Proteome changes in Oncidium sphacelatum (Orchidaceae) at different trophic stages of symbiotic germination

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.