Institute of Food Science and Technology, Fujian Academy of Agricultural Sciences, Wusi Road 247, Fuzhou, Fujian, 350003, P.R. China.
Key Laboratory of Subtropical Characteristic Fruits, Vegetables and Edible Fungi Processing (Co-construction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Fuzhou, Fujian, 350003, China.
BMC Plant Biol. 2024 Oct 10;24(1):944. doi: 10.1186/s12870-024-05667-4.
Spine grape (Vitis davidii) is a promising source of high-quality anthocyanins, with vast potential for application in food, pharmaceutical, and cosmetic industries. However, their availability is limited by resource constraints. Plant cell culture has emerged as a valuable approach for anthocyanin production and serves as an ideal model to investigate the regulation of anthocyanin biosynthesis. Elicitors are employed to achieve targeted enhancement of anthocyanin biosynthesis. The present study investigated the impact of 5-aminolevulinic acid (ALA) as an elicitor on the accumulation of anthocyanins and flavonoids during spine grape callus growth. Specifically, we examined the effects of ALA on anthocyanin and its component accumulation in callus, and biosynthetic anthocyanin gene expression.
ALA at 25 µg/L increased the biomass of spine grape callus. ALA induction enhanced the levels of flavonoids, anthocyanins and proanthocyanidins in callus, with maximum values reaching 911.11 mg/100 g DW, 604.60 mg/100 g DW, and 5357.00 mg/100 g DW, respectively, after callus culture for 45 days. Notably, those levels were 1.47-, 1.93- and 1.83-fold higher than controls. ALA induction modulated the flavonoid profile, and among 97 differential flavonoid metabolites differing from controls, 77 were upregulated and 20 were downregulated. Six kinds of anthocyanins, namely cyanidin (8), delphinidin (6), peonidin (5), malvidin (4), petunidin (3) and pelargonidin (3), were detected in callus, with peonidin most abundant. Compared with controls, anthocyanin components were increased in ALA-treated callus. The key genes PAL1, PAL2, PAL4, CHI, CHS3, F3'H, F3H, FLS, DFR, UFGT, MYBA1, LDOX, OMT3, GT1 and ACT involved in anthocyanin biosynthesis were upregulated following ALA treatment, resulting in anthocyanin accumulation.
This study revealed a novel mode of ALA-mediated promotion of plant anthocyanin biosynthesis and accumulation at the cellular level, and a strategy for enhancing anthocyanin content in spine grape callus. The findings advance commercial-scale production of anthocyanins via spine grape callus culture. we also explored the accumulation patterns of flavonoids and anthocyanins under ALA treatment. Augmentation of anthocyanins coincided with elevated expression levels of most genes involved in anthocyanin biosynthesis within spine grape callus following ALA treatment.
刺葡萄（Vitis davidii）是一种富含高质量花色苷的有前途的资源，在食品、制药和化妆品行业具有广泛的应用潜力。然而，其可用性受到资源限制的限制。植物细胞培养已成为生产花色苷的一种有价值的方法，并可作为研究花色苷生物合成调控的理想模型。诱导子用于实现花色苷生物合成的靶向增强。本研究调查了 5-氨基乙酰丙酸（ALA）作为诱导子对刺葡萄愈伤组织生长过程中花色苷和类黄酮积累的影响。具体而言，我们研究了 ALA 对愈伤组织中花色苷及其成分积累以及生物合成花色苷基因表达的影响。
25μg/L 的 ALA 增加了刺葡萄愈伤组织的生物量。ALA 诱导增强了愈伤组织中类黄酮、花色苷和原花色素的水平，在愈伤组织培养 45 天后，分别达到 911.11、604.60 和 5357.00mg/100g DW，分别比对照高 1.47、1.93 和 1.83 倍。ALA 诱导调节了类黄酮谱，与对照相比，97 种差异表达的类黄酮代谢物中有 77 种上调，20 种下调。在愈伤组织中检测到 6 种花色苷，即矢车菊素（8）、天竺葵素（6）、芍药素（5）、飞燕草素（4）、锦葵素（3）和锦葵素（3），其中飞燕草素含量最丰富。与对照相比，ALA 处理的愈伤组织中花色苷成分增加。参与花色苷生物合成的关键基因 PAL1、PAL2、PAL4、CHI、CHS3、F3'H、F3H、FLS、DFR、UFGT、MYBA1、LDOX、OMT3、GT1 和 ACT 被上调，导致花色苷积累。
本研究揭示了 ALA 介导的植物花色苷生物合成和积累的新机制，以及在刺葡萄愈伤组织中提高花色苷含量的策略。该研究推进了通过刺葡萄愈伤组织培养进行商业规模生产花色苷的进程。我们还探索了 ALA 处理下花色苷和花色苷的积累模式。在 ALA 处理后，刺葡萄愈伤组织中大多数参与花色苷生物合成的基因的表达水平升高，与花色苷的积累相一致。
