Cloning and Functional Study of SoMYB3 gene in Syringa oblata var. alba
ZHENG Zhen-Zhen1,2, ZHU Wen-Jie4, WANG Bin1,2, YANG Shuai4, LIU Bin4, YE Guang-Ji1,3,*
1 Academy of Agriculture and Forestry Sciences, Qinghai University, Xining 810016, China; 2 Qinghai Plateau Key Laboratory of Tree Genetics and Breeding, Xining 810016, China; 3 Key Laboratory of Tibet Plateau Biotechnology of the Ministry of Education/Laboratory for Research and Utilization of Tibet Plateau Germplasm Resources, Qinghai University, Xining 810016, China; 4 College of Agriculture and Animal Husbandry, Qinghai University, Xining 810016, China
Abstract：Syringa oblata var. alba is a white-flowered variant of Syringa oblata. The MYB transcription factor plays a crucial role in regulating anthocyanin biosynthesis.? In this study, the SoMYB3 transcription factor gene was cloned from the petals of S. oblata var. alba. The SoMYB3 gene, belonged to typical R2R3-MYB transcription factor, with an ORF length of 705 bp and encoding 234 amino acids. The phylogenetic analysis showed that SoMYB3 protein was closely related to grape (Vitis vinifera) VvMYBA1 and tea (Camellia sinensis) CsMYB113 protein. SoMYB3 was expressed in roots, stems, leaves, flower buds and flowers of S. oblata var. alba to varying degrees, and the highest expression level was in flowers. The heterologous expression of the SoMYB3 gene promoted the accumulation of anthocyanins in the roots, stems, leaves, flowers, capsules and seeds of tobacco (Nicotiana tabacum), by inducing the expression of anthocyanin 2 (NtAN2) (MYB transcription factor) and anthocyanin 1b (NtAN1b) (bHLH transcription factor), and upregulating the gene expression level of chalcone synthase (NtCHS), chalcone isomerase (NtCHI), dihydroflavonol 4-reductase (NtDFR), anthocyanidin synthase (NtANS), UDP-glucose:flavonoid 3-O-glucosyltransferase (NtUFGT), and anthocyanidin reductase (NtANR) in tobacco. In conclusion, the SoMYB3 transcription factor could induce anthocyanin synthesis, and the color of petal in S. oblata var. alba might be regulated by multiple transcription factors. This study provides a scientific basis for the directional improvement of Syringa oblata var. alba flower color.
郑贞贞, 朱文杰, 王彬, 杨帅, 刘斌, 叶广继. 白丁香SoMYB3基因克隆及功能研究[J]. 农业生物技术学报, 2026, 34(6): 1173-1183.
ZHENG Zhen-Zhen, ZHU Wen-Jie, WANG Bin, YANG Shuai, LIU Bin, YE Guang-Ji. Cloning and Functional Study of SoMYB3 gene in Syringa oblata var. alba. 农业生物技术学报, 2026, 34(6): 1173-1183.
[1] 陈敏, 杨清. 2013. 马铃薯花色苷生物合成与调控研究进展[J]. 中国马铃薯, 27(4):232-238. (Chen M, Yang Q.2013. Research advance on potato anthocyanins[J]. Chinese Potato Journal, 27(4): 232-238.) [2] 崔洪霞, 蒋高明, 臧淑英. 2004. 丁香属植物的地理分布及其起源演化[J]. 植物研究, 24(2): 141-145. (Cui H X, Jiang G M, Zang S Y.2004. The distribution, origin and evolution of Syringa[J]. Bulletin of Botanical Research, 24(2): 141-145.) [3] 贾羊毛加, 王芳, 叶广继, 等. 2019. 紫色马铃薯花青素StAN1基因的克隆及功能分析[J]. 西北植物学报, 39: 397-403. (Jia Y, Wang F, Ye G, et al.2019. Cloning and function analysis of anthocyanin StAN1 gene from purple potato[J]. Acta Botanica Boreali-Occidentalia Sinica, 39: 397-403.) [4] 李艳萍. 2021. 丁香新品种引种及观赏性评价[J]. 青海农林科技, (2): 76-78. (Li Y. 2021. Introduction and ornamental ovaluation of new varieties of Syringa[J]. Science and Technology of Qinghai Agriculture and Forestry, (2):76-78.) [5] 宋妮, 姜卫兵. 2006. 丁香的文化意蕴及其在园林绿化中的应用[J]. 河北林果研究, 21(3): 335-338. (Song N, Jiang W B.2006. Meanings of lilac and ways of application in garden greening[J]. Hebei Journal of Forestry and Orchard Research, 21(3): 335-338.) [6] 王蕊, 李彦慧, 郑健. 2018a. 紫丁香查尔酮异构酶基因SoCHI的克隆及表达分析[J]. 植物资源与环境学报, 27(3): 11-17. (Wang R, Li Y, Zheng J.2018. Cloning and expression analysis in chalcone isomerase gene SoCHI in Syringa oblata[J]. Journal of Plant Resources and Environment, 27(3): 11-17.) [7] 王蕊, 王羽, 李彦慧, 等. 2016. 华北紫丁香肉桂酸4-羟化酶(SoC4H)基因的克隆及表达分析[J]. 分子植物育种, 14(8): 2025-2030. (Wang R, Wang Y, Li Yi, et al.2016. Cloning and expression analysis of cinnamic acid 4-hydroxylase gene from Syringa oblata Lindl.[J]. Molecular Plant Breeding, 14(8): 2025-2030.) [8] 王蕊, 郑健, 李彦慧, 等. 2018b. 华北紫丁香黄烷酮3-羟化酶基因克隆及表达分析[J]. 分子植物育种, 16(12): 3863-3869. (Wang R, Zheng J, Li Y, et al.2018. Cloning and expression analysis of flavanone 3-hydroxylase gene from Syringa oblata Lindl.[J]. Molecular Plant Breeding, 16(12): 3863-3869.) [9] 王巍. 2013. 园林观赏植物丁香研究进展[J]. 国土与自然资源研究, 3: 84-85. (Wang W.2013. The research progress of ornamental plant Syringa[J]. Territory & Natural Resources Study, 3: 84-85.) [10] An J P, Zhang X W, You C X, et al.2019. MdWRKY40 promotes wounding-induced anthocyanin biosynthesis in association with MdMYB1 and undergoes MdBT2-mediated degradation[J]. New Phytologist, 224(1): 380-395. [11] Appelhagen I, Jahns O, Bartelniewoehner L, et al.2011. Leucoanthocyanidin dioxygenase in Arabidopsis thaliana: Characterization of mutant alleles and regulation by MYB-BHLH-TTG1 transcription factor complexes[J]. Gene, 484(1-2): 61-68. [12] Bovy A, Vos R, Kemper M, et al.2002. High-flavonol tomatoes resulting from the heterologous expression of the maize transcription factor genes LC and C1[J]. The Plant Cell, 14(10): 2509-2526. [13] Butelli E, Licciardello C, Zhang Y, et al.2012. Retrotransposons control fruit-specific, cold-dependent accumulation of anthocyanins in blood oranges[J]. The Plant Cell, 24(3): 1242-1255. [14] Chen L, Xia B, Li Z,et al.2022. Syringa oblata genome provides new insights into molecular mechanism of flower color differences among individuals and biosynthesis of its flower volatiles[J]. Frontiers in Plant Science, 13: 1078677. [15] Chiu L W, Zhou X, Burke S, et al.2010. The purple cauliflower arises from activation of a MYB transcription factor[J]. Plant Physiology, 154(3): 1470-1480. [16] Deluc L, Barrieu F, Marchive C, et al.2006. Characterization of a grapevine R2R3-MYB transcription factor that regulates the phenylpropanoid pathway[J]. Plant Physiology, 140(2): 499-511. [17] Espley R V, Hellens R P, Putterill J, et al.2007. Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10[J]. Plant Journal, 49(3): 414-427. [18] Holsters M, De Waele D, Depicker A, et al.1978. Transfection and transformation of Agrobacterium tumefaciens[J]. Molecular ＆ General Genetics, 163(2): 181-187. [19] Huang W, Sun W, Lv H, et al.2013. A R2R3-MYB transcription factor from Epimedium sagittatum regulates the flavonoid biosynthetic pathway[J]. PLOS ONE, 8(8): e70778. [20] Jin W, Wang H, Li M, et al.2016. The R2R3 MYB transcription factor PavMYB10.1 involves in anthocyanin biosynthesis and determines fruit skin colour in sweet cherry (Prunus avium L.)[J]. Plant Biotechnology Journal, 14(11): 2120-2133. [21] Lai B, Li X J, Hu B, et al.2014. LcMYB1 is a key determinant of differential anthocyanin accumulation among genotypes, tissues, developmental phases and ABA and light stimuli in Litchi chinensis[J]. PLOS ONE, 9(1): e86293. [22] Laitinen R A E, Ainasoja M, Broholm S K, et al.2008. Identification of target genes for a MYB-type anthocyanin regulator in Gerbera hybrida[J]. Journal of Experimental Botany, 59(13): 3691-3703. [23] 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. [24] Lim S H, Kim D H, Cho M C, et al.2022. Chili pepper AN2 (CaAN2): A visible selection marker for nondestructive monitoring of transgenic plants[J]. Plants, 11(6): 820. [25] Liu Y, Lin-Wang K, Espley R V, et al.2016. Functional diversification of the potato R2R3 MYB anthocyanin activators AN1, MYBA1, and MYB113 and their interaction with basic helix-loop-helix cofactors[J]. Journal of Experimental Botany, 67(8): 2159-2176. [26] Lloyd A M, Walbot V, Davis R W.1992. Arabidopsis and Nicotiana anthocyanin production activated by maize regulators R and C1[J]. Science, 258(5089): 1773-1775. [27] Ma B, Wu J, Shi T L, et al.2022. Lilac (Syringa oblata) genome provides insights into its evolution and molecular mechanism of petal color change[J]. Communications Biology, 5(1): 686. [28] Riaz B, Chen H, Wang J, et al.2019. Overexpression of maize ZmC1 and ZmR transcription factors in wheat regulates anthocyanin biosynthesis in a tissue-specific manner[J]. International Journal of Molecular Sciences, 20(22): 5806. [29] Schwinn K, Venail J, Shang Y, et al.2006. A small family of MYB-Regulatory genes controls floral pigmentation intensity and patterning in the genus Antirrhinum[J]. The Plant Cell, 18(4): 831-851. [30] Shui L, Li W, Yan M, et al.2023. Characterization of the R2R3 MYB transcription factor CsMYB113 regulates anthocyanin biosynthesis in tea plants (Camellia sinensis)[J]. Plant Molecular Biology Reporter, 41: 46-58. [31] Wang K L, Bolitho K, Grafton K, et al.2010. An R2R3 MYB transcription factor associated with regulation of the anthocyanin biosynthetic pathway in Rosaceae[J]. BMC Plant Biology 10: 50. [32] Wang Y, Dou Y, Wang R, et al.2017. Molecular characterization and functional analysis of chalcone synthase from Syringa oblata Lindl. in the flavonoid biosynthetic pathway[J]. Gene, 635: 16-23. [33] Yamagishi M, Shimoyamada Y, Nakatsuka T, et al.2010. Two R2R3-MYB genes, homologs of petunia AN2, regulate anthocyanin biosyntheses in flower tepals, tepal spots and leaves of Asiatic hybrid lily[J]. Plant and Cell Physiology, 51(3): 463-474. [34] Zhang Z, Chen C, Jiang C, et al.2024. VvWRKY5 positively regulates wounding-induced anthocyanin accumulation in grape by interplaying with VvMYBA1 and promoting jasmonic acid biosynthesis[J]. Horticulture Research, 11(5): uhae083. [35] Zheng J, Hu Z, Guan X, et al.2015. Transcriptome analysis of Syringa oblata Lindl. inflorescence identifies genes associated with pigment biosynthesis and scent metabolism[J]. PLOS ONE, 10(11): e0142542. [36] Zheng Z Z, Wang B, Zhu W J, et al.2025. Low expression of SrMYB1 leads to the white flower phenotype in Syringa reticulata subsp. amurensis (Ruprecht) P. S. Green & M. C. Chang[J]. All Life, 18(1): 2557265. [37] Zimmermann I M, Heim M A, Weisshaar B, et al.2004. Comprehensive identification of Arabidopsis thaliana MYB transcription factors interacting with R/B-like BHLH proteins[J]. The Plant Journal 40(1): 22-34.
