Salt stress is a major abiotic factor limiting wolfberry (
Lycium barbarum) growth. As a high-value medicinal and edible crop, wolfberry relies on its carotenoid content, a critical determinant of fruit quality and nutritional value. To elucidate the expression regulatory mechanisms of
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Salt stress is a major abiotic factor limiting wolfberry (
Lycium barbarum) growth. As a high-value medicinal and edible crop, wolfberry relies on its carotenoid content, a critical determinant of fruit quality and nutritional value. To elucidate the expression regulatory mechanisms of key genes in the carotenoid biosynthesis pathway under salt stress, this study systematically identified 17 structural genes within the
L. barbarum carotenoid pathway using genomic and transcriptomic approaches. Comprehensive analyses were conducted on gene structure, chromosomal distribution, conserved domains, and
cis-acting elements. The results revealed that these genes were clustered on chromosomes Chr08 and Chr10 and exhibit strong collinearity with tomato (18 syntenic pairs). Their promoters were enriched with light-responsive (G-box) and stress-responsive (ABRE, DRE) elements. Tissue-specific expression analysis demonstrated high expression in mid-to-late fruit developmental stages (
LbaPSY1,
LbaPDS) and in photoprotective genes (
LbaZEP,
LbaVDE) in leaves. Under 300 mM NaCl stress treatment, the genes exhibited a staged response: Early stage (1–3 h): upstream MEP pathway genes (
LbaDXS,
LbaGGPS) were rapidly induced to supply precursors. Mid-stage (6–12 h): midstream genes (
LbaPSY,
LbaPDS,
LbaZDS) were continuously upregulated, promoting lycopene synthesis and preferentially activating the β-branch (
LbaLCYB). Late stage (12–24 h): downstream xanthophyll cycle genes (
LbaBCH,
LbaZEP,
LbaVDE) were significantly enhanced, facilitating the accumulation of antioxidant compounds like violaxanthin and neoxanthin. This coordinated regulation formed a synergistic “precursor supply–antioxidant product” network. This study revealed the phased and coordinated regulatory network of carotenoid biosynthesis genes under salt stress in
L. barbarum. It also provided potential target genes for the new cultivar selection with enhanced salt tolerance and nutritional quality.
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