【摘要】：OVATE蛋白家族(Ovate Family Protein,OFP)是一類調(diào)控植物生長發(fā)育的轉(zhuǎn)錄抑制因子,在擬南芥、番茄和水稻等植物中均有研究報道,并鑒定出一些OFPs調(diào)控番茄、擬南芥果實形狀。果實形狀是葡萄一項重要的外觀品質(zhì),研究果形發(fā)育的分子機理對果形選育具有重要意義。本研究對葡萄OVATE轉(zhuǎn)錄因子家族進行了全面的鑒定和生物信息學(xué)分析,利用實時熒光相對定量PCR技術(shù)探究了該家族成員的表達模式,從'寶滿'葡萄中克隆得到兩個可能調(diào)控果形的相關(guān)基因VvOFP8和VvOFP17,并進行生物信息學(xué)分析和表達特性研究。主要研究結(jié)果如下:1葡萄OVATE轉(zhuǎn)錄因子家族包含17個成員,不均勻地分布在11條染色體上,均沒有內(nèi)含子結(jié)構(gòu),編碼115-444個氨基酸,等電點4.55-9.69,均為親水蛋白;所有蛋白均包含完整的OVATE保守結(jié)構(gòu)域,亞細胞定位主要在細胞核中,除VvOFP9外,其余蛋白均含有1-11個核輸出信號(Nuclear Export Signal,NES)保守位點,且多數(shù)基因僅含有1-2個保守位點。進化樹分析表明,葡萄和擬南芥OVATE蛋白家族可聚為六類,其中,Ⅱ、Ⅲ、Ⅴ類僅包含兩個基因,Ⅰ、Ⅳ、Ⅵ類中VvOFPs和AtOFPs相互交錯地聚類在一起;VvOFPs和AtOFPs共包含10個未知基序,保守元件1和2位于OVATE結(jié)構(gòu)域區(qū)域,此外,在OVATE結(jié)構(gòu)域外每個類別均包含特有基序。2葡萄OVATE基因家族具有組織表達特異性,多數(shù)VvOFPs在根、嫩莖和花中表達量較高,在嫩葉和果實中僅檢測到少數(shù)基因表達;VvOFPs在果實的不同發(fā)育期表達量存在差異,通常在開花前一周和開花期表達量較高,而花后4周果實中表達量較低。共有2對旁系同源基因(VvOFP1和VvOFP17,VvOFPV和VvOFP15)表達模式相似,4對旁系同源基因(VvOFP6和VvOFP10,VvOFP3和VvOFP7,VvOFP 和VvOFP11,VvOF,和VvOFP8)產(chǎn)生了新的表達模式;此外還發(fā)現(xiàn)一些成員的表達模式非常相似,如VvOFP6和VvOFP7在花前1周中的表達量較高,推測其可能含有冗余的功能。3 VvOFP8基因開放閱讀框(Open Reading Frame,ORF)長度為 1191bp,編碼 396個氨基酸,VvOFP17基因ORF長度為1209bp,編碼402個氨基酸,同NCBI上登錄序列相比,兩條序列中均存在位點突變。生物信息學(xué)分析表明,兩個蛋白均含有OVATE結(jié)構(gòu)域,二級結(jié)構(gòu)主要有無規(guī)則卷曲組成,VvOFP8蛋白定位在細胞質(zhì),在OVATE結(jié)構(gòu)域中預(yù)測含有1個卷曲螺旋結(jié)構(gòu),VvOFP17蛋白定位在內(nèi)質(zhì)網(wǎng)和細胞核,含有兩個核定位信號,在N端含有1個跨膜區(qū)。兩個蛋白和其他物種OVATE蛋白的進化關(guān)系與物種進化規(guī)律一致,功能預(yù)測表明VvOFP8和VvOFP17蛋白均具有轉(zhuǎn)錄調(diào)控作用。表達分析結(jié)果表明,VvOFP8在兩個品種的開花期表達量最高,且圓形果的表達量高于長形果;VvOFP17在圓形果品種花后2周表達量最高,而在長形果品種花期表達量最高,由此推測葡萄VvOFP8和VvOFP17基因的表達差異可能影響葡萄果實的形狀。
[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，

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