Morel Mélanie, Buée Marc, Chalot Michel, Brun Annick
IFR 110, UMR INRA/UHP 1136 Interactions Arbres Micro-organismes, Université Henri Poincaré- Nancy I, Faculté des Sciences et Techniques BP239, F-54506 Vandoeuvre-les-Nancy Cedex, France.
New Phytol. 2006;169(1):179-89. doi: 10.1111/j.1469-8137.2005.01556.x.
There is much controversy on the contribution of NADP-dependent glutamate dehydrogenase (NADP-GDH) in NH4+ assimilation in ectomycorrhizal (ECM) fungi and ectomycorrhizas. Experiments reported here provide information on the dispensability of NADP-GDH in various ectomycorrhizal isolates. Glutamate dehydrogenase and glutamine synthetase (GS) enzyme activities were measured on mycelia grown under various nitrogen (N) conditions. The contribution of GDH in ammonium assimilation was further estimated by following 15N incorporation from (15NH4)2SO4 into glutamate, when GS was inhibited by phosphinothricin. Finally, gene amplification on cDNA and genomic DNA was performed using degenerated primers. Two groups of fungi could be distinguished. The GDH+ fungi include Hebeloma cylindrosporum-like fungi, which possess a functional NADP-GDH. The GDH- fungi include Paxillus involutus-like fungi for which the NADP-GDH activity, as well as the GDHA transcripts, were not detected, whatever the growth condition. All the results are consistent with the dispensability of the NADP-GDH function in ECM fungi, suggesting a minor role in ammonium assimilation in ectomycorrhizal fungi. We hypothesize that the lack of a functional NADP-GDH could be an evolutive adaptation in relation to the ecological niche of ECM fungi, rather than a transitional regulation in response to changes in N contents of the extracellular medium.
关于烟酰胺腺嘌呤二核苷酸磷酸(NADP)依赖性谷氨酸脱氢酶(NADP-GDH)在外生菌根(ECM)真菌和外生菌根的铵同化作用中的贡献存在诸多争议。本文报道的实验提供了有关NADP-GDH在各种外生菌根分离物中的非必需性的信息。在不同氮(N)条件下生长的菌丝体上测定了谷氨酸脱氢酶和谷氨酰胺合成酶(GS)的酶活性。当谷氨酰胺合成酶被草丁膦抑制时,通过追踪(15NH4)2SO4中的15N掺入谷氨酸中来进一步估计谷氨酸脱氢酶在铵同化中的贡献。最后,使用简并引物对cDNA和基因组DNA进行基因扩增。可区分出两组真菌。GDH+真菌包括具有功能性NADP-GDH的柱状环柄菇样真菌。GDH-真菌包括卷缘桩菇样真菌,无论生长条件如何,均未检测到NADP-GDH活性以及GDHA转录本。所有结果均与NADP-GDH功能在外生菌根真菌中的非必需性一致,表明其在外生菌根真菌的铵同化中作用较小。我们推测,缺乏功能性NADP-GDH可能是与外生菌根真菌的生态位相关的一种进化适应,而不是对外界培养基中氮含量变化的一种过渡调节。