葡萄TPS基因家族的鉴定与表达分析

Identification and Expression Analysis of TPS Genes Family in Grape

为了明确TPS基因家族在葡萄非生物胁迫下的表达特性,在生物信息分析的基础上,以‘鄞红’葡萄(Vitis vinifera‘Yinhong’)扦插苗为试验材料,利用RT-qPCR技术检测了盐胁迫和光胁迫处理对葡萄TPS家族基因成员表达的影响。结果显示,葡萄TPS基因家族有9个成员,可分为3个亚族,分布在8条染色体上。通过氨基酸多重序列比对,发现葡萄(Vitis vinifera)、拟南芥(Arabidopsis thaliana)和水稻(Oryza sativa)TPS家族基因都存在G-X-FLH-X-PFPS-X-E-X5-P-X-R和X-F-X-T-X-D-X-ARHF-X2的特征序列。motif分析发现,VvTPS06和VvTPS07两个基因含有相同的氨基酸保守序列。亚细胞定位预测显示葡萄TPS基因主要分布在细胞质、叶绿体和细胞核中,其中VvTPS06、VvTPS07、VvTPS09定位于叶绿体中可能与植物光合作用相关。蛋白质二级结构表明,葡萄TPS蛋白主要以α-螺旋和无规则卷曲为主。基因芯片表达显示,VvTPS02、VvTPS03、VvTPS05、VvTPS07在葡萄不同时期不同组织均处于高表达状态,其中VvTPS07尤为明显。RT-qPCR分析结果表明,在盐胁迫中各VvTPS基因存在相似的表达模式,VvTPS05、VvTPS07和VvTPS09的表达量在高浓度盐胁迫(0.6%)时出现一定程度的升高,这表明VvTPS05、VvTPS07和VvTPS09基因与植物耐盐性关系密切,可为葡萄逆境胁迫机制研究提供参考。在光胁迫中,当遮光率达90%时,VvTPS02、VvTPS07和VvTPS09的表达量相比于对照组明显增加,推测其通过提高其表达量来促进光合作用,从而弥补光照的缺失。

In order to clarify the expression characteristics of TPS gene family in grape under abiotic stress, based on the analysis of biological information, the cuttings of Vitis vinifera 'Yinhong' grape were taken as the experimental material. The effects of salt and light stress on the expression of TPS family genes in grape were examined by RT-qPCR. The results showed that there were 9 members of the TPS gene family, which could be divided into3 subfamilies and distributed on 8 chromosomes. Multiple sequence alignments revealed that Vitis vinifera, Arabidopsis thaliana and Oryza sativa had a characteristic sequence(G-X-FLH-X-PFPS-X-E-X5-P-X-R and X-F-X-TX-D-X-ARHF-X2). Motif analysis showed that VvTPS06 and VvTPS07 genes contained the same conserved amino acid sequence. Subcellular localization predicted that grape TPS genes were mainly distributed in cytoplasm,chloroplast and nucleus. VvTPS06, VvTPS07 and VvTPS09 were distributed in chloroplast, we speculated that they may be related to photosynthesis. The protein secondary structure showed that grape TPS protein mainly consisted of alpha helix and random coil. The microarray data showed that VvTPS02, VvTPS03, VvTPS05 and VvTPS07 all have a high expression in different periods and tissues, especially VvTPS07. The results of RT-qPCR analysis showed that the expression patterns of VvTPS genes were similar under salt stress, and the expression levels of VvTPS05,VvTPS07 and VvTPS09 were increased to a certain extent under high salt stress(0.6%). This showed that VvTPS05,VvTPS07 and VvTPS09 were bound up with plant salt tolerance, and it would provide a reference for the research on abiotic stresses to grape. In shading stress, when shading rate was 90%, the expression of VvTPS02, VvTPS07 and VvTPS09 were increased significantly compared with control group. It is speculated that the expression levels of VvTPS02, VvTPS07 and VvTPS09 were increased to promote photosynthesis and make up for the lack of light.