AsGAD1 cloned from creeping bentgrass modulates cadmium tolerance of Arabidopsis thaliana by remodelling membrane lipids and cadmium uptake, transport and chelation

The gene GAD1 encodes a glutamate decarboxylase, which is a rate-limiting enzyme for the biosynthesis of endogenous gamma-aminobutyrate acid (GABA), but a potential role of GAD1 in regulating cadmium (Cd) tolerance needs to be further elucidated in plants. The objective of this study was to investigate Cd tolerance of creeping bentgrass (Agrostis stolonifera) and transgenic yeast (Saccharomyces cerevisiae) or Arabidopsis thaliana overexpressing AsGAD1. The Cd-tolerant creeping bentgrass cultivar LOFTSL-93 accumulated more endogenous GABA in relation to a significant upregulation of AsGAD1 in leaf and root than the Cd-sensitive W66569 in response to Cd stress. The overexpression of AsGAD1 significantly enhanced Cd tolerance of yeast or A. thaliana associated with improved endogenous GABA content, low oxidative damage, and high cell membrane stability and photochemical efficiency. Compared with wild type, AsGAD1-overexpressing plants or the atgad1 mutant maintained significantly lower or higher Cd content in leaf and root by down-regulating or up-regulating transcript levels of AtNRAMP1/2/3/4/5 and AtZIP1/2, respectively. Moreover, overexpression of AsGAD1 significantly up-regulated transcript levels of AtHMA1/3, contributing to better Cd compartmentalization from chloroplast into cytoplasm and then into vacuoles. AsGAD1 overexpression also induced expressions of AsMT1A/1B/1C/2/3, AsGSH1/2, and AsPCS1/2, indicating better capacity of Cd chelation in cytosol and vacuoles for Cd detoxification. Hence, AsGAD1-regulated detoxification mechanism of Cd could be related to Cd uptake, transport, and chelation. In addition, lipid contents (PC, PG, and DGDG) and the DGDG/MGDG and PC/PG ratios were improved by the AsGAD1 overexpression, which favors membrane stability and functionality under Cd stress. These findings provide new insight into the regulatory role of GAD1 in Cd tolerance in plants.