López-Pedrosa Agustín, González-Guerrero Manuel, Valderas Ascensión, Azcón-Aguilar Concepción, Ferrol Nuria
Departamento de Microbiología del Suelo y Sistemas Simbióticos, Estación Experimental del Zaidín, CSIC, Profesor Albareda 1, 18008 Granada, Spain.
Fungal Genet Biol. 2006 Feb;43(2):102-10. doi: 10.1016/j.fgb.2005.10.005. Epub 2005 Dec 28.
We report the cloning and characterization of the first NH(4)(+) transporter gene (GintAMT1) in an arbuscular mycorrhizal fungus. GintAMT1 encodes a polypeptide of 479 amino acids sharing high sequence similarity with previously characterized NH(4)(+) transporters from other fungi. Heterologous expression of GintAMT1 in the yeast triple mep mutant complemented the defect of this strain to grow in the presence of less than 1mM NH(4)(+). As revealed by [(14)C]methylammonium uptake experiments carried out in yeast, GintAMT1 encodes a high-affinity NH(4)(+) transporter. In mycelia developed in the presence of 0.9 m M NO(3)(-), GintAMT1 transcription was increased after the addition of 30 microM NH(4)(+) but decreased after the addition of 3 mM NH(4)(+). However, in mycelia grown in the presence of higher N concentrations, GintAMT1 transcripts decreased after the addition of NH(4)(+), irrespective of the concentration used. These data suggest that GintAMT1 is involved in NH(4)(+) uptake by the extraradical mycelia from the surrounding media when it is present at micromolar concentrations.
我们报道了丛枝菌根真菌中首个铵转运蛋白基因(GintAMT1)的克隆及特性分析。GintAMT1编码一个由479个氨基酸组成的多肽,与其他真菌中已鉴定的铵转运蛋白具有高度的序列相似性。GintAMT1在酵母三重mep突变体中的异源表达弥补了该菌株在铵浓度低于1mM时生长的缺陷。酵母中进行的[¹⁴C]甲基铵摄取实验表明,GintAMT1编码一种高亲和力的铵转运蛋白。在0.9mM硝酸根存在下生长的菌丝体中,添加30μM铵后GintAMT1转录增加,但添加3mM铵后转录减少。然而,在高氮浓度下生长的菌丝体中,添加铵后GintAMT1转录本减少,与所用浓度无关。这些数据表明,当微摩尔浓度的铵存在于周围介质中时,GintAMT1参与根外菌丝体对铵的吸收。