Isolation and characterization of chloroplastic glutamine synthetase gene (CsGS2) in tea plant Camellia sinensis

Abstract

Tea plant (Camellia sinensis) is an ammonium preferring plant species. However, little is known about the mechanism underlying this preference. Herein, a chloroplastic glutamine synthetase gene (CsGS2), which is vital for nitrogen assimilation in mesophyll tissue, was isolated from tea cultivar C. sinensis cv. ‘Longjing43’. The full length cDNA of CsGS2 was 1622 bp, having a 1299 bp open reading frame encoding a 432-amino acid protein. Homology search and sequence analysis demonstrated that CsGS2 protein carried the basic characteristics of a canonical GS2 domain and shared high identity with GS2s from other plant species. Subcellular localization and immunolocalization of CsGS2 revealed that it is localized in chloroplast. qRT-PCR and Western blot analyses showed that CsGS2 was expressed in a leaf-specific pattern, such that both CsGS2 and its protein were most abundant in mature leaves. Temporal expression patterns of CsGS2 showed minor differences in response to ammonium and nitrate nutrition. The transcript level of CsGS2 was significantly induced in mature leaves during the development of new shoots, whereas darkness inhibited this induction significantly. These results suggested that CsGS2 does not play a role in the differential utilization mechanisms of differing nitrogen forms in tea, and imply a light dependent transcription regulation in mature leaves during the development of new shoots.