牡丹早花基因PsELF3的克隆和表达特性分析  

Cloning and expression pattern analysis of EARLY FLOWERING gene PsELF3 in tree peony

作　　者：付祥梅 盖树鹏[1] 刘春英[1] 孙阳[1] 张玉喜[1] FU Xiangmei;GAI Shupeng;LIU Chunying;SUN Yang;ZHANG Yuxi(Key Lab of Plant Biotechnology in Universities of Shandong Province,College of Life Science,Qingdao Agricultural University,Qingdao,Shandong 266109,China)

机构地区：[1]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,山东青岛266109

出　　处：《植物生理学报》2020年第3期540-546,共7页Plant Physiology Journal

基　　金：国家自然科学基金(31872145和31672194);国家重点研发计划项目子课题(2018YFD100403);青岛农业大学高层次人才基金(6631116009)。

摘　　要：本研究利用BioEdit筛选了牡丹(Paeonia suffruticosa)花芽的第三代转录组数据库,得到一条早花基因3(ELF3)片段。利用3′cDNA末端快速扩增技术(RACE)扩增测序后与原序列进行序列拼接,得到了2 457 bp的全长cDNA,其中5′非翻译区(5′UTR)为122 bp, 3′UTR为274 bp;全长编码区为2 061 bp,编码687个氨基酸。同源性比对发现其编码蛋白(PsELF3)与葡萄(Vitis vinifera) VvELF3蛋白同源性最高,为58.48%。进化树分析结果显示PsELF3与葡萄VvELF3蛋白聚为一支,与同源性比对结果基本一致。实时定量PCR分析表明PsELF3基因在初花期不同组织中均有表达,但表达水平差异较大,其在心皮中表达水平最高,在根中最低。另外, PsELF3在花芽休眠解除过程中呈上调模式,在低温处理14 d后其表达水平显著升高, 28 d达到最高值。牡丹花芽在低温处理21 d后移入温室,实时定量PCR分析显示PsELF3从花芽膨大期到翘蕾期呈上调模式,其表达水平至翘蕾期达到最高值。研究结果拟对进一步研究PsELF3在牡丹发育、花芽内休眠解除进程及开花过程中的功能提供理论依据,为反季节催花提供候选基因。This study used BioEdit software to screen the third-generation transcriptome data of tree peony(Paeonia suffruticosa) flower buds, and an EARLY FLOWERING gene(ELF3) cDNA fragment was obtained. After 3′ RACE PCR amplification and sequencing, the original cDNA sequence was spliced with the sequence of 3′ RACE PCR amplification product, and a 2 457 bp full-length cDNA was obtained, of which the 5′ untranslated region(UTR) was 122 bp and the 3′ UTR was 274 bp. The full-length coding region was 2 061 bp, encoding 687 amino acids. The homology alignment revealed that the pupative PsELF3 protein has the highest homology(58.48%) with Vitis vinifera VvELF3 protein. The phylogenetic tree was constructed, showing PsELF3 protein was clustered with V. vinifera VvELF3, which is basically consistent with the result of homology alignment. Real-time quantitative PCR analysis showed that the PsELF3 gene expressed in different tissues at the early flowering stage, but the expression levels were different. The expression level was the highest in the carpel and the lowest in the root. In addition, PsELF3 showed up-regulated pattern during the bud dormancy release, and its expression level significantly increased after 14 d chilling treatment, reaching the highest level after 28 d chilling treatment. Tree peony was transferred to the greenhouse after 21 d of chilling treatment, the expression level of PsELF3 was up-regulated from flower bud emergence stage to the bud-sticking stage, and the expression level reached the highest at the bud-sticking stage. These results would provide new theoretical basis for further study of its function during the development, dormancy release and flowering stages in tree peony, and offer candidate gene for anti-seasonal culture.

关 键 词：牡丹 ELF3 克隆 休眠 表达特性 

分 类 号：S685.11[农业科学—观赏园艺]