Combining GWAS and RNA-seq approaches identifies the FtADH1 gene for drought resistance in Tartary buckwheat

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Drought is a major environmental constraint that significantly affects seedling emergence, yield, and quality of Tartary buckwheat, thereby hindering the development of its industry. However, the molecular mechanisms underlying drought tolerance genes in Tartary buckwheat remain largely unexplored. Alcohol dehydrogenase (ADH), an essential plant protein, plays a crucial role in growth, development, and stress responses; however, its specific role in drought resistance remains unclear. This study identifies an ADH gene, FtADH1, using a membership function value of drought tolerance (MFVD) combined with a genome-wide association study (GWAS) and transcriptomic profiles that confer drought tolerance in Tartary buckwheat. Our findings demonstrated that the overexpression of FtADH1 in Arabidopsis and Tartary buckwheat hairy roots enhances drought tolerance by promoting root elongation and mitigating elevated levels of reactive oxygen species (ROS). Our findings demonstrate that FtADH1 can enhance drought tolerance in Tartary buckwheat and Arabidopsis. This study identifies FtADH1 as a new regulator of Tartary buckwheat’s ROS levels and stress responses, functioning by regulating protective enzyme activities at a high level to scavenge ROS and modulating root growth under drought stress. Further, it identifies proteins interacting with FtADH1 through a prokaryotic expression pull-down assay combined with mass spectrometry, revealing that FtADH1 interacts explicitly with the S-adenosyl-L-methionine (SAM) synthetase protein, FtSAMS1. Overexpression of FtSAMS1 enhances ADH enzymatic activity, leading to increased SAM content in overexpressing Tartary buckwheat hairy roots under water-deficit conditions. Additionally, overexpression of FtSAMS1 induces a drought-resistant phenotype in Arabidopsis and Tartary buckwheat hairy roots under drought stress, revealing the biological function of FtADH1. Evolutionary analysis indicates that ADH1 in Fagopyrum species has undergone significant evolutionary events, including duplication and purifying selection, which may contribute to functional diversification and adaptive advantages such as drought resistance in cultivated buckwheat. In summary, this study suggests that FtADH1 is a key contributor to drought tolerance, and its interaction with FtSAMS1 offers promising potential for developing drought-resistant varieties in Tartary buckwheat and its relative species.

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