高丛蓝莓细胞色素CYP81家族的鉴定及表达分析  

Identification and Expression Analysis of CYP81 Family in Highbush Blueberry

作　　者：王卓依 邱佳琪 张颖[1] 俞蕾 刘良淼 李永强[1,2] 陈文荣[1,2] 郭卫东[1,2] 杨莉[1,2] Wang Zhuoyi;Qiu Jiaqi;Zhang Ying;Yu Lei;Liu Liangmiao;Li Yongqiang;Chen Wenrong;Guo Weidong;Yang Li(College of Life Sciences,Zhejiang Normal University,Jinhua,321004;Zhejiang Provincial Key Laboratory of Biotechnology on Specialty Economic Plants,Zhejiang Normal University,Jinhua,321004)

机构地区：[1]浙江师范大学生命科学学院,金华321004 [2]浙江师范大学,浙江省特色经济植物生物技术研究重点实验室,金华321004

出　　处：《分子植物育种》2025年第2期387-395,共9页Molecular Plant Breeding

基　　金：浙江省农业(果品)新品种选育重大科技专项(2021C02066-9);2020年省级大学生创新创业训练计划项目(202010345R129);金华市2021年度重大(重点)科学技术研究计划项目(2021-2-038)共同资助。

摘　　要：高丛蓝莓(Vaccinium corymbosum)是一种高经济价值作物,其果实因富含花青苷等次生代谢物,具有较高的营养价值与经济价值。CYP81是植物特有的A型细胞色素CYP450亚家族,广泛参与防御途径和化合物代谢过程,但有关蓝莓CYP81亚家族的成员数量、基因结构和表达模式等尚不清楚。本研究利用高丛蓝莓‘Draper’基因组测序数据鉴定CYP81家族基因,并对这些家族成员进行生物信息学分析;根据高丛蓝莓VcCYP81与拟南芥CYP81蛋白的序列比对,构建系统进化树;利用实验室前期获取的转录组数据分析VcCYP81基因在南高丛蓝莓‘奥尼尔’与‘蓝雨’坐果期的表达谱,以及利用实时荧光定量PCR(qPCR)技术分析差异表达基因在2个品种花果发育进程中的表达模式。基于拟南芥AtCYP81基因序列,检索‘Draper’基因组数据库获得43个Vc CYP81家族成员,分析发现这些VcCYP81家族成员分别包含2~8个外显子,且大部分成员包含6个排列顺序相同的保守基序。进化树分析将VcCYP81进一步分为VcCYP81D7-1~VcCYP81D7-7与VcCYP81K 8个亚家族,其中D7-1,D7-2,D7-4及D7-5亚家族成员间高度保守,而VcCYP81K亚家族成员与拟南芥同源序列聚类。结合转录组数据分析VcCYP81亚家族基因在‘奥尼尔’与‘蓝雨’幼果中的表达丰度,发现大部分家族成员表达丰度较低或不表达,而VcCYP81D7-2亚家族成员的表达水平极显著高于其他成员。进一步分析2个品种幼果期差异表达基因gene-18.47和gene-34.48在整个花芽膨大与果实发育时期的相对表达,发现2个基因在‘奥尼尔’与‘蓝雨’花芽与果实中的表达差异显著。本研究为进一步研究高丛蓝莓VcCYP81基因对果实生长发育及次生代谢物的生物合成与代谢等提供理论参考。Vaccinium corymbosum,also known as blueberry,is a crop of high economic value,and its fruit is rich in secondary metabolites such as anthocyanin,which has high nutritional and economic value.CYP81 is A plant-specific member of the A-type cytochrome CYP450 family,which is widely involved in defense pathways and compound metabolism.However,the number of members,gene structure and expression patterns of CYP81subfamily in blueberry remain unclear.In this study,CYP81 family genes were identified using genomic sequencing data of'Draper'in blueberry,and bioinformatics analysis was conducted for these family members.The phylogenetic tree was constructed based on the sequence comparison between CYP81 protein of highbush blueberry and Arabidopsis.The transcriptome data obtained in the laboratory were used to analyze the expression profiles of VcCYP81 gene in the fruit setting stage of'O'Neal'and'Bluerain'of southern highbush blueberry,and the expression patterns of differentially expressed genes in the fruit development of the two varieties were analyzed by RT-qPCR.Based on the AtCYP81 gene sequence of Arabidopsis,43 VcCYP81 family members were obtained by searching the'Draper'genome database.It was found that these VcCYP81 family members contained 2 to 8 exons respectively,and most of them contained 6 conserved momods with the same sequence.VcCYP81 was further divided into VcCYP81D7-1~VcCYP81D7-7 and Vc CYP81K subfamilies by evolutionary tree analysis.Among them,members of D7-1,D7-2,D7-4 and D7-5 subfamilies were highly conserved,while members of VcCYP81K subfamily were clustered with homologous sequences of Arabidopsis.The expression abundance of VcCYP81subfamily genes in'O'Neal'and'Bluerain'young fruits was analyzed with transcriptome data,and it was found that the expression abundance of most family members was low or no expression,while the expression level of VcCYP81D7-2 subfamily members was significantly higher than other members.Further analysis of the relative expression of the differentially expressed genes gene

关 键 词：蓝莓 VcCYP81基因 生物信息学分析 全基因组鉴定 表达分析 

分 类 号：S66[农业科学—果树学]