Hexitols as major intermediates of glucose assimilation by mycelium of Puccinia graminis

Abstract

Mycelium of Puccinia graminis was grown for 4 d on 200 mM D-[U-14C]glucose followed by a cold chase for 30 h. Analysis of cellular metabolites during the chase indicated significant turnover only in carbohydrates soluble in 80% (w/v) ethanol. A kinetic analysis of the depletion of [14C] in pools of free sugars and sugar alcohols indicated that the trehalose pools and a small proportion (12-16%) of the mannitol and glucitol pools did not turn over, whilst pools of glucose, fructose, and the remainder of the hexitols became totally,depleted of label during the chase. Because the [14C] was totally lost from the pools of glucose and fructose prior to the hexitols, it was deduced that both of these hexoses were precursors of the hexitols. Estimation of the carbon fluxes through pools indicated that 52, 36 and 16% of the carbon from glucose was assimilated via glucitol, fructose and mannitol respectively, demonstrating that glucitol could not have originated from fructose as sole precursor. After offering D-[U-14C]glucitol, [14C] was assimilated into trehalose phosphate, glucans and amino acids, but not into free glucose or fructose. These data indicate that hexitols are quantitatively important intermediates during the assimilation of glucose by Puccinia graminis.