γ辐照对单核增生李斯特菌细胞形态变化及毒力基因表达的影响

Hamouda Elabed
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引用次数: 0

摘要

射线辐照是食品工业中最常用的射线处理方法之一。它被用于控制各种产品中微生物的繁殖,但细菌对低剂量的反应尚不清楚。本研究主要研究了γ辐照后单核增生李斯特菌的形态变化和毒力基因表达。原子力显微镜(AFM)显示,0.5 kGy剂量对单核增生乳杆菌的膜形态没有影响。0.7 kGy处理后,细胞失去了原有的形态和光滑的细胞膜,1 kGy剂量完全破坏。此外,用色谱法分析了不同γ剂量后的膜脂肪酸组成。辐照菌株的脂肪酸组成发生了显著变化,显示膜脂的新合成:C12:0;C14:0;C15:0;C16:0和C18:0。此外,我们报道了饱和脂肪酸的增加,这是在胁迫条件下膜适应所必需的。采用实时荧光定量PCR技术,在相同条件下检测3个毒力基因(hlyA、fri和prfA)的表达水平。分析显示,伽玛治疗后prfA和fri基因均上调。prfA是一种调控基因,其诱导可能会影响其他控制菌株适应形式的基因的表达。本研究为进一步研究单核增生李斯特菌在亚致死辐照胁迫下适应性反应的调控机制开辟了前景。
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Morphological Alterations and Virulence Gene Expression of Listeria Monocytogenes Cells in Response to Gamma Irradiation Treatments
Gamma irradiation is one of the most popular ray treatments in food industry. It's used to control micro-organisms proliferation in a wide range of products however the response of bacteria to low doses is still unknown. In this study we mainly focus on morphological alteration and virulence gene expression in Listeria monocytogenes after gamma irradiation treatments. The atomic force micrographs (AFM) showed that 0.5 kGy dose has no effect on the membrane morphology of L. monocytogenes. However, after 0.7 kGy treatment, the cells lost their typical shape and smooth membrane and 1 kGy dose was totally destructive. Moreover, membrane fatty acid composition was analyzed by the chromatographic method after different gamma doses. Significant modifications on fatty acids composition were detected in the irradiated strain showing a novo synthesis of membrane lipids: C12:0; C14:0; C15:0; C16:0 and C18:0. In addition, we reported an increase of the saturated fatty acid, essential for membrane adaptation under stress conditions. The expression levels of three virulence genes (hlyA, fri and prfA) were studied in the same conditions using real-time PCR technique. The analysis revealed that both prfA and fri genes were up-regulated after gamma treatment. The induction of prfA, which is a regulator gene, may affect the expression other genes controlling the adaptive form in the treated strain. This study open prospects for further researches to explain the regulatory mechanisms of the adaptive response in Listeria monocytogenes when exposed to sublethal irradiation-stress.
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