香椿中3个水通道蛋白基因(TsAQP)的克隆和表达分析

Cloning and expression analysis of three aquaporin genes(TsAQP)in Toona sinensis

水通道蛋白(Aquaporins,AQP)能够促进水分和小分子物质在膜上的运输,维持细胞水分平衡。本研究利用香椿转录组数据,克隆到3个香椿水通道蛋白基因(TsAQP),即TsPIP1-5(GenBank登录号MT501152)、TsPIP2-5(GenBank登录号MT501153)和TsTIP2-1(GenBank登录号MT501154)。利用生物信息学和实时荧光定量PCR(qRT-PCR)技术,结合生理表型分析,研究TsPIP1-5、TsPIP2-5和TsTIP2-1蛋白性质、表达模式和在香椿芽采后贮藏中的功能。结果表明:从转录组数据中鉴定出的20个TsAQP可分为5类;TsPIP1-5、TsPIP2-5和TsTIP2-1是通过跨膜区定位于膜的疏水性稳定蛋白,含磷酸化位点,无信号肽;TsPIP1-5、TsPIP2-5和TsTIP2-1在根、茎和叶中均有表达且有一定的器官差异性;TsPIP1-5、TsPIP2-5和TsTIP2-1在香椿芽低温贮藏过程中表达量提高,暗示TsAQP在香椿芽采后脱水过程中有调控作用。本研究结果为进一步研究香椿中TsAQP基因的功能和作用机制奠定了基础。

Aquaporins(AQP)facilitate the transport of water and small molecules across membranes and maintain water balance of cells.Three aquaporin genes in Toona sinensis(TsAQP),TsPIP1-5(GenBank accession number MT501152),TsPIP2-5(GenBank accession number MT501153),and TsTIP2-1(GenBank accession number MT501154),were cloned based on transcriptome sequencing.Using a combination of bioinformatics and real-time fluorescence quantitative PCR(qRT-PCR),we studied the physiological phenotypes,protein properties,expression patterns,and function of TsPIP1-5,TsPIP2-5 and TsTIP2-1 during post harvest storage of Toona sinensis buds.The results indicated that 20 TsAQP proteins identified from the transcriptome data could be classified into five subfamilies.TsPIP1-5,TsPIP2-5 and TsTIP2-1 were hydrophobic stable proteins located on the membrane through the transmembrane region,containing phosphorylation sites and no signal peptide.Different expression levels of TsPIP1-5,TsPIP2-5,and TsTIP2-1 were detected in the roots,stems and leaves with certain organ differences.The expression levels of TsPIP1-5,TsPIP2-5,and TsTIP2-1 increased during cold storage of Toona sinensis buds,suggesting that they played a regulatory role in the process of post harvest dehydration.The results of this study lay a foundation for further research on function and regulatory mechanisms of TsAQPs in Toona sinensis.