Nutrient transport into germinating Trichoderma atroviride conidia and development of its driving force.

The exit from dormancy and the start of growth should be preceded or at least accompanied by the uptake of nutrients. In this work we studied changes in the transport of several nutrients into Trichoderma atroviride conidia. Germination started with a short period of isodiametric growth (conidial swelling), followed by polarized growth (germ tube formation) after about 8 h at 26 °C. The onset of isodiametric growth required the presence of external both phosphate and nitrate. At the same time, an increased uptake of precursors of macromolecules and phospholipids ((14)C- or (3)H-labelled valine, uracil, N-acetylglucosamine and choline) occurred. A low uptake of these precursors was observed also in non-germinating conidia. Concomitantly, this uptake developed an increased sensitivity to the uncoupler 3,3',4',5-tetrachlorosalicylanilide. Expression and activity of H(+)-ATPase started after completing isodiametric growth, suggesting that the proton-motive force (PMF) generated by H(+)-ATPase may be an accelerator of nutrient uptake and metabolism. (14)C-valine uptake was also measured into a mutant with disrupted pma1 gene. This mutant did not form conidia. The mutant also exhibited uncoupler sensitivity of (14)C-valine uptake. These observations showed that a PMF must have been generated by a mechanism(s) other than the H(+)-ATPase activity in the WT before H(+)-ATPase expression and in mycelia with disrupted H(+)-ATPase.

Changes in metabolome and in enzyme activities during germination of Trichoderma atroviride conidia.

The aim of this work was to study the metabolic changes during germination of Trichoderma atroviride conidia along with selected marker enzyme activities. The increase in proteinogenic amino acid concentrations together with the increase in glutamate dehydrogenase activity suggests a requirement for nitrogen metabolism. Even though the activities of tricarboxylic acid cycle enzymes also increased, detected organic acid pools did not change, which predisposes this pathway to energy production and supply of intermediates for further metabolism. The concentrations of many metabolites, including the main osmolytes mannitol and betaine, also increased during the formation of germ tubes. The activities of H(+)-ATPase and GDPase, the only marker enzymes that did not have detectable activity in non-germinated conidia, were shown with germ tubes.