本文研究了肉桂醛对酿酒酵母persister细胞形成的影响。通过96孔板微量法测定肉桂醛对酿酒酵母生长的最小抑制浓度(MIC)为0.4 m M;采用流式细胞仪以及梯度稀释滴平板计数法研究了肉桂醛处理后酿酒酵母persister细胞的形成情况。结果表明,肉桂醛可以抑制酿酒酵母生长,且能够诱导酿酒酵母细胞形成persister状态,该状态下的细胞对两性霉素B产生耐药性。进一步研究发现肉桂醛处理后可以使酿酒酵母细胞停滞在细胞周期的任何阶段,而雷帕霉素诱导的细胞自噬只能停留在G1期,所以酿酒酵母perisister与自噬状态存在区别。目前,对于Persister的研究集中在原核微生物,对真核生物persister的研究非常有限。由于persister群体通常占总群体极小一部分,这就给基因水平上研究persister的形成机制带来很大的挑战。本研究发现肉桂醛处理酿酒酵母细胞后,可以促使其大部分细胞形成persister。这就为从基因水平上认识真核生物persister的形成机制提供了方法,实验结果表明YGL基因也与酿酒酵母persister的形成有很大关系。
The effects of cinnamaldehyde(CA) on persister formation in Saccharomyces cerevisiae were investigated in this study.CA was found to effectively inhibit the growth of S.cerevisiae in a 96-well plate with a minimum inhibitory concentration value of 0.4 m M.The formation of persister cells in S.cerevisiae cells after exposure to cinnamaldehyde was further explored by flow cytometry and serial dilution-drop plate counting methods.The results showed that CA inhibited the growth of S.cerevisiae cells,and S.cerevisiae cells were induced to form the persister state showing amphotericin B resistance.Further studies showed that CA treatment led to growth arrest of S.cerevisiae cells at any stage in the cell cycle,whereas cells undergoing autophagy induced by rapamycin stopped during the G1 phase.Therefore,persisters in S.cerevisiae differed from cells in autophagy.Previous studies of persister mainly focused on prokaryotic pathogens rather than eukaryotic cells.Additionally,the natural persister population accounted for an extremely small proportion of the overall population,creating challenges in studies of the genetic mechanisms underlying their formation.The findings of this study showed that CA treatment induced persister formation in most S.cerevisiae cells,providing a method for understanding the genetic mechanism underlying persister formation in eukaryotes.Results also confirmed that the YGL gene was significantly associated with persister formation in S.cerevisiae.