牡丹PsSPL3基因的克隆和表达特性分析

发布时间：2018-02-28 01:57

  本文关键词： 牡丹 PsSPL RACE 实时定量PCR 表达模式　出处：《华北农学报》2017年04期 　论文类型：期刊论文

【摘要】：SPL(SQUAMOSA promoter-binding protein-like)转录因子在植物多个生理过程中发挥重要的生理功能,为了研究其在牡丹花芽休眠进程中的作用机理,采用RACE方法,从牡丹花芽中克隆得到了一个SPL基因。该基因全长cDNA 1 057 bp,完整开放阅读框为549 bp,编码182个氨基酸,Blast分析表明,编码的氨基酸序列中具有SBP-box基因家族所特有的SBP-box保守结构域。与拟南芥已知SPLs蛋白构建进化树结果表明,牡丹SPL蛋白与At SPL3聚为一支,该基因被命名为PsSPL3。生物软件预测PsSPL3蛋白分子量为20.349 4 kDa,理论等电点为9.62。同源性分析了牡丹PsSPL3与其他已知植物的SPL3,相似性为47.98%~69.46%,其中与白桦的SPL3蛋白相似性最高,为69.46%。实时定量PCR分析PsSPL3基因在初花期牡丹不同组织中和花芽休眠过程中的表达水平,结果表明,PsSPL3基因在不同组织中表达差异较大,其中在根中转录水平最高,在茎和花瓣中次之;PsSPL3基因在休眠进程中的转录水平呈先上升后下降趋势,其中低温处理14 d时,PsSPL3基因的表达量达到最高。低温7 d结合外源施加GA3的牡丹花芽中PsSPL3基因的表达量显著增加,推测PsSPL3基因的转录受GA3诱导来促进牡丹花芽内休眠解除的进程。
[Abstract]:SPL (SQUAMOSA promoter-binding protein-like) transcription factors play important physiological functions in multiple physiological processes in plants, in order to study the peony flower bud dormancy mechanism in the process of using the method of RACE, was cloned from the flower bud get a SPL gene. The gene was cDNA 1057 BP, the complete open reading frame of 549 BP, encoding 182 amino acids. Blast analysis showed that SBP-box has conserved domain specific SBP-box gene family encoding the amino acid sequence of the Arabidopsis SPLs protein known. And the construction of phylogenetic tree showed that, Dan SPL protein and At SPL3 were clustered into a, the gene was named PsSPL3. biological software to predict the molecular weight of PsSPL3 protein was 20.3494 kDa and the isoelectric point of 9.62. homology analysis of peony PsSPL3 and other known plant SPL3, similar to 47.98%~69.46%, and the birch SPL3 protein similarity The highest, the results showed that the PsSPL3 gene in different tissues and early flowering peony flower bud dormancy expression level in the process of 69.46%. real-time quantitative PCR analysis of PsSPL3 gene in different tissues of expression differences, including transcription in the root level highest in stems and petals of times; the transcriptional level of PsSPL3 gene in the course of dormancy in the form of a first increased and then decreased, the low temperature of 14 d, PsSPL3 expression was the highest. The low temperature of 7 d with PsSPL3 exogenous GA3 gene expression in peony flower bud increased significantly, suggesting that PsSPL3 gene transcription induced by GA3 to promote peony floral bud dormancy process.

【作者单位】： 青岛农业大学生命科学学院山东省高校植物生物技术重点实验室;
【基金】：国家自然科学基金面上项目(31471908;31372104;31672194)
【分类号】：Q943.2;S685.11
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本文编号：1545267