芍药C4H基因的克隆及其在钙处理下的表达特性分析

Cloning of C4H Gene in Herbaceous Peony and Expression Characteristic Analysis of It under Calcium Treatment

以芍药花茎RNA为模板,采用PCR技术对芍药肉桂酸-4-羟化酶基因(C4H)进行克隆与序列分析,并采用实时荧光定量PCR技术对C4H在钙处理下花茎中的表达特性进行分析。结果表明:芍药C4H基因长1 715 bp,具有完整的开放阅读框1 548 bp,共编码515个氨基酸,等电点为9.33,不具有信号肽、存在1个明显的跨膜结构域,属于不稳定蛋白和亲水性蛋白。花茎中C4H基因的表达水平随着芍药植株的发育而呈逐渐上升趋势,并且钙处理增强了C4H基因的表达。本研究有助于通过分子手段培育花茎挺直的芍药品种以及在生产上推广钙处理来提高芍药花茎挺直程度。

Taking RNA extracted from herbaceous peony inflorescence stem as template, the study used PCR method to clone cinnamate-4-hydroxylase gene（C4 H） and analyzed its sequence. Subsequently, real-time fluorescent quantitative PCR technology was applied to analyze its expression characteristic under calcium treatment. The results showed that herbaceous peony C4 H gene had 1 715 bp in length and a 1 548 bp open reading frame,encoding 515 amino acids, with the isoelectric point of 9.33, which had no signal peptide but an obvious transmembrane domain, belonging to the unstable and hydrophilic protein. The expression level of C4 H in herbaceous peony inflorescence stem increased with the development of herbaceous peony, and calcium treatment enhanced the expression of C4 H. This study would help to develop herbaceous peony cultivars with straight inflorescence stem by molecular methods and improve the straight level of herbaceous peony inflorescence stem by calcium treatment in production.