Determination of leghemoglobin components and xylem sap composition by capillary electrophoresis in hypernodulation soybean mutants cultivated in the field

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Abstract
The hypernodulation soybean mutant lines (NOD1-3, NOD2-4, NOD3-7) and their parent Williams, and the mutant En6500 and its parent Enrei were cultivated in a sandy dune field in Niigata, and the nodules and root bleeding xylem sap were sampled at 50, 70, 90, and 120 d after planting (DAP). The nodule size distribution patterns and concentration of leghemoglobin components were determined. The number of nodules of the hypernodulation mutant lines was about two to three times higher than that of the parent lines irrespective of the sampling date. At 50 DAP the nodule size was relatively smaller in the hypernodulation mutant lines, and the total dry weight of the nodules was almost the same in the mutant lines and their parents. At 70 DAP and 90 DAP, the size distribution of the hypernodulation mutant nodules became .almost the same as that of the parent lines, and both the number and total dry weight of the nodules were higher than those of the parent lines. The concentration of four Lb components was separately measured by capillary electrophoresis. The concentration of the Lb components in the hypernodulation mutant lines tended to be lower than in the parents, but the component ratios were not different between the hypernodulation mutants and their parents. Under field conditions, plant growth and nodulation characteristics were more similar between mutants and parents than in the hydroponic culture reported previously, although the mutants did exhibit hypernodulation traits. These findings suggest that the decrease in the Lb concentration and the different Lb components ratios in the mutants may be caused by secondary effects of excess nodulation, such as photosynthate deficiency, rather than by a genetic defect in mutation. The concentration of major nitrogenous compounds (allantoic acid, allantoin, asparagine, aspartic acid, and nitrate) in the xylem sap was also measured by capillary electrophoresis. The concentration of ureides and nitrate in xylem sap decreased with the plant age, but the asparagine concentration increased during the same period. The concentrations of ureides and asparagine were higher, and the nitrate concentration was lower in the mutant lines than in their parents, possibly due to the higher dependence on N2 fixation than N03 - utilization. In the xylem sap, nitrate was the major inorganic anion followed by phosphate, sulfate, and chloride, and potassium was the major cation followed by calcium or magnesium and sodium.