牡丹PsSPL3基因的克隆和表達特性分析
發(fā)布時間:2018-02-28 01:57
本文關鍵詞: 牡丹 PsSPL RACE 實時定量PCR 表達模式 出處:《華北農學報》2017年04期 論文類型:期刊論文
【摘要】:SPL(SQUAMOSA promoter-binding protein-like)轉錄因子在植物多個生理過程中發(fā)揮重要的生理功能,為了研究其在牡丹花芽休眠進程中的作用機理,采用RACE方法,從牡丹花芽中克隆得到了一個SPL基因。該基因全長cDNA 1 057 bp,完整開放閱讀框為549 bp,編碼182個氨基酸,Blast分析表明,編碼的氨基酸序列中具有SBP-box基因家族所特有的SBP-box保守結構域。與擬南芥已知SPLs蛋白構建進化樹結果表明,牡丹SPL蛋白與At SPL3聚為一支,該基因被命名為PsSPL3。生物軟件預測PsSPL3蛋白分子量為20.349 4 kDa,理論等電點為9.62。同源性分析了牡丹PsSPL3與其他已知植物的SPL3,相似性為47.98%~69.46%,其中與白樺的SPL3蛋白相似性最高,為69.46%。實時定量PCR分析PsSPL3基因在初花期牡丹不同組織中和花芽休眠過程中的表達水平,結果表明,PsSPL3基因在不同組織中表達差異較大,其中在根中轉錄水平最高,在莖和花瓣中次之;PsSPL3基因在休眠進程中的轉錄水平呈先上升后下降趨勢,其中低溫處理14 d時,PsSPL3基因的表達量達到最高。低溫7 d結合外源施加GA3的牡丹花芽中PsSPL3基因的表達量顯著增加,推測PsSPL3基因的轉錄受GA3誘導來促進牡丹花芽內休眠解除的進程。
[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.
【作者單位】: 青島農業(yè)大學生命科學學院山東省高校植物生物技術重點實驗室;
【基金】:國家自然科學基金面上項目(31471908;31372104;31672194)
【分類號】:Q943.2;S685.11
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本文編號:1545267
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