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    刘彬, 曹尚杰, 王营, 崔颖, 岳桦, 张彦妮. 过表达细叶百合LpNAC6基因增强烟草的耐盐性[J]. 北京林业大学学报, 2020, 42(4): 69-79. DOI: 10.12171/j.1000-1522.20190342
    引用本文: 刘彬, 曹尚杰, 王营, 崔颖, 岳桦, 张彦妮. 过表达细叶百合LpNAC6基因增强烟草的耐盐性[J]. 北京林业大学学报, 2020, 42(4): 69-79. DOI: 10.12171/j.1000-1522.20190342
    Liu Bin, Cao Shangjie, Wang Ying, Cui Ying, Yue Hua, Zhang Yanni. Overexpression of LpNAC6 gene in Lilium pumilum enhancing salt tolerance in transgenic tobacco[J]. Journal of Beijing Forestry University, 2020, 42(4): 69-79. DOI: 10.12171/j.1000-1522.20190342
    Citation: Liu Bin, Cao Shangjie, Wang Ying, Cui Ying, Yue Hua, Zhang Yanni. Overexpression of LpNAC6 gene in Lilium pumilum enhancing salt tolerance in transgenic tobacco[J]. Journal of Beijing Forestry University, 2020, 42(4): 69-79. DOI: 10.12171/j.1000-1522.20190342

    过表达细叶百合LpNAC6基因增强烟草的耐盐性

    Overexpression of LpNAC6 gene in Lilium pumilum enhancing salt tolerance in transgenic tobacco

    • 摘要:
      目的NAC转录因子是一类具有多种生物功能的新型转录因子,在植物抗逆响应中发挥着重要作用。本文旨在克隆并研究细叶百合LpNAC6基因在逆境胁迫下的表达模式,探究其在烟草中响应盐胁迫的功能。
      方法本研究采用同源克隆技术克隆得到细叶百合LpNAC6基因,利用生物信息学软件对LpNAC6基因进行分析;通过基因枪法对LpNAC6蛋白进行亚细胞定位;利用实时荧光定量PCR的方法分析LpNAC6基因在不同非生物胁迫和不同组织中的表达模式;构建植物表达载体pBI121-LpNAC6-GFP转化烟草,通过对转基因烟草进行盐胁迫处理验证LpNAC6基因的功能。
      结果LpNAC6基因长909 bp,编码302个氨基酸,存在一个高度保守的NAM结构域,属于NAC基因家族。LpNAC6为不稳定亲水性蛋白,无信号肽和跨膜结构域,有5个糖基化位点和20个磷酸化位点,亚细胞定位在细胞核。LpNAC6基因与黄褐棉的NAC转录因子进化关系最近。细叶百合中LpNAC6基因对ABA、干旱、低温及盐胁迫均有响应。盐胁迫下,过表达LpNAC6基因的转基因烟草其SOD、POD、CAT的活性和叶绿素、脯氨酸、可溶性蛋白的含量均显著高于野生型。
      结论细叶百合LpNAC6基因能够响应ABA、干旱、低温、盐胁迫等非生物胁迫,其过表达能够提高转基因烟草在盐胁迫下的代谢活力和抗氧化酶活性,从而增强烟草耐盐性。

       

      Abstract:
      ObjectiveNAC transcription factor is a new kind of transcription factor with many biological functions, which plays an important role in stress response. This paper aims to explore the expression pattern of LpNAC6 gene under stress and its response to salt stress in transgenic tobacco.
      MethodLpNAC6 gene from Lilium pumilum was cloned by homologous cloning and analyzed by bioinformatics softwares. Subcellular localization of LpNAC6 protein was performed by particle bombardment. The expression patterns of LpNAC6 gene in different abiotic stresses and different tissues were analyzed by RT-qPCR. The plant expression vector pBI121-LpNAC6-GFP was constructed, and tobacco was transformed for verifying the function of LpNAC6 gene under salt stress.
      ResultThe LpNAC6 gene was 909 bp in length and encoded 302 amino acids, it had a highly conserved NAM domain and belongs to the NAC gene family. LpNAC6 protein was a hydrophilic protein with no signal peptide and trans-membrane domain. It had 5 glycosylation sites, 20 phosphorylation sites and subcellular localization of LpNAC6 protein in nucleus. The evolutionary relationship between LpNAC6 gene and NAC transcription factor of Gossypium mustelinum was closest. LpNAC6 gene in L. pumilum responded to ABA, drought, low temperature and salt stress. Under salt stress, the activity of SOD, POD, CAT and the contents of chlorophyll, proline and soluble protein in transgenic tobacco overexpressing LpNAC6 gene were significantly higher than those in wild-type.
      ConclusionLpNAC6 gene of L. pumilum can respond to ABA, drought, low temperature and salt stress. Its overexpression can increase the metabolic activity and antioxidant enzyme activity of transgenic tobacco under salt stress, thus enhancing the salt tolerance of tobacco.

       

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