The nitrate transporter family protein LjNPF8.6 controls the N-fixing nodule activity

N-fixing nodules are new organs formed on legume roots as a result of the beneficial interaction with soil bacteria, rhizobia. The nodule functioning is still a poorly characterized step of the symbiotic interaction, as only a few of the genes induced in N-fixing nodules have been functionally characterized. We present here the characterization of a member of the Lotus japonicus nitrate transporter1/peptide transporter family, LjNPF8.6. The phenotypic characterization carried out in independent L. japonicus LORE1 insertion lines indicates a positive role of LjNPF8.6 on nodule functioning, as knockout mutants display N-fixation deficiency (25%) and increased nodular superoxide content. The partially compromised nodule functioning induces two striking phenotypes: anthocyanin accumulation already displayed 4 weeks after inoculation and shoot biomass deficiency, which is detected by long-term phenotyping. LjNPF8.6 achieves nitrate uptake in Xenopus laevis oocytes at both 0.5 and 30 mm external concentrations, and a possible role as a nitrate transporter in the control of N-fixing nodule activity is discussed

Multi-omics data integration provides insights into the post-harvest biology of a long shelf-life tomato landrace

In this study we investigated the transcriptome and epigenome dynamics of the tomato fruit during post-harvest in a landrace belonging to a group of tomatoes (Solanum lycopersicum L.) collectively known as "Piennolo del Vesuvio", all characterized by a long shelf-life. Expression of protein-coding genes and microRNAs as well as DNA methylation patterns and histone modifications were analysed in distinct post-harvest phases. Multi-omics data integration contributed to the elucidation of the molecular mechanisms underlying processes leading to long shelf-life. We unveiled global changes in transcriptome and epigenome. DNA methylation increased and the repressive histone mark H3K27me3 was lost as the fruit progressed from red ripe to 150 days post-harvest. Thousands of genes were differentially expressed, about half of which were potentially epi-regulated as they were engaged in at least one epi-mark change in addition to being microRNA targets in ~5% of cases. Down-regulation of the ripening regulator MADS-RIN and of genes involved in ethylene response and cell wall degradation was consistent with the delayed fruit softening. Large-scale epigenome reprogramming that occurred in the fruit during post-harvest likely contributed to delayed fruit senescence

LEGU-MED: Developing Biodiversity-Based Agriculture with Legume Cropping Systems in the Mediterranean Basin

Environmental degradation and the decrease of ecosystem service provision are currently of major concern, with current agricultural systems being a major driver. To meet our future environmental and sustainability targets a transformation of the agro-food systems and current agricultural value chain are crucial. One approach to redesign farming systems is the concept of biodiversity-based agriculture (BBA) which relies on sustainable diversification of biological components and their natural interactions in farming systems to maximize fertility, productivity, and resilience to external perturbations. Despite minimizing anthropogenic inputs, BBA is not yet able to meet all beneficial environmental objectives. BBA applied in the Mediterranean basin requires urgent innovation in approaches, methodologies, and models for small-holder traditional farming systems to ensure a stable provision of ecosystem services and better resilience to environmental stresses linked to climate change. Legumes are the backbone of the Mediterranean agro-ecosystems from ancient times, but their unique and wide biodiversity was not sufficiently valorized, especially by North-African countries. Here, we present LEGU-MED, a three-year international project funded by PRIMA initiative 2019. An international consortium was established involving five universities, 5 research institutes, and one private company from 8 countries: Italy, Germany, Spain, Algeria, Tunisia, Turkey, Lebanon, and Croatia. The main objective of this project is to put forward an international and well-integrated plan to valorize the legume agrobiodiversity of the Mediterranean in biodiversity-based farming systems and consequently enhance agro-ecosystem functions and services in the Mediterranean basin. The successful completion of LEGU-MED will have the following impacts on Mediterranean legume-based farming systems: (1) improve water use efficiency, (2) reduce the use of anthropogenic inputs through the maintenance of soil fertility, (3) enhance pollination and improve ecological connectivity with flora and fauna, (4) protect close-by wildland ecosystems, (5) enhance other ecosystem services (e.g., pest, disease, and weed suppression), and (6) provide healthier and safer protein-rich food. © 2022 by the authors. Licensee MDPI, Basel, Switzerland

The Lotus japonicus AFB6 Gene Is Involved in the Auxin Dependent Root Developmental Program

Auxin is essential for root development, and its regulatory action is exerted at different steps from perception of the hormone up to transcriptional regulation of target genes. In legume plants there is an overlap between the developmental programs governing lateral root and N2-fixing nodule organogenesis, the latter induced as the result of the symbiotic interaction with rhizobia. Here we report the characterization of a member of the L. japonicus TIR1/AFB auxin receptor family, LjAFB6. A preferential expression of the LjAFB6 gene in the aerial portion of L. japonicus plants was observed. Significant regulation of the expression was not observed during the symbiotic interaction with Mesorhizobium loti and the nodule organogenesis process. In roots, the LjAFB6 expression was induced in response to nitrate supply and was mainly localized in the meristematic regions of both primary and lateral roots. The phenotypic analyses conducted on two independent null mutants indicated a specialized role in the control of primary and lateral root elongation processes in response to auxin, whereas no involvement in the nodulation process was found. We also report the involvement of LjAFB6 in the hypocotyl elongation process and in the control of the expression profile of an auxin-responsive gene

Disruption of the Lotus japonicus transporter LjNPF2.9 increases shoot biomass and nitrate content without affecting symbiotic performances

BACKGROUND: After uptake from soil into the root tissue, distribution and allocation of nitrate throughout the whole plant body, is a critical step of nitrogen use efficiency (NUE) and for modulation of plant growth in response to various environmental conditions. In legume plants nitrate distribution is also important for the regulation of the nodulation process that allows to fix atmospheric N (N2) through the symbiotic interaction with rhizobia (symbiotic nitrogen fixation, SNF).[br/] RESULTS: Here we report the functional characterization of the Lotus japonicus gene LjNPF2.9, which is expressed mainly in the root vascular structures, a key localization for the control of nitrate allocation throughout the plant body. LjNPF2.9 expression in Xenopus laevis oocytes induces 15NO3 accumulation indicating that it functions as a nitrate importer. The phenotypic characterization of three independent knock out mutants indicates an increased shoot biomass in the mutant backgrounds. This phenotype is associated to an increased/decreased nitrate content detected in the shoots/roots. Furthermore, our analysis indicates that the accumulation of nitrate in the shoot does not affect the nodulation and N-Fixation capacities of the knock out mutants.[br/] CONCLUSIONS:[br/] This study shows that LjNPF2.9 plays a crucial role in the downward transport of nitrate to roots, occurring likely through a xylem-to-phloem loading-mediated activity. The increase of the shoot biomass and nitrate accumulation might represent a relevant phenotype in the perspective of an improved NUE and this is further reinforced in legume plants by the reported lack of effects on the SNF efficiency