species were grouped according to their putative light environment preference, the shade tolerant species were found to have a small but significantly higher efficiency of light capture for both diffuse and direct light as compared to the light demanding species. Within each grouping, however, there were few significant differences in light capture efficiency among species. The lower efficiencies of light demanding species was due to slightly higher self-shading and slightly lower angular efficiencies. Simulations of whole plant assimilation showed that light demanding species had greater daily assimilation in both direct and diffuse light due to the significantly greater light availability in the sites where light demanding species were found, as compared to those where shade tolerant species occurred. Among light demanding species, the above ground relative growth rate measured over a 1-year period by applying allometric equations for mass versus linear dimensions, was positively correlated with diffuse PFD and with mean daily assimilation estimated from Y-plant. For the shade tolerant plants, there was no significant correlation between RGR and mean daily assimilation or with any measure of light availability, probably because they occurred over a much narrower range of light environments. Overall, the results reveal a strong convergence in light capture efficiencies among the Psychotria species at lower values than previously observed in understory plants using similar approaches. Constraints imposed by other crown functions such as hydraulics and biomechanical support may place upper limits on light capture efficiency.

Photochemical efficiency of adult and young leaves of the neotropical understory shrub Psychotria limonensis (Rubiaceae) in response to changes in the light environment.

We explored the short-term adjustment in photochemical efficiency (Fv/Fm) in adult and young leaves of the understory neotropical shrub Psychotria limonensis Krause (Rubiaceae) in response to rapid changes in the light environment. Leaves were collected from 20 individual plants growing under sun and shade conditions on Gigante Peninsula, Barro Colorado Natural Monument (Republic of Panama), during the wet season of 1996. Leaves were distributed in four sequences of light treatments (AB leaves were expanded under sun and were transferred to shade, BA leaves experienced the opposite transfer, and the controls AA and BB leaves that were expanded and maintained under sun or shade conditions). Adult and young leaves did not differ in overall photochemical efficiency. Instead, differences were found among light environments, for which leaves transferred from shade to sun showed the lowest Fv/Fm ratios. There was no relationship between photochemical efficiency and leaf temperature. In P. limonensis, understory plants are susceptible of photoinhibition independently of the leaf ontogenetic stage. The approach utilized in this experiment allowed the rapid exploration of this capacity, and could be applied to poorly studied understory species.

Petiole twisting in the crowns of Psychotria liminesis: implications for light interception and daily carbon gain.

We used Y-plant, a computer-based model of crown architecture, to examine the implications of leaf reorientation resulting from petiole bending in Psychotria limonensis (Rubiaceae) seedlings. During this reorientation process, bending of the petioles of lower leaves that are potentially self-shaded by the upper leaves rotates the lamina around the stem's orthotropic axis so that self-shading is reduced. Simulations of daily light capture and assimilation revealed a 66% increase in daily C gain due to reorientation of the leaves as compared to simulations where the leaves remained in their characteristic opposite decussate pattern set by the phyllotaxy. This was due to enhanced carbon (C) gain of the lower leaves because of the reduction of shading by upper developing leaves in the same vertical plane. The light signal for this movement was experimentally examined by placing leaf-shaped filters above already fully expanded leaves and following the resulting shade-avoiding movements. The filters were either neutral density shade cloth that reduced the photon flux density (PFD) but did not alter the red to far red ratio (R:FR) or a film that reduced the PFD equivalently but also reduced the R:FR. Leaf reorientation was much more rapid and complete under the low R:FR as compared to the high R:FR indicating involvement of a phytochrome photosensory system that detected the presence of a shading leaf. Plants in gaps were found to lack a reorientation response indicating that the reorientation is specific to the shaded understory environment.