Genome-wide Identification of the MYB Transcription Factor Family in Camellia oleifera and Functional Analysis of CoMYB107 in Stamen Development

doi:10.13560/j.cnki.biotech.bull.1985.2026-0115

Abstract

Abstract:

Objective To identify members of the MYB gene family in Camellia oleifera and analyze the molecular regulatory network of its anther development, providing theoretical support for elucidating the reproductive development mechanism of C. oleifera. Method Based on the C. oleifera genome data, members of the MYB gene family were identified using bioinformatics methods, and their sequence characteristics, physicochemical properties, and evolutionary relationships were analyzed, and subcellular localization was predicted. Using transcriptome data and RT-qPCR technology, the expression patterns of candidate MYB genes in different tissues and at different developmental stages were investigated, key genes were screened, and heterologous overexpression of genes was performed using the tobacco leaf disc method to verify the functions of candidate genes. Result A total of 137 MYB genes were identified from the C. oleifera genome, with amino acid lengths ranging from 102 to 1 752 aa, distributed on 45 chromosomes, and the majority of members were predicted to localize in the nucleus. There were 55 segmental duplication events, involving 44 valid gene pairs. Among these, 38 gene pairs were under purifying selection, and 5 gene pairs were under positive selection. There were 95 collinear gene pairs with Arabidopsis. Phylogenetic analysis indicated that the MYB family in C. oleifera and Arabidopsis could be classified into 26 subfamilies. Promoter cis-acting elements were mainly associated with light response, hormone response, and stress response, with hormone-related cis-elements mainly including gibberellin, abscisic acid, and jasmonic acid elements. Transcriptome data and RT-qPCR expression pattern analysis showed that 8 genes exhibited specifically high expression in anthers. In addition, protein interaction network prediction suggested that CoMYB107, CoMYB55, and CoMYB136 may be involved in a specific protein interaction regulatory network; overexpression of CoMYB107 in tobacco significantly reduced pollen viability, elongated filament length, and delayed anther dehiscence. Conclusion CoMYB107 may regulate stamen development through the GA-JA signaling pathway, which manifests as promoting filament elongation, delaying anther dehiscence, and decreasing pollen viability.

Key words: Camellia oleifera, CoMYB107, transcriptome, anther development, expression analysis, MYB gene family, gibberellin, whole-genome identification

TAN Shu, SHEN Hong-jian, GAO Xiao-lei, YIN Qian, ZOU Feng. Genome-wide Identification of the MYB Transcription Factor Family in Camellia oleifera and Functional Analysis of CoMYB107 in Stamen Development[J]. Biotechnology Bulletin, 2026, 42(9): 196-209.

Figures/Tables 13

Fig. 1 The morphology of flower buds in different periods of C. oleiferaT1-T5 represent the dates of flower bud collection (2024). T1: September 5. T2: September 20. T3: October 5. T4: October 20. T5: November 4. The same below

Fig. 2 Structural analysis of C. oleiferaMYB genesA: Phylogenetic tree. B: Motif structure. C: Conserved domain. D: UTR and CDS

Fig. 3 Chromosomal localization of C. oleiferaMYB genes

Fig. 4 Collinearity analysis of MYB genesA: Collinearity analysis of C. Oleifera MYB genes. B: Collinearity analysis of A. thaliana MYB and C. oleifera MYB genes

Fig. 5 Analysis of C. oleiferaMYB gene promoters

Fig. 6 Phylogenetic tree of MYB proteins from A. thaliana and C. oleifera

Fig. 7 Heatmap of CoMYB gene expression levels in different periods

Fig. 8 Relative expression analysis of C. oleifera MYB genes in different tissueR: Root; S: stem; L: leaf; C: calyx; PE: petal; A: anther; PI: pistil. Data are presented as the mean ± standard error (SE) of three biological replicates (n=3). Different lowercase letters indicate significant differences (P<0.05). The same below

Fig. 9 Expression analysis of C. oleiferaMYB genes at different developmental stages

Fig. 10 Protein interaction network prediction

Fig. 11 Subcellular localization of CoMYB107 protein

Fig. 12 Phenotype of OE-CoMYB107Nicotiana tabacumA: Pollen viability of wild-type N. tabacum. B: Pollen viability of CoMYB107 transgenic N. tabacum. C: N. tabacum stamen phenotype (C1: Wild-type N. tabacum. C2: CoMYB107 transgenic N. tabacum). D: Statistics of pollen viability in wild-type and overexpression N. tabacum. ** P<0.01. The same below

Fig. 13 Expression analysis of 8 flower development-related genes in tobacco flowers

References 59

[36]	Lescot M, Déhais P, Thijs G, et al. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences [J]. Nucleic Acids Res, 2002, 30(1): 325-327.
[37]	Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 [J]. Genome Biol, 2014, 15(12): 550.
[38]	Reiser L, Bakker E, Subramaniam S, et al. The Arabidopsis information resource in 2024 [J]. Genetics, 2024, 227(1): iyae027.
[39]	Kumar S, Stecher G, Suleski M, et al. MEGA12: molecular evolutionary genetic analysis version 12 for adaptive and green computing [J]. Mol Biol Evol, 2024, 41(12): msae263.
[40]	Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap [J]. Evolution, 1985, 39(4): 783.
[41]	Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative CT method [J]. Nat Protoc, 2008, 3(6): 1101-1108.
[42]	宋志波, 刘敏, 贾宝光, 等. 油茶总RNA的提取与内参基因的初选 [J]. 经济林研究, 2014, 32(2): 93-98.
	Song ZB, Liu M, Jia BG, et al. RNA extraction and primary screening of reference genes in Camellia oleifera [J]. Nonwood For Res, 2014, 32(2): 93-98.
[43]	杨晨欣, 李梦秀, 姜唐, 等. 美人蕉R2R3-MYB基因家族的鉴定及与花青素相关成员的表达分析 [J]. 生物技术通报, 2025, 41(12): 201-213.
	Yang CX, Li MX, Jiang T, et al. Identification and expression analysis of anthocyanin-associated R2R3-MYB genes in Canna indica [J]. Biotechnol Bull, 2025, 41(12): 201-213.
[44]	Abdullah-Zawawi MR, Ahmad-Nizammuddin NF, Govender N, et al. Comparative genome-wide analysis of WRKY, MADS-box and MYB transcription factor families in Arabidopsis and rice [J]. Sci Rep, 2021, 11: 19678.
[45]	Chen XJ, Wang PJ, Gu MY, et al. R2R3-MYB transcription factor family in tea plant (Camellia sinensis): Genome-wide characterization, phylogeny, chromosome location, structure and expression patterns [J]. Genomics, 2021, 113(3): 1565-1578.
[46]	Li YT, Zhang J, Wang SJ, et al. Genome-wide identification of the Pyrus R2R3-MYB gene family and PhMYB62 regulation analysis in Pyrus hopeiensis flowers at low temperature [J]. Int J Biol Macromol, 2024, 257: 128611.
[47]	Du JH, Zhang QQ, Hou SJ, et al. Genome-wide identification and analysis of the R2R3-MYB gene family in Theobroma cacao [J]. Genes, 2022, 13(9): 1572.
[48]	Li J, Guo SK, Min Htwe Y, et al. Genome-wide identification, classification and expression analysis of MYB gene family in coconut (Cocos nucifera L.) [J]. Front Plant Sci, 2024, 14: 1263595.
[49]	Cannon SB, Mitra A, Baumgarten A, et al. The roles of segmental and tandem gene duplication in the evolution of large gene families in Arabidopsis thaliana [J]. BMC Plant Biol, 2004, 4: 10.
[50]	Nevill PG, Howell KA, Cross AT, et al. Plastome-wide rearrangements and gene losses in carnivorous Droseraceae [J]. Genome Biol Evol, 2019, 11(2): 472-485.
[51]	Battat M, Eitan AS, Rogachev I, et al. A MYB triad controls primary and phenylpropanoid metabolites for pollen coat patterning [J]. Plant Physiol, 2019, 180(1): 87-108.
[52]	Millar AA, Gubler F. The Arabidopsis GAMYB-Like genes, MYB33 and MYB65, are microRNA-regulated genes that redundantly facilitate anther development [J]. Plant Cell, 2005, 17(3): 705-721.
[53]	Lu JY, Xiong SX, Yin WZ, et al. MS1 a direct target of MS188 regulates the expression of key sporophytic pollen coat protein genes in Arabidopsis [J]. J Exp Bot, 2020, 71(16): 4877-4889.
[54]	Mitchum MG, Yamaguchi S, Hanada A, et al. Distinct and overlapping roles of two gibberellin 3-oxidases in Arabidopsis development [J]. Plant J, 2006, 45(5): 804-818.
[55]	Hsieh KT, Wu CC, Lee SJ, et al. Rice GA3ox1 modulates pollen starch granule accumulation and pollen wall development [J]. PLoS One, 2023, 18(10): e0292400.
[56]	马浩森, 朱婷, 王淑婷, 等. 普通小麦CONSTANS-LIKE(COL)基因家族的鉴定和表达分析 [J]. 麦类作物学报, 2024, 44(11): 1365-1376.
	Ma HS, Zhu T, Wang ST, et al. Identification and expression pattern analysis of CONSTANS-LIKE (COL) gene family in wheat (Triticum aestivum L.) [J]. J Triticeae Crops, 2024, 44(11): 1365-1376.
[1]	Ito M. Conservation and diversification of three-repeat Myb transcription factors in plants [J]. J Plant Res, 2005, 118(1): 61-69.
[2]	Dubos C, Stracke R, Grotewold E, et al. MYB transcription factors in Arabidopsis [J]. Trends Plant Sci, 2010, 15(10): 573-581.
[3]	Bhatt PA, Gurav TP, Kondhare KR, et al. MYB proteins: Versatile regulators of plant development, stress responses, and secondary metabolite biosynthetic pathways [J]. Int J Biol Macromol, 2025, 288: 138588.
[4]	Wu XY, Xia M, Su P, et al. MYB transcription factors in plants: a comprehensive review of their discovery, structure, classification, functional diversity and regulatory mechanism [J]. Int J Biol Macromol, 2024, 282: 136652.
[5]	Xu XF, Wang B, Feng YF, et al. AUXIN RESPONSE FACTOR17 directly regulates MYB108 for anther dehiscence [J]. Plant Physiol, 2019, 181(2): 645-655.
[6]	Xu Y, Iacuone S, Li SF, et al. MYB80 homologues in Arabidopsis, cotton and Brassica: regulation and functional conservation in tapetal and pollen development [J]. BMC Plant Biol, 2014, 14: 278.
[7]	Liu ZH, Bao WJ, Liang WQ, et al. Identification of gamyb-4 and analysis of the regulatory role of GAMYB in rice anther development [J]. J Integr Plant Biol, 2010, 52(7): 670-678.
[8]	Zhang Y, Zhang XL, Liu B, et al. A GAMYB homologue CsGAMYB1 regulates sex expression of cucumber via an ethylene-independent pathway [J]. J Exp Bot, 2014, 65(12): 3201-3213.
[9]	Ghelli R, Brunetti P, Napoli N, et al. A newly identified flower-specific splice variant of AUXIN RESPONSE FACTOR8 regulates stamen elongation and endothecium lignification in Arabidopsis [J]. Plant Cell, 2018, 30(3): 620-637.
[10]	李文超, 张艺, 赵淑青. 拟南芥RUS4基因沉默对花药药室内壁次生加厚的影响 [J]. 中国细胞生物学学报, 2019, 41(4): 619-626.
	Li WC, Zhang Y, Zhao SQ. Silencing of Arabidopsis RUS4 impairs anther endothecium secondary cell wall thickening [J]. Chin J Cell Biol, 2019, 41(4): 619-626.
[11]	Yadav S, Jalan K, Das S. Comparative functional characterization of nst1, nst2, and nst3 in Arabidopsis thaliana uncovers previously unknown functions in diverse developmental pathways beyond secondary wall formation [J]. Plant Mol Biol Rep, 2025, 43(1): 165-179.
[12]	Zhu J, Lou Y, Xu XF, et al. A genetic pathway for tapetum development and function in Arabidopsis [J]. J Integr Plant Biol, 2011, 53(11): 892-900.
[13]	Yao XZ, Hu W, Yang ZN. The contributions of sporophytic tapetum to pollen formation [J]. Seed Biol, 2022, 1(1): 1-13.
[14]	Ghelli R, Brunetti P, Marzi D, et al. The full-length Auxin Response Factor 8 isoform ARF8.1 controls pollen cell wall formation and directly regulates TDF1, AMS and MS188 expression [J]. Plant J, 2023, 113(4): 851-865.
[15]	Hua MY, Yin WZ, Fernández Gómez J, et al. Barley TAPETAL DEVELOPMENT and FUNCTION1 (HvTDF1) gene reveals conserved and unique roles in controlling anther tapetum development in dicot and monocot plants [J]. New Phytol, 2023, 240(1): 173-190.
[16]	田娟, 马容, 付万祥, 等. 钙锌硼叶面喷施对三个油茶品种生长和品质的影响 [J]. 植物生理学报, 2026, 62(1): 109-124.
	Tian J, Ma R, Fu WX, et al. Effects of foliar spraying of calcium, zinc and boron on growth and quality of three Camellia oleifera varieties [J]. Plant Physiol J, 2026, 62(1): 109-124.
[17]	Yang Y, Gao XL, Xiong H, et al. Cytological and transcriptomic analyses reveal the regulatory mechanisms of flowering and petaloid-anther development in Camellia oleifera [J]. Physiol Plant, 2025, 177(3): e70345.
[18]	严茂林, 郭丹, 周晓亮, 等. 中国油茶增产格局及其贡献因素分析 [J]. 中国油料作物学报, 2025, 47(1): 9-19.
	Yan ML, Guo D, Zhou XL, et al. Analysis on the pattern and contributing factors of increasing Camellia oleifera production in China [J]. Chin J Oil Crop Sci, 2025, 47(1): 9-19.
[19]	华余琴. 油茶生态化栽培技术经济效益研究 [J]. 现代农业科技, 2021(14): 130, 140.
	Hua YQ. Study on technical and economic benefits of ecological cultivation of Camellia oleifera [J]. Mod Agric Sci Technol, 2021(14): 130, 140.
[20]	刘兴慧, 胡英杰, 吴俊涛, 等. 栽植密度对湖南主栽油茶品种幼林生长及油脂品质的影响 [J]. 经济林研究, 2025, 43(1): 271-281.
	Liu XH, Hu YJ, Wu JT, et al. Effects of planting density on the growth and oil quality of main planted Camellia oleifera varieties in Hunan province [J]. Non Wood For Res, 2025, 43(1): 271-281.
[21]	Hu ZC, Yang KJ, Yu AL. The extraction technologies, nutritional compositions, and health benefits of Camellia oleifera seed oil A review [J]. J Oleo Sci, 2025, 74(10): 859-878.
[22]	Qin PJ, Shen JJ, Wei J, et al. A critical review of the bioactive ingredients and biological functions of Camellia oleifera oil [J]. Curr Res Food Sci, 2024, 8: 100753.
[23]	Shi H, Zhou XY, He XE, et al. Camellia oil enhances plasma antioxidant metabolism and improves plasma lipid metabolism in high-fat diet-fed rats [J]. Nat Prod Commun, 2022, 17(3): 1934578X221081368.
[24]	王悦. 侗族传统油茶产业的本土知识及其价值研究 [D]. 吉首: 吉首大学, 2022.
	Wang Y. Research on the local knowledge and value of traditional Camellia oleifera industry of Dong ethnic group [D]. Jishou: Jishou University, 2022.
[25]	Yang CY, Liu XM, Chen ZY, et al. Comparison of oil content and fatty acid profile of ten new Camellia oleifera cultivars [J]. J Lipds, 2016, 2016: 3982486.
[26]	陈展图, 马青霞, 卫艺佳, 等. 乡村振兴战略背景下非物质文化遗产产业化保护——以恭城油茶为例 [J]. 当代农村财经, 2023(10): 39-43.
	Chen ZT, Ma QX, Wei YJ, et al. Industrialization protection of intangible cultural heritage under the background of rural revitalization strategy—taking Gongcheng Camellia oleifera as an example [J]. Contemp Rural Finance Econ, 2023(10): 39-43.
[27]	朱阳. 非物质文化遗产瑶族油茶习俗的保护传承研究 [J]. 中国茶叶, 2024, 46(8): 76-81.
	Zhu Y. Research on the protection and inheritance of Yao people’s oil tea custom as intangible cultural heritage [J]. China Tea, 2024, 46(8): 76-81.
[28]	Yan JD, Liu Q, Guo PR, et al. Time-course transcriptome analysis unveils the CoFKF1-CoMYB4-CoFT1 regulatory module in flowering control of Camellia oleifera Abel [J]. Plant Cell Environ, 2025, 48(8): 6153-6169.
[29]	Zhu HG, Wang FQ, Xu ZP, et al. The complex hexaploid oil-Camellia genome traces back its phylogenomic history and multi-omics analysis of Camellia oil biosynthesis [J]. Plant Biotechnol J, 2024, 22(10): 2890-2906.
[30]	Wang YZ, Wu XY, Sun Q, et al. Genome-wide identification and transcriptomic analysis of MYB transcription factors in seashore Paspalum under salt stress [J]. Int J Mol Sci, 2026, 27(2): 1068.
[31]	Yang J, Sun SQ, Sun N, et al. HMMER-Extractor: an auxiliary toolkit for identifying genomic macromolecular metabolites based on Hidden Markov Models [J]. Int J Biol Macromol, 2024, 283: 137666.
[32]	Chauhan R, Prabhakaran S, Taj G, et al. Structural and functional characterization of an amino acid transporter in Amaranthus hypochondriacus using bioinformatics tools [J]. J Adv Biol Biotechnol, 2024, 27(8): 280-288.
[33]	Horton P, Park KJ, Obayashi T, et al. WoLF PSORT: protein localization predictor [J]. Nucleic Acids Res, 2007, 35(): W585-W587.
[34]	Chen CJ, Chen H, Zhang Y, et al. TBtools: an integrative toolkit developed for interactive analyses of big biological data [J]. Mol Plant, 2020, 13(8): 1194-1202.
[35]	Wang YP, Tang HB, DeBarry JD, et al. MCScanX: a toolkit for detection and evolutionary analysis of gene synteny and collinearity [J]. Nucleic Acids Res, 2012, 40(7): e49.
[57]	赵雪. 转录因子MYC2和MYB305对烟碱合成基因的表达调控研究 [D]. 北京: 中国农业科学院, 2020.
	Zhao X. Roles of MYC2 and MYB305 in regulating nicotine synthesis genes [D]. Beijing: Chinese Academy of Agricultural Sciences, 2020.
[58]	李文燕. 植物OPR基因家族系统发育分析及水稻OPR家族基因分子生物学功能研究 [D]. 广州: 中山大学, 2010.
	Li WY. Phylogenetic analysis of OPR gene family in plants and study of molecular biological functions of OPR family genes in rice [D]. Guangzhou: Sun Yat-sen University, 2010.
[59]	Gan H, Wang SY, Yang ZS, et al. Molecular decoding of phytohormone crosstalk: JA-mediated key regulatory nodes and signal integration [J]. Plants, 2025, 14(17): 2647.