山茶花SEPALLATASEP)家族基因克隆及表达分析

    Cloning and Expression Analysis of SEPALLATASEP)Gene Family in Camellia japonica

    • 摘要: 花型特征是山茶花重要的观赏性状之一,为了揭示山茶花SEP基因结构特征及其与山茶花重瓣花型的关系,采用同源克隆结合RACE方法进行山茶花SEP基因克隆,对其进行生物信息学分析,并采用实时荧光定量PCR检测SEP基因在不同花型山茶花品种中的表达特征。结果显示:从山茶花胭脂牡丹中克隆得到3个SEP基因CjSEP1a1235 bp)、CjSEP3(967 bp)、CjSEP4a1725 bp),GenBank登录号分别为OP172861、OP172863、OP172864,分别编码249、185和243个氨基酸。山茶花3个SEP基因的编码蛋白均呈不稳定、亲水、细胞核定位特征,具有MADS-box和K-box保守结构域,与同属物种茶树的SEP蛋白具有高度同源性。qPCR分析表明,完全重瓣型品种胭脂牡丹3个SEP基因的表达量显著高于其他品种,其次为半重瓣品种和单瓣型品种,3个特异重瓣花型品种的SEP表达量总体较低,因此,随着花型规则重瓣程度的加深,SEP基因的表达量呈上调表达。CjSEP基因与山茶花规则重瓣性状存在紧密关联性,不同CjSEP成员具有协同表达作用。

       

      Abstract: The traits of flower types are significant ornamental features of Camellia japonica. In order to reveal the structural characteristics of the SEP genes in C. japonica and its relationship with the double-petal flower type, homologous cloning combined with RACE method was used to clone the SEP genes, bioinformatics analysis was performed, and real time fluorescence quantitative PCR was used to detect the expression characteristics of the SEP genes in different flower types of C. japonica. Results showed that three SEP genes were cloned from C. japonica ‘Yanzhimudan’: CjSEP1a1235 bp), CjSEP3(967 bp), and CjSEP4a1725 bp), with their respective GenBank accession numbers OP172861, OP172863 and OP172864. These genes encoded proteins consisting of 249, 185 and 243 amino acids, respectively. All three SEP genes encoded unstable, hydrophilic proteins localized in the nucleus, featuring conserved domains of MADS-box and K-box, and exhibited high similarity to SEP proteins from C. sinensis. The qPCR analysis indicated that the expression levels of these CjSEP genes in the fully double-petal cultivar ‘Yanzhimudan’ were significantly upregulated compared to those in other cultivars, followed by the semi-double-petal cultivar and the single-petal cultivar. Interestingly, the expression levels of CjSEPs in three other specific double-petal cultivars were generally lower. Therefore, as the prominence of double-petal phenotype increases, the expression of SEP genes also rises. A strong relationship exists betwwn CjSEP genes and the characteristic double-petal trait in C. japonica, with CjSEP genes exhibiting synergistic effects in their expression.

       

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