Skip to main content

Advertisement

Log in

Overexpression of herbaceous peony miR156e-3p improves anthocyanin accumulation in transgenic Arabidopsis thaliana lateral branches

  • Original Article
  • Published:
3 Biotech Aims and scope Submit manuscript

Abstract

microRNAs (miRNAs) play critical regulatory roles in plant growth and development. In the present study, the function of herbaceous peony (Paeonia lactiflora Pall.) miR156e-3p in the regulation of color formation has been investigated. Firstly, P. lactiflora miR156e-3p precursor sequence (pre-miR156e-3p) was isolated. Subsequently, the overexpression vector of pre-miR156e-3p was constructed and transformed into Arabidopsis thaliana. Moreover, the medium screening, GUS staining, polymerase chain reaction (PCR) of the GUS region and real-time quantitative PCR (qRT-PCR) of miR156e-3p all confirmed that the purpose gene had been successfully transferred into Arabidopsis plants and expressed, which resulted in apparent purple lateral branches. And this change in color was caused by the improved anthocyanin accumulation. In addition, expression analysis had shown that the level of miR156e-3p transcript was increased, while transcription level of target gene squamosa promoter binding protein-like gene (SPL1), encoding SPL transcription factor that negatively regulated anthocyanin accumulation, was repressed in miR156e-3p-overexpressing transgenic plants, and its downstream gene dihydroflavonol 4-reductase gene (DFR) that was directly involved in anthocyanin biosynthesis was strongly expressed, which resulted in anthocyanin accumulation of Arabidopsis lateral branches. These findings would improve the understanding of miRNAs regulation of color formation in P. lactiflora.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Chen Z, Gao X, Zhang J (2015) Alteration of osa-miR156e expression affects rice plant architecture and strigolactones (SLs) pathway. Plant Cell Rep 34:767–781

    Article  CAS  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  CAS  Google Scholar 

  • Cui LG, Shan JX, Shi M, Gao JP, Lin HX (2014) The miR156-SPL9-DFR pathway coordinates the relationship between development and abiotic stress tolerance in plants. Plant J 80:1108–1117

    Article  CAS  Google Scholar 

  • Franco-Zorrilla JM, Valli A, Todesco M, Mateos I, Puga MI, Rubio-Somoza I, Leyva A, Weigel D, García JA, Paz-Ares J (2007) Target mimicry provides a new mechanism for regulation of microRNA activity. Nat Genet 39:1033–1037

    Article  CAS  Google Scholar 

  • Gou JY, Felippes FF, Liu CJ, Weigel D, Wang JW (2011) Negative regulation of anthocyanin biosynthesis in Arabidopsis by a miR156-targeted SPL transcription factor. Plant Cell 23:1512–1522

    Article  CAS  Google Scholar 

  • He H, Liang G, Li Y, Wang F, Yu D (2014) Two young microRNAs originating from target duplication mediate nitrogen starvation adaptation via regulation of glucosinolate synthesis in Arabidopsis thaliana. Plant Physiol 164:853–865

    Article  CAS  Google Scholar 

  • Hu G, Fan J, Xian Z, Huang W, Lin D, Li Z (2014) Overexpression of SlREV alters the development of the flower pedicel abscission zone and fruit formation in tomato. Plant Sci 229:86–95

    Article  CAS  Google Scholar 

  • Jia XY, Yu ZQ, Liang JP, Tang GL, Jin LH, Zhang L, He LH, Li RZ (2013) Cloning of Arabidopsis At-pri-miR828 gene and its genetic transformation into tomato. Acta Hortic Sin 40:2419–2428 (in Chinese)

    CAS  Google Scholar 

  • Jian XY, Zhang L, Li GL, Zhang L, Wang XJ, Cao XF, Fang X, Chen F (2010) Identification of novel stress-regulated microRNAs from Oryza sativa L. Genomics 95:47–55

    Article  CAS  Google Scholar 

  • Jiang JX (2014) Identification and functional characterization of microRNAs involved in pollen development in Brassica campestris ssp. Chinensis. Master Dissertation. Zhejiang University, Hangzhou, China (in Chinese)

  • Kozomara A, Griffiths-Jones S (2014) miRBase: annotating high confidence microRNAs using deep sequencing data. Nucleic Acids Res 42:68–73

    Article  Google Scholar 

  • Lee RC, Feinbaum RL, Ambros V (1993) The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 75:843–854

    Article  CAS  Google Scholar 

  • Lei KJ, Liu H (2016) Research advances in plant regulatory hub miR156 and targeted. Chem Life 36:13–20 (in Chinese)

    Google Scholar 

  • Li H, Deng Y, Wu T, Subramanian S, Yu O (2010) Misexpression of miR482, miR1512, and miR1515 increases soybean nodulation. Plant Physiol 153:1759–1770

    Article  CAS  Google Scholar 

  • Rhoades MW, Reinhart BJ, Lim LP, Burge CB, Bartel B, Bartel DP (2002) Prediction of plant microRNA targets. Cell 110:513–520

    Article  CAS  Google Scholar 

  • Rogers K, Chen X (2013) Biogenesis, turnover and mode of action of plant microRNAs. Plant Cell 25:2383–2399

    Article  CAS  Google Scholar 

  • Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 36:1101–1108

    Article  Google Scholar 

  • Schwab R, Palatnik JF, Riester M, Schommer C, Schmid M, Weigel D (2005) Specifc effects of microRNAs on the plant transcriptome. Dev Cell 8:517–527

    Article  CAS  Google Scholar 

  • Shen J (2015) The cloning and drought response of miR858 and the mechanism for it negatively regulates anthocyanin biosynthesis in tomato. Master Dissertation. Shanxi Agricultural University, Taigu, China (in Chinese)

  • Spanudakis E, Jackson S (2014) The role of microRNAs in the control of flowering time. J Exp Bot 65:365–380

    Article  CAS  Google Scholar 

  • Sunkar R, Zhu JK (2004) Novel and stress-regulated microRNAs and other small RNAs from Arabidopsis. Plant Cell 16:2001–2019

    Article  CAS  Google Scholar 

  • Wang JG, Zhang ZS (2005) Herbaceous peonies of China. China Forestry Publishing House, Beijing in Chinese)

    Google Scholar 

  • Wang L, Gu X, Xu D, Wang W, Wang H, Zeng M, Chang Z, Huang H, Cui X (2010a) miR396-targeted AtGRF transcription factors are required for coordination of cell division and differentiation during leaf development in Arabidopsis. J Exp Bot 62:761–773

    Article  Google Scholar 

  • Wang L, Mai YX, Zhang YC, Qian L, Yang HQ (2010b) MicroRNA171c-targeted SCL6-II, SCL6-III, and SCL6-IV genes regulate shoot branching in Arabidopsis. Mol Plant 3:794–806

    Article  Google Scholar 

  • Wang L, Zeng HQ, Song J, Feng SJ, Yang ZM (2015) miRNA778 and SUVH6 are involved in phosphate homeostasis in Arabidopsis. Plant Sci 238:273–285

    Article  CAS  Google Scholar 

  • Xu Q, Liu Y, Zhu A, Wu X, Ye J, Yu K, Guo W, Deng X (2010) Discovery and comparative profiling of microRNAs in a sweet orange red-flesh mutant and its wild type. BMC Genom 11:246

    Article  Google Scholar 

  • Yang F, Cai J, Yang Y, Liu Z (2013) Overexpression of microRNA828 reduces anthocyanin accumulation in Arabidopsis. Plant Cell Tiss Org Cult 2:159–167

    Article  Google Scholar 

  • Yao Y, Guo G, Ni Z, Sunkar R, Du J, Zhu JK, Sun Q (2007) Cloning and characterization of microRNAs from wheat (Triticum aestivum L.). Genome Biol 8:R96

    Article  Google Scholar 

  • Zhang BH, Pan XP, Anderson TA (2006) Identification of 188 conserved maize microRNAs and their targets. FEBS Lett 580:3753–3762

    Article  CAS  Google Scholar 

  • Zhao DQ, Jiang Y, Ning CL, Meng JS, Lin SS, Ding W, Tao J (2014) Transcriptome sequencing of a chimaera reveals coordinated expression of anthocyanin biosynthetic genes mediating yellow formation in herbaceous peony (Paeonia lactiflora Pall.). BMC Genom 15:689

    Article  Google Scholar 

  • Zhao DQ, Wei MR, Liu D, Tao J (2016) Anatomical and biochemical analysis reveal the role of anthocyanins in flower coloration of herbaceous peony. Plant Physiol Biochem 102:97–106

    Article  CAS  Google Scholar 

  • Zhao DQ, Wei MR, Shi M, Hao ZJ, Tao J (2017) Identification and comparative profiling of miRNAs in herbaceous peony (Paeonia lactiflora Pall.) with red/yellow bicoloured flowers. Sci Rep 7:44926

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Natural Science Foundation of China (31,400,592), the program of key members of Yangzhou University outstanding young teachers and the Priority Academic Program Development from Jiangsu Government.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jun Tao.

Ethics declarations

Conflict of interest

Author declares that there is no competing interest towards the publication of this manuscript.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 1205 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhao, D., Xia, X., Wei, M. et al. Overexpression of herbaceous peony miR156e-3p improves anthocyanin accumulation in transgenic Arabidopsis thaliana lateral branches. 3 Biotech 7, 379 (2017). https://doi.org/10.1007/s13205-017-1011-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s13205-017-1011-3

Keywords

Navigation