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An RT-qPCR approach to study the expression of genes responsible for sugar assimilation during rehydration of active dry yeast.
Food Microbiol. 2010 Sep; 27(6):802-8.FM

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 was the only one of the transporter genes studied that showed significant variations. These results suggest that during rehydration, expression is not simply regulated by the affinity to hexose but is also controlled by other mechanisms that allow the cell to bypass glucose control. Moreover, the expression of SUC2 showed little variation in media with sucrose, suggesting that other invertases and/or posttranscriptional controls exist.

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Authors+Show Affiliations

Vaudano E
CRA-Centro di Ricerca per l'Enologia, Via Pietro Micca, 35, 14100 Asti, Italy. biologia.molecolare@isenologia.it
Costantini A
No affiliation info available
Noti O
No affiliation info available
Garcia-Moruno E
No affiliation info available

MeSH

DNA, FungalFermentationFood MicrobiologyFungal ProteinsGene Expression Regulation, FungalGlucoseMonosaccharide Transport ProteinsRNA, FungalReverse Transcriptase Polymerase Chain ReactionSaccharomyces cerevisiaeSaccharomyces cerevisiae ProteinsWaterWinebeta-Fructofuranosidase

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20630323

Citation

Vaudano, Enrico, et al. "An RT-qPCR Approach to Study the Expression of Genes Responsible for Sugar Assimilation During Rehydration of Active Dry Yeast." Food Microbiology, vol. 27, no. 6, 2010, pp. 802-8.
Vaudano E, Costantini A, Noti O, et al. An RT-qPCR approach to study the expression of genes responsible for sugar assimilation during rehydration of active dry yeast. Food Microbiol. 2010;27(6):802-8.
Vaudano, E., Costantini, A., Noti, O., & Garcia-Moruno, E. (2010). An RT-qPCR approach to study the expression of genes responsible for sugar assimilation during rehydration of active dry yeast. Food Microbiology, 27(6), 802-8. https://doi.org/10.1016/j.fm.2010.04.010
Vaudano E, et al. An RT-qPCR Approach to Study the Expression of Genes Responsible for Sugar Assimilation During Rehydration of Active Dry Yeast. Food Microbiol. 2010;27(6):802-8. PubMed PMID: 20630323.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An RT-qPCR approach to study the expression of genes responsible for sugar assimilation during rehydration of active dry yeast. AU - Vaudano,Enrico, AU - Costantini,Antonella, AU - Noti,Olta, AU - Garcia-Moruno,Emilia, Y1 - 2010/04/27/ PY - 2009/10/20/received PY - 2010/03/30/revised PY - 2010/04/20/accepted PY - 2010/7/16/entrez PY - 2010/7/16/pubmed PY - 2010/10/13/medline SP - 802 EP - 8 JF - Food microbiology JO - Food Microbiol VL - 27 IS - 6 N2 - 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 was the only one of the transporter genes studied that showed significant variations. These results suggest that during rehydration, expression is not simply regulated by the affinity to hexose but is also controlled by other mechanisms that allow the cell to bypass glucose control. Moreover, the expression of SUC2 showed little variation in media with sucrose, suggesting that other invertases and/or posttranscriptional controls exist. SN - 1095-9998 UR - https://www.unboundmedicine.com/medline/citation/20630323/An_RT_qPCR_approach_to_study_the_expression_of_genes_responsible_for_sugar_assimilation_during_rehydration_of_active_dry_yeast_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0740-0020(10)00084-5 DB - PRIME DP - Unbound Medicine ER -