ABSTRACT In vitro rearing protocols already established for honeybees are currently being adapted to assess the risk of pesticides, and to conduct comparative developmental biology studies on stingless bees. However, differences in critical life-history traits (development time and the type of larval nutrition leading to caste differentiation process) among social bees require the development of an in vitro rearing protocol for each species and caste. We generated a protocol to produce workers of Frieseomelitta varia (Lepeletier, 1836), a non-endangered and highly eusocial pollinator species with wide geographical distribution. We tested the viability of using either the eggs or the first instar larvae as the starting point for in-vitro transfer. In vitro rearing was performed in acrylic plates at 30 °C and 99% relative humidity during the larval feeding phase. The humidity was subsequently reduced to 75% during the following days of development. The experimental larvae were offered either 25 µL or 27 µL of larval food. The development time, emergence and mortality rates, and morphological parameters of the emerged workers were assessed. In the process of validating the protocol, the adults that emerged after in vitro rearing were compared with colony-reared adults. In our results, 27 µL of larval food allowed 90% of workers to emerge. No significant differences were found between the emerging workers reared in vitro and those reared in the colony. The described protocol is a useful method for rearing F. varia workers in vitro, which can be used for diverse types of experimental approaches.

Abstract Interactions between plant and pollinators are associated with the origin and maintenance of species diversity, as well as ecosystem functioning. The potential of pollination as an ecosystem service is evidenced by its association with food production. Understanding pollination at the landscape scale is essential for characterizing the pollination service for several crops that depend on pollinators for fruit and seed set that make up the human diet. Our aim was to carry out a literature review of studies and projects funded by BIOTA/FAPESP to illustrate the main research approaches developed in the field of Pollination Biology, especially related to plant-pollinator interactions. Plant-pollinator interactions in the Atlantic forest were leveraged as a result of this long-term research program, during which several papers were published in international journals. Pollination by bees (melittophily) was the most representative pollination system studied. In addition to melittophily, other interactions were studied such as pollination by hawkmoths (sphingophily), by hummingbirds (ornithophily) and by bats (chiropterophily). The specific mutualistic relationships between fig trees and fig wasps were also subject of studies within the Program. At the beginning of the BIOTA/FAPESP Program, there were many gaps in basic information about pollination and breeding systems of Brazilian native plant species. Thus, the Program was fundamental to fuel research on the natural history of plants and pollinators from the Atlantic forest. Overall, the Program funded studies that investigated themes such as functional pollination ecology, pollinator effectiveness, plant population genetics, structure and dynamics of plant-pollinator interaction networks, as well as geographic distribution and macroevolution of pollination systems, as well as genetic and molecular studies of native plant populations focusing on pollen flow and genetic structure of populations. Additionally, studies on pollination in the context of landscape ecology had the aim of assessing the effects of forest fragmentation on the functioning of plant populations and their interactions with pollinators and the relationships between landscape structure and ecological processes, biodiversity, and ecosystem service. Therefore, the Program had a prominent role in producing basic data with great implications for understanding the ecology and promoting the conservation of plant-pollinator interactions.

Resumo A interação planta-polinizador está associada à origem e manutenção da diversidade de espécies de plantas e ao funcionamento dos ecossistemas. O potencial da polinização como serviço ecossistêmico é destacado quando associado à produção de alimentos. Compreender esta interação na escala da paisagem é essencial para caracterizar o serviço de polinização para muitos cultivos que dependem dos polinizadores para a formação de frutos e sementes que integram a dieta humana. O objetivo deste trabalho foi realizar uma revisão bibliográfica de estudos e projetos financiados pelo BIOTA/FAPESP para ilustrar as principais abordagens de pesquisa desenvolvidas no campo da Biologia da Polinização, especialmente relacionadas à interação planta-polinizador. As interações planta-polinizador na Mata Atlântica foram alavancadas como resultado desse programa de pesquisa de longo prazo, durante o qual vários artigos foram publicados em revistas internacionais. A polinização por abelhas (melitofilia) foi o sistema de polinização mais representativo estudado. Além da melitofilia, outras interações foram estudadas, como a polinização por mariposas (esfingofilia), por beija-flores (ornitofilia) e por morcegos (quiropterofilia). As relações mutualísticas específicas entre figueiras e vespas do figo também foram objeto de estudos no âmbito do Programa. No início do Programa BIOTA/FAPESP, havia muitas lacunas sobre informações básicas sobre polinização e sistemas de reprodução de espécies vegetais nativas brasileiras. Assim, o Programa foi fundamental para desenvolver pesquisas sobre a história natural de plantas e polinizadores da Mata Atlântica. No geral, o Programa financiou estudos que investigaram temas como ecologia funcional da polinização, eficácia de polinizadores, genética de populações de plantas, estrutura e dinâmica de redes de interação planta-polinizador, bem como distribuição geográfica e macroevolução dos sistemas de polinização, além de estudos genéticos e moleculares de populações de plantas nativas com foco no fluxo de pólen. Adicionalmente, estudos sobre polinização no contexto da ecologia da paisagem tiveram como objetivo avaliar os efeitos da fragmentação florestal no funcionamento das populações de plantas e suas interações com os polinizadores e as relações entre a estrutura da paisagem e os processos ecológicos, biodiversidade e serviços ecossistêmicos. Portanto, o Programa teve um papel de destaque na produção de dados básicos com grandes implicações para o entendimento da ecologia e promoção da conservação das interações planta-polinizador.

ABSTRACT Toxic metals have become some of the main abiotic agents that influence the stress of organisms due to their numerous agricultural and industrial uses that result in bioaccumulation and toxicity at different levels, depending on the type and concentration in the environment. Aluminum and lead are present as a result of anthropogenic activities and cause soil contamination and, consequently, pollution effects on producers such as plants. In this review, we used meta-analysis techniques to assess the effect of aluminum and lead on plant physiology. We calculated the overall effect size for the response variables photosynthesis, maximum quantum efficiency of photosystem II and plastid pigments (chlorophyll a, chlorophyll b and carotenoids). Moreover, it was possible to test the the effect of each metal and the phylogenetic relationship for the probability of Fabaceae species to respond to the pollution caused by these heavy metals, due to the number of studies found using this family. Overall, exposure to the toxic metals at high magnitudes affects photosynthesis and chlorophyll b (by reducing its amount). When considering the effect of each metal separately, aluminum reduced the content of the chlorophyll a and chlorophyll b. However, lead reduced photosynthesis, maximum quantum efficiency of photosystem II and chlorophyll b. When considering the Fabaceae species, there was no reduction of the parameters analyzed under stress by toxic metals. Therefore, our results confirm that these toxic metals (aluminum and lead) are toxic to plant physiology, mainly affecting the efficiency of the photosynthetic system and the plastid content.

Abstract Current phylogenetic hypotheses support that ancestral Poales were animal-pollinated and that subsequent shifts to wind pollination have occurred. Ten of the 16 Poales families are widely distributed in the Neotropics, however a comprehensive understanding of their pollination systems' diversity is still lacking. Here we surveyed studies on pollination biology of Neotropical species of Poales. Poaceae, Cyperaceae and Juncaceae are predominantly wind-pollinated but insect pollination also occurs. Thurniaceae and Thyphaceae fit on anemophily but empirical data are missing. Pollen flowers with poricidal anthers have evolved independently in Mayacaeae and Rapateaceae. Pollen- and nectar-flowers occur in Xyridaceae, which are mainly pollinated by bees. Eriocaulaceae flowers secrete minute quantity of nectar and are pollinated by "diverse small insects". Pollination of Bromeliaceae is carried out by a great variety of animal groups, mainly hummingbirds, and includes anemophily. The diversity in floral forms is very high within the order but more constant within the families. This trend indicates that many events of species diversification may have occurred without divergence in the pollination mode. Still, parallel shifts in pollination modes are found, including possible reversals to wind- or animal-pollination, changes in the type of pollinators (e.g. from hummingbirds to bee or bats) and the arising of ambophily.

Es importante determinar la eficacia de las áreas de conservación cuando se están implementando diferentes categorías de manejo, y una forma de hacerlo es conociendo si se mantiene una reproducción sexual efectiva en las especies de plantas y sus interacciones con grupos de animales. Para evaluar esta cuestión, se comparó la fenología, el éxito reproductivo, la polinización y la pre-dispersión de semillas depredadas de la leguminosa Senna multijuga en dos áreas protegidas sometidas a diferente tipos de manejo en el sureste de Brasil: el Parque Nacional de Itatiaia y el Área de Protección Ambiental de Serrinha de Alambari, de diciembre 2007 a diciembre 2008. La fenodinámica vegetativa y reproductiva fue registrada mensualmente en 80 individuos; otras evaluaciones incluyeron 104 horas de observación de la polinización (marzo-mayo 2008) en 51 inflorescencias, además del conteo de frutos, fecundidad y depredación de semillas. S. multijuga tiene una reproducción sexual que depende de la polinización de abejas grandes (Bombus, Centris, Epicharis y Xylocopa) y se considera una especie auto-incompatible. Las semillas son depredadas por especies de Bruchidae de los géneros Acanthoscelides y Sennius. La fenodinámica vegetativa y reproductiva difirió entre sitios. Nuestros resultados indican que las interacciones ecológicas fueron menores en el área protegida, pero los procesos reproductivos de S. multijuga no se vieron interrumpidos ni degradados de manera crítica. Esto refuerza la idea de que las zonas de paisaje con niveles intermedios de protección, tales como las áreas de protección ambiental, son adecuadas como zonas de amortiguamiento, y por lo tanto, relevantes para la conservación de los procesos ecológicos cuando se asocian con áreas en las que se da una protección más estricta

One important subject is to determine the effectiveness of conservation areas, where different management categories are being applied, to maintain effective sexual reproduction in plants and their interactions with animal groups. To evaluate this issue, we compared the phenology, reproductive success, pollination and pre-dispersal seed predation of the legume tree Senna multijuga in two differently managed protected areas in Southeastern Brazil: the Itatiaia National Park and the Environmental Protection Area of Serrinha do Alambari, from December 2007 to December 2008. Vegetative and reproductive phenodinamycs were registered monthly in 80 individuals; other evaluations included 104 observation hours for pollination (March-May 2008) in 51 inflorescences; besides, fruit counts, fecundity and seed predation. Sexual reproduction of S. multijuga depends on the transfer of pollen by large bees (Bombus, Centris, Epicharis and Xylocopa), as the species is self-incompatible. Bruchidae species of the genus Acanthoscelides and Sennius predate seeds. Vegetative and reproductive phenodynamics differed among sites. Our results indicated that ecological interactions were lower at the protected area, but the reproductive processes in S. multijuga were not ruptured or critically degraded. This reinforces the idea that landscape areas with intermediate levels of protection, such as environmental protection areas, are suitable as buffer zones, and thus, relevant to the conservation of ecological processes when associated with more strictly protected areas. Rev. Biol. Trop. 59 (4): 1939-1948. Epub 2011 December 01

Resumo O conhecimento da biologia reprodutiva e polinização de árvores neotropicais vêm aumentando nas últimas décadas. No entanto, ainda é incipiente para a Mata Atlântica e muitas questões persistem, por exemplo, sobre a prevalência de autoincompatibilidade. A biologia floral, o sistema reprodutivo e os visitantes florais de Senna multijuga foram estudados, com o objetivo de verificar a ocorrência de autoincompatibilidade e determinar seus polinizadores. A espécie apresenta características florais típicas do gênero, como flores assimétricas, enantiostilia, heteranteria e deiscência poricida. A antese é diurna e a viabilidade do pólen não diferiu entre os morfos de anteras. Os polinizadores foram abelhas dos gêneros Bombus, Epicharis, Centris e Xylocopa e abelhas menores atuaram como pilhadores de pólen. A reprodução sexual em S. multijuga depende da transferência de pólen pelos polinizadores, pois não foram formados frutos por apomixia ou autopolinização. Este último resultado pode ser devido à depressão por endogamia ou a um mecanismo de autoincompatibilidade de ação tardia, uma vez que houve aborto uniforme de pistilos e penetração de tubos polínicos nos óvulos após autopolinização. Diferença na fecundidade após polinização manual e em condições naturais indica que a limitação polínica afeta o sucesso reprodutivo feminino, assim como a disponibilidade de recursos.

Abstract Knowledge of reproductive biology and pollination of Neotropical tree species has been increasing in recent decades. However, this is still incipient in the Atlantic Forest, and many questions remain, for instance, about the prevalence of self-incompatibility. We studied the floral biology, breeding system and floral visitors of Senna multijuga in order to verify the occurrence of self-incompatibility and determine its pollinators. This species bears the typical floral traits of the genus, such as asymmetry, enantiostily, heteranthery and poricid anthers. Anthesis is diurnal and pollen viability did not differ among the morphs of anthers. Pollinators were bees of the genera Bombus, Epicharis, Centris and Xylocopa and smaller bees acted as pollen thieves. Sexual reproduction of Senna multijuga depends on the transfer of pollen by pollinators, as no fruit was formed by either apomixis or self-pollination. The latter result may be due to inbreeding depression or by a mechanism of late-acting self-incompatibility, as we observed after self-pollination both penetration of pollen tubes in the ovules and uniform abortion of pistils. Differences on fertility after manual crossings and under natural conditions indicate that female fitness is affected by pollen limitation, as well as by the availability of resources.