Skip to main content
SpringerLink
Log in

A R2R3-type MYB transcription factor gene from soybean, GmMYB12, is involved in flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis

  • Original Article
  • Published:
Plant Biotechnology Reports Aims and scope Submit manuscript

Abstract

The R2R3-type MYB transcription factors have been shown to increase flavonoids accumulation by regulating the expression of key enzyme genes related to flavonoid biosynthesis pathway. However, the roles and underlying mechanisms of the soybean GmMYB12 gene in regulation of flavonoids accumulation and tolerance to abiotic stresses are rarely known. In the present study, the GmMYB12 gene was isolated and its function was characterized. Sequence and yeast one-hybrid analyses showed that GmMYB12 contained two MYB domains and belonged to R2R3-MYB protein with transactivation activity. Subcellular localization analysis in onion epidermal cells indicated that GmMYB12 was localized to the nucleus. Overexpression of GmMYB12 increased the production of downstream flavonoids and the expression of related genes in the flavonoid biosynthesis pathway. It also improved resistance to salt and drought stresses during seed germination, root development, and growing stage. Further component and enzymatic analyses showed significant increases of proline content, pyrroline-5-carboxylate synthase (P5CS), superoxide dismutase (SOD), and peroxidase (POD) activities, as well as significant reduction of H2O2 and malonaldehyde (MDA) content under salt and drought stresses in transgenic plants. Meanwhile, the expression level of AtP5CS, AtSOD and AtPOD genes was up-regulated against salt and drought stresses. Together, our finding indicated that changing the expression level of GmMYB12 in plants alters the accumulation of flavonoids and regulates plantlet tolerance to abiotic stress by regulating osmotic balance, protecting membrane integrity and maintaining ROS homeostasis. The GmMYB12 gene has the potential to be used to increase the content of valuable flavonoids and improve the tolerance to abiotic stresses in plants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12

Similar content being viewed by others

References

  • Albert NW, Lewis DH, Zhang H, Irving LJ, Jameson PE, Davies KM (2009) Light-induced vegetative anthocyanin pigmentation in Petunia. J Exp Bot 60:2191–2202

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Alia, Mohanty P, Matysik J (2001) Effect of proline on the production of singlet oxygen. Amino Acids 21:195–200

    Article  CAS  PubMed  Google Scholar 

  • Balasundram N, Sundram K, Samman S (2006) Phenolic compounds in plants and agri-industrial by-products: antioxidant activity, occurrence, and potential uses. Food Chem 99:191–203

    Article  CAS  Google Scholar 

  • Baudry A, Heim MA, Dubreucq B, Caboche M, Weisshaar B, Lepiniec L (2004) TT2, TT8, and TTG1 synergistically specify the expression of BANYULS and proanthocyanidin biosynthesis in Arabidopsis thaliana. Plant J 39:366–380

    Article  CAS  PubMed  Google Scholar 

  • Ben-Amor M, Flores B, Latche A, Bouzayen M, Pech JC, Romojaro F (1999) Inhibition of ethylene biosynthesis by antisense ACC oxidase RNA prevents chilling injury in Charentais cantaloupe melons. Plant Cell Environ 22:1579–1586

    Article  CAS  Google Scholar 

  • Bohnert HJ, Nelson DE, Jensen RG (1995) Adaptations to environmental stresses. Plant Cell 7:1099–1111

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Buer CS, Imin N, Djordjevic MA (2010) Flavonoids: new roles for old molecules. J Integr Plant Biol 52:98–111

    Article  CAS  PubMed  Google Scholar 

  • Butelli E, Titta L, Giorgio M, Mock HP, Matros A, Peterek S, Schijlen EG, Hall RD, Bovy AG, Luo J, Martin C (2008) Enrichment of tomato fruit with health-promoting anthocyanins by expression of select transcription factors. Nat Biotechnol 26:1301–1308

    Article  CAS  PubMed  Google Scholar 

  • Czemmel SL, Stracke R, Weisshaar B, Cordon N, Harris NN, Walker AR, Robinson SP, Bogs J (2009) The grapevine R2R3-MYB transcription factor VvMYBF1 regulates flavonol synthesis in developing grape berries. Plant Physiol 151:1513–1530

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Deshmukh R, Sonah H, Patil G, Chen W, Prince S, Mutava R, Vuong T, Valliyodan B, Nguyen HT (2014) Integrating omic approaches for abiotic stress tolerance in soybean. Front Plant Sci 5:244

    Article  PubMed  PubMed Central  Google Scholar 

  • Dixon RA, Liu C, Jun JH (2013) Metabolic engineering of anthocyanins and condensed tannins in plants. Curr Opin Biotechnol 24:329–335

    Article  CAS  PubMed  Google Scholar 

  • Fan WJ, Zhang M, Zhang HX, Zhang P (2012) Improved tolerance to various abiotic stresses in transgenic sweetpotato (Ipomoea batatas) expressing spinach betaine aldehyde dehydrogenase. PLoS One 7:e37344

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Feller A, Machemer K, Braun EL, Grotewold E (2011) Evolutionary and comparative analysis of MYB and bHLH plant transcription factors. Plant J 66:94–116

    Article  CAS  PubMed  Google Scholar 

  • Gao JJ, Zhang Z, Peng RH, Xiong AS, Xu J, Zhu B, Yao QH (2011) Forced expression of Mdmyb10, a myb transcription factor gene from apple, enhances tolerance to osmotic stress in transgenic Arabidopsis. Mol Biol Rep 38:205–211

    Article  CAS  PubMed  Google Scholar 

  • Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Bioch 48:909–930

    Article  CAS  Google Scholar 

  • Grotewold E (2006) The genetics and biochemistry of floral pigments. Annu Rev Plant Biol 57:761–780

    Article  CAS  PubMed  Google Scholar 

  • Guo J, Zhou W, Lu Z, Li H, Li H, Gao F (2015) Isolation and functional analysis of chalcone isomerase gene from purple-fleshed sweetpotato. Plant Mol Biol Rep 33:1451–1463

    Article  CAS  Google Scholar 

  • Harborne JB, Williams CA (2000) Advances in flavonoid research since 1992. Phytochemistry 55:481–504

    Article  CAS  PubMed  Google Scholar 

  • Hichri I, Barrieu F, Bogs J, Kappel C, Delrot S, Lauvergeat V (2011) Recent advances in the transcriptional regulation of the flavonoid biosynthetic pathway. J Exp Bot 62:2465–2483

    Article  CAS  PubMed  Google Scholar 

  • Hollman PCH, Katan MB (1999) Dietary flavonoids: intake, health effects and bioavailability. Food Chem Toxicol 37:937–942

    Article  CAS  PubMed  Google Scholar 

  • Hong Z, Lakkineni K, Zhang ZM, Verma DPS (2000) Removal of feedback inhibition of ∆1-pyrroline-5-carboxylate synthetase results in increased proline accumulation and protection of plants from osmotic stress. Plant Physiol 122:1129–1136

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kang C, Zhai H, Xue LY, Zhao Z, He SZ, Liu QC (2018) A lycopene β-cyclase gene, IbLCYB2, enhances carotenoid contents and abiotic stress tolerance in transgenic sweetpotato. Plant Sci 272:243–254

    Article  CAS  PubMed  Google Scholar 

  • Khan N, Adhami VM, Mukhtar H (2010) Apoptosis by dietary agents for prevention and treatment of prostate cancer. Endocr Relat Cancer 17:R39–R52

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Koca H, Ozdemir F, Turkan I (2006) Effect of salt stress on lipid peroxidation and superoxide dismutase and peroxidase activities of Lycopersicon esculentum and L. pennellii. Biol Plant 50:745–748

    Article  CAS  Google Scholar 

  • Krasensky J, Jonak C (2012) Drought, salt, and temperature stress-induced metabolic rearrangements and regulatory networks. J Exp Bot 63:1593–1608

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kulcheski FR, de Oliveira LFV, Molina LG, Almerão MP, Rodrigues FA, Marcolino J, Barbosa JF, Stolf-Moreira R, Nepomuceno AL, Marcelino-Guimarães FC, Abdelnoor RV, Nascimento LC, Carazzolle MF, Pereira GAG, Margis R (2011) Identification of novel soybean microRNAs involved in abiotic and biotic stresses. BMC Genom 12:307

    Article  CAS  Google Scholar 

  • Lai Y, Li H, Yamagishi M (2013) A review of target gene specificity of flavonoid R2R3-MYB transcription factors and a discussion of factors contributing to the target gene selectivity. Front Biol 8:577–598

    Article  CAS  Google Scholar 

  • Lännenpää M (2014) Heterologous expression of AtMYB12 in kale (Brassica oleracea var. acephala) leads to high flavonol accumulation. Plant Cell Rep 33:1377–1388

    Article  CAS  PubMed  Google Scholar 

  • Lepiniec L, Debeaujon I, Routaboul JM, Baudry A, Pourcel L, Nesi N, Caboche M (2006) Genetics and biochemistry of seed flavonoids. Annu Rev Plant Biol 57:405–430

    Article  CAS  PubMed  Google Scholar 

  • Li XW, Li JW, Zhai Y, Zhao Y, Zhao X, Zhang HJ, Su LT, Wang Y, Wang QY (2013) A R2R3-MYB transcription factor, GmMYB12B2, affects the expression levels of flavonoid biosynthesis genes encoding key enzymes in transgenic Arabidopsis plants. Gene 532(1):72–79

    Article  CAS  PubMed  Google Scholar 

  • Li R, Kang C, Song XJ, Yu L, Liu DG, He SZ, Zhai H, Liu QC (2017) A ζ-carotene desaturase gene, IbZDS, increases β-carotene and lutein contents and enhances salt tolerance in transgenic sweetpotato. Plant Sci 262:39–51

    Article  CAS  PubMed  Google Scholar 

  • Liu DG, He SZ, Zhai H, Wang LJ, Zhao Y, Wang B, Li RJ, Liu QC (2014) Overexpression of IbP5CR enhances salt tolerance in transgenic sweetpotato. Plant Cell Tiss Org 117:1–16

    Article  CAS  Google Scholar 

  • Liu Y, Ji X, Nie X, Qu M, Zheng L, Tan Z, Zhao H, Huo L, Liu S, Zhang B, Wang Y (2015) Arabidopsis AtbHLH112 regulates the expression of genes involved in abiotic stress tolerance by binding to their E-box and GCG-box motifs. New Phytol 207:692–709

    Article  CAS  PubMed  Google Scholar 

  • Lou XM, Yao QH, Zhang Z, Peng RH, Xiong AS, Wang KK (2007) Expression of human hepatitis B virus large surface antigen gene in transgenic tomato. Clin Vaccine Immunol 14:464–469

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Luo J, Butelli E, Hill L, Parr A, Niggeweg R, Bailey P, Weisshaar B, Martin C (2008) AtMYB12 regulates caffeoyl quinic acid and flavonol synthesis in tomato: expression in fruit results in very high levels of both types of polyphenol. Plant J 56:316–326

    Article  CAS  PubMed  Google Scholar 

  • Manavalan LP, Guttikonda SK, Tran L-SP, Nguyen HT (2009) Physiological and molecular approaches to improve drought resistance in soybean. Plant Cell Physiol 50:1260–1276

    Article  CAS  PubMed  Google Scholar 

  • Mehrtens F, Kranz H, Bednarek P, Weisshaar B (2005) The Arabidopsis transcription factor MYB12 is a flavonol-specific regulator of phenylpropanoid biosynthesis. Plant Physiol 138:1083–1096

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Petroni K, Tonelli C (2011) Recent advances on the regulation of anthocyanin synthesis in reproductive organs. Plant Sci 181:219–229

    Article  CAS  PubMed  Google Scholar 

  • Qiu J, Gao F, Shen G, Li C, Han X, Zhao Q, Zhao D, Hua X, Pang Y (2013) Metabolic engineering of the phenylpropanoid pathway enhances the antioxidant capacity of Saussurea involucrata. PLoS One 8:e70665

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Quattrocchio F, Wing JF, van derWoude K, Mol JNM, Koes R (1998) Analysis of bHLH and MYB domain proteins: species specific regulatory differences are caused by divergent evolution of target anthocyanin genes. Plant J 13:475–488

    Article  CAS  PubMed  Google Scholar 

  • Routaboul JM, Kerhoas L, Debeaujon I, Pourcel L, Caboche M, Einhorn J, Lepiniec L (2006) Flavonoid diversity and biosynthesis in seed of Arabidopsis thaliana. Planta 224:96–107

    Article  CAS  PubMed  Google Scholar 

  • RoyChoudhury A, Roy C, Sengupta DN (2007) Transgenic tobacco plants overexpressing the heterologous lea gene Rab16A from rice during high salt and water deficit display enhanced tolerance to salinity stress. Plant Cell Rep 26:1839–1859

    Article  CAS  PubMed  Google Scholar 

  • Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108

    Article  CAS  PubMed  Google Scholar 

  • Smirnoff N, Cumbes QJ (1989) Hydroxyl radical scavenging activity of compatible solutes. Phytochemistry 28:1057–1060

    Article  CAS  Google Scholar 

  • Stracke R, Werber M, Weisshaar B (2001) The R2R3-MYB gene family in Arabidopsis thaliana. Curr Opin Plant Biol 4:447–456

    Article  CAS  PubMed  Google Scholar 

  • Stracke R, Favory JJ, Gruber H, Bartelniewoehner L, Bartels S, Binkert M, Funk M, Weisshaar B, Ulm R (2010) The Arabidopsis bZIP transcription factor HY5 regulates expression of the PFG1/MYB12 gene in response to light and ultraviolet-B radiation. Plant Cell Environ 33:88–103

    CAS  PubMed  Google Scholar 

  • Tahara S (2007) A journey of 25 years through the ecological biochemistry of flavonoids. Biosci Biotechnol Biochem 71:1387–1404

    Article  CAS  PubMed  Google Scholar 

  • Tattini M, Galardi C, Pinelli P, Massai R, Remorini D, Agati G (2004) Differential accumulation of flavonoids and hydroxycinnamates in leaves of Ligustrum vulgare under excess light and drought stress. New Phytol 163:547–561

    Article  CAS  Google Scholar 

  • Walker AR, Davison PA, Bolognesiwinfield AC, James CM, Srinivasan N, Blundell TL, Esch JJ, David Marks M, Gray JC (1999) The TRANSPARENT TESTA GLABRA1 locus, which regulates trichome differentiation and anthocyanin biosynthesis in Arabidopsis, encodes a WD40 repeat protein. Plant Cell 11:1337–1349

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang H, Fan W, Li H, Yang J, Huang J, Zhang P (2013) Functional characterization of dihydroflavonol-4-reductase in anthocyanin biosynthesis of purple sweet potato underlies the direct evidence of anthocyanins function against abiotic stresses. PLoS One 8:e78484

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang FB, Kong WL, Wong G, Fu LF, Peng RH, Li ZJ, Yao QH (2016a) AtMYB12 regulates flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis thaliana. Mol Genet Genomics 291:1545–1559

    Article  CAS  PubMed  Google Scholar 

  • Wang FB, Tong WJ, Zhu H, Kong WL, Peng RH, Liu QC, Yao QH (2016b) A novel Cys2/His2 zinc finger protein gene from sweetpotato, IbZFP1, is involved in salt and drought tolerance in transgenic Arabidopsis. Planta 243:783–797

    Article  CAS  PubMed  Google Scholar 

  • Wang FB, Zhu H, Chen DH, Li ZJ, Peng RH, Yao QH (2016c) A grape bHLH transcription factor gene, VvbHLH1, increases the accumulation of flavonoids and enhances salt and drought tolerance in transgenic Arabidopsis thaliana. Plant Cell Tiss Org 125:387–398

    Article  CAS  Google Scholar 

  • Wang FB, Zhu H, Kong WL, Peng RH, Liu QC, Yao QH (2016d) The Antirrhinum AmDEL gene enhances flavonoids accumulation and salt and drought tolerance in transgenic Arabidopsis. Planta 244:59–73

    Article  CAS  PubMed  Google Scholar 

  • Wang FB, Ren GL, Li FS, Qi ST, Xu Y, Wang BW, Yang YL, Ye YX, Zhou Q, Chen XH (2018) A chalcone synthase gene AeCHS from Abelmoschus esculentus regulates flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis. Acta Physiol Plant 40:97

    Article  CAS  Google Scholar 

  • Wong CC, Li HB, Cheng KW, Chen F (2006) A systematic survey of antioxidant activity of 30 Chinese medicinal plants using the ferric reducing antioxidant power assay. Food Chem 97:705–711

    Article  CAS  Google Scholar 

  • Xiong H, Li J, Liu P, Duan J, Zhao Y, Guo X, Li Y, Zhang H, Ali J, Li Z (2014) Overexpression of OsMYB48-1, a novel MYB-related transcription factor, enhances drought and salinity tolerance in rice. PLoS One 9:e92913

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yan J, Wang B, Jiang Y, Cheng L, Wu T (2014) GmFNSII-controlled soybean flavone metabolism responds to abiotic stresses and regulates plant salt tolerance. Plant Cell Physiol 55:74–86

    Article  CAS  PubMed  Google Scholar 

  • Zhai H, Wang FB, Si ZZ, Huo JX, Xing L, An YY, He SZ, Liu QC (2016) A myo-inositol-1-phosphate synthase gene, IbMIPS1, enhances salt and drought tolerance and stem nematode resistance in transgenic sweetpotato. Plant Biotechnol J 14:592–602

    Article  CAS  PubMed  Google Scholar 

  • Zhang X, Henriques R, Lin SS (2006) Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method. Nat Protoc 1:641–646

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Han B, Wang T, Chen S, Li H, Zhang Y, Dai S (2012) Mechanisms of plant salt response: insights from proteomics. J Proteome Res 11:49–67

    Article  CAS  PubMed  Google Scholar 

  • Zhao Q, Zhang H, Wang T, Chen SX, Dai SJ (2013) Proteomics-based investigation of salt-responsive mechanisms in plant roots. J Proteomics 82:230–253

    Article  CAS  PubMed  Google Scholar 

  • Zhu JK (2001) Plant salt tolerance. Trends Plant Sci 6:66–71

    Article  CAS  PubMed  Google Scholar 

  • Zhu JK (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53:247–273

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was funded by the Natural Science Fund for Colleges and Universities in Jiangsu Province of China (17KJB210001), the Talent Introduction Research Project of Huaiyin Institute of Technology (Z301B16534) and the College Student Practice Innovation Program of Jiangsu Province of China (201811049014YJ and 201811049086X).

Author information

Author notes

  1. Feibing Wang, Xuqin Ren, Fan Zhang and Mingyang Qi have contributed equally to this work.

Authors and Affiliations

  1. School of Life Science and Food Engineering, Huaiyin Institute of Technology, Huai’an, 223003, Jiangsu, People’s Republic of China

    Feibing Wang, Xuqin Ren, Mingyang Qi, Huiyun Zhao, Xinhong Chen, Yuxiu Ye, Yi Zhang, Yuan Niu & Qing Zhou

  2. Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014, Jiangsu, People’s Republic of China

    Fan Zhang

  3. Huaiyin Institute of Agricultural Sciences of Xuhuai Region, Huai’an, 223001, Jiangsu, People’s Republic of China

    Jiayin Yang & Shuguang Li

Authors
  1. Feibing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xuqin Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mingyang Qi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Huiyun Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xinhong Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yuxiu Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jiayin Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Shuguang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Yi Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Yuan Niu
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Qing Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Feibing Wang.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Table S1

Primers used in this study (DOCX 24 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, F., Ren, X., Zhang, F. et al. A R2R3-type MYB transcription factor gene from soybean, GmMYB12, is involved in flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis. Plant Biotechnol Rep 13, 219–233 (2019). https://doi.org/10.1007/s11816-019-00530-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11816-019-00530-7

Keywords

Search

Navigation