【作者】 赵巍巍；

【导师】 宗成文； 曹后男；

【作者基本信息】 延边大学 ， 果树学， 2010， 硕士

【摘要】 葡萄是世界上最重要的果树之一,其种植面积和产量都在世界前列。研究花发育的机理对果树栽培、育种工作的发展以及促进果品生产都具有重要的意义。本试验以藤稔葡萄的花序为试材,应用电子克隆技术从葡萄EST数据库中拼接到两个MADS-box基因,并用RT-PCR方法扩增得到了该基因的编码区全长序列,分别命名为VvSVP和VvMADS12。结果如下：VvSVP基因序列编码区全长684 bp,编码227个氨基酸残基,与PtMADS1、MdJOINTLESS、PkMADS、SVP分别有84.1%、81.7%、78.9%和74.9%的同源性。该序列具有MADS-box基因典型的MADS-box和K-box结构域。系统发育分析同样将VvSVP基因归入MADS-box基因家族的STMADS11进化系。电子表达谱分析显示,该基因在葡萄的芽、叶片、花序及果实中表达,在花、花梗、茎、根中不表达,推测该基因与葡萄花序及果实发育有关。将VvSVP mRNA与基因组序列比对发现,VvSVP基因由8个外显子,7个内含子组成,与SVP基因的结构不完全相同。VvMADS12基因序列编码区全长630 bp,编码209个氨基酸残基,与PdMADS-box、AmDEFH7、MODEFH7、SlTDR8、CsMADS-box、PrMADS-box、ZmMADS15分别有70%、63%、62%、60%、66%、45%和50%的同源性。该序列具有MADS-box基因典型的MADS-box和K-box结构域。系统发育分析将VvMADS12基因归入MADS-box基因家族的TM3进化系。分析VvMADS12 mRNA的表达谱,结果表明：VvMADS12基因在葡萄的花序、花梗、茎中表达,在花、果实、叶、根中没有检测到,说明VvMADS12与葡萄花序发育有关。将VvSVP mRNA与基因组序列比对发现,VvSVP基因由8个外显子,7个内含子组成。成功构建了VvSVP和VvMADS12基因的植物双元表达载体。更多还原

【Abstract】 Grape is one of the most important fruit trees in the world, its culture area and total yields are advanced in the world. it is a great significance to study the mechanism of flower development on fruit trees cultivation, breeding and promoting the development of fruit production.A SHORT VEGETATIVE PHASE (SVP) homologue was isolated from Vitis vinifera x V. labrusca’Fujiminori’inflorescence by bioformatic analysis and RT-PCR. It is 684 bp, coding a polypeptide of 227 amino acids, named as VvSVP. Homology analysis showed that the deduced VvSVP protein was highly homologous to the MADS-box genes, PMADS1, MdJOINTLESS、PkMADS、SVP, the identity are 84.1%、81.7%、78.9% and74.9%, respectively. Phylogenetic analysis also indicated that VvSVP belongs to the STMADS11 lineage in MADS-box gene family. Expression pattern analysis showed that VvSVP expressed in buds, leaves, inflorescenes, fruits, but not expressed in flowers, pedicels, stems and roots. Blast of VvSVP with a shotgun genome sequence indicated that VvSVP genome is consist of 8 exons and 7 introns, the number and length of exon is not the same as SVP.Electronic VvMADS12 homologue was isolated from Vitis vinifera×V. labrusca ’Fujiminori’inflorescence by bioformatic analysis and RT-PCR. It is 630 bp, coding a polypeptide of 209 amino acids, named as VvMADS12. Homology analysis showed that the deduced VvMADS12 protein was highly homologous to the MADS-box genes, Pd MADS-box、AmDEFH7、MODEFH7、SlTDR8、CsMADS-box、PrMADS-box、ZmMADS15the identity are 70%、63%、62%、60%、66%、45%、and50%, respectively. Phylogenetic analysis also indicated that VvMADS12 belongs to the TM3 lineage in MADS-box gene family. Expression pattern analysis showed that VvMADS12 expressed in buds, leaves, inflorescenes, fruits, but not expressed in flowers, pedicels, stems and roots. Blast of VvMADS12 with a shotgun genome sequence indicated that VvMADS12 genome is consist of 8 exons and 7 introns.We constructed plant overexpression vector of VvSVP and VvMADS12.更多还原