Departamento de Microbiología del Suelo y Sistemas Simbióticos, Estación Experimental del Zaidín, CSIC, Profesor Albareda 1, 18008, Granada, Spain.
Mycorrhiza. 2012 Aug;22(6):485-91. doi: 10.1007/s00572-012-0452-0. Epub 2012 Jul 1.
The kinetics and energetics of (15)NH (4) (+) uptake by the extraradical mycelium of the arbuscular mycorrhizal fungus Rhizophagus irregularis were investigated. (15)NH (4) (+) uptake increased with increasing substrate concentration over the concentration range of 0.002 to 25 mM. Eadie-Hofstee plots showed that ammonium (NH (4) (+) ) uptake over this range was biphasic. At concentrations below 100 μM, NH (4) (+) uptake fits a Michaelis-Menten curve, typical of the activity of a saturable high-affinity transport system (HATS). At concentrations above 1 mM, NH (4) (+) influx showed a linear response typical of a nonsaturable low-affinity transport system (LATS). Both transport systems were dependent on external pH. The HATS and, to a lesser extent, the LATS were inhibited by the ionophore carbonylcyanide m-chlorophenylhydrazone (CCCP) and the ATP-synthesis inhibitor 2,4-dinitrophenol. These data indicate that the two NH (4) (+) transport systems of R. irregularis are dependent on metabolic energy and on the electrochemical H(+) gradient. The HATS- and the LATS-mediated (15)NH (4) (+) influxes were also regulated by acetate. This first report of the existence of active high- and low-affinity NH4(+) transport systems in the extraradical mycelium of an arbuscular mycorrhizal fungus and provides novel information on the mechanisms underlying mycosymbiont uptake of nitrogen from the soil environment.
(15)NH(4)(+)被丛枝菌根真菌粗糙无梗囊霉(Rhizophagus irregularis)外生菌根的动力学和能量学进行了研究。(15)NH(4)(+)的摄取随着基质浓度在 0.002 至 25 mM 的范围内的增加而增加。Eadie-Hofstee 图表明,在该范围内,铵(NH(4)(+))的摄取呈双相。在低于 100 μM 的浓度下,NH(4)(+)的摄取符合米氏方程,这是一种饱和高亲和力运输系统(HATS)的典型活性。在高于 1 mM 的浓度下,NH(4)(+)的流入呈现出典型的非饱和低亲和力运输系统(LATS)的线性响应。这两种运输系统都依赖于外部 pH 值。HATS 以及在较小程度上,LATS 受到离子载体羰基氰化物 m-氯代苯腙(CCCP)和 ATP 合成抑制剂 2,4-二硝基苯酚的抑制。这些数据表明,R. irregularis 的两种 NH(4)(+)运输系统依赖于代谢能量和电化学 H(+)梯度。HATS 和 LATS 介导的(15)NH(4)(+)流入也受到乙酸盐的调节。这是首次报道在丛枝菌根真菌的外生菌根中存在主动的高亲和力和低亲和力 NH4(+)运输系统,并提供了关于真菌共生体从土壤环境中吸收氮的机制的新信息。