Gibberellin and miRNA156-targeted SlSBP genes synergistically regulate tomato floral meristem determinacy and ovary patterning.

Many developmental processes associated with fruit development occur at the floral meristem (FM). Age-regulated microRNA156 (miR156) and gibberellins (GAs) interact to control flowering time, but their interplay in subsequent stages of reproductive development is poorly understood. Here, in tomato (Solanum lycopersicum), we show that GA and miR156-targeted SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE (SPL or SBP) genes interact in the tomato FM and ovary patterning. High GA responses or overexpression of miR156 (156OE), which leads to low expression levels of miR156-silenced SBP genes, resulted in enlarged FMs, ovary indeterminacy and fruits with increased locule number. Conversely, low GA responses reduced indeterminacy and locule number, and overexpression of a S. lycopersicum (Sl)SBP15 allele that is miR156 resistant (rSBP15) reduced FM size and locule number. GA responses were partially required for the defects observed in 156OE and rSBP15 fruits. Transcriptome analysis and genetic interactions revealed shared and divergent functions of miR156-targeted SlSBP genes, PROCERA/DELLA and the classical WUSCHEL/CLAVATA pathway, which has been previously associated with meristem size and determinacy. Our findings reveal that the miR156/SlSBP/GA regulatory module is deployed differently depending on developmental stage and create novel opportunities to fine-tune aspects of fruit development that have been important for tomato domestication.

Antibacterial and Toxic Activity of Geopropolis Extracts from Melipona subnitida (Ducke, 1910) (Hymenoptera: Apidae) and Scaptotrigona depilis (Moure, 1942) (Hymenoptera: Apidae).

Bacteria are associated with many infections that affect humans and present antibiotic resistance mechanisms, causing problems in health organisations and increased mortality rates. Therefore, it is necessary to find new antibacterial agents that can be used in the treatment of these microorganisms. Geopropolis is a natural product from stingless bees, formed by a mixture of plant resins, salivary secretions, wax and soil particles, the chemical composition of this natural product is diverse. Thus, this study aimed to evaluate antibacterial activity, antibiotic modulation and the toxicity of geopropolis extracts from the stingless bees, Melipona subnitida (Ducke, 1910) and Scaptotrigona depilis (Moure, 1942) against standard and multi-resistant Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa bacteria. Geopropolis samples were collected in a meliponary located in Camaragibe, Pernambuco, Brazil. To determine the Minimum Inhibitory Concentration (MIC) and antibiotic modulation we performed broth microdilution tests. Mortality tests were used to verify extract toxicity in the model Drosophila melanogaster. The microbiological tests showing that the M. subnitida extracts had better inhibitory effects compared to S. depilis, presenting direct antibacterial activity against standard and multi-resistant strains. The extracts potentialized antibiotic effects, suggesting possible synergy and did not present toxicity in the model used. The information obtained in this study highlights extracts as promising antibacterial agents and is the first study to evaluate bacterial activity in these extracts, in addition to verifying their modulating effects and determining toxicity in the model used.

Temporal Changes in Gut Microbiota Composition and Pollen Diet Associated with Colony Weakness of a Stingless Bee.

Compared to honeybees and bumblebees, the effect of diet on the gut microbiome of Neotropical corbiculate bees such as Melipona spp. is largely unknown. These bees have been managed for centuries, but recently an annual disease is affecting M. quadrifasciata, an endangered species kept exclusively by management in Southern Brazil. Here we report the results of a longitudinal metabarcoding study involving the period of M. quadrifasciata colony weakness, designed to monitor the gut microbiota and diet changes preceding an outbreak. We found increasing amounts of bacteria associated to the gut of forager bees 2 months before the first symptoms have been recorded. Simultaneously, forager bees showed decreasing body weight. The accelerated growth of gut-associated bacteria was uneven among taxa, with Bifidobacteriaceae dominating, and Lactobacillaceae decreasing in relative abundance within the bacterial community. Dominant fungi such as Candida and Starmerella also decreased in numbers, and the stingless bee obligate symbiont Zygosaccharomyces showed the lowest relative abundance during the outbreak period. Such changes were associated with pronounced diet shifts, i.e., the rise of Eucalyptus spp. pollen amount in forager bees' guts. Furthermore, there was a negative correlation between the amount of Eucalyptus pollen in diets and the abundance of some bacterial taxa in the gut-associated microbiota. We conclude that diet and subsequent interactions with the gut microbiome are key environmental components of the annual disease and propose the use of diet supplementation as means to sustain the activity of stingless bee keeping as well as native bee pollination services.

Climate-induced distribution dynamics of Plebeia flavocincta, a stingless bee from Brazilian tropical dry forests.

AIM: The objective of this study is to estimate the current potential geographic distribution of Plebeia flavocincta and to evaluate the influence of climate on the dynamics of suitable habitat availability in the past and in the future. LOCATION: Northeast region of Brazil and dry forest areas. METHODS: The habitat suitability modeling was based on two algorithms, two global circulation models, and six different scenarios. We used this tool to estimate the areas of occurrence in the past (Last Interglacial and Last Glacial Maximum), in the present, and in the future (years 2050 and 2070). RESULTS: According to the models, P. flavocincta had great dynamics in the availability of suitable habitats with periods of retraction and expansion of these areas in the past. Our results suggest that this taxon may benefit in terms of climate suitability gain in Northeast Brazil in the future. In addition, we identified high-altitude areas and the eastern coast as climatically stable. CONCLUSION: The information provided can be used by decision makers to support actions toward protecting and sustainably managing this taxon. Protection measures for this taxon are particularly important because this insect contributes to the local flora and, although our results indicate that the climate may favor this taxon, other factors can negatively affect it, such as high levels of habitat loss due to anthropogenic activities.

Landscape genomics to the rescue of a tropical bee threatened by habitat loss and climate change.

Habitat degradation and climate change are currently threatening wild pollinators, compromising their ability to provide pollination services to wild and cultivated plants. Landscape genomics offers powerful tools to assess the influence of landscape modifications on genetic diversity and functional connectivity, and to identify adaptations to local environmental conditions that could facilitate future bee survival. Here, we assessed range-wide patterns of genetic structure, genetic diversity, gene flow, and local adaptation in the stingless bee Melipona subnitida, a tropical pollinator of key biological and economic importance inhabiting one of the driest and hottest regions of South America. Our results reveal four genetic clusters across the species' full distribution range. All populations were found to be under a mutation-drift equilibrium, and genetic diversity was not influenced by the amount of reminiscent natural habitats. However, genetic relatedness was spatially autocorrelated and isolation by landscape resistance explained range-wide relatedness patterns better than isolation by geographic distance, contradicting earlier findings for stingless bees. Specifically, gene flow was enhanced by increased thermal stability, higher forest cover, lower elevations, and less corrugated terrains. Finally, we detected genomic signatures of adaptation to temperature, precipitation, and forest cover, spatially distributed in latitudinal and altitudinal patterns. Taken together, our findings shed important light on the life history of M. subnitida and highlight the role of regions with large thermal fluctuations, deforested areas, and mountain ranges as dispersal barriers. Conservation actions such as restricting long-distance colony transportation, preserving local adaptations, and improving the connectivity between highlands and lowlands are likely to assure future pollination services.

Pleistocene climate changes shaped the population structure of Partamona seridoensis (Apidae, Meliponini), an endemic stingless bee from the Neotropical dry forest.

Partamona seridoensis is an endemic stingless bee from the Caatinga, a Neotropical dry forest in northeastern Brazil. Like other stingless bees, this species plays an important ecological role as a pollinator. The aim of the present study was to investigate the genetic structure and evolutionary history of P. seridoensis across its current geographic range. Workers from 84 nests from 17 localities were analyzed for COI and Cytb genic regions. The population structure tests (Bayesian phylogenetic inference, AMOVA and haplotype network) consistently characterized two haplogroups (northwestern and eastern), with little gene flow between them, generating a high differentiation between them as well as among the populations within each haplogroup. The Mantel test revealed no isolation by distance. No evidence of a potential geographic barrier in the present that could explain the diversification between the P. seridoensis haplogroups was found. However, Pleistocene climatic changes may explain this differentiation, since the initial time for the P. seridoensis lineages diversification took place during the mid-Pleistocene, specifically the interglacial period, when the biota is presumed to have been more associated with dry conditions and had more restricted, fragmented geographical distribution. This event may have driven diversification by isolating the two haplogroups. Otherwise, the climatic changes in the late Pleistocene must not have drastically affected the population dynamics of P. seridoensis, since the Bayesian Skyline Plot did not reveal any substantial fluctuation in effective population size in either haplogroup. Considering its importance and the fact that it is an endemic bee from a very threatened Neotropical dry forest, the results herein could be useful to the development of conservation strategies for P. seridoensis.

Bees for development: Brazilian survey reveals how to optimize stingless beekeeping.

Stingless bees are an important asset to assure plant biodiversity in many natural ecosystems, and fulfill the growing agricultural demand for pollination. However, across developing countries stingless beekeeping remains an essentially informal activity, technical knowledge is scarce, and management practices lack standardization. Here we profited from the large diversity of stingless beekeepers found in Brazil to assess the impact of particular management practices on productivity and economic revenues from the commercialization of stingless bee products. Our study represents the first large-scale effort aiming at optimizing stingless beekeeping for honey/colony production based on quantitative data. Survey data from 251 beekeepers scattered across 20 Brazilian States revealed the influence of specific management practices and other confounding factors over productivity and income indicators. Specifically, our results highlight the importance of teaching beekeepers how to inspect and feed their colonies, how to multiply them and keep track of genetic lineages, how to harvest and preserve the honey, how to use vinegar traps to control infestation by parasitic flies, and how to add value by labeling honey containers. Furthermore, beekeeping experience and the network of known beekeepers were found to be key factors influencing productivity and income. Our work provides clear guidelines to optimize stingless beekeeping and help transform the activity into a powerful tool for sustainable development.

Pollination biology in the dioecious orchid Catasetum uncatum: How does floral scent influence the behaviour of pollinators?

Catasetum is a neotropical orchid genus that comprises about 160 dioecious species with a remarkable sexual dimorphism in floral morphology. Flowers of Catasetum produce perfumes as rewards, which are collected only by male euglossine bees. Currently, floral scents are known to be involved in the selective attraction of specific euglossine species. However, sexual dimorphism in floral scent and its eventual role in the pollination of Catasetum species have never been investigated. Here, we have investigated the pollination of Catasetum uncatum and asked: (1) Is floral scent a sexual dimorphic trait? (2) Does pollinarium removal/deposition affect scent emission? (3) Does sexual dimorphism in floral scent and changed scent emission have implications with regard to the behaviour of the pollinators? The frequency and behaviour of floral visitors were observed in non-manipulated flowers (both flower sexes) and in manipulated flowers (pistillate only) in which pollinaria were deposited. Scents of staminate and pistillate flowers (both manipulated and non-manipulated) were collected by using dynamic headspace methods and analysed chemically. Electrophysiological analyses were performed to detect compounds triggering antennal depolarisation in the euglossine species. C. uncatum is pollinated mainly by males of Euglossa nanomelanotricha. Pollinators were more frequent in pistillate than in staminate inflorescences. Bees approaching staminate flowers frequently flew away without visiting them, a behavioural pattern not observed in pistillate flowers. In the chemical analyses, we recorded 99 compounds, 31 of which triggered antennal depolarisation in pollinators. Multivariate analyses with the electrophysiological-active compounds did not detect differences between the scent composition of staminate and pistillate flowers. Pollinarium removal or deposition resulted in diminished scent emission within 24h in staminate and pistillate flowers, respectively. Surprisingly, bees discriminated pollinated from non-pollinated pistillate flowers as early as 2h after pollination. The rapid loss in the attractiveness of flowers following pollinarium removal/deposition can be interpreted as a strategy to direct pollinators to non-pollinated flowers. We have found no evidence that euglossine males discriminate staminate from pistillate flowers by means of floral scent. Instead, we speculate that bees use visual cues, such as sex dimorphic traits, to discriminate flowers of different sexes. Together, our results provide interesting insights into the evolution of floral signals in gender-dimorphic species and into its significance in plant reproductive biology.

An aromatic volatile attracts oligolectic bee pollinators in an interdependent bee-plant relationship.

Chemical signals emitted by the plant frequently mediate host-plant localization in specialized animal - plant associations. Studying the interdependent highly specialized association of the narrowly oligolectic bee pollinator Protodiscelis palpalis (Colletidae, Neopasiphaeinae) with Hydrocleys martii (Alismataceae) in ephemeral aquatic water bodies in semi-arid Caatinga of Brazil, we asked if specific volatile compounds produced by the flowers mediate pollinator attraction. The yellow Hydrocleys flowers are the sole pollen and nectar resources, and mating sites for the bees. We analyzed the floral scents of this species and of the closely related H. nymphoides, which is not visited by P. palpalis, and tested the main volatile compounds of both species under field conditions to evaluate their attractiveness to bees of P. palpalis. Methoxylated aromatics were the dominant floral scent components in both species, but each species exhibited a characteristic scent profile. Dual choice bioassays using artificial flowers made of yellow and blue adhesive paper clearly revealed that ρ-methylanisole alone, the dominant volatile of H. martii, attracted significantly more bees than unbaited flowers. This compound represents an olfactory communication channel used by the plant that lures its effective oligolectic pollinators to its flowers. Yellow artificial flowers baited significantly more bees than blue ones. Our study reinforces the recent findings that specific compounds in complex floral scent bouquets are crucial for host-plant location in oligolectic bees.

Nocturnal bees are attracted by widespread floral scents.

Flower localization in darkness is a challenging task for nocturnal pollinators. Floral scents often play a crucial role in guiding them towards their hosts. Using common volatile compounds of floral scents, we trapped female nocturnal Megalopta-bees (Halictidae), thus uncovering olfactory cues involved in their search for floral resources. Applying a new sampling method hereby described, we offer novel perspectives on the investigation of nocturnal bees.