溶藻弧菌细胞密度相关sRNA的鉴定及其对毒力的调控作用  

Identification of cell-density-related sRNA in Vibrio alginolyticus and its regulation on virulence

作　　者：李莹[1] 李印可 周欣 李莹玉 杨文娟[1] 王浩[1] 何佩云 刘欢[1,2] LI Ying;LI Yin-ke;ZHOU Xin;LI Ying-yu;YANG Wen-juan;WANG Hao;HE Pei-yun;LIU Huan(School of Food Science and Engineering,Shaanxi University of Science&Technology,Xi′an 710021,China;Shaanxi Research Insititute of Agricultural Products Processing Technology,Xi′an 710021,China)

机构地区：[1]陕西科技大学食品科学与工程学院,陕西西安710021 [2]陕西农产品加工技术研究院,陕西西安710021

出　　处：《陕西科技大学学报》2023年第1期52-58,共7页Journal of Shaanxi University of Science & Technology

基　　金：国家自然科学基金项目(32070129);陕西省重点研发计划项目(2020NY-121);陕西省西安市未央区科技计划项目(201937);陕西科技大学博士科研启动基金项目(BJ12-24);陕西科技大学大学生创新创业训练计划项目(S202110708124)。

摘　　要：溶藻弧菌作为四大致病弧菌之一,可引起鱼虾的系统性疾病以及人类肠道和创伤性伤口感染,其毒力因子的表达及致病性与细胞密度紧密相关.sRNA是存在于细菌中的非编码小RNA,作为转录后调控因子广泛参与细菌的毒力、环境应激、生长代谢等过程.利用转录组测序等手段,在溶藻弧菌中筛选出了显著差异表达的细胞密度相关非编码RNA分子,sRNA0087,并对其二级结构以及靶标mRNA进行预测.通过构建其突变株及回补株,探究了sRNA0087对溶藻弧菌毒力因子的调控作用.实验结果表明,溶藻弧菌在高细胞密度(9 h)相较于低细胞密度共有82个差异表达sRNA,其中47个表达上调,35个表达下调.sRNA0087下调水平最为显著,具有典型RNA二级结构,包含4个茎环结构以及3’端poly(U)结构,其在溶藻弧菌基因组中存在多个潜在靶标mRNA分子.成功构建Δ0087缺失株和0087^(+)回补株,sRNA0087对溶藻弧菌的运动性、生物被膜形成以及胞外蛋白酶的分泌具有显著的促进作用.Vibrio alginolyticus is one of the″big four pathogenic Vibrio″,which can cause systemic diseases of fish and shrimp and human intestinal and traumatic wound infection.The expression of the virulence factors are closely related to the cell density.sRNA is a non-coding small RNA in bacteria and widely involved in the bacterial virulence,environmental stress,growth and metabolism as a post-transcriptional regulator.In this paper,a differentially-expressed cell density related small non-coding RNA,sRNA0087,was characterized in V.alginolyticus by the transcriptome sequencing and other methods.The secondary structure and target mRNA of sRNA0087 were predicted.Then the regulation of sRNA0087 on various virulence factors of V.alginolyticus was explored after constructing its mutant and complementary strains.The results showed that there were 82 differentially expressed sRNAs in high cell density(9 h)compared with low cell density,of which 47 were up-regulated and 35 were down-regulated.The down-regulation level of sRNA0087 was the most significant,with typical RNA secondary structure,including four stem-loop structures and 3′-terminal poly(U)structure.There were multiple potential target mRNA molecules in the genome of V.alginolyticus.TheΔ0087 deletion strain and 0087^(+)complement strain were successfully constructed.The sRNA0087 had a significant role in promoting the motility,biofilm formation and extracellular protease secretion of V.alginolyticus.

关 键 词：溶藻弧菌 SRNA 毒力调控 

分 类 号：TS201.3[轻工技术与工程—食品科学]