菊花CgF3′H基因克隆及表达特性分析

Cloning and Expression Profiles of CgF3'H in Chrysanthemum

根据菊花花器官表达序列标签序列设计引物,采用PCR和5'-RACE技术对类黄酮3'-羟化酶(F3'H)基因进行全长cDNA克隆。克隆获得F3'HcDNA全长为1760bp,其开放阅读框(ORF)为1524bp,编码一条包含508个氨基酸残基的多肽,GenBank登录号为AB523844。预测该基因编码的蛋白分子式为C2535H4022N684O707S18,分子量为55.97kD,理论等电点pI为7.23。其氨基酸序列与蓝眼菊属(ABB29899.1)、紫茎泽兰(ABM46853.1)、欧亚种葡萄(ACN38268.1)和高粱(ABG54320.1)等植物的F3'H氨基酸序列的同源性分别为83.27%、80.51%、75.05%和59.96%。系统进化分析表明其与蓝眼菊属的亲缘关系最近。荧光定量PCR分析发现F3'H基因在切花菊‘H5’各个器官中均有表达,花托中表达量最高,而叶片中表达量最低。

Primers was designed according to the available expressed sequence tags of F3＇H, RT-PCR and 5＇-RACE PCR cloning were employed to clone the full length of F3＇H. The full length cDNA is 1 760 bp, with an open reading frame （ORF） of 1 524 bp in length. The GenBank accession number of CgF3＇H is AB523844. The predicted molecular weight is 55.97 kD, with a theoretical pI of 7.23, and the molecular formula is C2535H4022N684O707S18. The identity between amino acid sequence of CgF3＇H and those from Osteospermum （ABB29899.1）, Ageratina adenophora （ABM46853.1）, Vitis vinifera （ACN38268.1） and Sorghum bicolor （ABG54320.1） are 83.27%, 80.51%, 75.05% and 59.96%, respectively. Phylogeny analysis revealed that CgF3＇H is closest to that from Osteospermum. Real-time quantitative PCR analysis showed the CgF3＇H gene expressed in all organs of cut chrysanthemum cultivar ‘H5’, the expression level of CgF3＇H is highest in receptacles but least in leaves.