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牡丹PsELF1基因的克隆及表达分析

Cloning and Expression Analysis of PsELF1 Gene from Tree Peony(Paeonia suffruticosa)
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摘要 本研究以已获得的早花基因的EST序列为模板设计引物,通过RACE技术扩增全长,使用生物学软件和q RT-PCR技术分别进行生物信息学分析和表达模式分析,获得牡丹早花基因Ps ELF1的全长及其表达模式,为后续研究其功能提供条件。结果显示,扩增得到7 107 bp的Ps ELF1全长cDNA,其中,包括3'UTR485 bp,5'UTR 464 bp和6 258 bp的编码区,共编码2 085个氨基酸,分子量为238.43 kD。Ps ELF1包括4个可能的磷酸化位点,且均位于N端。二级结构预测显示,Ps ELF1含α-螺旋和无规卷曲较多。同源性分析结果表明,Ps ELF1与杨树ELF1同源性最高。表达特性分析表明,Ps ELF1在花芽休眠解除过程中Ps ELF1的表达水平在0~14 d下调,在14~21 d上调,然后在21~28 d下调。而在低温21 d移入温室后的开花过程中呈下调模式。本研究结果对培育牡丹早花或晚花品种、延长牡丹的观赏期、提高牡丹观赏质量和经济价值具有重要意义。 The paper was aimed to get the full length of cDNA and the analysis of the expression level for early flower gene PsELF1,which would lay a foundation for the function study of PsELF1.In this study,Based on the EST of ELFs from our laboratory,the full length of cDNA for PsELF1 was obtained by RACE amplification and the analysis of the expression level was fulfilled by qRT-PCR.Meanwhile,bioinformatics analysis was performed by bioinformatics software.The result shows that the length of cDNA for PsELF1 was 7107 bp including 3'UTR of 485 bp,5'UTR of 464 bp and 6258 bp encoding frame,which encoded 2085 amino acids and had a molecular weight of 238.43 kD.The predication of phosphorylation sites showed that the amino acid sequence of PsELF1 had 4 possible phosphorylation sites and all the phosphorylation sites were at the N-terminal sequence of the Ps-%ELF1.The secondary structures of PsELF1 were mainlyα-helix and random coil.The phylogenetic tree indicated that PsELF1 was highly conserved with that of Populus trichocarpa ELF1.The expression level of PsELF1 showed down-regulation at 0~14 d,and then up-regulation at 14~21 d,and next down-regulation at 21~28 d during the bud dormancy release.In addition,tree peonies were transferred to the greenhouse after 21 d chilling treatment and the down-regulation pattern in the expression level of PsELF1 was indicated during the course of floral organ develop-ment from flower bud emergence stage to the bud-sticking stage.The results of this study lay a foundation for breeding the cultivars of early blossoms or late ones and prolonging florescence,which were the key ways for improvement of ornamental quality and economic value in tree peony.
作者 张艳萍 张英昊 付祥梅 张玉喜 刘春英 Zhang Yanping;Zhang Yinghao;Fu Xiangmei;Zhang Yuxi;Liu Chunying(Key Lab of Plant Biotechnology in Universities of Shandong Province,College of Life Sciences,Qingdao Agricultural University,Qingdao,266109)
出处 《分子植物育种》 CAS CSCD 北大核心 2020年第16期5247-5251,共5页 Molecular Plant Breeding
基金 国家自然基金面上项目(31872145) 青岛农业大学高层次人才科研基金(6631116009)共同资助。
关键词 牡丹(Paeonia suffruticosa) Ps ELF1 RACE扩增 表达分析 Paeonia suffruticosa PsELF1 RACE amplification Expression analysis
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