An RT-qPCR approach to study the expression of genes responsible for sugar assimilation during rehydration of active dry yeast

Food Microbiology
Enrico VaudanoEmilia Garcia-Moruno

Abstract

A short reactivation period in aqueous media is required for active dry yeast (ADY) to be utilised in winemaking. Rehydration restores the active metabolic conditions necessary for good fermentative and competitive abilities. We used a reverse transcription-quantitative PCR (RT-qPCR) method with relative quantification to investigate the expression of seven hexose transporter genes (HXT1-7) and one invertase-encoding gene (SUC2) during ADY rehydration in water with or without sucrose. For this, seven candidate reference genes were evaluated, and the three most stably expressed genes were selected and used for mRNA normalisation. The results show that, during the rehydration in the presence of sucrose, yeast cells are able to immediately hydrolyse this sugar into glucose and fructose as soon as they are introduced in the medium. Subsequently, differential glucose/fructose uptake occurs, which is mediated by hexose transporters. At the transcriptomic level, there is a strong induction of the high-affinity transporters, HXT2 and HXT4, and the low-affinity transporters, HXT3 and HXT1, when ADY is rehydrated with sucrose, while HXT5 and HXT6/7 are expressed at high levels with a moderate tendency to decrease. In water, the HXT2 gene...Continue Reading

References

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Apr 12, 2006·Applied Microbiology and Biotechnology·Tristan RossignolBruno Blondin
Jul 16, 2008·International Journal of Food Microbiology·B Rodríguez-PorrataR Cordero Otero
Dec 9, 2008·International Journal of Food Microbiology·Enrico VaudanoEmilia Garcia-Moruno

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Citations

Aug 5, 2011·Applied Microbiology and Biotechnology·Antonella CostantiniEmilia Garcia-Moruno

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