Responses of important virulence factors of Aeromonas veronii to environmental conditions
-
摘要: 鞭毛蛋白 (Flagellin)、密度感应系统 (Quorum sensing, QS) 和Ⅲ型分泌系统 (Type Ⅲ secretion system, T3SS) 是与维氏气单胞菌 (Aeromonas veronii) 致病性相关的重要因子，且受多种环境条件的共同调控。为研究外界环境因子变化对维氏气单胞菌致病因子的影响，以2株维氏气单胞菌的ascF、fliE和luxR致病因子为研究对象，通过实时荧光定量PCR技术从转录水平上探究了ascF、fliE和luxR基因对温度、pH、无机盐离子和培养转速等环境因子的响应。结果表明，维氏气单胞菌的3个基因对偏酸性环境 (pH 6.5~7.0)、中低培养转速 (150~210 r·min−1) 和锌离子 (Zn2+)、镁离子 (Mg2+) 为正响应；2株不同分子分型的维氏气单胞菌的3个基因的响应模式各有不同，说明了2株不同分子分型的维氏气单胞菌的致病机制可能有所差异；另外，维氏气单胞菌的重要致病因子受环境条件的调控，而呈现不同的变化规律，为进一步分析维氏气单胞菌的发病机制提供参考。Abstract: As important pathogenic factors of Aeromonas veronii, flagellin, quorum sensing and Type III secretion system (T3SS) are closely related to their pathogenicity and regulated by various environmental conditions. In order to study the effect of variation in external environmental factors on the pathogenic factors of A. veronii, we used the ascF, fliE and luxR factors of A. veronii as objects by real-time PCR method. The response of ascF, fliE and luxR genes to environmental factors such as temperature, pH, rotating speed and ions were explored at the transcription level. The results show that the three genes of A. veronii had a positive response to acidic environment (pH 6.5−7.0), low and medium speed (150−210 r·min−1), Zn2+ and Mg2+. The response patterns of the three genes of the two strains of A. veronii with different molecular types were different, indicating that the pathogenic mechanisms of A. veronii might be different. Besides, the important virulence factors of A. veronii were regulated by environmental conditions and showed different regular change. The results provide references for further research on the pathogenesis of A. veronii.
-
Key words:
- Aeromonas veronii /
- Virulence factors /
- Real-time PCR /
- Temperature /
- pH /
- Rotating speed
-
图 1 维氏气单胞菌ascF、fliE、luxR毒力基因对温度的响应
注：*. 与对照组相比，具有显著差异 (P<0.05)；**. 与对照组相比，具有极显著差异 (P<0.01)。后图同此。
Figure 1. Responses of virulence factors (ascF, fliE, luxR) of A. veronii to temperature
Note: *. Significant difference compared with the control group (P<0.05); **. Very significant difference compared with the control group (P<0.01). The same case in the following figures.
图 2 维氏气单胞菌ascF、fliE、luxR毒力基因对pH的响应
Figure 2. Response of virulence factors (ascF, fliE, luxR) of A. veronii to pH
图 3 维氏气单胞菌ascF、fliE、luxR毒力基因对无机盐离子的响应
Figure 3. Response of virulence factors (ascF, fliE, luxR) of A. veronii to ions
图 4 维氏气单胞菌ascF、fliE、luxR毒力基因对转速的响应
Figure 4. Response of virulence factors (ascF, fliE, luxR) of A. veronii to rotating speed
表 1 引物序列
Table 1. Primer sequence
基因
Gene引物
Primer引物序列 (5'—3')
Primer sequence (5'–3')扩增长度
Amplification length/bp退火温度
Annealing temperature/℃16S rDNA 16S-F CCTACGGGAGGCAGCAG 101 16S-R ATTACCGCGGCTGCTGG ascF ascF-F GCAGCACAAGATCAACAAATGG 60 62 ascF-R GCCCGAGTCACGGTGGAGT fliE fliE-F GCAACGCCGCTGGCACAA 121 65 fliE-R CGAAACGGGTACGCAGGTCA luxR luxR-F AGCAATCTGGGGAAGTTGGT 109 58.5 luxR-R GCAAAACCGGCTCAATGAAC 
下载: 导出CSV
-
[1] RAMSAMY Y, MLISANA K P, AMOAKO D G, et al. Comparative pathogenomics of Aeromonas veronii from pigs in South Africa: dominance of the novel ST657 clone[J]. Microorganisms, 2020, 8(12): 2008-2024. doi: 10.3390/microorganisms8122008 [2] ZHAO X, WU G, CHEN H, et al. Analysis of virulence and immunogenic factors in Aeromonas hydrophila: towards the development of live vaccines[J]. J Fish Dis, 2020, 43(7): 747-755. doi: 10.1111/jfd.13174 [3] DHANAPALA P M, KALUPAHANA R, KALUPAHANA A W, et al. Characterization and antimicrobial resistance of environmental and clinical Aeromonas species isolated from fresh water ornamental fish and associated farming environment in Sri Lanka[J]. Microorganisms, 2021, 9(10): 2106. doi: 10.3390/microorganisms9102106 [4] DWORACZEK K, DRZEWIECKA D, PEKALA S A, et al. Structural and serological studies of the O6-related antigen of Aeromonas veronii bv. sobria strain K557 isolated from Cyprinus carpio on a polish fish farm, which contains l-perosamine (4-amino-4, 6-dideoxy-L-mannose), a unique sugar characteristic for Aeromonas serogroup O6[J]. Mar Drugs, 2019, 17(7): 399. doi: 10.3390/md17070399 [5] BHOWMICK U D, BHATTACHARJEE S. Bacteriological, clinical and virulence aspects of Aeromonas-associated diseases in humans[J]. Pol J Microbiol, 2018, 67(2): 137-149. doi: 10.21307/pjm-2018-020 [6] FU M Q, KUANG R, WANG W C, et al. Hepcidin protects yellow catfish (Pelteobagrus fulvidraco) against Aeromonas veronii-induced ascites disease by regulating iron metabolism[J]. Antibiotics, 2021, 10(7): 848. doi: 10.3390/antibiotics10070848 [7] 王宝屯. 花鲈维氏气单胞菌致病性分析及灭活疫苗和中草药效果评价[D]. 上海: 上海海洋大学, 2021: 1. [8] 胡安东, 杨霞, 张飘, 等. 水生动物维氏气单胞菌病概述[J]. 贵州畜牧兽医, 2019, 43(2): 39-42. doi: 10.3969/j.issn.1007-1474.2019.02.012 [9] YANG B, CHEN C, SUN Y, et al. Comparative genomic analysis of different virulence strains reveals reasons for the increased virulence of Aeromonas veronii[J]. J Fish Dis, 2020, 44(1): 13262. [10] LIU Y G, HAO C X, SHI S Y, et al. Transcriptome analysis of the immunomodulation by Arctium lappa L. polysaccharides in the Chinese mitten crab Eriocheir sinensis against Aeromonas hydrophila[J]. Aquaculture, 2021, 534(2): 736255. [11] 赵泽林, 单晓枫, 田磊, 等. 维氏气单胞菌TH0426株ompW基因的克隆、生物信息学分析及其原核表达[J]. 中国兽医学报, 2019, 39(8): 1545-1550. [12] LI T, RAZA S H A, YANG B T, et al. Aeromonas veronii infection in commercial freshwater fish: a potential threat to public health[J]. Animals, 2020, 10(4): 608. doi: 10.3390/ani10040608 [13] 杨超, 董浚键, 刘志刚, 等. 大口黑鲈源维氏气单胞菌的分离鉴定[J]. 南方水产科学, 2021, 17(3): 54-61. [14] 宋明芳, 张冬星, 张海鹏, 等. 维氏气单胞菌毒力因子的研究进展[J]. 中国兽医科学, 2018, 48(8): 1038-1042. [15] ANA F B, MARIA J F. An update on the genus Aeromonas: taxonomy, epidemiology, and pathogenicity[J]. Microorganisms, 2020, 8(1): 129. doi: 10.3390/microorganisms8010129 [16] PREDIGER K D C, DALLAGASSA C B, MORIEL B, et al. Virulence characteristics and antimicrobial resistance of Aeromonas veronii biovar sobria 312M, a clinical isolate[J]. Braz J Microbiol, 2020, 51(2): 511-518. doi: 10.1007/s42770-019-00180-5 [17] YUWONO C, WEHRHAHN M C, LIU F, et al. The isolation of Aeromonas species and other common enteric bacterial pathogens from patients with gastroenteritis in an Australian population[J]. Microorganisms, 2021, 9(7): 1440. doi: 10.3390/microorganisms9071440 [18] 高彩霞, 任燕, 王庆, 等. 草鱼源致病性维氏气单胞菌的分离鉴定及药物敏感性分析[J]. 安徽农业大学学报, 2018, 45(3): 409-415. [19] 覃初斌. 干酪乳杆菌对斑马鱼抵御气单胞菌感染的分子机制研究[D]. 杭州: 浙江大学, 2017: 9. [20] SHA J, WANG S, SUAREZ G, et al. Further characterization of a type III secretion system (T3SS) and of a new effector protein from a clinical isolate of Aeromonas hydrophila-Part I[J]. Microb Pathog, 2007, 43(4): 127-146. doi: 10.1016/j.micpath.2007.05.002 [21] 丁晓燕. 环境因子对副溶血弧菌T3SS基因表达的影响及养殖环境细菌多样性分析[D]. 烟台: 烟台大学, 2016: 39-50 . [22] SONG H, ZHANG S, YANG B, et al. Effects of four different adjuvants separately combined with Aeromonas veronii inactivated vaccine on haematoimmunological state, enzymatic activity, inflammatory response and disease resistance in crucian carp[J]. Fish Shellfish Immunol, 2022, 120: 658-673. doi: 10.1016/j.fsi.2021.09.003 [23] TIAN J X, KANG Y H, CHU G S, et al. Oral administration of Lactobacillus casei expressing flagellin A protein confers effective protection against Aeromonas veronii in common carp, Cyprinus carpio[J]. Int J Mol Sci, 2019, 21(1): 33. doi: 10.3390/ijms21010033 [24] TAN J, ZHANG X, WANG X, et al. Structural basis of assembly and torque transmission of the bacterial flagellar motor[J]. Cell, 2021, 184(10): 2665-2679. doi: 10.1016/j.cell.2021.03.057 [25] HU R M, YANG T C, YANG S H, et al. Deduction of upstream sequences of Xanthomonas campestris flagellar genes responding to transcription activation by FleQ[J]. Biochem Biophys Res Commun, 2005, 335(4): 1035-1043. doi: 10.1016/j.bbrc.2005.07.171 [26] REED K A, HOBERT M E, KOLENDA C E, et al. The Salmonella typhimurium flagellar basal body protein FliE is required for flagellin production and to induce a proinflammatory response in epithelial cells[J]. J Biol Chem, 2002, 277(15): 13346-13353. doi: 10.1074/jbc.M200149200 [27] 孙晓佳, 李婷婷, 赫彬彬, 等. 环境条件对温和气单胞菌群体感应基因luxI/luxR表达的影响[J]. 中国食品学报, 2021, 21(4): 71-78. [28] KAI P, BASSLER B L, et al. Quorum sensing signal-response systems in Gram-negative bacteria[J]. Nat Rev Microbiol, 2016, 14(9): 576-588. doi: 10.1038/nrmicro.2016.89 [29] 励建荣, 李婷婷, 王当丰. 微生物群体感应系统及其在现代食品工业中应用的研究进展[J]. 食品科学技术学报, 2020, 38(1): 1-11. [30] BAZHENOV S, NOVOYATLOVA U, SCHEGLOVA E, et al. Influence of the luxR regulatory gene dosage and expression level on the sensitivity of the whole-cell biosensor to acyl-homoserine lactone[J]. Biosensors, 2021, 11(6): 166. doi: 10.3390/bios11060166 [31] AWAN F, DONG Y, WANG N, et al. The fight for invincibility: environmental stress response mechanisms and Aeromonas hydrophila[J]. Microb Pathog, 2018, 116: 135-145. doi: 10.1016/j.micpath.2018.01.023 [32] ROY P K, HA J W, MIZAN M, et al. Effects of environmental conditions (temperature, pH, and glucose) on biofilm formation of Salmonella enterica serotype Kentucky and virulence gene expression[J]. Poultry Sci, 2021, 100(7): 101209. doi: 10.1016/j.psj.2021.101209 [33] 胡靖, 李爱华, 胡成钰, 等. 温度和pH值对嗜水气单胞菌毒力基因表达的影响[J]. 南京理工大学学报(自然科学版), 2006, 30(3): 375-380. [34] 庞欢瑛, 周泽军, 丁燏, 等. 溶藻弧菌 vscO基因在不同环境下的表达差异[J]. 吉首大学学报 (自然科学版), 2014, 35(4): 59-63. [35] WOLFGANG M C, LEE V T, GILMORE M E, et al. Coordinate regulation of bacterial virulence genes by a novel adenylate cyclase-dependent signaling pathway[J]. Dev Cell, 2003, 4(2): 253-263. doi: 10.1016/S1534-5807(03)00019-4 [36] 孔西曼, 张公亮, 王佳莹, 等. 环境因素对即食海参蜂房哈夫尼菌群体感应的影响[J]. 现代食品科技, 2017, 33(1): 87-92,105. [37] 赵丹丹, 刘琳, 王迪, 等. pH和NaCl质量浓度对发酵鱼糜中腐败菌Aeromonas veronii群体感应的影响[J]. 食品与发酵工业, 2018, 44(2): 33-39. -
下载:
点击查看大图
图(4) / 表(1)
计量
- 文章访问数: 611
- HTML全文浏览量: 181
- PDF下载量: 26
- 被引次数: 0
粤公网安备 44010502001741号
