Cloning,subcellular localization and expression analysis of aquaporin LoPIP2;6 gene of lily anther
In order to further explore the role of members of the PIP2 subfamily of aquaporins in anther development and dehiscence,lily anthers were taken as experimental materials.The LoPIP2;6 gene was obtained from lily anthers using homologous cloning method.Protein structure and evolutionary tree analysis were conducted using DNAMAN and MEGA software.Subcellular localization was detected using tobacco transient expression,and gene expression was detected using real-time fluorescence quantitative PCR.The results were as follows:The open reading frame of LoPIP2;6 gene contained 849 bp encoding 282 amino acids.The LoPIP2;6 was a hydrophobic protein that contained 6 typical transmembrane helical domains and 2 NPA motifs.LoPIP2;6 protein had closer relationship with Phalaenopsis equestris and Dendrobium catenatum,and the protein was localized to the plasma membrane and endoplasmic reticulum.The LoPIP2;6 gene displayed the highest expression level at the tetrad stage,followed by the independent microspore stage.It was speculated that LoPIP2;6 might play a role in the cell growth during the early stage of anther development,as well as in the water transfer of anther connective tissue and crack zone cells during the late stage.In conclusion,this study provides a theoretical reference for further exploring the regulation of lily anther development and dehiscence by aquaporins.