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采用RT-PCR方法从‘凤丹’牡丹(Paeonia ostii‘Fengdan’)种子中克隆得到1个与ABA信号途径相关的转录因子基因PobZIP1,并利用超高效液相色谱—串联质谱(UPLC–MS/MS)的方法测定了不同发育时期种子的ABA含量。PobZIP1 cDNA全长957 bp,编码318个氨基酸,其编码的蛋白C末端含有bZIP转录因子的典型结构域,GenBank登录号为KJ009392。PobZIP1蛋白与拟南芥bZIP转录因子聚类分析显示,PobZIP1转录因子属于拟南芥bZIP转录因子的A亚族,与AtbZIP39/ABI5聚在一起。与其他物种bZIP蛋白的系统进化分析表明,牡丹PobZIP1与蔷薇科的苹果bZIP亲缘关系最近。SqRT-PCR结果表明:PobZIP1在‘凤丹’牡丹根、茎、叶和花芽中均有表达,其中花芽中的表达量最低,根、茎、叶中的表达量相近。在种子发育过程中,PobZIP1的表达呈现出先增加后降低的趋势。ABA含量测定结果显示,随着种子的发育ABA含量迅速上升,达到最大值后缓慢下降并保持在较高水平。推测种子发育后期,较高含量的ABA和表达相对较高的PobZIP1可能是诱导种子休眠形成的原因。
A transcription factor gene PobZIP1 related to the ABA signaling pathway was cloned from the seeds of Paeonia ostii’Fengdan ’by RT-PCR and analyzed by UPLC-MS / MS. MS) method to determine the ABA content of seeds at different developmental stages. The full-length cDNA of PobZIP1 is 957 bp in length and encodes a protein of 318 amino acids. The Coding-site of PobZIP1 contains the typical domain of bZIP transcription factor. GenBank accession number is KJ009392. Cluster analysis of the PzZIP1 protein and Arabidopsis bZIP transcription factor revealed that the PobZIP1 transcription factor belongs to the A subfamily of Arabidopsis bZIP transcription factor and is clustered with AtbZIP39 / ABI5. Phylogenetic analysis of bZIP protein from other species showed that peony PobZIP1 has the closest genetic relationship with bZIP from Rosaceae. The results of SqRT-PCR showed that PobZIP1 was expressed in roots, stems, leaves and flower buds of ’Fengdan’. The expression level of PobZIP1 was the lowest in flower buds and similar in roots, stems and leaves. During seed development, the expression of PobZIP1 showed the first increase and then decrease. The results of ABA assay showed that the content of ABA increased rapidly with the development of seeds, then decreased slowly and remained at a high level. It is speculated that late stage of seed development, higher levels of ABA and higher expression of PobZIP1 may be responsible for inducing seed dormancy.