日本沼虾桩蛋白基因的克隆与镉胁迫对其表达的影响  

Cloning of paxillin gene from Macrobrachium nipponense and effect of cadmium stress on its expression

作　　者：彭佳诚 吴越 徐洁皓 夏美文 齐天鹏 徐海圣[1,3] PENG Jiacheng;WU Yue;XU Jiehao;XIA Meiwen;QI Tianpeng;XU Haisheng(College of Animal Sciences,Zhejiang University,Hangzhou 310058,China;College of Biological and Environmental Sciences,Zhejiang Wanli University,Ningbo 315100,Zhejiang,China;Zhejiang University Huzhou South Taihu Lake Modern Agricultural Science and Technology Promotion Center,Huzhou 313000,Zhejiang,China)

机构地区：[1]浙江大学动物科学学院,浙江杭州310058 [2]浙江万里学院生物与环境科学学院,浙江宁波315100 [3]浙江大学湖州市南太湖现代农业科技推广中心,浙江湖州313000

出　　处：《浙江农业学报》2024年第2期247-253,共7页Acta Agriculturae Zhejiangensis

基　　金：国家科技重大专项(2017ZX07401-002);宁波市海洋生物资源高价值利用公共服务平台(NBHY-2017-P2);湖州市农业科学与技术研究院技术创新团队(2019HN02)。

摘　　要：镉(Cd)是蓄积性和毒性较强的生物非必需重金属元素之一,会对水生生物造成较大的危害。为更好地了解镉对日本沼虾的分子毒性作用,为水环境镉污染的生态风险预警和评估提供生物标志物,克隆了日本沼虾(Macrobrachium nipponenes)桩蛋白(paxillin)基因MnPXN,并检测了镉胁迫对其表达的影响。MnPXN基因全长为3655 bp,包含1491 bp的编码区,编码496个氨基酸;MnPXN蛋白具有6个低复杂区域和3个保守的LIM结构域,与凡纳滨对虾(Litopenaeus vannamei)聚成一支,与家蚕(Bombyx mori)、赤拟谷盗(Tribolium castaneum)等昆虫的亲缘关系较近,与线虫(Nematoda)亲缘关系最远。实时荧光定量PCR(qRT-PCR)结果显示,MnPXN在日本沼虾的肝胰腺、胃、鳃中表达水平较高,在心脏和肌肉中表达水平较低。在0.2427 mg·L^(-1)CdCl_(2)中暴露24 h,肝胰腺中MnPXN基因表达水平较对照组均显著升高,0.0200 mg·L^(-1)CdCl_(2)胁迫6 h后MnPXN基因表达水平显著升高,24 h表达量达到最大值,96 h恢复至起始水平。RNA干扰能显著(P<0.05)降低MnPXN在肝胰腺中的表达水平,显著(P<0.001)增加日本沼虾的死亡率,表明MnPXN基因能够降低镉胁迫对日本沼虾的毒性作用,其作用机理尚需进一步研究。Cadmium(Cd),with strong accumulation and toxicity,is one of the non-essential heavy metal elements,which could cause great harm to aquatic organisms.To better understand the molecular toxicity of cadmium to Macrobrachium nipponense,and provide biomarkers for ecological risk warning and assessment of cadmium pollution in water environment.In this paper,paxillin gene(MnPXN)of M.nipponense was cloned,and the effect of cadmium stress on its expression was examined.The total length of MnPXN gene is 3655 bp,including 1491 bp coding region,encoding 496 amino acids;6 low complexity regions and 3 conservative LIM domains are included in MnPXN,forming a branch with Litopenaeus vannamei.It is closely related to insects such as silkworm and Tribolium castaneum,and is farthest related to nematodes.The results of quantitative reverse transcriptase-mediated PCR(qRT-PCR)showed that the expression level of MnPXN was high in hepatopancreas,stomach and gill,but low in heart and muscle.The expression level of MnPXN gene in hepatopancreas that after 24 h of treatment with 0.2427 mg·L^(-1)CdCl_(2)was significantly higher than that in control group;After 6 h of 0.0200 mg·L^(-1)CdCl_(2)stress,the expression level of MnPXN gene increased significantly,reaching the maximum at 24 h,and gradually recovered to the initial level at 96 h.Besides,RNA interference could significantly reduce the expression level of MnPXN in hepatopancreas and significantly increase the mortality of M.nipponense,indicating that MnPXN could reduce the toxic effect of cadmium stress on M.nipponense,and its mechanism needs further study.

关 键 词：镉胁迫 日本沼虾 桩蛋白 基因表达 RNA干扰 

分 类 号：S917.4[农业科学—水产科学]