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| Study on the Regulation of Anthocyanin Accumulation in Lycoris albiflora by Transcription Factor LaMYB56 |
| SUN Ming-Xia, ZHANG Lin-Jing, LIN Xin-Yu, GAO Yan-Hui* |
| National Key Laboratory for the Development and Utilization of Forest Food Resources/Zhejiang Key Laboratory of Forest Genetics and Breeding, Zhejiang A& F University, Hangzhou 311300, China |
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Abstract The new variety Lycoris albiflora 'Astro Girl' with a unique color and a temporal variation pattern, has extremely high ornamental value and is an ideal material for studying the mechanism of plant color gradient. To explore flower color formation mechanism in L. albiflora, R2R3-MYB transcription factor LaMYB56 gene associated with anthocyanin accumulation was cloned by RT-PCR. The regulation of anthocyanin accumulation was validated by molecular biology techniques. The results showed that the cDNA sequence of LaMYB56 with length of 1 031 bp and 723 bp ORF was obtained. LaMYB56 protein contains 2 conserved SANT domains at N-terminus. Phylogenetic analysis indicated that LaMYB56 was grouped with S20 subfamily in Arabidopsis thaliana. The expression level of LaMYB56 peaked on the first flowering day and gradually declined over time. The expression of LrF3'H, LrANS, LrGST genes were significantly decreased after silencing of LaMYB56 in the petals of L. radiata (P<0.05). Overexpression of LaMYB56 in Arabidopsis significantly increased the expression levels of AtUFGT and AtF3'H (P<0.05), leading to elevated anthocyanin content. The dual luciferase assay report and yeast one hybrid result indicated that LaMYB56 could bind to the promoter region of LaGST7 to regulate its expression and affect anthocyanin accumulation. The results demonstrated that LaMYB56 transcription factor was involved in the anthocyanins accumulation in L. albiflora. This study provides a theoretical found for the genetic improvement and molecular breeding of flower color in Lycoris genus and ornamental plants.
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Received: 01 September 2025
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Corresponding Authors:
*gaoyanhui408@126.com
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[1] 高燕会, 黄春红, 朱玉球, 等. 2012. 植物花青素苷生物合成及调控的研究进展[J]. 中国生物工程杂志, 32(8):94-99. (Gao Y H, Huang C H, Zhu Y Q, et al.2012. Progress on plant anthocyanin biosynthesis and regulation[J]. China Biotechnology, 32(8):94-99.) [2] 蒋婷婷, 高燕会, 童再康. 2015. 石蒜属植物实时荧光定量 PCR 内参基因的选择[J]. 园艺学报, 42(6):1129-1138. (Jiang T T, Gao Y H, Tong Z K.2015. Selection of reference genes for quantitative real-time PCR in Lycoris[J]. Acta Horticulturae Sinica, 42(6):1129-1138.) [3] 任毅, 符木, 高亦珂. 2013. 石蒜属植物育种研究进展[J]. 中国园艺文摘, 29(10):15-19. (Ren Y, Fu M, Gao Y K.2013. A review of the breeding research in genus Lycoris[J]. Chinese Horticulture Abstracts, 29(10):15-19.) [4] 萨日娜, 蔡军火, 魏绪英, 等. 2025. 红花石蒜种子脱水过程中的形态与生理变化[J]. 浙江农林大学学报, 42(3):572-580. (Sa R N, Cai J H, Wei X Y, et al.2025. Morphological and physiological changes during the dehydration process of Lycoris radiata seeds[J]. Journal of Zhejiang A&F University, 42(3):572-580.) [5] 王江昱, 田淑婷, 张涵, 等. 2022. 单子叶观赏植物花青素苷呈色机制研究进展[J]. 扬州大学学报 (农业与生命科学版), 43(5):101-114. (Wang J Y, Tian S T, Zhang H, et al.2022. Coloring mechanism of anthocyanin in monocot ornamental plants[J]. Journal of Yangzhou University (Agricultural and Life Science Edition), 43(5):101-114.) [6] 薛惠敏, 周洋丽, 高燕会. 2022. 换锦花花青素合成酶基因(LsANS)的克隆及启动子功能分析[J]. 农业生物技术学报, 30(8):1468-1479. (Xue H M, Zhou Y L, Gao Y H.2022. Cloning and promoter function analysis of the anthocyanin synthase gene (LsANS) in Lycoris sprengeri[J]. Journal of Agricultural Biotechnology, 30(8):1468-1479.) [7] 殷丽青, 邵雅东, 李青竹, 等. 2021. 石蒜属植物组织培养及其植物生长调节剂应用的研究进展[J]. 上海农业学报, 37(4):147-154. (Yin L Q, Shao Y D, Li Q Z, et al.2021. Research progress on tissue culture and application of plant growth regulators in Lycoris[J]. Acta Agriculturae Shanghai, 37(4):147-154.) [8] 于冰, 田烨, 李海英, 等. 2019. 植物 bHLH 转录因子的研究进展[J]. 中国农学通报, 35(9):75-80. (Yu B, Tian Y, Li H Y, et al.2019. Research progress of plant bHLH transcription factor[J]. Chinese Agricultural Science Bulletin, 35(9):75-80.) [9] 张中华, 张会灵, 张菊平, 等. 2024. MYB 基因调控植物花青素合成的研究进展[J]. 分子植物育种, 22(7):2184-2193. (Zhang Z H, Zhang H L, Zhang J P, et al.2024. Recent advances of MYB gene regulating the anthocyanin synthesis in plants[J]. Molecular Plant Breeding, 22(7):2184-2193.) [10] 赵梦婧, 殷素雅, 高燕会. 2025. 石蒜属植物种间杂交种染色体制片体系优化和细胞遗传学分析[J]. 浙江农林大学学报, 42(3):581-591. (Zhao M J, Yin S Y, Gao Y H.2025. Chromosome preparation system optimization and cytogenetic analysis of interspecific hybrids of Lycoris[J]. Journal of Zhejiang A&F University, 42(3):581-591.) [11] 郑正权, 赵梦婧, 高燕会. 2024. 换锦花 LsMYB7 基因克隆与功能研究[J]. 浙江农林大学学报, 41(3):586-596. (Zheng Z Q, Zhao M J, Gao Y H.2024. Cloning and function analysis of LsMYB7 gene in Lycoris sprengeri[J]. Journal of Zhejiang A&F University, 41(3):586-596.) [12] 中国科学院中国植物志编辑委员会. 1985. 中国植物志[M]. 北京:科学出版社, pp. 209-217. (The Editorial Committee of Flora Reipublicae Popularis Sinicae of Chinese Academy of Sciences. 1985. Flora Reipublicae Popularis Sinicae[M]. Science Press, Beijing, China, pp. 209-217.) [13] 周洋丽, 侯朔, 郑正权, 等. 2020. 基于 VIGS 基因沉默体系的换锦花 LsMYBs 基因功能研究[J]. 农业生物技术学报, 28(6):974-983. (Zhou Y L, Hou S, Zheng Z Q, et al.2020. Study on LsMYBs gene function in Lycoris sprengeri based on VIGS gene silencing system[J]. Journal of Agricultural Biotechnology, 28(6):974-983.) [14] Anwar M, Wang G, Wu J, et al.2018. Ectopic overexpression of a novel R2R3-MYB, NtMYB2 from Chinese Narcissus represses anthocyanin biosynthesis in tobacco[J]. Molecules, 23(4):781. [15] Borevitz J O, Xia Y, Blount J, et al.2000. Activation tagging identifies a conserved MYB regulator of phenylpropanoid biosynthesis[J]. The Plant Cell, 12(12):2383-2393. [16] Cabrita L, Fossen T, Andersen Ø M.2000. Colour and stability of the six common anthocyanidin 3-glucosides in aqueous solution[J]. Food Chemistry, 68(1):101-107. [17] Chen P, Takatsuka H, Takahashi N, et al.2017. Arabidopsis R1R2R3-Myb proteins are essential for inhibiting cell division in response to DNA damage[J]. Nature Communications, 8(1):635-647. [18] Fang S, Lin M, Ali M M, et al.2023. LhANS-rr1, LhDFR, and LhMYB114 regulate anthocyanin biosynthesis in flower buds of Lilium 'Siberia'[J]. Genes, 14(3):559. [19] Gerats A G M, Vrijlandt E, Wallroth M, et al.1985. The influence of the genes an1, an2, and an4 on the activity of the enzyme UDP-glucose:Flavonoid 3-o-glucosyltransferase in flowers of Petunia hybrida[J]. Biochemical Genetics, 23(7-8):591-602. [20] Gonzalez A, Zhao M, Leavitt J M, et al.2008. Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings[J]. The Plant Journal, 53(5):814-827. [21] Grotewold E.2006. The genetics and biochemistry of floral pigments[J]. Annual Review of Plant Biology, 57(1):761-780. [22] Lai Y S, Shimoyamada Y, Nakayama M, et al.2012. Pigment accumulation and transcription of LhMYB12 and anthocyanin biosynthesis genes during flower development in the asiatic hybrid lily (Lilium spp.)[J]. Plant Science, 193-194:14-24. [23] Li A, Du Z, Liao M, et al.2019. Discovery and characterisation of lycorine-type alkaloids in Lycoris spp (Amaryllidaceae) using UHPLC-QTOF-MS[J]. Phytochemical Analysis, 30(3):268-277. [24] Liu X F, Feng C, Zhang M M, et al.2013. The MrWD40-1 gene of Chinese bayberry (Myrica rubra) interacts with MYB and bHLH to enhance anthocyanin accumulation[J]. Plant Molecular Biology Reporter, 31(6):1474-1484. [25] Lotkowska M E, Tohge T, Fernie A R, et al.2015. The Arabidopsis transcription factor MYB112 promotes anthocyanin formation during salinity and under high light stress[J]. Plant Physiology, 169(3):1862-1880. [26] Nishihara M, Nakatsuka T, Yamamura S.2005. Flavonoid components and flower color change in transgenic tobacco plants by suppression of chalcone isomerase gene[J]. FEBS Letters, 579(27):6074-6078. [27] Ren Z, Wang J, Huang N, et al., 2026. Meiosis in bulbous flower species Lycoris:Dances underground[J]. Frontiers in Plant Science, 16:1691599. [28] Sun X T, Zhang D Y, Guo R F, et al.2024. Molecular insights into TT2-type MYB regulators illuminate the complexity of floral flavonoids biosynthesis in Freesia hybrida[J]. Horticulture Research, 11:uhae352. [29] Tanaka Y, Sasaki N, Ohmiya A.2008. Biosynthesis of plant pigments:Anthocyanins, betalains and carotenoids[J]. The Plant Journal, 54(4):733-749. [30] This P, Lacombe T, Cadle-Davidson M, et al.2007. Wine grape (Vitis vinifera L.) color associates with allelic variation in the domestication gene VvmybA1[J]. Theoretical & Applied Genetics, 114(4):723-730. [31] Wang X X, Dong J J, HuY T, et al.2024. Identification and characterization of the glutathione S-transferase gene family in blueberry (Vaccinium corymbosum) and their potential roles in anthocyanin intracellular transportation[J]. Plants, 13(10):1316. [32] Xie S, Lei Y, Chen H, et al.2020. R2R3-MYB transcription factors regulate anthocyanin biosynthesis in grapevine vegetative tissues[J]. Frontiers in Plant Science, 11:527. [33] Xu H F, Yang G X, Zhang J, et al.2018. Overexpression of a repressor MdMYB15L negatively regulates anthocyanin and cold tolerance in red-fleshed callus[J]. Biochemical and Biophysical Research Communications, 499(1):405-410. [34] Xue L, Huang X, Zhang Z, et al.2022. An anthocyanin-related glutathione S-transferase, MrGST1, plays an essential role in fruit coloration in Chinese bayberry (Morella rubra)[J]. Frontiers in Plant Science, 13:903333. [35] Xue L, Li X, Li S, et al.2025 Integrated transcriptome and metabolome analysis reveals the mechanism for unique dynamic change in the flower color of Lycoris albiflora[J]. Plant Physiology and Biochemistry, 299:110373. [36] Xue Q, Wang B, Feng J, et al.2024. Lycorine (Lycoris radiata)-a unique natural medicine on breast cancer[J]. Journal of Cellular and Molecular Medicine, 28(16):11. [37] Zhang X, Henriques R, Lin S S, et al.2006. Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method[J]. Nature Protocols, 1(2):641-646. [38] Zhu Z G, Li G R, Liu L, et al.2018. A R2R3-MYB transcription factor, VvMYBC2L2, functions as a transcriptional repressor of anthocyanin biosynthesis in grapevine (Vitis vinifera L.)[J]. Molecules, 24(1):92. |
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