This review systematically elucidated the complete "root-to-shoot" metabolic pathway regulated by key genes and transporters of L-theanine, and summarized stress-adaptive mechanisms where L-theanine serves as both nitrogen reservoir and stress protectant, and further constructed a comprehensive analytical for L-theanine-derived flavor compound formation during tea processing. By bridging fundamental discoveries in plant physiology with practical applications in food chemistry, this work provides new insights into innovative strategies for developing climate-resilient tea varieties through molecular breeding and optimizing processing technologies to enhance product quality.