Although previous studies on N2-fixing legumes have demonstrated the contribution of acid phosphatases to their phosphorus (P) use efficiency under P-deficient growth conditions, localization of these enzymes in bean nodules has not been demonstrated. In this study, phosphoenol pyruvate phosphatase (PEPase) gene transcripts were localized within the nodule tissues of two recombinant inbred lines, RIL115 (P-deficiency tolerant) and RIL147 (P-deficiency sensitive), of Phaseolus vulgaris. Nodules were induced by Rhizobium tropici CIAT899 under hydroaeroponic conditions with a sufficient versus a deficient P supply. The results indicated that PEPase transcripts were particularly abundant in the nodule infected zone and cortex of both RILs. Analysis of fluorescence intensity indicated that nodule PEPase was induced under conditions of P deficiency to a significantly higher extent in RIL147 than in RIL115, and more in the inner cortex (91%) than in the outer cortex (71%) or the infected zone (79%). In addition, a significant increase (39%) in PEPase enzyme activity in the P-deficient RIL147 correlated with an increase (58%) in the efficiency of use in rhizobial symbiosis. It was concluded that nodule PEPase is upregulated under conditions of P deficiency in the P-deficiency-sensitive RIL147, and that this gene may contribute to adaptation of rhizobial symbiosis to low-P environments.
Materials and methods
Plant material and growth conditionsFour Moroccan alfalfa populations, Tafilalet 1 (Taf 1), Tafilalet 2 (Taf 2), Demnate (Dem), and Tata, were used in this study. Seeds were supplied by National Institute for
Although recent studies have addressed the effects of phosphorus (P) deficiency on nodule O 2 permeability, little attention has been given to the relationship between nodule P status and nodule permeability. To study these traits, four recombinant inbred lines, namely RILs 34, 83, 115, 147 and one local variety (Concesa) of common bean (Phaseolus vulgaris) were inoculated with RhM11 (a native rhizobial strain from Haouz area of Marrakesh), and grown in hydroaeroponic culture under Psufficiency (250 μmol P plant −1 week −1 ) versus P-deficiency (75 μmol P plant −1 week −1 ) conditions. At the flowering stage, the biomass of plants and nodules and their P contents was determined after measuring O 2 uptake by nodulated roots (Conr) and nodule conductance to O 2 diffusion (g n ).The results showed that P-deficiency significantly decreased plant growth and nodulation, though there were differences between bean genotypes. P-deficiency also induced a decrease in nodule P content (31%) in both sensitive (83, 147) and tolerant lines (34, 115), a 42 and 27% reduction in shoots of sensitive and tolerant lines, respectively. These decreases were associated with significant variations in nodule surface and O 2 permeability among bean genotypes and P-nutrition. Under P-deficiency, g n increased more for the sensitive (39%) than for the tolerant lines (27%). This increase was linked with a rise both in the P levels in nodules and shoots, as well as in the efficiency of symbiotic nitrogen fixation as determined by nodule-dependent biomass production for the sensitive lines. Furthermore, positive correlations were found between O 2 permeability, g n and P content both in nodules and shoots (r 2 =0.94** and r 2 =0.96**). We conclude that nodule variations in Conr and g n are related to nodule P content, and may contribute to the adaptation of energy metabolism in N 2 -fixing bean nodules to P-deficiency.
The effect of phosphorus (P) deficiency on phosphatases activities in N2-fixing legumes has been widely studied in hydroponic culture. However, the response of acid phosphatase (APase) and phytase in rhizosphere, nodules and seeds of Phaseolus vulgaris to low soil’s P-availability is not yet fully understood. In this study, six genotypes of N2-fixing P. vulgaris were grown under contrasting soil P-availabilities; i.e., low (4.3 mg P kg−1) and sufficient (16.7 mg P kg−1) in the Haouz region of Morocco. At flowering and maturity stages, plants were harvested and analyzed for their phosphatases activities, growth and P content. Results show that, low P decreased nodulation, growth, P uptake and N accumulation in all the genotypes, but to a greater extent in the sensitive recombinant inbreed line 147. In addition, while seed P content was slightly reduced under low P soil; a higher P was noticed in the Flamingo and Contender large seeded-beans (6.15 to 7.11 mg g−1). In these latter genotypes, high APase and phytase activities in seeds and nodules were associated with a significant decline in rhizosphere’s available P. APase activity was mainly stimulated in nodules, whereas phytase activity was highly induced in seeds (77%). In conclusion, the variations of APase and phytase activities in nodules and seeds depend on genotype and can greatly influence the internal utilization of P, which might result in low P soil tolerance in N2-fixing legumes
This study compared growth, nodulation, nitrogen fixation, and nodular enzyme activities in response to salinity in some common bean-rhizobia symbiotic combinations. Seeds of Paulista and Efequince, two varieties of the common bean (Phaseolus vulgaris) were germinated and seedlings were transferred to pots containing vermiculite inoculated with the reference Rhizobium strain CIAT899 or with RhM11 or RhM14, two local strains. Plants were grown in a temperature-controlled glasshouse at 28°C and irrigated with a nutrient solution without NaCl (control) or supplemented with 25 mM NaCl (stressed). Plants were harvested at the flowering stage. The results showed that in controls, inoculation with RhM11 improved plant and nodule growth compared with those inoculated with RhM14 and CIAT 899. NaCl treatment generally had a negative affect on plant and nodule growth. Under the saline treatment, symbiotic nitrogen fixation was not significantly affected in the CIAT899-Paulista, CIAT899-Efequince and RhM11-Paulista combinations. Plant mineral nutrition was negatively affected under salt treatment for all of the tested symbiotic combinations. Inoculation with CIAT899 and RhM11 conferred more plant tolerance to salinity than inoculation with RhM14. The nodular phosphoenolpyruvate carboxylase (PEPC) and malate dehydrogenase (MDH) exhibited higher activities and were less affected by salinity in plants inoculated with the reference strain CIAT899 than those inoculated with local strains. We conclude that plants inoculated with CIAT899 and RhM11 showed more salinity stress tolerance than those inoculated with RhM14.
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