Frankia alni Carbonic Anhydrase Regulates Cytoplasmic pH of Nitrogen-Fixing Vesicles.

A phyloprofile of Frankia genomes was carried out to identify those genes present in symbiotic strains of clusters 1, 1c, 2 and 3 and absent in non-infective strains of cluster 4. At a threshold of 50% AA identity, 108 genes were retrieved. Among these were known symbiosis-associated genes such as nif (nitrogenase), and genes which are not know as symbiosis-associated genes such as can (carbonic anhydrase, CAN). The role of CAN, which supplies carbonate ions necessary for carboxylases and acidifies the cytoplasm, was thus analyzed by staining cells with pH-responsive dyes; assaying for CO2 levels in N-fixing propionate-fed cells (that require a propionate-CoA carboxylase to yield succinate-CoA), fumarate-fed cells and N-replete propionate-fed cells; conducting proteomics on N-fixing fumarate and propionate-fed cells and direct measurement of organic acids in nodules and in roots. The interiors of both in vitro and nodular vesicles were found to be at a lower pH than that of hyphae. CO2 levels in N2-fixing propionate-fed cultures were lower than in N-replete ones. Proteomics of propionate-fed cells showed carbamoyl-phosphate synthase (CPS) as the most overabundant enzyme relative to fumarate-fed cells. CPS combines carbonate and ammonium in the first step of the citrulline pathway, something which would help manage acidity and NH4+. Nodules were found to have sizeable amounts of pyruvate and acetate in addition to TCA intermediates. This points to CAN reducing the vesicles' pH to prevent the escape of NH3 and to control ammonium assimilation by GS and GOGAT, two enzymes that work in different ways in vesicles and hyphae. Genes with related functions (carboxylases, biotin operon and citrulline-aspartate ligase) appear to have undergone decay in non-symbiotic lineages.

Traceability of Croatian extra virgin olive oils to the provenance soils by multielement and carbon isotope composition and chemometrics.

A capacity to determine the provenance of high-value food products is of high scientific and economic interest. With the aim to develop a tool for geographical traceability of Croatian extra virgin olive oils (EVOO), multielement composition and 13C/12C isotope ratio in EVOO as well as the geochemistry of the associated soils were analysed in samples collected from three regions along the Croatian Adriatic coast. Soil geochemistry was shown to influence the transfer and elemental composition of EVOO. The most discriminating variables to distinguish EVOO from different regions were S, Mo, Rb, Mg, Pb, Mn, Sn, K, V and δ13C. The predictive models achieved high sensitivity and specificity, especially when carbon isotope composition was added. The results suggest that interregional geographical traceability of Croatian EVOO is possible based on matching their multielement composition with that of the soils in the provenance area.

Physiological characterization of a pepper hybrid rootstock designed to cope with salinity stress.

In pepper crops, rootstocks that tolerate salt stress are not used because available commercial rootstocks offer limited profits. In this context, we obtained the hybrid NIBER®, a new salinity-tolerant rootstock that has been tested under real salinity field conditions for 3 years with 32%-80% higher yields than ungrafted pepper plants. This study aimed to set up the initial mechanisms involved in the salinity tolerance of grafted pepper plants using NIBER® as a rootstock to study root-shoot behavior, a basic requirement to develop efficient rootstocks. Gas exchange, Na+/K+, antioxidant capacity, nitrate reductase activity, ABA, proline, H2O2, phenols, MDA concentration and biomass were measured in ungrafted plants of cultivar Adige (A), self-grafted (A/A), grafted onto NIBER® (A/N) and reciprocal grafted plants (N/A), all exposed to 0 mM and 70 mM NaCl over a 10-day period. Salinity significantly and quickly decreased photosynthesis, stomatal conductance and nitrate reductase activity, but to lower extent in A/N plants compared to A, A/A and N/A. A/N plants showed decreases in the Na+/K+ ratio, ABA content and lipid peroxidation activity. This oxidative damage alleviation in A/N was probably due to an enhanced H2O2 level that activates antioxidant capacity to cope salinity stress, and acts as a signal molecule rather than a damaging one by contributing a major increase in phenols and, to a lesser extent, in proline concentration. These traits led to a minor impact on biomass in A/N plants under salinity conditions. Only the plants with the NIBER® rootstock controlled the scion by modulating responses to salinity.