The use of MES buffer in early nodulation studies with annual Medicago species

Abstract

The usefulness of 2-(N-morpholino)ethanesulfonic acid (MES) to stabilize the pH of solutions in the range pH 5.5 to 7.0 during early growth and nodulation of annual Medicago (medic) species was investigated. In the first experiment, effects of MES concentrations (0, 1, 2, 5, 10 mM) on growth and nodulation of Medicago polymorpha L. and the stability of solution pH were investigated. In the second experiment we assessed the effect of MES on the growth and nodulation of three medic species (M. truncatula Gaernt., M. polymorpha and M. murex Willd.) at a range of starting pH levels (5.5, 6.0, 6.5 and 7.0) with different concentrations of MES buffer (0, 0.5, 10 mM). In a third experiment, the effects of the concentration of extra cations and the species (K-ion or Na-ion), used as hydroxide to bring solutions containing different MES concentrations to target pH were investigated. MES had no effect on growth at MES concentrations as high as 10 mM but nodule numbers were increased by concentrations of MES of 2 mM or greater. The presence of extra K-ion or Na-ion in solutions had no effect on plant growth or nodulation. With low MES concentrations (0.5 mM), solution pH stability was improved by daily titration and the use of a high nitrate to ammonium ratio (9 : 1) in solution. This combination of strategies maintained pH within a narrow range without effects on annual medic growth or nodulation.