葡萄SVP类MADS-box基因的克隆及表达分析

摘要

从无核白葡萄(Vitis vinifera cv.Thompson seedless)中克隆获得一个胚珠发育相关MADS-box基因.该基因cDNA全长1248bp,ORF为729 bp,编码一个合有243个氨基酸的蛋白.氨基酸多序列比对和系统发育分析显示,该基因属于SVP类MADS-box基因.采用半定量技术进行表达分析表明,该基因在葡萄的根、茎、叶、花蕾、盛花和胚珠中均有表达,但是有强弱差异.实时定量PCR技术分析表明在无核葡萄品种无核白胚珠发育过程中基因表达量先升后降,而在有核葡萄品种黑比诺胚珠发育过程中表达量一直呈下降趋势.%A MADS-box gene associated with ovule development was cloned from Vitis. vinifera cv. Thompson seedless. The full-length of cDNA gene was 1 248 bp, including a 729 bp open reading frame, and encoding 243 amino acids. The homology and phylogenetic analysis showed that this gene belonged to SVP-like MADS-box genes. The gene was expressed in all of the grape tissue type examined by semi-quantity RT-PCR, including root, stem, leaf, bud. full bloom and ovules, but varied widely in the level of expression. Real-time PCR analysis revealed that gene expression level during ovule development in Thompson increased firstly and then decreased. In contrast to this, gene expression level in Pinot showed downward trend in all stages of development.