白菜BrYUCCA5.1基因表达和生物信息学及功能分析

Expression and bioinformatics and function analysis of BrYUCCA5.1 gene in Chinese cabbage

[目的]本文旨在研究白菜YUCCA家族基因BrYUCCA5.1在结球白菜和不结球白菜叶片中的表达模式,并探索其在白菜叶片发育中的作用。[方法]利用生物信息学软件对BrYUCCA5.1基因的进化关系、编码蛋白质的保守结构域、蛋白质的理化性质等特性进行分析;利用实时荧光定量PCR检测结球白菜W30和不结球白菜082及其F2代植株不同部位BrYUCCA5.1表达量的差异;利用芜菁黄化花叶病毒(TYMV)介导的病毒诱导的基因沉默(VIGS)技术沉默不结球白菜082植株的BrYUCCA5.1,研究该基因的功能。[结果]结球白菜BrYUCCA5.1所编码的蛋白与拟南芥、野生甘蓝、甘蓝型油菜、萝卜的同源性较高;保守结构域分析结果表明BrYUCCA5.1蛋白属于FMO类超家族。该基因在结球白菜W30叶片基部和中部以及F2代结球单株叶片基部的表达量极显著高于其他不结球单株。利用VIGS技术沉默不结球白菜的BrYUCCA5.1,沉默效率达84%,且与对照组相比,BrYUCCA5.1-pTY发病株叶片出现大面积病毒侵染症状,叶片卷曲皱缩,表现出远轴化的形态。[结论]BrYUCCA5.1基因参与调控白菜的叶片发育,且在结球白菜叶球发育过程中具有重要作用。

[Objectives]The paper aimed to study the expression pattern of YUCCA family gene BrYUCCA5.1 in heading and non-heading Chinese cabbage and explore its function during the leaf development of Chinese cabbage.[Methods]To gain insightful information of the putative YUCCA protein,the phylogenetic relationship,conserved domain and its chemical properties were conducted by bioinformatics analysis software.Quantitative real-time PCR(RT-qPCR)was performed to explore the expression level of BrYUCCA5.1 gene in different segments of heading Chinese cabbage W30,non-heading Chinese cabbage 082 and its F2 generation.Tumip yellow mosaic virus(TYMV)-derived virus induced gene silencing(VIGS)technology was employed to knock down the expression of BrYUCCA5.1 gene in non-heading Chinese cabbage 082,and study the function of this gene.[Results]The protein encoded by BrYUCCA5.1 gene of heading cabbage had high homology with Arabidopsis thaliana,Brassica oleracea,Brassica napus and Raphanus sativus.The core regions showed high conservation with structures of FMO-like protein.Compared with other individual non-heading plants,BrYUCCA5.1 was highly expressed in the middle and basal region of the leaves in heading Chinese cabbage W30,as well as the middle region of F2 generation.Using VIGS technology to silence BrYUCCA5.1 of non-heading Chinese cabbage,the silencing efficiency reached 84%.Compared with the control group,BrYUCCA5.1-pTY susceptible plants showed symptoms of virus infection on the leaves of a large area,the leaves curled and shrunk,and the morphology was teleaxialization.[Conclusions]BrYUCCA5.1 gene participated in the regulation of leaf development of cabbage and played important role in the development of leaf bulbs of heading Chinese cabbage.