Stem Coloration in Alfalfa: Anthocyanin Accumulation Patterns and Nutrient Profiles of Red- and Green-Stemmed Variants
2025
Zhengfeng Cao | Jiaqing Li | Chuanjie Wang | Xueyang Min | Zhenwu Wei
Anthocyanins, crucial flavonoids in plants, enhance stress tolerance in alfalfa and are attracting attention due to their antioxidant properties. This study analyzed red- and green-stemmed alfalfa using spectrophotometry, frozen sections, and LC-MS/MS. Anthocyanins were concentrated in stem vascular cambium, with red stems peaking at 61.08 mg g&minus:1 DW during the bud stage. Seven anthocyanidins were identified, with their corresponding aglycones including cyanidin, peonidin, and malvidin. At early flowering, red-stemmed alfalfa contained 35 classes of anthocyanins compared to 17 in green-stemmed varieties, with cyanidin-3-O-glucoside levels significantly higher in red stems (4.423 &mu:g g&minus:1, p <: 0.05). Red-stemmed alfalfa also showed higher contents of acid detergent fiber, crude fat, Cu, Fe, and Zn (p <: 0.05), especially Zn (p <: 0.01). Correlation analysis revealed a strong positive link between cyanidin and crude fat (Spearman&rsquo:s &rho: = 0.93, p <: 0.01) and a significant negative correlation with neutral detergent fiber (&rho: = &minus:0.88, p <: 0.05). Cyanidin and peonidin are associated with stem color differentiation, with cyanidin contributing predominantly to red pigmentation, whereas zinc and crude fat exhibit a synergistic correlation with anthocyanin accumulation. These findings may inform breeding strategies to develop anthocyanin-enriched alfalfa.
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