热带海洋学报

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低温胁迫下红榄李(Lumnitzera littorea (Jack) Voigt)DEAD-box RNA解旋酶基因的表达分析

郝露露1,2,柯明思2,朱奕秀2,许燕敏2,张颖2,3,郑春芳1*   

  1. 1. 温州大学生命与环境科学学院,浙江 温州 325035

    2. 岭南师范学院生命科学与技术学院, 广东 湛江 524048

    3. 海南省林业科学研究(海南省红树林研究院),海南 海口 571129

  • 收稿日期:2022-03-29 修回日期:2022-05-23 出版日期:2022-05-31 发布日期:2022-05-31
  • 通讯作者: 郑春芳
  • 基金资助:
    海南省林业科学研究院(海南省红树林研究院)基础性科研工作(KYYS-2021-04);海南省科研院所技术创新专项(KYYS-2021-13,KYYS-2021-22); 海南省科研院所技术创新专项基础性科研工作项目(jcxk202003); 国家自然科学基金(32071503)

Expression of DEAD-box RNA helicase enzyme genes in Lumnitzera littorea (Jack) Voigt under low temperature stress

HAO Lulu1,2, KE Mingsi2, ZHU Yixiu2, XU Yanmin2, ZHANG Ying2,3, ZHENG Chunfang1*   

  1. 1. College of Life and Environmental Science, Wenzhou University, Wenzhou, Zhejiang 325035, China

    2. School of Life Sciences and Technology, Lingnan Normal University, Zhanjiang, Gaungdong 524048, China

    3. Forestry Science Research of Hainan Province (Hainan Mangrove Research Institute), Haikou, Hainan 571129, China

  • Received:2022-03-29 Revised:2022-05-23 Online:2022-05-31 Published:2022-05-31
  • Contact: chun-fang ZHENG
  • Supported by:

     Basic scientific research work of Hainan Forestry Research Institute (Hainan Mangrove Research Institute) (KYYS-2021-04);Technological Innovation Special project of Hainan Scientific Research Institute(KYYS-2021-13,KYYS-2021-22);Special Basic Research Work Project for Technological Innovation in Hainan Research Institutes (jcxk202003);The National Natural Science Foundation of China(32071503)

摘要: 转录组分析发现DEAD-box RNA解旋酶家族参与红榄李(Lumnitzera littorea (Jack) Voigt) 对低温胁迫的响应。本文对4个低温显著差异表达基因:LlDEAD12,LlDEAD32,LlDEAD43和LlDEAD65的生物信息学特性,组织特异性,以及在不同温度处理下红榄李幼苗中的差异表达进行研究。结果表明:LlDEAD12、LlDEAD32、LlDEAD43和LlDEAD65均属于疏水性蛋白,二级结构均由α-螺旋、β-转角、延伸链和无规则卷曲组成,具有多个糖基化位点和磷酸化位点,且不含有跨膜结构域和信号肽。亚细胞定位分析表明,LlDEAD12和LlDEAD32分别定位于细胞质和细胞核,而LlDEAD43和LlDEAD65则定位于线粒体上。通过蛋白质氨基酸序列的比对分析发现LlDEAD12、LlDEAD32和LlDEAD43分别与马尾松(Pinus massoniana Lamb.)、巨桉 (Eucalyptus grandis Hill)和石榴 (Punica granatum L.)的DEAD-box RNA解旋酶有较近的亲缘关系。荧光定量PCR技术分析发现,LlDEAD12和LlDEAD32在叶中高表达,LlDEAD43和LlDEAD65在茎和花中高表达,但这4个基因在根中都不表达,说明这4个基因对红榄李生长发育的调控主要集中在叶、茎和花器官。低温胁迫对红榄李幼苗中这4个基因的表达均有显著的抑制,说明他们参与了红榄李在低温环境下的分子响应,其中LlDEAD12和LlDEAD32可能参与了叶绿体的发育,LlDEAD43和LlDEAD65可能参与到了维持线粒体功能的稳定。以上结果为红榄李抗冷性苗木的培育提供科学依据。

关键词: 红树林, 红榄李, DEAD-box解旋酶, 低温胁迫, 生物信息学分析

Abstract: Transcriptome differential analysis revealed that the DEAD-box RNA helicase family plays an important role in the response of Lumnitzera littorea (Jack) Voigt to low temperature stress. In this study, we analyzed the bioinformatic properties, tissue-specific expression and differential expression of four genes, LlDEAD12, LlDEAD32, LlDEAD43 and LlDEAD65, in L. littorea seedlings under low temperature treatments. The results showed that LlDEAD12, LlDEAD32, LlDEAD43, and LlDEAD65 are hydrophobic proteins with secondary structures consisting of α-helix, β-turn, extended chain and irregular coiling, with many glycosylation sites and phosphorylation sites, and do not contain transmembrane structural domains or signal peptides. Subcellular localization analysis showed that LlDEAD12 and LlDEAD32 localized to the cytoplasm and nucleus, respectively, while LlDEAD43 and LlDEAD65 localized to the mitochondria. Comparative analysis of protein amino acid sequences indicated that LlDEAD12, LlDEAD32 and LlDEAD43 had similar affinities to DEAD-box RNA helicase enzymes of Pinus massoniana Lamb., Eucalyptus grandis Hill and Punica granatum L. , respectively. Analysis of the tissue expression specificity of the four genes by fluorescence quantitative PCR revealed that LlDEAD12 and LlDEAD32 highly expressed in leaves, while LlDEAD43 and LlDEAD65 highly expressed in stems and flowers, and none of the four genes expressed in roots, indicating that the regulation of growth and development of L. littorea by these four genes was mainly concentrated in leaf, stem and flower organs. The expression of these four genes was significantly suppressed in L. littorea seedlings under low temperature stress, indicating that these four genes are involved in the molecular response of L. littorea under cold temperature environment, among which LlDEAD12 and LlDEAD32 may be involved in chloroplast development, and LlDEAD43 and LlDEAD65 may be involved in maintaining the stability of mitochondrial function. Our results provide a scientific foundation for the breeding of cold-resistant seedlings of L. littorea.

Key words: mangrove, Lumnitzera littorea (Jack) Voigt, DEAD-box RNA helicase enzyme, cold stress, bioinformatics analysis