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The phyllosphere of the Brazilian Atlantic Forest has been estimated to contain several million bacterial species that are associated with approximately 20000 plant species. Despite the high bacterial diversity in the phyllosphere, the function of these microorganisms and the mechanisms driving their community assembly are largely unknown. In this study, we characterized the bacterial communities in the phyllospheres of four tree species of the Atlantic Forest (Mollinedia schottiana, Ocotea dispersa, Ocotea teleiandra, and Tabebuia serratifolia) and their metaproteomes to examine the basic protein functional groups expressed in the phyllosphere. Bacterial community analyses using 16S rRNA gene sequencing confirmed prior observations that plant species harbor distinct bacterial communities and that plants of the same taxon have more similar communities than more distantly related taxa. Using LC-ESI-Q-TOF, we identified 216 nonredundant proteins, based on 3503 peptide mass spectra. Most protein families were shared among the phyllosphere communities, suggesting functional redundancy despite differences in the species compositions of the bacterial communities. Proteins involved in glycolysis and anaerobic carbohydrate metabolism, solute transport, protein metabolism, cell motility, stress and antioxidant responses, nitrogen metabolism, and iron homeostasis were among the most frequently detected. In contrast to prior studies on crop plants and Arabidopsis, a low abundance of OTUs related to Methylobacterium and no proteins associated with the metabolism of one-carbon molecules were detected in the phyllospheres of the tree species studied here. Our data suggest that even though the phyllosphere bacterial communities of different tree species are phylogenetically diverse, their metaproteomes are functionally convergent with respect to traits required for survival on leaf surfaces.

Mutualistic symbioses between plants and fungi are a widespread phenomenon in nature. Particularly in orchids, association with symbiotic fungi is required for seed germination and seedling development. During the initial stages of symbiotic germination, before the onset of photosynthesis, orchid protocorms are fully mycoheterotrophic. The molecular mechanisms involved in orchid symbiotic germination and development are largely unknown, but it is likely that changes in plant energy metabolism and defense-related responses play a central role in these processes. We have used 2D-LC-MS/MS coupled to isobaric tagging for relative and absolute quantification to identify proteins with differential accumulation in Oncidium sphacelatum at different stages of mycorrhizal protocorm development (achlorophyllous and green protocorms) after seed inoculation with a Ceratobasidium sp. isolate. We identified and quantified 88 proteins, including proteins putatively involved in energy metabolism, cell rescue and defense, molecular signaling, and secondary metabolism. Quantitative analysis showed that the expected changes in carbon metabolism in green protocorms were accompanied by enhanced accumulation of proteins involved in the modulation of reactive oxygen species homeostasis, defense-related responses, and phytoalexins and carotenoid biosynthesis. Our results suggest profound metabolic changes in orchid protocorms during the switch from the fully mycoheterotrophic to the photosynthetic stage. Part of these changes may be also related to the obligatory nature of the interaction with the endomycorrhizal fungus.

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