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碱性螺旋-环-螺旋(basic helix-loop-helix,bHLH)转录因子家族在植物的生长发育、响应非生物胁迫等方面发挥着重要的调控作用。为了进一步丰富丹参(Salvia miltiorrhiza)中该家族成员的研究,寻找新的参与调控丹参次生代谢产物合成相关的bHLH成员,本研究基于丹参转录组和基因组数据库,分别从cDNA和gDNA水平克隆获得SmbHLH37基因序列。该基因序列长1 506 bp,包含1个27 bp的内含子序列,1 479 bp完整的CDS(GenBank登录号:KP257470.1),编码492个氨基酸。Blast P分析结果显示,SmbHLH37与芝麻(Sesamum indicum)等植物的bHLH3同源性较高;SmbHLH37蛋白与拟南芥(Arabidopsis thaliana)bHLH转录因子家族的分子进化树结果显示,SmbHLH37与拟南芥bHLH转录因子At JAM3(A.thaliana jasmonate-associated MYC2-like3)亲缘关系最近。亚细胞定位结果显示,SmbHLH37蛋白主要定位在细胞核中。qRT-PCR分析结果显示,SmbHLH37在叶中的表达量最高,花中最低;用脱落酸(abscisic acid,ABA)和机械诱导处理后,该基因的表达短时间呈上调趋势;同时,SmbHLH37在Sm MYC2过表达株系OE-8和OE-12中的表达量显著上调(P<0.05)。研究结果为进一步开展SmbHLH37功能研究提供了基础资料。
The basic helix-loop-helix (bHLH) transcription factor family plays an important regulatory role in plant growth and development, in response to abiotic stresses. In order to further enrich the family members of Salvia miltiorrhiza and find new bHLH members involved in the regulation of the secondary metabolites of Salvia miltiorrhiza, SmbHLH37 was cloned from cDNA and gDNA based on the Salvia miltiorrhiza transcriptome and genomic database respectively. gene sequence. The gene was 1 506 bp in length and contained a 27 bp intron sequence and a complete 1 479 bp CDS (GenBank accession number KP257470.1) encoding 492 amino acids. Blast P analysis showed that the homology of bHLH3 between SmbHLH37 and Sesamum indicum was high. The molecular phylogenetic tree of SmbHLH37 protein and bHLH transcription factor family of Arabidopsis thaliana showed that SmbHLH37 and Arabidopsis thaliana bHLH The transcription factor At JAM3 (A.thaliana jasmonate-associated MYC2-like3) has the closest genetic relationship. Subcellular localization results showed that SmbHLH37 protein mainly located in the nucleus. The results of qRT-PCR analysis showed that SmbHLH37 had the highest expression in leaves and the lowest in flowers. After treated with abscisic acid (ABA) and mechanical induction, the expression of SmbHLH37 was up-regulated in a short time. Meanwhile, The expression of MYC2 over-expression lines OE-8 and OE-12 were significantly up-regulated (P <0.05). The results provide the basic information for further study on the function of SmbHLH37.