茉莉花JsNEL/LINS基因的克隆与表达分析
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福建农林大学园艺学院/茶学福建省高校重点实验室

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Cloning and Expression Analysis of JsNEL/LINS from Jasminum sambac
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    摘要:

    本研究基于茉莉花花瓣转录组测序结果,以茉莉花花瓣为材料,分别采用RT-PCR和染色体步移技术,克隆获得NES/LINS基因的全长cDNA序列及其启动子序列,应用SPEC-GC-MS技术和RT-qPCR技术检测茉莉花开放过程中芳樟醇化合物的相对含量及JsNES/LINS基因的表达水平,并分析JsNES/LINS在植物激素处理后的表达水平。结果表明,获得的JsNES/LINS基因(登录号为:MH513857.1)cDNA全长为1843bp,开放阅读框(ORF)全长为1755bp,编码584个氨基酸的蛋白质,为亲水性稳定蛋白。NCBI-BLAST分析结果表明,JsNES/LINS与油橄榄OeNES、金鱼草AmNES/LINS1和AmNES/LINS 2以及山茶花CsLINS的相似性分别为56%、54%、54%和50%;获得了JsNES/LINS起始密码子上游1141bp的启动子序列,经PlantCare预测结果显示,该序列包含多种与植物激素和光照等响应相关的顺式作用元件;GC-MS分析结果表明,芳樟醇化合物的相对含量在茉莉花未开放18:00时最低,半开放22:00时达到最大,随后呈下降趋势;RT-qPCR结果分析显示,JsNES/LINS的相对表达量变化与芳樟醇化合物的相对含量变化趋势一致;经IAA、GA、ABA及MeJA处理后JsNES/LINS的表达量受到不同程度的诱导表达,其中MeJA的诱导作用最为明显,诱导作用由大至小分别为:MeJA>GA>ABA>IAA;推测JsNES/LINS在芳樟醇化合物的生物合成过程中具有重要条件作用,且受植物激素调控表达。

    Abstract:

    In this study,based on the transcriptome results of jasmine petals, the full length cDNA and promoter sequence of NEL/LINS were cloned using RT-PCR and Genome Walking Technique, respectively. The content of linalool was detected by GC-MS, and the expression level of JsNEL/LINS during the opening process and after treatment of IAA, GA, ABA and MeJA. The results showed that the full length cDNA of JsNEL/LINS (GenBank accession number:MH513857) was 1843bp,contained 1755 bp open reading frame (ORF),encoding a protein of 584 amino acids, and belonged to hydrophilic and stable protein. The results of NCBI-BLAST showed that JsNES/LINS had 56%,54%,54% and 50% similarities with Olea europaea OeNES,Antirrhinum majus AmNES/LINS1,AmNES/LINS 2 and Camellia saluenensis CsLINS,respectively. The result of Subcellular localization showed that JsNEL/LINS was located on the cytoplasm. 1141bp promoter sequence of the JsNES/LINS was isolated, and the result predicted that it contained several cis-acting elements involved in plant hormones and light response,indicating that the expression of JsNAL/LINS was regulated by plant hormones and light. The results of GC-MS analysis showed that the relative content of linalool was the lowest when jasmine was not opened at 18:00, and reached the maximum when it was half open at 22:00, and then decreased. The analysis of RT-qPCR showed that the relative expression level of JsNES/LINS was consistent with the change of the relative content of linalool compounds during the flowering process. The expression of JsNES/LINS was induced by IAA, GA, ABA and MeJA, and the induction of MeJA was the most obvious, and the induction from high to low were MeJA>GA>ABA>IAA. It is speculated that JsNES/LINS plays an important role in the biosynthesis of linalool compounds and is regulated by plant hormones.

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  • 收稿日期:2019-04-24
  • 最后修改日期:2019-07-06
  • 录用日期:2019-07-08
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