葡萄OVATE转录因子家族的鉴定、表达分析及VvOFP8和VvOFP17基因的克隆
发布时间：2019-06-10 14:52
【摘要】：OVATE蛋白家族(Ovate Family Protein,OFP)是一类调控植物生长发育的转录抑制因子,在拟南芥、番茄和水稻等植物中均有研究报道,并鉴定出一些OFPs调控番茄、拟南芥果实形状。果实形状是葡萄一项重要的外观品质,研究果形发育的分子机理对果形选育具有重要意义。本研究对葡萄OVATE转录因子家族进行了全面的鉴定和生物信息学分析,利用实时荧光相对定量PCR技术探究了该家族成员的表达模式,从'宝满'葡萄中克隆得到两个可能调控果形的相关基因VvOFP8和VvOFP17,并进行生物信息学分析和表达特性研究。主要研究结果如下:1葡萄OVATE转录因子家族包含17个成员,不均匀地分布在11条染色体上,均没有内含子结构,编码115-444个氨基酸,等电点4.55-9.69,均为亲水蛋白;所有蛋白均包含完整的OVATE保守结构域,亚细胞定位主要在细胞核中,除VvOFP9外,其余蛋白均含有1-11个核输出信号(Nuclear Export Signal,NES)保守位点,且多数基因仅含有1-2个保守位点。进化树分析表明,葡萄和拟南芥OVATE蛋白家族可聚为六类,其中,Ⅱ、Ⅲ、Ⅴ类仅包含两个基因,Ⅰ、Ⅳ、Ⅵ类中VvOFPs和AtOFPs相互交错地聚类在一起;VvOFPs和AtOFPs共包含10个未知基序,保守元件1和2位于OVATE结构域区域,此外,在OVATE结构域外每个类别均包含特有基序。2葡萄OVATE基因家族具有组织表达特异性,多数VvOFPs在根、嫩茎和花中表达量较高,在嫩叶和果实中仅检测到少数基因表达;VvOFPs在果实的不同发育期表达量存在差异,通常在开花前一周和开花期表达量较高,而花后4周果实中表达量较低。共有2对旁系同源基因(VvOFP1和VvOFP17,VvOFPV和VvOFP15)表达模式相似,4对旁系同源基因(VvOFP6和VvOFP10,VvOFP3和VvOFP7,VvOFP 和VvOFP11,VvOF,和VvOFP8)产生了新的表达模式;此外还发现一些成员的表达模式非常相似,如VvOFP6和VvOFP7在花前1周中的表达量较高,推测其可能含有冗余的功能。3 VvOFP8基因开放阅读框(Open Reading Frame,ORF)长度为 1191bp,编码 396个氨基酸,VvOFP17基因ORF长度为1209bp,编码402个氨基酸,同NCBI上登录序列相比,两条序列中均存在位点突变。生物信息学分析表明,两个蛋白均含有OVATE结构域,二级结构主要有无规则卷曲组成,VvOFP8蛋白定位在细胞质,在OVATE结构域中预测含有1个卷曲螺旋结构,VvOFP17蛋白定位在内质网和细胞核,含有两个核定位信号,在N端含有1个跨膜区。两个蛋白和其他物种OVATE蛋白的进化关系与物种进化规律一致,功能预测表明VvOFP8和VvOFP17蛋白均具有转录调控作用。表达分析结果表明,VvOFP8在两个品种的开花期表达量最高,且圆形果的表达量高于长形果;VvOFP17在圆形果品种花后2周表达量最高,而在长形果品种花期表达量最高,由此推测葡萄VvOFP8和VvOFP17基因的表达差异可能影响葡萄果实的形状。
[Abstract]:OVATE protein family (Ovate Family Protein,OFP is a class of transcriptional inhibitors that regulate plant growth and development. It has been reported in Arabidopsis, tomato and rice plants, and some OFPs regulation of tomato and Arabidopsis fruit shape has been identified. Fruit shape is an important appearance quality of grape. It is of great significance to study the molecular mechanism of fruit shape development for fruit shape breeding. In this study, the OVATE transcription factor family of grape was comprehensively identified and bioinformatics analyzed, and the expression pattern of grape OVATE transcription factor family was investigated by real-time fluorescence relative quantitative PCR technique. Two genes VvOFP8 and VvOFP17, which may regulate fruit shape, were cloned from 'Baoman' grape and analyzed by bioinformatics and their expression characteristics were studied. The main results are as follows: (1) the grape OVATE transcription factor family consists of 17 members, which are unevenly distributed on 11 chromosomes with no intron structure, encoding 115 鈮，
本文编号：2496532
[Abstract]:OVATE protein family (Ovate Family Protein,OFP is a class of transcriptional inhibitors that regulate plant growth and development. It has been reported in Arabidopsis, tomato and rice plants, and some OFPs regulation of tomato and Arabidopsis fruit shape has been identified. Fruit shape is an important appearance quality of grape. It is of great significance to study the molecular mechanism of fruit shape development for fruit shape breeding. In this study, the OVATE transcription factor family of grape was comprehensively identified and bioinformatics analyzed, and the expression pattern of grape OVATE transcription factor family was investigated by real-time fluorescence relative quantitative PCR technique. Two genes VvOFP8 and VvOFP17, which may regulate fruit shape, were cloned from 'Baoman' grape and analyzed by bioinformatics and their expression characteristics were studied. The main results are as follows: (1) the grape OVATE transcription factor family consists of 17 members, which are unevenly distributed on 11 chromosomes with no intron structure, encoding 115 鈮，
本文编号：2496532
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