【摘要】 为探索SWEET基因家族在葡萄果实发育中的表达与功能,以本实验室完成的酿酒葡萄品种赤霞珠Ⅰ期和Ⅲ期果实的转录组数据为基础,筛选出在Ⅰ期和Ⅲ期表达量存在显著差异的9个SWEET基因家族成员,利用生物信息学工具,对这9个基因的基因结构、蛋白的基本理化性质、二级结构、亚细胞定位、保守基序和序列同源性等进行预测分析,并利用实时荧光定量PCR(qPCR)技术对分析结果进行验证。基因组定位结果发现这9个SWEET基因分布在7条染色体上,蛋白序列可分成4个亚族。不同成员间氨基酸数目、氨基酸序列间的疏水性存在一定的差异;二级结构预测结果显示,这9个SWEET基因的氨基酸序列以α-螺旋和无规则卷曲为主要组成部分;基因结构分析表明,除VvSWEET4含有4个内含子,其余8个均含有5个内含子;对9个SWEET基因家族成员蛋白的亚细胞定位预测分析表明:它们大多定位在质膜、叶绿体类囊体膜和液泡膜上;QPCR结果表明,在Ⅰ期和Ⅲ期差异表达的9个SWEET基因家族成员中,5个基因上调表达,4个基因下调表达,与转录组分析结果一致。更多还原

【Abstract】 To explore SWEET gene family expression and function in grape berry,we 9 SWEETs,which were differentially expressed between grape berry development stageⅠ and Ⅲ by RNA-Seq data analyzes of Vitis vinifera L.cv.Cabernet Sauvignon were selected.By using bioinformatics approaches,their gene structure,protein physicochemical properties,secondary structures,subcellular localization,conserved motifs as well as sequence homology predicted and analyzed and real-time fluorescence quantitative PCR was used to validate some genes.Genetic mapping for SWEET genome localization found that they was distributed on seven chromosomes,9 VvSWEETs sequences was divided into four sub-families.Differences existed in the amino acid number and hydrophobic character among the 9 VvSWEETs.Secondary structure prediction revealed that the major composition of the amino acid sequences of the 9 VvSWEETs were alpha helix and random coil.Structural analysis of exons and introns showed that only VvSWEET4 has four introns,others have five introns.Protein subcellular localization analysis indicated that 9VvSWEETs were localized in plasma membrane,chloroplast thylakoid membrane and the vacuole membrane.QPCR results indicated that the 9 SWEETs with differentially expression between grape berry development stage I and III,5 of which were up-regulated,and 4 down-regulated,which were consisting with transcriptome sequencing analysis.更多还原