牡丹<i>PlSEP1</i>基因的鉴定与调控开花时间功能研究

doi:10.7668/hbnxb.20193099

摘要/Abstract

摘要：

SEPALLATA1(SEP1)基因是重要的MADS-box家族基因,在植物开花时间调控中发挥重要作用。为明确牡丹中PlSEP1基因在牡丹开花过程中的生物学功能及表达特性,进一步了解其参与开花时间调控的机制,基于前期得到的牡丹二次开花品种海黄花芽发育过程转录组数据中筛选到的一个MADS-box家族基因PlSEP1,应用实时荧光定量PCR检测PlSEP1基因在牡丹花芽发育过程中的相对表达量,通过在拟南芥中过量表达PlSEP1基因探究其发挥的功能。结果显示,获得的PlSEP1基因具有735 bp的CDS序列,共编码244个氨基酸。经生物信息学分析得知,PlSEP1编码的蛋白具有稳定的亲水性,并且不含信号肽。进化分析显示,PlSEP1与拟南芥、大豆亲缘关系较近;亚细胞定位发现,PlSEP1蛋白在细胞核中表达;实时荧光定量PCR结果表明,PlSEP1在花芽中表达量最高,且在分化期达到最高。拟南芥中过量表达PlSEP1会导致开花时间提前,表明PlSEP1参与调控开花过程。

关键词: 牡丹, PlSEP1, 花芽发育, 开花时间, 亚细胞定位

Abstract:

SEPALLATA1(SEP1) gene is a member of MADS-box gene family which plays an important role in flowering time regulation.In order to clarify the biological function and expression characteristics of PlSEP1 gene in tree peony during flowering,and further understand the mechanism of PlSEP1 gene involved in the regulation of flowering time.A MADS-box family gene,PlSEP1,was screened from the transcriptome data of floral bud development in the reblooming cultivar of tree peony,High noon,the relative expression of PlSEP1 gene in the floral bud development of tree peony was detected by quantitative Real-time PCR,and the function of PlSEP1 gene was explored through over expression in Arabidopsis thaliana.The results showed that,PlSEP1 encoded 244 amino acid with a ORF of 735 bp.The bioinformative analysis showed that PlSEP1 encoded a stable hydrophilic protein,which contained none signal peptide.Phylogenetic tree showed that the PlSEP1 was close with its homologs in Arabidopsis thaliana,Glycine max.Subcellular location assay showed that PlSEP1 protein was located in the nucleus.PlSEP1 expressed highest in flower buds,and reached the highest in the differentiation stage.The phenotype of early flowering in over-expression transgene plant suggested that PlSEP1 regulate flowering time.

Key words: Tree peony, PlSEP1, Floral bud development, Flowering time, Subcellular location

常艳婷, 张闻博, 夏梦思, 范可可, 胡晓萌, 王淑华, 张雪, 白一苇, 张娜, 胡陶, 江泽慧. 牡丹PlSEP1基因的鉴定与调控开花时间功能研究[J]. 华北农学报, 2023, 38(2): 51-56. doi: 10.7668/hbnxb.20193099.

CHANG Yanting, ZHANG Wenbo, XIA Mengsi, FAN Keke, HU Xiaomeng, WANG Shuhua, ZHANG Xue, BAI Yiwei, ZHANG Na, HU Tao, JIANG Zehui. Identification and Function Study in Flowering Time Regulation of PlSEP1 Gene in Tree Peony[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(2): 51-56. doi: 10.7668/hbnxb.20193099.

http://www.hbnxb.net/CN/Y2023/V38/I2/51

图/表 7

表1 引物信息列表

Tab.1 Primer information list

引物Primer	引物序列Primer sequence
PlSEP1-F	GCTCTAGAATGGGAAGAAAAAAAGTGGAGATCAAGCGG
PlSEP1-R	TCCCCCCGGGGAGATCTACACTTACACCATCGACATTCGC
PlSEP1-qpcr-F	CTCTGTGATGCTGATGTGGCTCTCA
PlSEP1-qpcr-R	GCTCTAGCTCCTTCGAATCTAAAGGCC
TUBULIN-qpcr-F	GATGTGTGCTGCTGATCCTCGACAC
TUBULIN-qpcr-R	CTCCTGGATCGAAGTCGAGTTGCC

图1 PlSEP1氨基酸序列与其他物种氨基酸的多序列比对 NoSEP1.香睡莲;NnSEP1.中国莲;AtSEP1.拟南芥;GMSEP1.大豆;PlSEP1.牡丹;TgSEP1.郁金香;PcSEP1.同色兜兰;AcSEP1.中华猕猴桃;DcSEP1.鸽石斛。

Fig.1 Multiple sequence alignment of PlSEP1 with other species NaSEP1.Nymphaea odorata;NnSEP1.Nelumbo nucifera;TgSEP1.Tulipa gesneriana;AtSEP1.Arabidopsis thaliana;AcSEP1.Actinidia chinensis; GMSEP1.Glycine max;PcSEP1.Paphiopedilum concolor;DcSEP1.Dendrobium crumenatum.

图2 PlSEP1与其他物种SEP1的进化

Fig.2 Phylogenetic tree of PlSEP1 and SEP1 proteins from other species

图3 PlSEP1的亚细胞定位

Fig.3 Subcellular localization of PlSEP1

图4 PlSEP1在不同组织中的表达量分析不同小写字母代表差异显著(P<0.05)。图5-6同。

Fig.4 Relative expression level of PlSEP1 in different tissue Different lowercase letters above bars means significant difference(P<0.05).The same as Fig.5-6.

图5 PlSEP1在花芽发育过程中的表达量分析

Fig.5 Relative expression level of PlSEP1 in floral bud development

图6 过表达PlSEP1拟南芥的早花表型

Fig.6 Early flowering phynotype of OE-PlSEP1 in Arabidopsis

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牡丹PlSEP1基因的鉴定与调控开花时间功能研究

Identification and Function Study in Flowering Time Regulation of PlSEP1 Gene in Tree Peony

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