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| Polymorphism Analysis and Functional Study of Anthocyanidin Synthase Gene (LiANS) Among Different Flower-color Varieties of Lagerstroemia indica |
| LU Di1, YUAN Tian-Yi1, HOU Xiao-Yu1, YU Can-Nan1, ZHAO Kai-Zuo1, LIN Jing-Jing1, GUO Zheng-Hao1, ZHANG Jian1,2, YU Chun-Mei1,2,* |
1 School of Life Science, Nantong University, Nantong 226019, China;
2 Nantong Key Laboratory of Landscape Plant Genetics and Breeding, Nantong 226019, China |
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Abstract Anthocyanidin synthase (ANS) catalyzes the conversion of colorless proanthocyanidins into colored anthocyanidins. This study used 4 crape myrtle (Lagerstroemia indica) cultivars with white, pink, purple, and red petals to investigate the characteristics of the LiANS gene. Through multiple sequence alignment, phylogenetic tree construction, homology modeling, detection of the activity of prokaryotically expressed fusion protein LiANS-His6, qPCR, and promoter cloning, the characteristics of LiANS in different cultivars were analyzed. The results showed that the ORF of the LiANS gene was 1 068 bp, encoding 355 amino acids. According to the differences among LiANS, it was found that there were 4 LiANS allelic variants in 4 cultivars. Homology modeling and enzyme activity assays of the 3 purified LiANS-His6 variants showed that these variations had no effect on enzyme activity. qPCR analysis revealed that the LiANS gene was expressed constitutively in leaves from upper, middle, and lower part of shoots, and the flower at 5 developmental stages, but the expression patterns were different. The overall anthocyanin content peaked at the S3 stage of petal development. Among the samples, the red-flowered cultivar 'Dynamite' maintained relatively high expression levels during the full bloom and senescence stages (S3 and S4). The expression levels of the LiANS gene were not always positively correlated among the 4 varieties. Promoter sequence analysis indicated that 3 allelic variants of the promoter existed among the 4 cultivars, featuring different G-box elements and MYB binding sites. Notably, the white-flowered cultivar 'Pure white' exhibited a 195 bp deletion in the proANS region. This study provides genetic resources for molecular breeding of crape myrtle cultivars with different flower colors and lays a foundation for further elucidation of its regulatory mechanisms.
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Received: 15 February 2026
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Corresponding Authors:
*ychmei@ntu.edu.cn
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[1] 陈可欣, 王晓明, 曾慧杰, 等. 2021. 紫薇LiANS基因的克隆和表达及与花青素含量的关系[J]. 分子植物育种, 19(12): 3915-3922.
(Chen K X, Wang X M, Zeng H J, et al.2021. Cloning and expression analysis of LiANS gene and its correlations with anthocyanin accumulation in Lagerstroemia indica[J]. Molecular Plant Breeding, 19(12): 3915-3922.)
[2] 代军. 2015. 大肠杆菌感受态细胞制备及转化条件优化[J]. 江苏农业科学, 43(04): 53-54.
(Dai J.2015. Optimization of preparation and transformation conditions for Escherichia coli competent cells[J]. Jiangsu Agricultural Sciences, 43(4): 53-54.)
[3] 高子龙. 2025. 紫薇花色物质类黄酮合成基因LiDFR和LiFLSs的克隆与功能分析[D]. 硕士学位论文, 河南农业大学, 导师: 李永华, 陈卓梅, 34-54.
(Gao Z L.2025. Cloning and expression analysis of flavonoid synthesis genes LiDFR and LiFLSs in Lagerstroemi indica[D]. Thesis for M.S., Henan Agricultural University, Supervisor: Li YH, Chen ZM, 34-54.)
[4] 高子龙, 王金凤, 陈卓梅, 等. 2025. 紫薇LiUFGT1基因克隆与表达分析[J]. 北方园艺, (06): 18-25.
(Gao Z L, Wang J F, Chen Z M, et al. 2025. Cloning and expression analysis of LiUFGT1 gene in Lagerstroemia indica[J]. Northern Horticulture, (6): 18-25.)
[5] 衡蒙, 孔祥莹, 赵海平, 等. 2023. 华丽龙胆花青素合成酶GsANS基因的克隆及其表达分析[J]. 分子植物育种, 21(11): 3498-3504.
(Heng M, Kong X Y, Zhao H P, et al.2023. Cloning and expression analysis of GsANS gene in Gentiana sino-ornata[J]. Molecular Plant Breeding, 21(11): 3498-3504.)
[6] 黄嘉可, 洪思丹, 徐海华, 等. 2025. 紫薇花色呈色机制研究进展[J]. 园艺学报, 52(12): 3412-3426.
(Huang J K, Hong S D, Xu H H, et al.2025. Progress on the formation mechanism of Lagerstroemia indica flower color[J]. Acta Horticulturae Sinica, 52(12): 3412-3426.)
[7] 黄玲, 胡先梅, 梁泽慧, 等. 2022. 郁金香花青素合成酶基因TgANS的克隆与功能鉴定[J]. 园艺学报, 49(9): 1935-1944.
(Huang L, Hu X M, Liang Z H, et al.2022. Cloning and function identification of anthocyanidin synthase gene TgANS in Tulipa gesneriana[J]. Acta Horticulturae Sinica, 49(9): 1935-1944.)
[8] 蒋宝鑫, 杨国霞, 吕思佳, 等. 2023. 比利时杜鹃花花青素合成酶基因(RhANS)克隆及其功能分析[J]. 核农学报, 37(03): 449-460.
(Jiang B X, Yang G X, Lv S J, et al.2023. Cloning and functional analysis of anthocyanin synthase gene (RhANS) in Rhododendron hybridum Hort[J]. Journal of Nuclear Agricultural Sciences, 37(3): 449-460.)
[9] 李金璐, 王硕, 于婧, 等. 2013. 一种改良的植物DNA提取方法[J]. 植物学报, 48(1): 72-78.
(Li J L, Wang S, Yu J, et al.2013. A modified CTAB protocol for plant DNA extraction[J]. Chinese Bulletin of Botany, 48(1): 72-78.)
[10] 卢帅杰, 尹若勇, 秦欣如, 等. 2025. 5个紫薇品种花色呈色的生理研究[J]. 华南农业大学学报, 46(4): 577-586.
(Lu S J, Yin R Y, Qin X R, et al.2025. Physiological study on the flower color formation of five varieties of Lagerstroemia indica[J]. Journal of South China Agricultural University, 46(4): 577-586.)
[11] 袁美静, 马誉, 巫瑞, 等. 2024. 影响月季花瓣呈色的理化因子及花色苷组分分析[J]. 西北植物学报, 44(2): 255-269.
(Yuan M J, Ma Y, Wu R, et al.2024. Physicochemical characteristics and anthocyanin components affecting the color of rose petals[J]. Acta Botanica Boreali-Occidentalia Sinica, 44(2): 255-269.)
[12] 张启翔. 1991. 紫薇品种分类及其在园林中的应用(英文)[J]. 北京林业大学学报, 13(04):57-66.
(Zhang Q X.1991. Studies on cultivars of crape-myrtle (Lagerstroemia indica) and their uses in urban greening[J]. Journal of Beijing Forestry University, 13(04):57-66.)
[13] An J, Yao J, Xu R, et al.2018. Apple bZIP transcription factor MdbZIP44 regulates abscisic acid-promoted anthocyanin accumulation[J]. Plant Cell and Environment, 41(11):2678-2692.
[14] Barros J, Dixon R A.2020. Plant phenylalanine/tyrosine ammonia-lyases[J]. Trends in Plant Science, 25(1): 66-79.
[15] Bi M, Liang R, Wang J, et al.2023. Multifaceted roles of LhWRKY44 in promoting anthocyanin accumulation in Asiatic hybrid lilies (Lilium spp.)[J]. Horticulture Research, 10(9): uhad167.
[16] Chen C, Wu Y, Li J, et al.2023. TBtools-II: A "one for all, all for one" bioinformatics platform for biological big-data mining[J]. Molecular Plant, 16(11): 1733-1742.
[17] Feng L, Shen P, Chi X, et al.2024. The anthocyanin formation of purple leaf is associated with the activation of LfiHY5 and LfiMYB75 in crape myrtle[J]. Horticultural Plant Journal, 10(5): 1230-1246.
[18] Feng Y, Tian X, Liang W, et al.2023. Genome-wide identification of grape ANS gene family and expression analysis at different fruit coloration stages[J]. BMC Plant Biology, 23(1): 632.
[19] Gao Z, Chen Z, Wang J, et al.2024. Integrated transcriptome and metabolome analysis revealed mechanism underlying anthocyanin biosynthesis during flower color formation in Lagerstroemia indica[J]. Horticulturae, 10(11): 1229.
[20] Gonda I, Abu-Abied M, Adler C, et al.2023. Two independent loss-of-function mutations in anthocyanidin synthase homeologous genes are responsible for the all-green phenotype of sweet basil[J]. Physiologia Plantarum, 175(1): e13870.
[21] Gu C, Hong S, Wang J, et al.2024. Identification and expression analysis of the bZIP and WRKY gene families during anthocyanins biosynthesis in Lagerstroemia indica L[J]. Horticulture, Environment, and Biotechnology, 65(1): 169-180.
[22] Hu X, Liang Z, Sun T, et al.2024. The R2R3-MYB transcriptional repressor TgMYB4 negatively regulates anthocyanin biosynthesis in Tulips (Tulipa gesneriana L.)[J]. International Journal of Molecular Sciences, 25(1):563.
[23] Imaizumi R, Mameda R, Takeshita K, et al.2021. Crystal structure of chalcone synthase, a key enzyme for isoflavonoid biosynthesis in soybean[J]. Proteins: Structure, Function, and Bioinformatics, 89(1): 126-131.
[24] Jia Y H, He F, Shen Z L, et al.2023. Molecular characterization of a chalcone synthase gene RhCHS from Rhododendron × hybridum Hort[J]. Gene, 857: 147176.
[25] Khoo H E, Azlan A, Tang ST, et al.2017. Anthocyanidins and anthocyanins: Colored pigments as food, pharmaceutical ingredients, and the potential health benefits[J]. Food & Nutrition Research, 61(1): 1361779.
[26] Li C, Wu J, Hu K D, et al.2020. PyWRKY26 and PybHLH3 cotargeted the PyMYB114 promoter to regulate anthocyanin biosynthesis and transport in red-skinned pears[J]. Horticulture Research, 7: 37.
[27] Li X, Cao L, Jiao B, et al.2022. The bHLH transcription factor AcB2 regulates anthocyanin biosynthesis in onion (Allium cepa L.)[J]. Horticulture Research, 9: uhac128.
[28] Lin C, Xing P, Jin H, et al.2022. Loss of anthocyanidin synthase gene is associated with white flowers of Salvia miltiorrhiza Bge. f. alba, a natural variant of S. miltiorrhiza[J]. Planta, 256(1): 15.
[29] Liu H, Shu Q, Lin-Wang K, et al.2021. The PyPIF5-PymiR156a-PySPL9-PyMYB114/MYB10 module regulates light-induced anthocyanin biosynthesis in red pear[J]. Molecular Horticulture, 1(1): 14.
[30] Lv F, Yang R T, Wang Q, et al.2024. Multi-omics analysis uncovers novel gene regulatory networks of flower coloration in Lagerstroemia indica[J]. Industrial Crops and Products, 216: 118710.
[31] Ma X, Liang G, Xu Z, et al.2025. CaMYBA-CaMYC-CaTTG1 complex activates the transcription of anthocyanin synthesis structural genes and regulates anthocyanin accumulation in pepper (Capsicum annuum L.) leaves[J]. Frontiers in Plant Science, 16: 1538607.
[32] Morita Y, Ishiguro K, Tanaka Y, et al.2015. Spontaneous mutations of the UDP-glucose:flavonoid 3-O-glucosyltransferase gene confers pale- and dull-colored flowers in the Japanese and common morning glories[J]. Planta: An International Journal of Plant Biology, 242(3): 575-587.
[33] Ni L, Wang J, Zhou F, et al.2025. Integrated multi-omics reveals Li-miR828z-LiMYB114 regulatory module controlling anthocyanin biosynthesis during flower color development in Lagerstroemia indica[J]. Industrial Crops and Products, 234: 121524.
[34] Saigo T, Wang T, Watanabe M, et al.2020. Diversity of anthocyanin and proanthocyanin biosynthesis in land plants[J]. Current Opinion in Plant Biology, 55: 93-99.
[35] Tang Q, Wang X, Ma S, et al.2024. Molecular mechanism of abscisic acid signaling response factor VcbZIP55 to promote anthocyanin biosynthesis in blueberry (Vaccinium corymbosum)[J]. Plant Physiology and Biochemistry, 210: 108611.
[36] Xin T, Zhang Z, Zhang Y, et al.2025. Recessive epistasis of a synonymous mutation confers cucumber domestication through epitranscriptomic regulation[J]. Cell, 188(17): 4517-4529.e15.
[37] Yu C, Lian B, Fang W, et al.2021. Transcriptome-based analysis reveals that the biosynthesis of anthocyanins is more active than that of flavonols and proanthocyanins in the colorful flowers of Lagerstroemia indica[J]. Biologia Futura, 72(4): 473-488.
[38] Yu C, Liu G, Qin J, et al.2024a. Genomic and transcriptomic studies on flavonoid biosynthesis in Lagerstroemia indica[J]. BMC Plant Biology, 24(1):171.
[39] Yu M, Bai M, Chen M, et al.2024b. Identification of bHLH transcription factors and screening of anthocyanin-related genes in Lagerstroemia indica[J]. Genetica, 152(4-6): 179-197.
[40] Yu S I, Kim H, Yun D J, et al.2019. Post-translational and transcriptional regulation of phenylpropanoid biosynthesis pathway by Kelch repeat F-box protein SAGL1[J]. Plant Molecular Biology, 99(1-2): 135-148.
[41] Zhang X, Jiang X, He Y, et al.2019. AtHB2, a class II HD-ZIP protein, negatively regulates the expression of CsANS, which encodes a key enzyme in Camellia sinensis catechin biosynthesis[J]. Physiologia Plantarum, 166(4): 936-945.
[42] Zhou Y, Zheng T, Cai M, et al.2023. Genome assembly and resequencing analyses provide new insights into the evolution, domestication and ornamental traits of crape myrtle[J]. Horticulture Research, 10(9): uhad146. |
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