No abstract

A beneficial effect of B and Ca application on symbiotic interaction between legume and rhizobia under saline conditions has recently been shown, suggesting conventional agricultural practices to increase crop salt tolerance. However, nothing is known about application of both nutrients on early events of legume development under salt stress, prior to the establishment of a symbiotic interaction. Therefore, the effects of different levels of B (from 9.3 to 93 µM B) and Ca (from 0.68 to 5.44 mM Ca) on seed germination, root elongation, plant development, and mineral composition of pea (Pisum sativum L. cv. Argona) grown under 0 to 150 mM NaCl, were analysed. Development of plants previously germinated in the presence of salt was more impaired than that of plants put under salt stress once seeds were germinated. A NaCl concentration of 75 mM and 150 mM inhibited pea seed germination and seedling growth. The addition of either extra B or extra Ca to the germination solution prevented the reduction caused by 75 mM NaCl but not that of 150 mM NaCl. However, root elongation and plant development under salt stress (75 mM NaCl) was enhanced only by addition of both B and Ca. When plants were cultivated in the absence of external N, N content in roots and shoots originating from seeds was diminished by salt and enhanced by B and Ca, suggesting a role of these nutrients in remobilisation of seed nutrient stores. Salinity also led to an extremely high concentration of Na + ions, and to a decrease of B and Ca concentrations. This can be overcome by addition of both nutrients, increasing salt tolerance of developing pea plants. The necessity of nutritional studies to increase crop production in saline soils is discussed and proposed.

Because boron (B) and calcium (Ca 2+ + + + ) seem to have a strong effect on legume nodulation and nitrogen fixation, rhizobial symbiosis with leguminous plants, grown under varying concentrations of both nutrients, was investigated. The study of early pre-infection events included the capacity of root exudates to induce nod genes, and the degree of adsorption of bacteria to the root surface. Both phenomena were inhibited by B deficiency, and increased by addition of Ca 2+ , resulting in an increase of the number of nodules. The infection and invasion steps were investigated by fluorescence microscopy in pea nodules harbouring a Rhizobium leguminosarum strain that constitutively expresses green fluorescent protein. High Ca 2+ + + + enhanced cell and tissue invasion by Rhizobium , which was highly inhibited after B deficiency. This was combined with an increased B concentration in nodules of plants grown on B-free medium and supplemented with high Ca 2+ + + + concentrations, and that can be attributed to an increased B import to the nodules. Histological examination of indeterminate (pea) and determinate (bean) nodules showed an altered nodule anatomy at low B content of the tissue. The moderate increase in nodular B due to additional Ca 2+ + + + was not sufficient to prevent the abnormal cell wall structure and the aberrant distribution of pectin polysaccharides in B-deficient treatments. Overall results indicate that the development of the symbiosis depends of the concentration of B and Ca 2+ + + + , and that both nutrients are essential for nodule structure and function.

The effects of modifying boron (B) and calcium (Ca2+) concentrations on the establishment and development of rhizobial symbiosis in Pisum sativum plants grown under salt stress were investigated. Salinity almost completely inhibited the nodulation of pea plants by Rhizobium leguminosarum bv. viciae 3841. This effect was prevented by addition of Ca2+ during plant growth. The capacity of root exudates derived from salt‐treated plants to induce Rhizobium nod genes was not significantly decreased. However, bacterial adsorption to roots was highly inhibited in plants grown with 75 mM NaCl. Moreover, R. leguminosarum 3841 did not grow in minimal media containing such salt concentration. High Ca2+ levels enhanced both rhizobial growth and adsorption to roots, and increased nodule number in the presence of high salt. Nevertheless, the nodules developed were not functional unless the B concentration was also increased. Because B has a strong effect on infection and cell invasion, these processes were investigated by fluorescence microscopy in pea nodules harbouring a R. leguminosarum strain that expresses green fluorescent protein. Salt‐stressed plants had empty nodules and only those treated with high B and high Ca2+ developed infection threads and exhibited enhanced cell and tissue invasion by Rhizobium. Overall, the results indicate that Ca2+ promotes nodulation and B nodule development leading to an increase of salt tolerance of nodulated legumes.