Plastocyanin, which requires copper (Cu) as a cofactor, is an electron carrier in the thylakoid lumen and essential for photoautotrophic growth of plants. The Cu microRNAs, which are expressed during Cu deprivation, down-regulate several transcripts that encode for Cu proteins. Since plastocyanin is not targeted by the Cu microRNAs, a cofactor economy model has been proposed in which plants prioritize Cu for use in photosynthetic electron transport. However, defects in photosynthesis are classic symptoms of Cu deprivation, and priorities in Cu cofactor delivery have not been determined experimentally. Using hydroponically grown Populus trichocarpa (clone Nisqually-1), we have established a physiological and molecular baseline for the response to Cu deficiency. An integrated analysis showed that Cu depletion strongly reduces the activity of several Cu proteins including plastocyanin, and consequently, photosynthesis and growth are decreased. Whereas plastocyanin mRNA levels were only mildly affected by Cu depletion, this treatment strongly affected the expression of other Cu proteins via Cu microRNA-mediated transcript down-regulation. Polyphenol oxidase was newly identified as Cu regulated and targeted by a novel Cu microRNA, miR1444. Importantly, a spatiotemporal analysis after Cu resupply to previously depleted plants revealed that this micronutrient is preferentially allocated to developing photosynthetic tissues. Plastocyanin and photosynthetic electron transport efficiency were the first to recover after Cu addition, whereas recovery of the other Cu-dependent activities was delayed. Our findings lend new support to the hypothesis that the Cu microRNAs serve to mediate a prioritization of Cu cofactor use. These studies also highlight poplar as an alternative sequenced model for spatiotemporal analyses of nutritional homeostasis.

Intensive agricultural practices are leading to loss of soil fertility and overexploitation of natural resources which cause nutrients imbalance and further impair ecosystem services. Organic farming (OF), also coupled with minimum tillage and crop rotations, represents one of the strategies to limit this process and maintain soil functions. In a two-year field trial, organic farming practices, including a set of fertilizations combined with crop rotations and association with nitrogen fixing cover crops, were compared. The aim of this research was to assess in the short-term the effects on soil organic carbon, aggregate stability, and soil enzymes activities of using a combination of promising management practices in the delta region of the Po river. Results did not show improvements in organic carbon content and soil aggregate stability. Conversely, enzymatic activities were always significantly higher in OF treatments than the conventional one. Crop rotation and associated legumes were effective in enhancing β-glucosidase and P fixation through phosphatases activities. The present work suggests that an effective choice of crop species coupled with legumes can enhance biological activity re-starting main mechanisms of microbial development even without a contemporary increase of organic matter.