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脊尾白虾甘露糖结合凝集素(MBL)基因在抗镉胁迫中的生物学功能分析
徐莞媛1, 马杭柯1, 孙金秋1, 段健诚1, 邓高威1, 高焕1,2,3,4, 阎斌伦1,2,3,4
1.江苏省海洋生物资源与生态环境重点实验室 江苏省海洋生物技术重点实验室 淮海工学院 连云港 222005;2.江苏省海洋生物产业技术协同创新中心 连云港 222005;3.江苏省农业种质资源保护与利用平台 南京 210014;4.江苏省海洋资源开发研究院(连云港) 连云港 222005
摘要:
为研究Cd2+胁迫下脊尾白虾(Exopalaemon carinicauda)甘露糖集合凝集素(MBL)在肝胰腺中表达量的变化特征,本研究利用重金属镉(Cd2+)对脊尾白虾进行96 h急性毒性实验。实验共设置5组Cd2+浓度胁迫(0、0.01、0.0175、0.021和0.0278 mmol/L),分别在Cd2+胁迫后0、3、6、12、24、48、72和96 h共8个时间点取样。实时荧光定量结果显示,高浓度Cd2+(0.0175、0.021和0.0278 mmol/L)胁迫下,脊尾白虾MBL基因的表达量先呈上升趋势,在72 h达到峰值后随后下降,但各时间点表达量均与对照组存在显著性差异(P<0.01);Cd2+浓度为0.01 mmol/L时,脊尾白虾MBL基因的表达量整体呈下降趋势。进一步采用RNA技术干扰该基因表达后,发现Cd2+胁迫后的脊尾白虾个体死亡率显著增加。本研究表明,脊尾白虾MBL在应答Cd2+胁迫过程中可对机体起到一定的保护作用。
关键词:  脊尾白虾  MBL    RNA干扰技术  急性毒性实验
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Biological Functional Analysis of MBL Gene in Resistance to Cadmium Stress in Exopalaemon carinicauda
XU Wanyuan1, MA Hangke1, SUN Jinqiu1, DUAN Jiancheng1, DENG Gaowei1, GAO Huan1,2,3,4, YAN Binlun1,2,3,4
1.Jiangsu Key Laboratory of Marine Bioresources and Environment, Jiangsu Key Laboratory of Marine Biotechnology, Huaihai Institute of Technology, Lianyungang 222005;2.Co-Innovation Center of Jiangsu Marine Bio-Industry Technology, Huaihai Institute of Technology, Lianyungang 222005;3.Jiangsu Provincial Infrastructure for Conservation and Utilization of Agricultural Germplasm, Nanjing 210014;4.Marine Resource Development Institute of Jiangsu (Lianyungang), Lianyungang 222005
Abstract:
In order to study the expression profile of mannose-binding lectin (MBL) in response to the stress of heavy metal cadmium (Cd2+) in the ridgetail white prawn, Exopalaemon carinicauda, changes of MBL expression level during 96 h of stress were analyzed in the tissue of hepatopancreas. Five Cd2+ stress concentrations (0, 0.01, 0.0175, 0.021 and 0.0278 mmol/L) were set up in the experiment, and the samples were taken at 8 time points, i.e. 0, 3, 6, 12, 24, 48, 72 and 96 hours after Cd2+ stress. The real-time fluorescence quantitative results showed that under the stress of high concentration of Cd2+ (0.0175, 0.021 and 0.0278 mmol/L), the expression level of MBL gene in the ridgetail white prawn increased first, then decreased after reaching the peak at 72 h. There are significant differences between experimental group and control group at all the stress time points. When the cadmium ion was 0.01 mmol/L, a lower concentration, the expression level of MBL gene in the ridgetail white prawn tends to decrease from the beginning. Further interfering down the expression level of MBL using the RNA technology, it was discovered that the mortality rate of Cd2+ stressed individuals were higher than those of the control group. This study showed that MBL potentially took part in the protection of E. carinicada from Cd2+ stress.
Key words:  Exopalaemon carinicauda  MBL  Cadmium  RNA interference  Acute toxicity test