Abstract
The genome of Aspergillus nidulans accommodates two glycerol 3-phosphate dehydrogenase genes, gfdA and gfdB. Previous studies confirmed that GfdA is involved in the osmotic stress defence of the fungus. In this work, the physiological role of GfdB was characterized via the construction and functional characterization of the gene deletion mutant ΔgfdB. Unexpectedly, ΔgfdB strains showed oxidative stress sensitivity in the presence of a series of well-known oxidants including tert-butyl-hydroperoxide (tBOOH), diamide as well as hydrogen peroxide. Moderate sensitivity of the mutant towards the cell wall stress inducing agent CongoRed was also observed. Hence, both Gfd isoenzymes contributed to the environmental stress defence of the fungus but their functions were stress-type-specific. Furthermore, the specific activities of certain antioxidant enzymes, like catalase and glutathione peroxidase, were lower in ΔgfdB hyphae than those recorded in the control strain. As a consequence, mycelia from ΔgfdB cultures accumulated reactive species at higher levels than the control. On the other hand, the specific glutathione reductase activity was higher in the mutant, most likely to compensate for the elevated intracellular oxidative species concentrations. Nevertheless, the efficient control of reactive species failed in ΔgfdB cultures, which resulted in reduced viability and, concomitantly, early onset of programmed cell death in mutant hyphae. Inactivation of gfdB brought about higher mannitol accumulation in mycelia meanwhile the erythritol production was not disturbed in unstressed cultures. After oxidative stress treatment with tBOOH, only mannitol was detected in both mutant and control mycelia and the accumulation of mannitol even intensified in the ΔgfdB strain.
Highlights
Fungi encounter abiotic and biotic environmental stresses frequently in their natural habitats and, they possess an effective stress response system to adapt to and survive under such harsh and randomly varying circumstances (Miskei et al, 2009; Gasch, 2007)
Glycerol can be produced in two steps including the enzyme glycerol 3phosphate dehydrogenase (G3PDH), which catalyses the conversion of dihydroxyacetone phosphate (DHAP) into glycerol 3phosphate (G3P), and G3P is subsequently converted into glycerol by a phosphatase (Fillinger et al, 2001)
Mycelia from 0.5 ml culture aliquots of cultures were transferred into test tubes containing 0.5 ml fresh media supplemented with 10 mg mlÀ1 MTT
Summary
Fungi encounter abiotic and biotic environmental stresses frequently in their natural habitats and, they possess an effective stress response system to adapt to and survive under such harsh and randomly varying circumstances (Miskei et al, 2009; Gasch, 2007). Fungal stress response systems rely on both enzymatic and non-enzymatic elements to combat environmental challenges (Pocsi et al, 2004; Duran et al, 2010; Gasch and WernerWashburne, 2002). The production of glycerol - which is one of the main osmoregulatory solutes in fungal cells - plays an important role in the prevention of hyperosmotic stress (Fillinger et al, 2001; Hagiwara et al, 2016; Saito and Posas, 2012).
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