Chemical similarity and local community assembly in the species rich tropical genus Piper.

Community ecologists have strived to find mechanisms that mediate the assembly of natural communities. Recent evidence suggests that natural enemies could play an important role in the assembly of hyper-diverse tropical plant systems. Classic ecological theory predicts that in order for coexistence to occur, species differences must be maximized across biologically important niche dimensions. For plant-herbivore interactions, it has been recently suggested that, within a particular community, plant species that maximize the difference in chemical defense profiles compared to neighboring taxa will have a relative competitive advantage. Here we tested the hypothesis that plant chemical diversity can affect local community composition in the hyper-diverse genus Piper at a lowland wet forest location in Costa Rica. We first characterized the chemical composition of 27 of the most locally abundant species of Piper. We then tested whether species with different chemical compositions were more likely to coexist. Finally, we assessed the degree to which Piper phylogenetic relationships are related to differences in secondary chemical composition and community assembly. We found that, on average, co-occurring species were more likely to differ in chemical composition than expected by chance. Contrary to expectations, there was no phylogenetic signal for overall secondary chemical composition. In addition we found that species in local communities were, on average, more phylogenetically closely related than expected by chance, suggesting that functional traits other than those measured here also influence local assembly. We propose that selection by herbivores for divergent chemistries between closely related species facilitates the coexistence of a high diversity of congeneric taxa via apparent competition.

Timing of pollen release and stigma receptivity period of Piper vicosanum: New insights into sexual reproduction of the genus.

PREMISE OF THE STUDY: Dichogamy is a common characteristic among angiosperms, including Piper species. In this genus, the tiny flowers are morphologically similar and have an asynchronous stamen development. However, there is no information on the duration of stigma receptivity and whether it overlaps with pollen release. To better understand mechanisms of floral function in Piper vicosanum, we provide a detailed characterization of the timing of pollen release from the four stamens and the period of stigma receptivity and exposure mode of the receptive areas. METHODS: We investigated plants of a natural population in a semideciduous seasonal forest (Viçosa, Minas Gerais State, southeastern Brazil), based on chemical tests, light microscopy, and scanning electron microscopy analyses. KEY RESULTS: Incomplete protogyny-a mechanism that favors outcrossing-was recorded. The period of stigma receptivity was long (14 d), and the sequential exposure and senescence of stigmatic papillae occurred gradually and in a basipetal direction. Pollen release began 2-6 d after the beginning of the pistillate phase, with an average pollen viability of 87.7%, during the bisexual flower phase. Pollen was released for up to 6 d and occurred in one stamen at a time. The fruit set observed in tests of self-pollination indicated self-compatibility. CONCLUSIONS: The gradual and sequential exposure of stigmatic papillae in P. vicosanum flowers is described here as the mechanism for the long duration of receptivity. Anther development and pollen release were also sequential. These findings are yet unreported reproductive characteristics of the genus and offer new perspectives for future studies on the floral biology of other Piper species.

Intra- and interspecific diversity in a tropical plant clade alter herbivory and ecosystem resilience.

Declines in biodiversity generated by anthropogenic stressors at both species and population levels can alter emergent processes instrumental to ecosystem function and resilience. As such, understanding the role of biodiversity in ecosystem function and its response to climate perturbation is increasingly important, especially in tropical systems where responses to changes in biodiversity are less predictable and more challenging to assess experimentally. Using large-scale transplant experiments conducted at five neotropical sites, we documented the impacts of changes in intraspecific and interspecific plant richness in the genus Piper on insect herbivory, insect richness, and ecosystem resilience to perturbations in water availability. We found that reductions of both intraspecific and interspecific Piper diversity had measurable and site-specific effects on herbivory, herbivorous insect richness, and plant mortality. The responses of these ecosystem-relevant processes to reduced intraspecific Piper richness were often similar in magnitude to the effects of reduced interspecific richness. Increased water availability reduced herbivory by 4.2% overall, and the response of herbivorous insect richness and herbivory to water availability were altered by both intra- and interspecific richness in a site-dependent manner. Our results underscore the role of intraspecific and interspecific richness as foundations of ecosystem function and the importance of community and location-specific contingencies in controlling function in complex tropical systems.

Directed seed dispersal of Piper by Carollia perspicillata and its effect on understory plant diversity and folivory.

Directed dispersal occurs when seeds are differentially deposited to sites where offspring survivorship is higher than at randomly chosen sites. Traditionally, characteristics of the dispersal target sites that could increase survivorship of the dispersed plants are thought to be intrinsic to the sites. If directed dispersal is constant over extended periods of time, however, it is likely that nonrandom patterns of dispersal could modify the ecological characteristics of the target site in ways that could increase survivorship and fitness of the dispersed plants. Here we report patterns of Piper diversity (richness, equitability, and similarity) and Piper folivory within plots near natural or artificial roosts of Carollia perspicillata vs. similar plots without bat roosts. Plots with bat roosts, both natural and artificial, had significantly higher Piper species diversity. Additionally, we found that plots with a higher Piper species diversity showed less specialist folivory, higher generalist folivory, and lower total herbivore leaf damage than plots with low Piper diversity. Finally, plots with bat roosts also showed less specialist folivory, lower generalist folivory, and lower total folivory when compared to plots without roosts. We propose that long-lasting nonrandom patterns of seed dispersal can change the local ecological characteristics of target sites via changes in plant diversity, and that these changes are likely to reduce the local rates of folivory and, therefore, increase seed and adult plant survivorship.

Long-term dynamics of tropical walking sticks in response to multiple large-scale and intense disturbances.

Understanding the effects of disturbance and secondary succession on spatio-temporal patterns in the abundance of species is stymied by a lack of long-term demographic data, especially in response to infrequent and high intensity disturbances, such as hurricanes. Moreover, resistance and resilience to hurricane-induced disturbance may be mediated by legacies of previous land use, although such interactive effects are poorly understood, especially in tropical environments. We address these central issues in disturbance ecology by analyzing an extensive dataset, spanning the impacts of Hurricanes Hugo and Georges, on the abundance of a Neotropical walking stick, Lamponius portoricensis, in tabonuco rainforest of Puerto Rico during the wet and dry seasons from 1991 to 2007. By synthesizing data from two proximate sites in tabonuco forest, we show that resistance to Hurricane Hugo (97% reduction in abundance) was much less than resistance to Hurricane Georges (21% reduction in abundance). Based on a powerful statistical approach (generalized linear mixed-effects models with Poisson error terms), we documented that the temporal trajectories of abundance during secondary succession (i.e., patterns of resilience) differed between hurricanes and among historical land use categories, but that the effects of hurricanes and land use histories were independent of each other. These complex results likely arise because of differences in the intensities of the two hurricanes with respect to microclimatic effects (temperature and moisture) in the forest understory, as well as to time-lags in the response of L. portoricensis to changes in the abundance and distribution of preferred food plants (Piper) in post-hurricane environments.

Herbivores on a dominant understory shrub increase local plant diversity in rain forest communities.

Indirect effects of trophic interactions on biodiversity can be large and common, even in complex communities. Previous experiments with dominant understory Piper shrubs in a Costa Rican rain forest revealed that increases in herbivore densities on these shrubs caused widespread seedling mortality as a result of herbivores moving from Piper to seedlings of many different plant genera. We tested components of the Janzen-Connell hypothesis by conducting focused studies on the effects of specialist and generalist Piper herbivores on local seedling diversity. Whereas specialist herbivores are predicted to increase mortality to neighboring seedlings that are closely related to the source plant, true generalists moving from source plants may cause density-dependent mortality of many species, and possibly increase richness if new species replace abundant species that have been thinned by herbivores. Therefore, we hypothesized that seedling richness would be greater in understory control plots created in patches of Piper that had normal densities of generalist herbivores compared to plots from which we removed generalist herbivores manually from all Piper shrubs. After 15 months, generalist-herbivore-removal plots had > 40% fewer seedlings, > 40% fewer species, and 40% greater seedling evenness, on average, than control plots with generalist herbivores intact. Using a complementary approach in unmanipulated plots in four forests, we used path analysis to test for a positive association between seedling diversity and herbivore damage on Piper species. In unmanipulated plots, for both generalist and specialist herbivores, our data were significant fits to the causal model that Piper herbivores decrease evenness and increase plant species richness, corroborating the experimental results. Because herbivores changed how individuals were apportioned among the species and families present (lower evenness), one interpretation of these associations between herbivores on Piper shrubs and local seedling richness is that high seedling mortality in dominant families allowed the colonization or survival of less common species. If interspecific or apparent competition allowed for a relative increase in species richness, then the Janzen-Connell hypothesis may extend its predictions to generalist seedling predators. We speculate that apparent competition may explain some of the deviations from neutral model predictions, especially at small scales.

Synergistic effects of amides from two piper species on generalist and specialist herbivores.

Plants use a diverse mix of defenses against herbivores, including multiple secondary metabolites, which often affect herbivores synergistically. Chemical defenses also can affect natural enemies of herbivores via limiting herbivore populations or by affecting herbivore resistance to parasitoids. In this study, we performed feeding experiments to examine the synergistic effects of imides and amides (hereafter "amides") from Piper cenocladum and P. imperiale on specialist (Eois nympha, Geometridae) and generalist (Spodoptera frugiperda, Noctuidae) lepidopteran larvae. Each Piper species has three unique amides, and in each experiment, larvae were fed diets containing different concentrations of single amides or combinations of the three. The amides from P. imperiale had negative synergistic effects on generalist survival and specialist pupal mass, but had no effect on specialist survival. Piper cenocladum amides also acted synergistically to increase mortality caused by parasitoids, and the direct negative effects of mixtures on parasitoid resistance and pupal mass were stronger than indirect effects via changes in growth rate and approximate digestibility. Our results are consistent with plant defense theory that predicts different effects of plant chemistry on generalist versus adapted specialist herbivores. The toxicity of Piper amide mixtures to generalist herbivores are standard bottom-up effects, while specialists experienced the top-down mediated effect of mixtures causing reduced parasitoid resistance and associated decreases in pupal mass.

Characterizing the cauline domatia of two newly discovered ecuadorian ant plants in piper: an example of convergent evolution.

The stems of some myrmecophytes in Piper are used as domatia by resident ant colonies. Hollow, ant-occupied stems were previously known only in four species of southern Central American Piper, all members of Section Macrostachys. Here we present two additional, unrelated, hollow-stemmed myrmecophytes from Ecuador: P. immutatum and P. pterocladum (members of sections Radula and Peltobryon, respectively). Although similar superficially, stem cavities of the Ecuadorian Piper species differ morphologically and developmentally from those of Central American taxa. The stem cavities of P. immutatum, and possibly P. pterocladum, are formed during stem development, and begin forming only a few millimeters behind the apical meristem. This mode of cavity formation differs markedly from myrmecophytes in section Macrostachys, where the stems remain solid unless excavated by the specialized ant partner Pheidole bicornis. The stems of P. immutatum and P. pterocladum do not produce wound-response tissue around the cavity, unlike the stems in section Macrostachys. The entrance holes in stems of P. immutatum are formed through apoptotic processes and are located at each node below the petiole, whereas those in section Macrostachys are excavated by the ants in the leaf axil. This study documents convergent evolution of ant-plant associations in Piper, and emphasizes the need for careful comparison of apparently homologous, ant-associated structures in specialized myrmecophytes.

Proteomic profile of Piper tuberculatum (Piperaceae).

Piper tuberculatum (Piperaceae) is a species that accumulates especially amides as secondary metabolites and several biological activities was previously reported. In this article, we report a proteomic study of P. tuberculatum. Bidimensional electrophoresis (2D SDS-PAGE) and mass spectrometry (ESI-Q-TOF) were used in this study. Over a hundred spots and various peptides were identified in this species and the putative functions of these peptides related to defense mechanism as biotic and abiotic stress were assigned. The information presented extend the range of molecular information of P. tuberculatum.

Origin of the subepidermal tissue in Piper L. leaves.

Studies on the anatomy of Piper leaves demonstrate the presence of a subepidermal tissue distinct from the adjacent epidermis, which cells show thin walls and hyaline contents. Some authors consider such cells a hypodermal tissue, while others refer to them as components of a multiple epidermis. In this study, the nature of this subepidermal tissue was investigated through the analysis of leaf ontogeny in three Piper species. The analysis showed that the referred tissue originates from the ground meristem and, thus, should be considered a hypodermis. The studied species suggests that the role of the hypodermis would be to protect the photosynthetic apparatus from excess light, regulating the intensity of light reaching the chlorophyll parenchyma.

Proteomic profile of Piper tuberculatum (Piperaceae) / Perfil proteômico e metabólico de Piper tuberculatum (Piperaceae)

Abstract Piper tuberculatum (Piperaceae) is a species that accumulates especially amides as secondary metabolites and several biological activities was previously reported. In this article, we report a proteomic study of P. tuberculatum. Bidimensional electrophoresis (2D SDS-PAGE) and mass spectrometry (ESI-Q-TOF) were used in this study. Over a hundred spots and various peptides were identified in this species and the putative functions of these peptides related to defense mechanism as biotic and abiotic stress were assigned. The information presented extend the range of molecular information of P. tuberculatum.

Resumo Piper tuberculatum (Piperaceae) é uma espécie que acumula especialmente amidas como metabólitos secundários e diversas atividades biológicas dessa espécie foram relatadas anteriormente. No presente artigo, relatamos um estudo proteômico dessa espécie. Eletroforese bidimensional (2D SDS-PAGE) e espectrometria de massas (ESI-Q-TOF) foram utilizadas nesse estudos. Mais de cem spots e vários peptídeos foram identificados nesta espécie e as funções putativas desses peptídeos relacionadas a mecanismo de defesa como estresse biótico e abiótico foram atribuídos. As informações apresentadas ampliam a gama de informações moleculares dessa espécie.

Origin of the subepidermal tissue in Piper L. leaves / Origem do tecido subepidérmico em folhas de Piper L.

Studies on the anatomy of Piper leaves demonstrate the presence of a subepidermal tissue distinct from the adjacent epidermis, which cells show thin walls and hyaline contents. Some authors consider such cells a hypodermal tissue, while others refer to them as components of a multiple epidermis. In this study, the nature of this subepidermal tissue was investigated through the analysis of leaf ontogeny in three Piper species. The analysis showed that the referred tissue originates from the ground meristem and, thus, should be considered a hypodermis. The studied species suggests that the role of the hypodermis would be to protect the photosynthetic apparatus from excess light, regulating the intensity of light reaching the chlorophyll parenchyma.

Os estudos de anatomia foliar de espécies de Piper revelam a presença de um tecido subepidérmico distinto da epiderme e cujas células apresentam paredes finas e conteúdo hialino. Estas células são referenciadas por alguns autores como um tecido hipodérmico e por outros, como sendo constituintes de uma epiderme múltipla. Nesse estudo verificou-se a natureza deste tecido subepidérmico a partir da análise da ontogênese foliar de três espécies de Piper. A análise revelou que o referido tecido tem origem do meristema fundamental e, portanto, deve ser denominado de hipoderme. Para as espécies avaliadas, sugere-se que a hipoderme teria função de, proteger o aparato fotossintético do excesso de luminosidade, regulando a intensidade luminosa que atinge o parênquima clorofiliano.

Inter- and intraspecific comparisons of antiherbivore defenses in three species of rainforest understory shrubs.

Plants defend themselves against herbivores and pathogens with a suite of morphological, phenological, biochemical, and biotic defenses, each of which is presumably costly. The best studied are allocation costs that involve trade-offs in investment of resources to defense versus other plant functions. Decreases in growth or reproductive effort are the costs most often associated with antiherbivore defenses, but trade-offs among different defenses may also occur within a single plant species. We examined trade-offs among defenses in closely related tropical rain forest shrubs (Piper cenocladum, P. imperiale, and P. melanocladum) that possess different combinations of three types of defense: ant mutualists, secondary compounds, and leaf toughness. We also examined the effectiveness of different defenses and suites of defenses against the most abundant generalist and specialist Piper herbivores. For all species examined, leaf toughness was the most effective defense, with the toughest species, P. melanocladum, receiving the lowest incidence of total herbivory, and the least tough species, P. imperiale, receiving the highest incidence. Although variation in toughness within each species was substantial, there were no intraspecific relationships between toughness and herbivory. In other Piper studies, chemical and biotic defenses had strong intraspecific negative correlations with herbivory. A wide variety of defensive mechanisms was quantified in the three Piper species studied, ranging from low concentrations of chemical defenses in P. imperiale to a complex suite of defenses in P. cenocladum that includes ant mutualists, secondary metabolites, and moderate toughness. Ecological costs were evident for the array of defensive mechanisms within these Piper species, and the differences in defensive strategies among species may represent evolutionary trade-offs between costly defenses.

Farmacognosia, farmacobotánica, farmacogeografía y farmacoetimología del platanillo de Cuba (Piper aduncum subespecie ossanum) / Pharmacognosy, pharmacobotany, pharmacogeography, and pharmacoetimology of platanillo de Cuba (Piper aduncum subspecie ossanum)

Introducción: los estudios farmacognósticos son de vital importancia en la investigación de plantas medicinales, así como en la producción de medicamentos herbarios y fitofármacos. La especie Piper aduncum L (Piperaceae), posee un gran aval de uso tradicional en Cuba y otras regiones del mundo. Objetivo: estudiar desde el punto de vista farmacognóstico la droga de Piper aduncum en cuanto a botánica, geografía y etimología. Métodos: se realizó el estudio botánico, de distribución geográfica y etimología de esta droga, mediante el empleo de literatura especializada y la determinación de caracteres macromorfológicos de hojas de esta planta colectada en la provincia Ciego de Ávila, Cuba. Resultados: existen 2 categorías infraespecíficas bien localizadas geográficamente de la planta que en Cuba se llama por lo común platanillo de Cuba, guayuyo o canilla de muerto, una de las cuales -Piper aduncum subespecie ossanum (C.DC.) Saralegui-, endémica, es la que se investiga. Las características macromorfológicas de la droga coinciden con las descritas en la literatura. Se hallaron aspectos etimológicos y de distribución geográfica de interés relacionados con esta planta. Conclusiones: en las investigaciones farmacognósticas los estudios botánicos, geográficos y etimológicos pueden ser de gran significación, como en el caso de esta planta. Para cualquier investigación con esta especie resulta imprescindible abordar su tratamiento taxonómico en el país, porque incluye 2 categorías infraespecíficas en las cuales la ubicación geográfica es fundamental

Introduction: pharmacognostic studies are essential on medicinal plant research, as well as in herbal medicine and phytomedicine production. Piper aduncum L. (Piperaceae) specie known as "platanillo de Cuba", has wide recognition due to its traditional use in Cuba and other regions. Objective: to research the drug derived from Piper aduncum (platanillo de Cuba) from the pharmacognostic viewpoint in terms of pharmacobotany, pharmacogeography and pharmacoetymology. Methods: the pharmacobotanical, geographical distribution and etymological study of this drug was conducted by using specialized literature and determination of macromorphologic characteristics of leaves collected in Ciego de Ávila province, Cuba. Results: there are two geographically located infraspecific taxa of the plant commonly called "platanillo de Cuba", "guayuyo" o "canilla de muerto" in Cuba. One of them -Piper aduncum subspecie ossanum (C.DC.) Saralegui- is endemic and the topic of this research. Macromorphologic characteristics of the drug are comparable to those described in literature. Geographical distribution and etymologic aspects of interest were found. Conclusions: in the pharmacognostic research, pharmacobotanical, pharmacogeographical and pharmacoetymological studies could be very signicant, as in the case of this plant. For any further research on this species, it is fundamental to approach its taxonomic treatment in the country, since it includes two infraspecific taxa in which geographical location is essential.

High-frequency plant regeneration through cyclic secondary somatic embryogenesis in black pepper (Piper nigrum L.).

A high-frequency plantlet regeneration protocol was developed for black pepper (Piper nigrum L.) through cyclic secondary somatic embryogenesis. Secondary embryos formed from the radicular end of the primary somatic embryos which were originally derived from micropylar tissues of germinating seeds on growth regulator-free SH medium in the absence of light. The process of secondary embryogenesis continued in a cyclic manner from the root pole of newly formed embryos resulting in clumps of somatic embryos. Strength of the medium and sucrose concentration influenced the process of secondary embryogenesis and fresh weight of somatic embryo clumps. Full-strength SH medium supplemented with 1.5% sucrose produced significantly higher fresh weight and numbers of secondary somatic embryos while 3.0 and 4.5% sucrose in the medium favored further development of proliferated embryos into plantlets. Ontogeny of secondary embryos was established by histological analysis. Secondary embryogenic potential was influenced by the developmental stage of the explanted somatic embryo and stages up to "torpedo" were more suitable. A single-flask system was standardized for proliferation, maturation, germination and conversion of secondary somatic embryos in suspension cultures. The system of cyclic secondary somatic embryogenesis in black pepper described here represents a permanent source of embryogenic material that can be used for genetic manipulations of this crop species.

Short dry spells in the wet season increase mortality of tropical pioneer seedlings.

Variation in plant species performance in response to water availability offers a potential axis for temporal and spatial habitat partitioning and may therefore affect community composition in tropical forests. We hypothesized that short dry spells during the wet season are a significant source of mortality for the newly emerging seedlings of pioneer species that recruit in treefall gaps in tropical forests. An analysis of a 49-year rainfall record for three forests across a rainfall gradient in central Panama confirmed that dry spells of > or = 10 days during the wet season occur on average once a year in a deciduous forest, and once every other year in a semi-deciduous moist and an evergreen wet forest. The effect of wet season dry spells on the recruitment of pioneers was investigated by comparing seedling survival in rain-protected dry plots and irrigated control plots in four large artificially created treefall gaps in a semi-deciduous tropical forest. In rain-protected plots surface soil layers dried rapidly, leading to a strong gradient in water potential within the upper 10 cm of soil. Seedling survival for six pioneer species was significantly lower in rain-protected than in irrigated control plots after only 4 days. The strength of the irrigation effect differed among species, and first became apparent 3-10 days after treatments started. Root allocation patterns were significantly, or marginally significantly, different between species and between two groups of larger and smaller seeded species. However, they were not correlated with seedling drought sensitivity, suggesting allocation is not a key trait for drought sensitivity in pioneer seedlings. Our data provide strong evidence that short dry spells in the wet season differentially affect seedling survivorship of pioneer species, and may therefore have important implications to seedling demography and community dynamics.

The influence of a neotropical herbivore (Lamponius portoricensis) on nutrient cycling and soil processes.

The role of phytophagous insects in ecosystem nutrient cycling remains poorly understood. By altering the flow of litterfall nutrients from the canopy to the forest floor, herbivores may influence key ecosystem processes. We manipulated levels of herbivory in a lower montane tropical rainforest of Puerto Rico using the common herbivore, Lamponius portoricensis (Phasmatidea), on a prevalent understory plant, Piper glabrescens (Piperaceae), and measured the effects on nutrient input to the forest floor and on rates of litter decomposition. Four treatment levels of herbivory generated a full range of leaf area removal, from plants experiencing no herbivory to plants that were completely defoliated (>4,000 cm(2) leaf area removed during the 76-day study duration). A significant (P<0.05) positive regression was found between all measures of herbivory (total leaf area removed, greenfall production, and frass-related inputs) and the concentration of NO (3) (-) in ion exchange resin bags located in the litter layer. No significant relationship was found between any of the herbivory components and resin bag concentrations of NH (4) (+) or PO (4) (-) . Rates of litter decay were significantly affected by frass-related herbivore inputs. A marginally significant negative relationship was also found between the litter mass remaining at 47 days and total leaf area removed. This study demonstrated a modest, but direct relationship between herbivory and both litter decomposition and NO (3) (-) transfer to the forest floor. These results suggest that insect herbivores can influence forest floor nutrient dynamics and thus merit further consideration in discussions on ecosystem nutrient dynamics.

[Reproductive structures of Piper amalago var. medium Linnaeus (Piperaceae)]. / Estruturas de reprodução de Piper amalago var. medium Linnaeus (Piperaceae).

Flowers, fruits, seeds and subterranean organ of Piper amalago var. medium Linnaeus were analyzed structurally. Flowers are hermaphrodite lacking perianth, with two stamens and a tricarpelate gynoecea. Tetrasporangiate anthers shows epidermis, endothecium, one or two middle layers and secretory tapetum. Ovary has a simple structure, with ventral meristem. There is one orthotropous, bitegmic and crassinucellate ovule. Fruits are drupes. Seeds are endotegmic with copious perisperm. Putamen is composed of a sclerified inner mesocarp and endocarp. Individuals of the species can be interlinked by radicular subterranean organ, that can spread vegetatively.