灯盏花Eb14-3-3基因的克隆及表达分析  

Cloning and Expression Analysis of Eb14-3-3 Gene in Erigeron breviscapus

作　　者：宋婉玲 卢迎春[1,2] 李锐 范伟[1,2,3] 龙光强 刘冠泽[1,2] 张广辉 杨生超[1,2] 赵艳[1,2,3] Song Wanling;Lu Yingchun;Li Rui;Fan Wei;Long Guangqiang;Liu Guanze;Zhang Guanghui;Yang Shengchao;Zhao Yan(National&Local Joint Engineering Research Center on Germplasms Utilization&Innovation of Chinese Medicinal Materials in Southwest,Yunnan Agricultural University,Kunming,650201;Yunnan Provincial Key Laboratory of Medicinal Plant Biology,Yunnan Agricultural University,Kunming,650201;College of Agriculture and Biotechnology,Yunnan Agricultural University,Kunming,650201)

机构地区：[1]云南农业大学西南中药材种质创新与利用国家地方联合工程研究中心,昆明650201 [2]云南农业大学云南省药用植物生物学重点实验室,昆明650201 [3]云南农业大学农学与生物技术学院,昆明650201

出　　处：《分子植物育种》2021年第20期6653-6661,共9页Molecular Plant Breeding

基　　金：国家自然科学基金项目(81760694);云南农业大学自然科学青年基金资助项目(2015ZR15);云南省重大专项(2017ZF002)共同资助。

摘　　要：为探究灯盏花Eb14-3-3基因表达特征,本研究从灯盏花基因组数据库中克隆了拟南芥同源基因灯盏花14-3-3υ序列信息,利用生物信息学方法对该蛋白的功能和活性位点进行预测,通过荧光定量PCR分析在非生物胁迫条件下该基因的表达变化。结果表明,Eb14-3-3的ORF序列长804 bp,编码267个氨基酸;该蛋白的分子量和理论等电点分别为65296 k D和5.16。Eb14-3-3蛋白序列中富含磷酸化位点,为一个非分泌型蛋白,并且在其结构中无跨膜结构域。二级结构分析显示有156个α-螺旋,47个片层延伸,9个β转角,55个无规则卷曲,属于14-3-3蛋白家族成员。RT-qPCR表达模式分析表明,5%PEG处理后,灯盏花叶、花和茎中Eb14-3-3基因的表达量在24 h明显上升,但在处理48 h后,Eb14-3-3的表达量大幅度下降;Eb14-3-3基因在灯盏花根部的表达量,在5%PEG处理10 h后,其表达量显著上升,并在处理后48 h呈现显著下降趋势。本研究将为培育新的灯盏花抗逆种质资源提供研究理论基础。To explore Eb14-3-3 gene expression characteristics in Erigeron breviscapus,the sequence information of 14-3-3υencoding homologous gene from E.breviscapus was obtained by searching genome database and Eb14-3-3 was cloned.Using bioinformatics methods to predict the function of the protein and active site,and analyzed gene expression changes in abiotic stress by using RT-qPCR.The results showed that the ORF sequence of Eb14-3-3 was 804 bp,encoding 267 amino acids.The molecular weight and theoretical isoelectric point of the protein were 65.296 kD and 5.16,respectively.Eb14-3-3 protein sequence is rich in phosphorylation sites,is a non-secretory protein,and has no transmembrane domain.Secondary structure analysis revealed 156α-helix,47 lamellar extensions,9β-turn,and 55 random coil,belonging to the 14-3-3 protein family.RT-qPCR analysis showed that,the expression level of Eb14-3-3 gene significantly increased at 24 h after 5%PEG treatment,however,expression level of Eb14-3-3 gene significantly decreased at 48 h in leaf,flower and stem.The expression level of Eb14-3-3 gene significantly increased at 10 h in the root in E.breviscapus after 5%PEG treatment,and decreased significantly at 48 h.This study will provide a theoretical basis for further research that cultivating new resistant germplasm resources.

关 键 词：灯盏花 Eb14-3-3 生物信息学 非生物胁迫 

分 类 号：S567.239[农业科学—中草药栽培]