普通白菜PRR5的克隆、表达及功能验证  

Cloning,Expression and Functional Identification of PRR5 Gene in Pakchoi

作　　者：侯瑞泽 鲍悦 陈启亮 毛桂玲 韦博霖 侯雷平[1] 李梅兰[1] HOU Rui-ze;BAO Yue;CHEN Qi-liang;MAO Gui-ling;WEI Bo-lin;HOU Lei-ping;LI Mei-lan(College of Horticulture,Shanxi Agricultural University,Jinzhong 030800)

机构地区：[1]山西农业大学园艺学院,晋中030800

出　　处：《生物技术通报》2023年第10期128-135,共8页Biotechnology Bulletin

基　　金：山西省应用基础研究计划项目(20210302123421);山西农业大学2021年生物育种工程项目(YZGC121);山西省重点研发计划项目(20220214060005)。

摘　　要：克隆BrcPRR5,并研究其在不同组织和不同发育时期的表达模式及功能,为了解PRR5对普通白菜成花转变的影响奠定基础。运用RT-PCR方法克隆PRR5的同源基因BrcPRR5,并对其进行生物信息学分析,利用荧光定量PCR测定该基因在不同组织部位和不同发育时期的相对表达量,构建过表达载体并转化拟南芥进行基因的功能验证。结果表明,BrcPRR5的CDS全长为1 701 bp,编码566个氨基酸,通过与其他物种的同源蛋白进行氨基酸序列多重比对,确定得到的序列属于普通白菜PRR5同源基因。BrcPRR5在茎和花中的表达量高于叶和果荚中的表达量;在S0茎尖中表达量最高,说明其在普通白菜成花转变过程中表达量上调可能促进成花转变。超表达转基因植株开花期提前,株高和茎粗明显优于野生型。BrcPRR5可以促进植株提早抽薹开花。Cloning BrcPRR5 and studying its expression pattern and function in different tissues and different developmental stages may lay a foundation for understanding the effect of PRR5 on flowering transformation of pakchoi.The homologous gene BrcPRR5 of PRR5 was cloned by RT-PCR and analyzed by bioinformatics.The relative expressions of the gene in different tissue sites and different developmental stages were determined by fluorescence quantitative PCR.The overexpression vector was constructed and transformed into Arabidopsis for functional verification.The results showed that the full-length CDS of BrcPRR5 was 1 701 bp,encoding 566 amino acids.Through multiple amino acid sequence alignment with homologous proteins of other species,the obtained sequence belonged to PRR5 homologous gene of pak choi.The expression of BrcPRR5 in the stem and flower was higher than that in the leaf and pod,and the highest expression was in S0 stem tip,indicating that the up-regulated expression of S0 promoted floral transformation during flowering transition of pakchoi.The flowering stage of overexpressed transgenic plants was earlier,and the plant height and stem diameter of transgenic plants were significantly better than those of wild type.BrcPRR5 may promote plant bolting and flowering earlier.

关 键 词：普通白菜 BrcPRR5 成花转变 基因克隆 功能验证 

分 类 号：S634[农业科学—蔬菜学]