Stem Coloration in Alfalfa: Anthocyanin Accumulation Patterns and Nutrient Profiles of Red- and Green-Stemmed Variants

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...

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Main Authors: Zhengfeng Cao, Jiaqing Li, Chuanjie Wang, Xueyang Min, Zhenwu Wei
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Agronomy
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Online Access:https://www.mdpi.com/2073-4395/15/4/862
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author Zhengfeng Cao
Jiaqing Li
Chuanjie Wang
Xueyang Min
Zhenwu Wei
author_facet Zhengfeng Cao
Jiaqing Li
Chuanjie Wang
Xueyang Min
Zhenwu Wei
author_sort Zhengfeng Cao
collection DOAJ
description 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<sup>−1</sup> 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-<i>O</i>-glucoside levels significantly higher in red stems (4.423 μg g<sup>−1</sup>, <i>p <</i> 0.05). Red-stemmed alfalfa also showed higher contents of acid detergent fiber, crude fat, Cu, Fe, and Zn (<i>p <</i> 0.05), especially Zn (<i>p <</i> 0.01). Correlation analysis revealed a strong positive link between cyanidin and crude fat (Spearman’s ρ = 0.93, <i>p <</i> 0.01) and a significant negative correlation with neutral detergent fiber (ρ = −0.88, <i>p</i> < 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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spelling doaj-art-8ba386a0fc9f4e83b2a735f5c3fd263b2025-08-20T02:24:42ZengMDPI AGAgronomy2073-43952025-03-0115486210.3390/agronomy15040862Stem Coloration in Alfalfa: Anthocyanin Accumulation Patterns and Nutrient Profiles of Red- and Green-Stemmed VariantsZhengfeng Cao0Jiaqing Li1Chuanjie Wang2Xueyang Min3Zhenwu Wei4College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaCollege of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaCollege of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaCollege of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaCollege of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaAnthocyanins, 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<sup>−1</sup> 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-<i>O</i>-glucoside levels significantly higher in red stems (4.423 μg g<sup>−1</sup>, <i>p <</i> 0.05). Red-stemmed alfalfa also showed higher contents of acid detergent fiber, crude fat, Cu, Fe, and Zn (<i>p <</i> 0.05), especially Zn (<i>p <</i> 0.01). Correlation analysis revealed a strong positive link between cyanidin and crude fat (Spearman’s ρ = 0.93, <i>p <</i> 0.01) and a significant negative correlation with neutral detergent fiber (ρ = −0.88, <i>p</i> < 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.https://www.mdpi.com/2073-4395/15/4/862anthocyaninalfalfacyanidin-3-<i>O</i>-glucosideLC-MS/MS targeted analysisstem colorzinc
spellingShingle Zhengfeng Cao
Jiaqing Li
Chuanjie Wang
Xueyang Min
Zhenwu Wei
Stem Coloration in Alfalfa: Anthocyanin Accumulation Patterns and Nutrient Profiles of Red- and Green-Stemmed Variants
Agronomy
anthocyanin
alfalfa
cyanidin-3-<i>O</i>-glucoside
LC-MS/MS targeted analysis
stem color
zinc
title Stem Coloration in Alfalfa: Anthocyanin Accumulation Patterns and Nutrient Profiles of Red- and Green-Stemmed Variants
title_full Stem Coloration in Alfalfa: Anthocyanin Accumulation Patterns and Nutrient Profiles of Red- and Green-Stemmed Variants
title_fullStr Stem Coloration in Alfalfa: Anthocyanin Accumulation Patterns and Nutrient Profiles of Red- and Green-Stemmed Variants
title_full_unstemmed Stem Coloration in Alfalfa: Anthocyanin Accumulation Patterns and Nutrient Profiles of Red- and Green-Stemmed Variants
title_short Stem Coloration in Alfalfa: Anthocyanin Accumulation Patterns and Nutrient Profiles of Red- and Green-Stemmed Variants
title_sort stem coloration in alfalfa anthocyanin accumulation patterns and nutrient profiles of red and green stemmed variants
topic anthocyanin
alfalfa
cyanidin-3-<i>O</i>-glucoside
LC-MS/MS targeted analysis
stem color
zinc
url https://www.mdpi.com/2073-4395/15/4/862
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AT chuanjiewang stemcolorationinalfalfaanthocyaninaccumulationpatternsandnutrientprofilesofredandgreenstemmedvariants
AT xueyangmin stemcolorationinalfalfaanthocyaninaccumulationpatternsandnutrientprofilesofredandgreenstemmedvariants
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