Effect of Carbon and Nitrogen Availability on Metabolism of Amino Acids in Germinating Spores of Arbuscular Mycorrhizal Fungi

Abstract

The effects of carbon (C) and nitrogen (N) sources on N utilization and biosynthesis of amino acids were examined in the germinating spores of the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith after exposure to various N substrates, CO 2, glucose, and/or root exudates. The N uptake and de novo biosynthesis of amino acids were analyzed using stable isotopic labeling with mass spectrometric detection. High-performance liquid chromatography-based analysis was used to measure amino acid levels. In the absence of exogenous N sources and in the presence of 25 mL L −1 CO 2, the germinating AM fungal spores utilized internal N storage as well as C skeletons derived from the degradation of storage lipids to biosynthesize the free amino acids, in which serine and glycine were produced predominantly. The concentrations of internal amino acids increased gradually as the germination time increased from 0 to 1 or 2 weeks. However, asparagine and glutamine declined to the low levels; both degraded to provide the biosynthesis of other amino acids with C and N donors. The availability of exogenous inorganic N (ammonium and nitrate) and organic N (urea, arginine, and glutamine) to the AM fungal spores using only CO 2 for germination generated more than 5 times more internal free amino acids than those in the absence of exogenous N. A supply of exogenous nitrate to the AM fungal spores with only CO 2 gave rise to more than 10 times more asparagine than that without exogenous N. In contrast, the extra supply of exogenous glucose to the AM fungal spores generated a significant enhancement in the uptake of exogenous N sources, with more than 3 times more free amino acids being produced than those supplied with only exogenous CO 2. Meanwhile, arginine was the most abundant free amino acid produced and it was incorporated into the proteins of AM fungal spores to serve as an N storage compound.