许瑞瑞1,2，李睿2，王小非2，郝玉金2
1潍坊学院生物与农业工程学院/山东省高校生物化学与分子生物学重点实验室，山东潍坊 261061；2山东农业大学园艺科学与工程学院/国家苹果工程技术研究中心，山东泰安 271018
Identification and Expression Analysis Under Abiotic Stresses of OFP Gene Family in Apple
XU RuiRui1,2, LI Rui2, WANG XiaoFei2, HAO YuJin2
1College of Biological and Agricultural Engineering, Weifang University/Key Laboratory of Biochemistry and Molecular Biology in Universities of Shandong, Weifang 261061, Shandong; 2College of Horticulture Science and Technology, Shandong Agricultural University/National Research Center for Apple Engineering and Technology, Tai’an 271018, Shandong
摘要 【目的】从苹果全基因组中鉴定OFP（OVATE family protein）家族蛋白成员，对其进行基因结构特征、组织表达及非生物逆境等系统分析，为研究苹果OFP的潜在功能提供理论基础。【方法】利用生物信息学手段，在苹果基因组数据库中筛选鉴定OFP基因家族成员；利用MEGA5.0软件进行系统进化树分析；通过MapDraw和GSDS等生物信息学工具分析基因结构及染色体定位；根据已有的苹果芯片数据库结果进行OFP基因表达谱分析；利用实时荧光定量PCR技术检测13个MdOFP的组织表达和诱导表达情况。【结果】苹果OFP基因家族包含28个成员，根据系统进化关系将其分为4组，分别包含13、6、4和5个成员；苹果中13条染色体上均有OFP基因分布，其中第12条染色体最多，有6个MdOFP成员，该基因家族的分布具有广泛性；芯片表达谱分析结果表明该类基因家族在花、果实和叶中的表达量较高，qRT-PCR验证结果较一致；经NaCl和PEG处理后，苹果根部与地上部呈现出不同程度的响应差异，NaCl处理明显诱导两组织中MdOFP04和MdOFP20的表达，MdOFP01、MdOFP12和MdOFP18的表达在根部与地上部组织则相反；温度胁迫明显影响MdOFPs的表达量，其中MdOFP04和MdOFP17经高温和低温胁迫处理后均明显上调。【结论】苹果OFP基因家族共有28个成员，分布于13条染色体上，该家族成员呈现出不同的组织表达模式和胁迫响应模式。
关键词 ： 苹果, OFP基因家族, 系统进化, 胁迫, 基因表达
Abstract：【Objective】 Identification of the OFP (OVATE family protein) genes from apple genome and analysis of gene characteristic, tissue expression pattern and response to abiotic stresses of OFP family genes in apple will be useful to the functional analysis of plant OFP genes. 【Method】 Based on apple genome database, the OFP gene family members were identified and the genes were analyzed using bioinformatics methods. A phylogenetic tree was created using the MEGA5.0 program. Gene structure and chromosomes location were carried out by MapDraw and GSDS separately. Expression pattern analysis of OFP genes in different tissues was done based on the existing microarray database and qRT-PCR. The expression of 13 MdOFPs genes was also analyzed under various stress conditions using qRT-PCR. 【Result】 A total of 28 OFP genes was systematically identified from apple genome and classified into 4 groups including 13, 6, 4 and 5 members according to the gene structure and conserved domain phylogeny relationship. All OFP genes are distributed on 13 apple chromosomes with the largest number six MdOFP on Chr12, suggesting that they have an extensive distribution on the apple chromosomes. Most of the OFP genes have distinctive expression patterns in tissues and response to NaCl and PEG treatment stresses in root and shoot, respectively. MdOFP04 and MdOFP20 were up-regulated obviously in root and shoot, while MdOFP01, MdOFP12 and MdOFP18 have opposite expression pattern in root and shoot under NaCl stress. Temperature stresses significantly regulate the expression of MdOFP and the expression of MdOFP04 and MdOFP17 were significantly increased after high temperature and low temperature stresses. 【Conclusion】 Twenty-eight OFP genes in apple were identified by genome-wide screening. They are classified into four groups and distributed on 13 chromosomes with different tissue patterns and different stress response patterns. These results will be helpful to the functional analysis of OFP genes in apple.
Key words： apple OFP gene family phylogeny analysis stress gene expression
收稿日期: 2017-09-20
基金资助:国家自然科学基金青年基金（31400225）、潍坊市科学技术发展计划（2016GX010）
通讯作者: 许瑞瑞，Tel：0536-8785288；E-mail：xuruirui2006@163.com。郝玉金，Tel：0538-8246692；E-mail：haoyujin@sdau.edu.cn
作者简介: 许瑞瑞，Tel：0536-8785288；E-mail：xuruirui2006@163.com
