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    李智博, 董世满, 李淑霞, 等. 木薯SR45亚家族基因鉴定及表达[J]. 华南农业大学学报, 2022, 43(5): 20-28. doi: 10.7671/j.issn.1001-411X.202111032
    引用本文: 李智博, 董世满, 李淑霞, 等. 木薯SR45亚家族基因鉴定及表达[J]. 华南农业大学学报, 2022, 43(5): 20-28. doi: 10.7671/j.issn.1001-411X.202111032
    shu
    LI Zhibo, DONG Shiman, LI Shuxia, et al. Identification and expression of SR45 subfamily gene in Manihot esculenta[J]. Journal of South China Agricultural University, 2022, 43(5): 20-28. doi: 10.7671/j.issn.1001-411X.202111032
    Citation: LI Zhibo, DONG Shiman, LI Shuxia, et al. Identification and expression of SR45 subfamily gene in Manihot esculenta[J]. Journal of South China Agricultural University, 2022, 43(5): 20-28. doi: 10.7671/j.issn.1001-411X.202111032
    shu

    木薯SR45亚家族基因鉴定及表达

    Identification and expression of SR45 subfamily gene in Manihot esculenta

      摘要
    • 摘要:
      目的  精氨酸和丝氨酸富集蛋白(Arginine/serine-rich proteins，SR)为剪接复合体的主要成员，不仅参与植物前体mRNA可变剪接过程，在植物非生物胁迫中也具有相当重要的作用。SR45基因为SR基因亚家族成员之一。本研究旨在分析木薯SR45亚家族成员蛋白结构特征与表达模式，为进一步了解该亚家族基因在木薯中的功能提供理论支持。
      方法  利用生物信息学技术重新构建木薯SR基因家族进化树，对木薯SR45亚家族蛋白理化性质、基因结构、保守结构域进行分析，同时利用转录组数据分析低温与干旱胁迫下SR基因表达变化，运用实时荧光定量PCR(RT-qPCR)研究各基因成员在不同组织中的特异性表达以及对低温胁迫的响应。
      结果  木薯SR基因家族共7个大类26个成员，SR45亚家族共有5个成员；SR45亚家族基因编码蛋白长度为135~423 aa，相对分子质量为14970~47210，等电点为5.19~12.34；预测其主要定位于细胞核和叶绿体；SR45编码RS结构域和RRM结构域，具有Motif 3、Motif 6、Motif 2和Motif 1保守基序。转录组数据和RT-qPCR分析表明，SR45基因均响应木薯低温胁迫，且MeSR45-2显著上调表达，MeSR45-4、MeSR45-5在木薯根和叶中有较高表达量。
      结论  木薯SR45亚家族基因显著响应低温胁迫；将MeSR45-2基因列为调控低温逆境变化的候选基因，将根和叶列为研究SR45基因亚类成员的主要组织。本研究为探索木薯SR45基因奠定了理论基础，并为进一步研究木薯SR45基因逆境胁迫应答过程指明了方向。

       

      Abstract:
      Objective  As major members of the splicing complex, arginine/serine-rich proteins (SR) not only participate in the process of alternative splicing of plant precursor mRNA, but also play an important role in abiotic stress. SR45 gene is a member of SR gene subfamily, this study was aimed to analyze the structural characteristics and expression patterns of SR45 subfamily proteins in cassava, and provide a theoretical support for further understanding the functions of SR45 subfamily genes in cassava.
      Method  The evolutionary tree of cassava SR gene family was reconstructed by bioinformatics, and the physical and chemical properties, gene structure and conserved domain of cassava SR45 subfamily protein were analyzed. At the same time, transcriptome data were used to analyze the changes of SR gene expression under low temperature and drought stress, quantitative real-time PCR (RT-qPCR) was used to study the specific expression of SR gene members in different tissues and their responses to low temperature stress.
      Result  The cassava SR gene family consisted of 26 members in seven categories, and the SR45 subfamily consisted of five members. The length of proteins encoded by SR45 subfamily genes were 135–417 aa, with relative molecular mass being 14970–47210, PI being 5.19–12.34. It was predicted that they were mainly located in the nucleus and chloroplast. The RS domain and RRM domain encoded by SR45 had Motif 3, Motif 6, Motif 2 and Motif 1 conserved motifs. Transcriptional data and RT-qPCR analysis showed that all SR45 genes were responsive to low temperature stress in cassava, MeSR45-2 was significantly up-regulated, MeSR45-4and MeSR45-5 were highly expressed in roots and leaves of cassava.
      Conclusion  SR45 subfamily genes of cassava significantly respond to low temperature stress. MESR45-2 gene is selected as the candidate gene for regulating low temperature stress, root and leaf are the main tissues for SR45 subfamily gene research. This study lays a theoretical foundation for the exploration of cassava SR45 gene, and points out a direction for further research on cassava SR45 gene responsive to stress.

       

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