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Journal of Integrative Agriculture  2023, Vol. 22 Issue (8): 2335-2345    DOI: 10.1016/j.jia.2023.04.046
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Systematic analysis of MYB transcription factors and the role of LuMYB216 in regulating anthocyanin biosynthesis in the flowers of flax (Linum usitatissimum L.)
XIE Dong-wei1, LI Jing1, ZHANG Xiao-yu2, DAI Zhi-gang2, ZHOU Wen-zhi3, SU Jian-guang2, SUN Jian1#

1 School of Life Science, Nantong University, Nantong 226019, P.R.China

2 Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410221, P.R.China

3 Jiangsu Sanshu Biotechnology Co. Ltd., Nantong 226001, P.R.China

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摘要  

花色苷是影响植物颜色和营养品质的重要色素。MYBs在植物花色苷的合成和积累中起着重要作用。然而,MYB转录因子在亚麻花的花色苷合成中的调节作用尚不清楚。在本研究中,从亚麻基因组中共鉴定出402个MYB转录因子,这些MYB成员在15条染色体上分布不均。其中,R2R3-LuMYB成员被分为32个亚家族。实时荧光定量(qRT-PCR)分析显示,进化树相邻亚家族中的7个R2R3-LuMYB基因具有相似的表达模式,其中LuMYB216在不同颜色的花瓣中高度表达。此外,对LuMYB216在亚麻中进行基因编辑后,观察到突变体植物的花瓣颜色、花药颜色和种皮颜色明显浅于野生型植株。测定lumyb216突变体植株的花瓣和种皮的花色苷,发现含量较野生型显著降低。相关分析表明,LuMYB216与其上游调控因子bHLH30共表达,且显著正相关。本研究系统分析了亚麻中的MYB基因家族,为研究LuMYB216在亚麻花的花色苷合成中的调控奠定了基础。



Abstract  

Anthocyanin is an important pigment that affects plant color and nutritional quality.  MYBs play an important role in plant anthocyanin synthesis and accumulation.  However, the regulatory function of MYB transcription factors in anthocyanin synthesis in flax flowers is still unclear.  In this study, 402 MYB transcription factors were identified in the flax genome.  These MYB members are unevenly distributed on 15 chromosomes.  The R2R3-LuMYB members were divided into 32 phylogenetic subfamilies.  qRT-PCR analysis showed that seven R2R3-LuMYB genes in the adjacent subfamily of the evolutionary tree had similar expression patterns, among which LuMYB216 was highly expressed in the petals of different colors.  Moreover, gene editing of LuMYB216 in flax showed that the petal color, anther color and seed coat color of mutant plants were significantly lighter than those of wild-type plants, and the anthocyanin content of lumyb216 mutant plants was significantly reduced.  Correlation analysis indicated that LuMYB216 was significantly positively correlated with the upstream regulator bHLH30.  This study systematically analyzed the MYB gene family in flax, laying a foundation for studying the regulation of LuMYB216 in flax flower anthocyanin synthesis.

Keywords:  flax        MYB gene family        anthocyanin biosynthesis        LuMYB216        functional characterization  
Received: 23 January 2023   Accepted: 10 April 2023
Fund: This work was supported by the National Natural Science Foundation of China (31801409), the Innovation and Entrepreneurship Training Program for College Students, Jiangsu Province, China (202210304103Y), and the Science and Technology Project of Nantong City, Jiangsu Province, China (JC2020156). We thank all the people who participated in this research.
About author:  XIE Dong-wei, E-mail: xiedongwei@163.com; #Correspondence SUN Jian, Tel: +86-513-85012818, E-mail: sunjian8416@163.com

Cite this article: 

XIE Dong-wei, LI Jing, ZHANG Xiao-yu, DAI Zhi-gang, ZHOU Wen-zhi, SU Jian-guang, SUN Jian. 2023. Systematic analysis of MYB transcription factors and the role of LuMYB216 in regulating anthocyanin biosynthesis in the flowers of flax (Linum usitatissimum L.). Journal of Integrative Agriculture, 22(8): 2335-2345.

Alexander A D, Ghislaine H, Céline R, Mengin V, Ollat N, Bordenave L, Decroocq S, Delaunay J C, Delrot S, Merillon J M, Monti J P, Gomes E, Richard T. 2012. Anthocyanin identification and composition of wild Vitis spp. accessions by using LC-MS and LC-NMR. Analytica Chimica Acta, 732, 145–152.

Belwal T, Singh G, Jeandet P, Pandey A, Giri L, Ramola S, Bhatt I D, Venskutonis P R, Georgiev M I, Clement C, Luo Z S. 2020. Anthocyanins, multi-functional natural products of industrial relevance: recent biotechnological advances. Biotechnology Advances, 43, 107600–107613.

Chen Z, Lu X, Li Q, Li T, Zhu L, Ma Q, Wang J, Lan W, Ren J. 2021. Systematic analysis of MYB gene family in Acer rubrum and functional characterization of ArMYB89 in regulating anthocyanin biosynthesis. Journal of Experiental Botany, 72, 6319–6335.

Chiou C Y, Yeh K W. 2008. Differential expression of MYB gene (OgMYB1) determines color patterning in floral tissue of oncidium gower ramsey. Plant Molecular Biology, 66, 379–388.

Dubos C, Stracke R, Grotewold E, Weisshaar B, Martin C, Lepiniec L. 2010. MYB transcription factors in Arabidopsis. Trends in Plant Science, 15, 573–581.

Espley R V, Hellens R P, Putterill J, Stevenson D E, Kutty-Amma S, Allan A C. 2007. Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. Plant Journal, 49, 414–427.

Ezzat S M, Shouman S A, Elkhoely A, Attia Y M, Elsesy M S, El A S, Choucry M A, El S G, El A S, Sattar E A, Tanbouly N E. 2018. Anticancer potentiality of lignan rich fraction of six flaxseed cultivars. Scientific Reports, 8, 544.

Gu Z Y, Zhu J, Hao Q, Yuan Y W, Duan Y W, Men S Q, Wang Q Y, Hou Q Z, Liu Z A, Shu Q Y, Wang L H. 2019. A novel R2R3-MYB transcription factor contributes to petal blotch formation by regulating organspecific expression of PsCHS in tree peony (Paeonia suffruticosa). Plant Cell Physiology, 60, 599–611.

Heppel S C, Jaffé F W, Takos A M, Schellmann S, Bogs J. 2013. Identification of key amino acids for the evolution of promoter target specificity of anthocyanin and proanthocyanidin regulating MYB factors. Plant Molecular Biology, 82, 457–471.

Jie L, Butelli E, Hill L, Parr A, Niggeweg R, Bailey P, Weisshaar B, Martin C. 2010. AtMYB12 regulates caffeoyl quinic acid and flavonol synthesis in tomato: expression in fruit results in very high levels of both types of polyphenol. Plant Journal, 56, 316–326.

Kallam K, Appelhagen I, Luo J, Albert N, Zhang H, Deroles S, Hill L, Findlay K, Andersen O M, Davies K, Martin C. 2017. Aromatic decoration determines the formation of anthocyanic vacuolar inclusions. Current Biology, 27, 945–957.

Kobayashi S, Ishimaru M, Hiraoka K C. 2002. MYB-related genes of the Kyoho grape (Vitis labruscana) regulate anthocyanin biosynthesis. Planta, 215, 924–933.

Kranz H D, Denekamp M, Greco R, Weisshaar B. 1998. Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana. Plant Journal, 16, 263–276.

Lee W J, Jeong C Y, Kwon J, Kien V V, Lee D, Hong S W, Lee H. 2016. Drastic anthocyanin increase in response to PAP1 overexpression in fls1 knockout mutant confers enhanced osmotic stress tolerance in Arabidopsis thaliana. Plant Cell Reports, 35, 2369–2379.

Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2–∆∆CT method. Methods, 25, 402–408.

Matus J T, Aquea F, Arce-johnson P. 2008. Analysis of the grape MYB R2R3 subfamily reveals expanded wine qualityrelated clades and conserved gene structure organization across Vitis and Arabidopsis genomes. BMC Plant Biology, 8, 83.

Millard P S, Kragelund B B, Burow M. 2019. R2R3 MYB transcription factors - Functions outside the DNA-binding domain. Trends in Plant Science, 24, 934–946.

Naing A H, Kim C K. 2018. Roles of R2R3-MYB transcription factors in transcriptional regulation of anthocyanin biosynthesis in horticultural plants. Plant Molecular Biology, 98, 1–18.

Nesi N, Jond C, Debeaujon I, Caboche M, Lepiniec L. 2001. The Arabidopsis TT2 gene encodes an R2R3 MYB domain protein that acts as a key determinant for proanthocyanidin accumulation in developing seed. Plant Cell, 13, 2099–2114.

Ogata K, Kanei-ishii C, Sasaki M, Hatanaka H, Nagadoi A, Enari M, Nakamura H, Nishimura Y, Ishii S, Sarai A. 1996. The cavity in the hydrophobic core of MYB DNA-binding domain is reserved for DNA recognition and trans-activation. Nature Structural and Molecular Biology, 3, 178–187.

Reiter F, Wienerroither S, Stark A. 2017. Combinatorial function of transcription factors and cofactors. Current Opinion in Genetics & Development, 43, 73–81.

Romero I, Fuertes A, Benito M J, Malpica J M, Leyva A, Paz-Ares J. 1998. More than 80 R2R3-MYB regulatory genes in the genome of Arabidopsis thaliana. Plant Journal, 14, 273–284.

Stracke R, Werber M, Weisshaar B. 2001. The R2R3-MYB gene family in Arabidopsis thaliana. Current Opinion in Plant Biology, 4, 447–456.

Tian J, Peng Z, Zhang J, Song T T, Wan H H, Zhang M L, Yao Y C. 2015. McMYB10 regulates coloration via activating McF3’H and later structural genes in ever-red leaf crabapple. Plant Biotechnology Journal, 13, 948–961.

Xie D, Yang X, He R, Huo H, Ye Z, Ren X, Yuan H, Dai Z, Sun J, Su J. 2022. Comprehensive analysis of the UDP glycosyltransferase gene family in flax [Linum usitatissimum L.] and functional verification of the role of LuUGT175 in the regulation of lignin biosynthesis. Industrial Crops and Products, 188, 115720.

Yamasaki K, Kigawa T, Inoue M, Watanabe S, Tateno M, Seki M, Shinozaki K, Yokoyama S. 2008. Structures and evolutionary origins of plant-specific transcription factor DNA-binding domains. Plant Physiology & Biochemistry, 46, 394–401.

Yan H, Pei X, Zhang H, Li X, Zhang X, Zhao M, Chiang V L, Sederoff R R, Zhao X. 2021. MYB-mediated regulation of anthocyanin biosynthesis. International Journal of Molecular Sciences, 22, 3103–3131.

Yan M, Zhou S R, Xue H W. 2015. CRISPR primer designer: Design primers for knockout and chromosome imaging CRISPR-Cas system. Journal of Integrative Plant Biology, 57, 613–617.

Zhou M L, Yu T, Zhang K X, Li F L, Yang P Y, Tang Y X, Wu Y M, Shao J R. 2013. Identification of TT2 gene from floral transcriptome in Fagopyrum tataricum. Food Research International, 54, 1331–1333.

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