Functional analysis of the nitrogen metabolism-related gene CsGS1 in cucumber

Abstract Glutamine synthetase (GS) plays an important role in nitrogen (N) metabolism in cucumber. In this study, we cloned and sequenced the CsGS1 gene, and analyzed the expression patterns and subcellular localization of the GS1 protein in response to different N conditions in order to determine its role in low-nitrogen (LN) tolerance. CsGS1 was abundantly expressed in the leaves of the low N-requiring cultivar D0328, while the high N-requiring cultivar D0422 showed similar expression levels across different tissues including leaves, shoots and roots. Furthermore, the GS1 protein was primarily localized in the cytoplasm of plant cells. Both cultivars were then transformed with the CsGS1 coding sequence or antisense sequence via Agrobacterium tumefaciens in order to overexpress and silence GS1 expression, respectively. Overexpression of CsGS1 significantly improved LN tolerance and photosynthetic parameters, and increased chlorophyll b content, biomass, plant height, root length, N accumulation and GS activity under LN condition compared to the control. CsGS1 silencing on the other hand significantly reduced the above indices. Taken together, CsGS1 is crucial for maintaining N metabolism in cucumber plants during N deprivation, and is a promising target for generating novel transgenic breeds with increasing nitrogen utilization efficiency.