革胡子鲶感染维氏气单胞菌后β-防御素基因的时空表达分析  

Spatio-temporal Expression Profiles ofβ-defensin Gene in African Catfish Clarias garepanius Challenged with Aeromonas veronii

作　　者：冯苗苗 王晓梅 李转转 顾晨刊 陈成勋 FENG Miaomiao;WANG Xiaomei;LI Zhuanzhuan;GU Chenkan;CHEN Chengxun(Tianjin Key Laboratory of Aqua-Ecology and Aquaculture,College of Fisheries,Tianjin Agricultural University,Tianjin 300384,China)

机构地区：[1]天津农学院水产学院,天津市水产生态及养殖重点实验室,天津300384

出　　处：《水产学杂志》2021年第6期18-23,共6页Chinese Journal of Fisheries

基　　金：天津市应用基础与前沿技术研究计划重点项目(15JCZDJC33500).

摘　　要：给体质量(63.84±5.91)g的革胡子鲶(Clarias garepanius)腹腔注射0.2 mL浓度为4×10^(8) CFU/mL的维氏气单胞菌(Aeromonas veronii),对照组注射同等剂量的生理盐水。注射后3 h、6 h、12 h、24 h、36 h、48 h、72 h和96 h时,感染组和对照组各取鱼15尾,用200 mg/L的MS-222麻醉后,应用实时荧光定量PCR技术,分析感染维氏气单胞菌前后β-防御素基因在革胡子鲶脾脏、肾脏、头肾、鳃、肠道和皮肤中的表达。结果显示:β-防御素基因在健康革胡子鲶体内(对照组)上述6种组织中均有表达,在脾脏中表达量显著高于其余5种组织(P<0.05)。感染维氏气单胞菌后3~96 h,6种组织中β-防御素基因的时序表达谱不同。攻菌3 h头肾、皮肤和鳃中该基因表达量显著上调,高于对照组以及感染后的其他各时间点,分别为对照组的6.48倍、3.86倍和1.80倍(P<0.05);脾脏在24 h表达量达到峰值,为对照组的2.76倍(P<0.05);肾脏和肠道也在3 h表达量最大,但与对照组差异不显著(P>0.05)。组织间比较得出:在感染后3 h和6 h,脾脏和头肾间β-防御素基因的表达量差异不显著,但头肾和脾脏的表达量分别在3 h和6 h显著高于其余4种组织,之后各感染时间点均是脾脏的表达量最高,且在24 h差异最大(P<0.05)。上述结果说明:检测的6种组织都参与革胡子鲶对维氏气单胞菌感染的免疫应答反应,但在感染早期(3 h和6 h)头肾和脾脏防御作用较强,之后脾脏在病原菌侵染中起更为重要的防御作用。African catfish(Clarias garepanius)weighing(63.84±5.91)g was intraperitoneally injected with 0.2 mL of Aeromonas veronii at a concentration of 4×10^(8) CFU/mL,and African catfish in the control group was injected with the same dose of normal saline.Fifteen fish were sampled from the infected group and the control group at 3 h,6 h,12 h,24 h,36 h,48 h,72 h,and 96 h post injection(hpi),and anesthetized by MS-222 at a dose of 200 mg/L MS-222.Then the spleen,kidney,head kidney,gill,intestine and skin of African catfish were dissected and the real-time fluorescent quantitative PCR technology was used to investigate the expression profiles of theβ-defensin gene in the tissues above of African catfish before and after artificial infection of A.veronii.The results showed thatβ-defensin gene was constitutively expressed in the all examined tissues of healthy(control)individuals,significantly higher expression level in spleen than that in other five tissues.The expression levels in head kidney,skin and gill were shown to be peaked in African catfish at 3 hpi,as 6.41-fold,3.86-fold and 1.80-fold as those in the control(P<0.05),respectively,with the maximum expression in spleen reached at 24 hpi as 2.76-fold as that in the control(P<0.05).The peak expression levels in kidney and intestine were also observed at 3 hpi,without significant difference from the control.There were no significant differences in the expression ofβ-defensin gene between spleen and head kidney among examined tissues at 3 and 6 hpi,significantly higher at 3 hpi in head kidney and at 6 hpi in spleen compared to other four tissues;from 12~96 hpi,the expression in spleen was the maximal with top difference at 24 hpi(P<0.05).These results indicated that all of examined six tissues involved in the immune response to the pathogen A.veronii infection.However,both the head kidney and spleen had a strong defense function in the early stage of infection(3 hpi and 6 hpi),afterwards,the spleen played a more robust role in the defense against A.veronii.

关 键 词：革胡子鲶 维氏气单胞菌 Β-防御素 基因表达 实时荧光定量PCR 

分 类 号：S917[农业科学—水产科学] S965.128