Evidence for viable but nonculturable yeasts in botrytis-affected wine
Evidence for viable but nonculturable yeasts in botrytis-affected wine
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DOI：
10.1111/j.1365-2672.2005.02578.x
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发表时间：
2005-01-01
影响因子：
4
通讯作者：
Lonvaud-Funel, A
中科院分区：
文献类型：
--
作者：
Divol, B;Lonvaud-Funel, A
目的：在受葡萄孢霉病影响的葡萄酒中，添加高浓度的二氧化硫，以阻止酒精发酵，稳定葡萄酒。在桶中成熟或瓶装陈化过程中，有时会发生不必要的参考。然而，葡萄酒样品在成熟初期通常的平板计数结果表明葡萄酒微生物的稳定性。方法与结果：采用直接荧光技术，观察了添加SO2后酵母细胞的行为，并与平板计数结果进行了比较。采用聚合酶链式反应-限制性片段长度多态性和聚合酶链式反应-变性梯度凝胶电泳法对宿存酵母菌进行鉴定。经鉴定为酿酒酵母和星状假丝酵母，成熟几个月后，出现了其他腐败酵母，如粘性红酵母和百合酵母菌。结论：这些酵母的所有特征都导致了酵母在葡萄酒中以一种活的、但不能培养的状态(VBNC)存在。抑制游离SO2的作用并没有导致细胞复苏；然而，另一种方法证明了细胞退出类VBNC状态的能力。研究的意义和影响：本研究允许表征葡萄酒中存在类VBNC酵母。这种“参照”现象可能是由于VBNC状态的退出所致。
Aims: In Botrytis- affected wine, high concentrations of SO2 are added to stop the alcoholic fermentation and to stabilize the wine. During maturing in barrels or bottle- ageing, an unwanted refermentation can sometimes occur. However, results of the usual plate count of wine samples at the beginning of maturing suggest wine microbial stability. The aim of this study was to investigate the mode of yeasts survival after the addition of SO2 and to identify surviving yeasts.Methods and Results: Using direct epifluorescence technique, we observed the behaviour of cells after SO2 addition and compared the cell number determined by this method with the result of plate counts. The persistent yeast species were identified using two methods: polymerase chain reaction ( PCR)- restriction fragment length polymorphism and PCR- denaturing gradient gel electrophoresis. They were identified as Saccharomyces cerevisiae and Candida stellata, and after few months of maturing, other spoiling yeasts appeared, like Rhodotorula mucilaginosa or Zygosaccharomyces bailii.Conclusions: All characteristics of the cells lead to the conclusion that yeast persisted in wine in a viable but nonculturable- like state ( VBNC). Suppression of the effect of free- SO2 did not lead to the resuscitation of the cells; however, another method proved the capacity of the cells to exit from the VBNC- like state.Significance and Impact of the Study: This study permits the characterization of the presence of VBNC- like yeasts in wine. The 'refermentation' phenomenon is probably due to the exit of the VBNC state.