海马齿NHX基因家族的鉴定及表达分析

    Identification and Expression Analysis of NHX Gene Family of Sesuvium portulacastrum

    • 摘要:
      目的 海马齿(Sesuvium portulacastrum L.)是典型的海岸植物和红树林伴生植物,能够固沙护岸,在海水中可以正常生长,具有极强的耐盐性。Na+/H+逆向转运蛋白(Na+/H+ exchange,NHX)在植物耐盐和生长发育中起关键作用,为了解NHX在海马齿耐盐过程中的作用,对海马齿NHX基因家族进行鉴定和分析。
      方法 采用生物信息学方法从海马齿全长转录组数据中鉴定NHXs成员,对其蛋白理化性质、保守基序和进化关系进行分析,并利用荧光定量PCR技术研究盐胁迫下NHXs成员的表达模式。
      结果 从海马齿中共鉴定出12个SpNHX基因,命名为SpNHX1~SpNHX12SpNHXs基因编码的氨基酸长度为276~554 aa,分子量为31.22~61.21 kD,等电点为5.55~8.64,不稳定指数为32.14~50.54,均为疏水性蛋白。系统发育分析表明,SpNHX蛋白可分为2个亚组,同一亚组具有相似的保守基序。多序列比对结果发现,SpNHX均具有Na+/H+ exchange保守结构域。转录组数据和荧光定量PCR结果显示,SpNHXs基因在盐胁迫下均受到不同程度的诱导,其中SpNHX1SpNHX9SpNHX10SpNHX11SpNHX12在盐胁迫下显著上调表达。
      结论 本研究明确了海马齿NHX基因家族在高盐胁迫下的表达模式,表明SpNHX1SpNHX9SpNHX10SpNHX11SpNHX12可能参与海马齿盐胁迫响应,为进一步研究NHX在海马齿耐盐过程中的作用提供理论基础。

       

      Abstract:
      Objective Sesuvium portulacastrum, a typical coastal and mangrove associate plant, is capable of retaining sand and protecting banks. It can grow normally in seawater and has a strong salt tolerance. Na+/H+ exchange (NHX) plays a key role in salt tolerance and plant growth with development. In order to understand the role of NHX in salt tolerance of S. portulacastrum, the NHX genes were identified and analyzed.
      Method In this study, the S. portulacastrum NHX genes were identified by using informatic method through its full length transcriptome data. Their physicochemical properties, conserved motifs and evolutionary relationships were analyzed, and the expression patterns of NHX genes under s alt stress were studied by quantative real-time PCR (qRT-PCR).
      Result The results showed that 12 SpNHX genes were identified from S. portulacastrum and named as SpNHX1 to SpNHX12. The number of amino acids ranged from 276 to 554, with the molecular weight ranged from 31.22 to 61.21 kD, isoelectric point from 5.55 to 8.64, and instability ind ex from 32.14 to 50.54. All the proteins were hydrophobic. Phylogenetic analysis showed that SpNHX proteins can be divided into two subgroups. Similar conserved motifs composition was found in each subgroup. Multiple sequence alignment results showed that all SpNHX proteins had Na+/H+ exchange conserved domains. RNA-seq data and qRT-PCR results showed that SpNHX genes were induced to different degrees under salt stress, and SpNHX1, SpNHX9, SpNHX10, SpNHX11 and SpNHX12 were significantly up-regulated under salt stress.
      Conclusion This study clarified the expression patterns of NHXs in S. portulacastrum under high salt stress, indicating that SpNHX1, SpNHX9, SpNHX10, SpNHX11 and SpNHX12 may be involved in the response of S. portulacastrum to salt stress, which provided a theoretical basis for further research on the role of NHX in the process of salt tolerance of S. portulacastrum.

       

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