华丽龙胆GsF3′5′H和GsFNS基因的克隆及表达分析

Cloning and Expression Analysis of GsF3′5′H and GsFNS Genes from Gentiana sino-ornata

该研究以华丽龙胆(Gentiana sino-ornata)5个不同开放阶段(H1~H5)的蓝色花冠为试材,利用RT-PCR技术克隆GsF3′5′H和GsFNS全长序列,并进行生物信息学分析,比较GsF3′5′H和GsFNS在不同组织和不同开放阶段的基因表达模式。结果显示:(1)所克隆的GsF3′5′H和GsFNS基因分别包含1560 bp和1590 bp开放阅读框(OFR),并分别编码520和529个氨基酸。(2)结构分析显示,GsF3′5′H和GsFNS均具有典型的F3′5′H和FNSⅡ蛋白保守结构域。(3)系统进化树分析表明,GsF3′5′H和GsFNS亲缘关系最近的物种是三花龙胆(Gentiana triflora)。(4)qRT-PCR结果显示,GsF3′5′H和GsFNS基因在根、茎、叶和花冠中均表达,其中GsF3′5′H基因在花冠H3阶段表达量最高。GsFNS在根中表达量最高,其次在花冠H4阶段两基因的表达均较高。研究推测,GsF3′5′H基因表达产生的飞燕草素苷和GsFNS表达产生的黄酮共着色作用可能使华丽龙胆的花冠呈更稳定艳丽的蓝色,为蓝色花分子育种提供重要的基因资源。

To investigate the function of GsF3′5′H and GsFNS in Gentiana sino-ornata,with blue corolla during 5 different flowering stages(H1-H5)as the test material,we cloned the full-length cDNA by using RT-PCR techniques.The results showed that:(1)GsF3′5′H and GsFNS contain 1560 bp and 1590 bp open reading frame(OFR),which encode 520 and 529 amino acids,respectively.(2)Structural analysis revealed that both GsF3′5′H and GsFNS had typical conserved domains of F3′5′H and FNSⅡ.(3)Phylogenetic analysis indicated that Gentiana triflora was the closest relative species between GsF3′5′H and GsFNS.(4)qRT-PCR showed that GsF3′5′H and GsFNS were expressed in roots,stems,leaves and corolla,and the expression level of GsF3′5′H was the highest in corolla H3 stage,the expression level of GsFNS was the highest in roots,which followed by the corolla H4 stage.It showed that the co-pigmentation effect of delphinin and flavone expressed by GsF3′5′H and GsFNS may make the corolla of Gentiana sino-ornata more stable and bluer,which providing important genetic resources for the molecular breeding of blue flowers.