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EjMYB4 is a transcriptional activator of 4-Coumarate:coenzyme A ligase involved in lignin biosynthesis in loquat (Eriobotrya japonica)

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Abstract

Lignin is one of the major structural compounds for plants and the transcriptional regulatory mechanisms governing its production have been extensively studied in model plants. However, the underlying mechanisms may differ between organs and information is still limited for fruit lignification. Loquat accumulates lignin not only in vegetative organs but also in fruit flesh. To gain insight into the transcriptional regulation of loquat lignification, yeast one hybrid library screening was conducted with the 4-Coumarate:coenzyme A ligase (Ej4CL1) promoter and four proteins (EjNF-YC1, EjNF-YC2, EjMYB4 and EjGTE1) were identified with binding affinity. Dual-luciferase assays showed that EjMYB4 and EjGTE1 selectively trans-activated promoters of lignin biosynthesis genes from both loquat and Arabidopsis, while EjNF-YC1 and EjNF-YC2 didn’t have the trans-activation effects. Gene expression analysis indicated that EjMYB4 was higher expressed in vegetative tissues compared with reproductive tissues, whereas EjGTE1 expression was not correlated with lignin content in different tissues. Subcellular localization results indicated that EjMYB4 was located mainly in the nucleus and electrophoretic mobility-shift assay further validated that EjMYB4 directly binds to AC-element of Ej4CL1 promoter. EjMYB4 transient over-expression in tobacco leaves led to an increase in lignin content and up-regulation of tobacco endogenous lignin related genes. These results revealed the function of EjMYB4 in regulating lignin biosynthesis in loquat.

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References

  • Airoldi CA, Rovere FD, Falasca G, Marino G, Kooiker M, Altamura MM, Citterio S, Kater MM (2010) The Arabidopsis BET bromodomain factor GTE4 is involved in maintenance of the mitotic cell cycle during plant development. Plant Physiol 152:1320–1334

    Article  CAS  Google Scholar 

  • Baudin M, Laloum T, Lepage A, Rã­Podas C, Ariel F, Frances L, Crespi M, Gamas P, Blanco FA, Zanetti ME, Carvalho-Niebel F, Niebel A (2015) A phylogenetically conserved group of nuclear factor-Y transcription factors interact to control nodulation in legumes. Plant Physiol 169:2761–2773

    CAS  PubMed  PubMed Central  Google Scholar 

  • Boerjan W, Ralph J, Baucher M (2003) Lignin biosynthesis. Annu Rev Plant Biol 54:519–546

    Article  CAS  Google Scholar 

  • Bomal C, Bedon F, Caron S, Mansfield SD, Levasseur C, Cooke JEK, Blais S, Tremblay L, Morency MJ, Pavy N, Grima-Pettenati J, Segui A, MacKay J (2008) Involvement of Pinus taeda MYB1 and MYB8 in phenylpropanoid metabolism and secondary cell wall biogenesis: a comparative in planta analysis. J Exp Bot 59:3925–3939

    Article  CAS  Google Scholar 

  • Cai C, Xu CJ, Li X, Ferguson IB, Chen KS (2006a) Accumulation of lignin in relation to change in activities of lignification enzymes in loquat fruit flesh after harvest. Postharvest Biol Technol 40:163–169

    Article  CAS  Google Scholar 

  • Cai C, Li X, Chen KS (2006b) Acetylsalicylic acid alleviates chilling injury of postharvest loquat (Eriobotrya japonica Lindl.) fruit. Eur Food Res Technol 223:533–539

    Article  CAS  Google Scholar 

  • Cai C, Xu CJ, Shan LL, Li X, Zhou CH, Zhang WS, Ferguson I, Chen KS (2006c) Low temperature conditioning reduces postharvest chilling injury in loquat fruit. Postharvest Biol Technol 41:252–259

    Article  CAS  Google Scholar 

  • Cai C, Chen KS, Xu WP, Zhang WS, Li X, Ferguson I (2006d) Effect of 1-MCP on postharvest quality of loquat fruit. Postharvest Biol Technol 40:155–162

    Article  CAS  Google Scholar 

  • Cai X, Ballif J, Endo S, Davis E, Liang M, Chen D, DeWald D, Kreps J, Zhu T, Wu Y (2007) A putative CCAAT-binding transcription factor is a regulator of flowering timing in Arabidopsis. Plant Physiol 145:98–105

    Article  CAS  Google Scholar 

  • Cao SF, Zheng YH, Wang KT, Rui HJ, Tang SS (2008) Effect of methyl jasmonate on cell wall modification of loquat fruit in relation to chilling injury after harvest. Food Chem 118:641–647

    Article  Google Scholar 

  • Chua YL, Channelière S, Mott E, Gray JC (2005) The bromodomain protein GTE6 controls leaf development in Arabidopsis by histone acetylation at ASYMMETRIC LEAVES1. Genes Dev 19:2245–2254

    Article  CAS  Google Scholar 

  • Duque P, Chua NH (2003) IMB1, a bromodomain protein induced during seed imbibition, regulates ABA- and phyA-mediated responses of germination in Arabidopsis. Plant J 35:787–799

    Article  CAS  Google Scholar 

  • Espley RV, Hellens RP, Putterill J, Stevenson DE, Kutty-Amma S, Allan AC (2007) Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. Plant J 49:414–427

    Article  CAS  Google Scholar 

  • Fornale S, Shi XH, Cha CL, Encina A, Irar S, Capellades M, Fuguet E, Torres JL, Rovira P, Puigdomenech P, Rigau J, Grotewold E, Gray J, Caparrós-Ruiz D (2010) ZmMYB31 directly represses maize lignin genes and redirects the phenylpropanoid metabolic flux. Plant J 64:633–644

    Article  CAS  Google Scholar 

  • Ge H, Zhang J, Zhang YJ, Li X, Yin XR, Grierson D, Chen KS (2017) EjNAC3 transcriptionally regulates chilling-induced lignification of loquat fruit via physical interaction with an atypical CAD-like gene. J Exp Bot 68:5129–5136

    Article  CAS  Google Scholar 

  • Goicoechea M, Lacombe E, Legay S, Mihaljevic S, Rech P, Jauneau A, Lapierre C, Pollet B, Verhaegen D, Chaubet-Gigot N, Grima-Pettenati J (2005) EgMYB2, a new transcriptional activator from Eucalyptus xylem, regulates secondary cell wall formation and lignin biosynthesis. Plant J 43:553–567

    Article  CAS  Google Scholar 

  • Guo HY, Wang YC, Wang LQ, Hu P, Wang YM, Jia YY, Zhang CR, Zhang Y, Zhang YM, Wang C (2017) Expression of the MYB transcription factor gene BplMYB46 affects abiotic stress tolerance and secondary cell wall deposition in Betula platyphylla. Plant Biotechnol J 15:107–121

    Article  CAS  Google Scholar 

  • He Y, Fu JL, Yu CL, Wang XM, Jiang QS, Hong J, Lu KX, Xue GP, Yan CQ, James A, Xu LG, Chen JP, Jiang DA (2015) Increasing cyclic electron flow is related to Na+ sequestration into vacuoles for salt tolerance in soybean. J Exp Bot 66:6877–6889

    Article  CAS  Google Scholar 

  • Huang C, Hu G, Li F, Li Y, Wu J, Zhou X (2013) NbPHAN, a MYB transcriptional factor, regulates leaf development and affects drought tolerance in Nicotiana benthamiana. Physiol Plant 149:297–309

    CAS  PubMed  Google Scholar 

  • Jin HL, Cominelli E, Bailey P, Parr A, Mehrtens F, Jones J, Tonelli C, Weisshaar B, Martin C (2000) Transcriptional repression by AtMYB4 controls production of UV-protecting sunscreens in Arabidopsis. EMBO J 19:6150–6161

    Article  CAS  Google Scholar 

  • Kosma DK, Murmu J, Razeq FM, Santos P, Bourgault R, Molina I, Rowland O (2014) AtMYB41 activates ectopic suberin synthesis and assembly in multiple plant species and cell types. Plant J 80:216–229

    Article  CAS  Google Scholar 

  • Kumimoto RW, Adam L, Hymus GJ, Repetti PP, Reuber TL, Marion CM, Hempel FD, Ratcliffe OJ (2008) The Nuclear Factor Y subunits NF-YB2 and NF-YB3 play additive roles in the promotion of flowering by inductive long-day photoperiods in Arabidopsis. Planta 228:709–723

    Article  CAS  Google Scholar 

  • Kumimoto RW, Zhang Y, Siefers N, Holt BF (2010) NF-YC3, NF-YC4 and NF-YC9 are required for CONSTANS-mediated, photoperiod-dependent flowering in Arabidopsis thaliana. Plant J 63:379–391

    Article  CAS  Google Scholar 

  • Laloum T, De MS, Gamas P, Baudin M, Niebel A (2013) CCAAT-box binding transcription factors in plants: Y so many? Trends Plant Sci 18:157–166

    Article  CAS  Google Scholar 

  • Laporte P, Lepage A, Fournier J, Catrice O, Moreau S, Jardinaud MF, Mun JH, Larrainzar E, Cook DR, Gamas P, Niebel A (2014) The CCAAT box-binding transcription factor NF-YA1 controls rhizobial infection. J Exp Bot 65:481–494

    Article  CAS  Google Scholar 

  • Legay S, Sivadon P, Blervacq AS, Pavy N, Baghdady A, Tremblay L, Levasseur C, Ladouce N, Lapierre C, Seguin A, Hawkins S, Mackay J, Grima-Pettenati J (2010) EgMYB1, an R2R3 MYB transcription factor from eucalyptus negatively regulates secondary cell wall formation in Arabidopsis and poplar. New Phytol 188:774–786

    Article  CAS  Google Scholar 

  • Li CF, Wang XQ, Lu WX, Liu R, Tian QY, Sun YM, Luo KM (2014) A poplar R2R3-MYB transcription factor, PtrMYB152, is involved in regulation of lignin biosynthesis during secondary cell wall formation. Plant Cell Tissue Org 119:553–563

    Article  CAS  Google Scholar 

  • Li R, Reddy VA, Jin JJ, Rajan C, Wang Q, Yue GH, Lim CH, Chua NH, Ye J, Sarojam R (2017a) Comparative transcriptome analysis of oil palm flowers reveals an EAR-motif-containing R2R3-MYB that modulates phenylpropene biosynthesis. BMC Plant Biol 17:219

    Article  Google Scholar 

  • Li X, Zang C, Ge H, Zhang J, Donald G, Yin XR, Chen KS (2017b) Involvement of PAL, C4H, and 4CL in chilling injury-induced flesh lignification of loquat fruit. HortScience 52:127–131

    Article  Google Scholar 

  • Lippold F, Sanchez DH, Musialak M, Schlereth A, Scheible WR, Hincha DK, Udvardi MK (2009) AtMYB41 regulates transcriptional and metabolic responses to osmotic stress in Arabidopsis. Plant Physiol 149:1761–1772

    Article  CAS  Google Scholar 

  • Liu J, Osbourn A, Ma P (2015) MYB transcription factors as regulators of phenylpropanoid metabolism in plants. Mol Plant 8:689–708

    Article  CAS  Google Scholar 

  • Mantovani R (1999) The molecular biology of the CCAAT-binding factor NF-Y. Gene 239:15–27

    Article  CAS  Google Scholar 

  • McCarthy RL, Zhong RQ, Fowler S, Lyskowski D, Piyasena H, Carleton K, Spicer C, Ye ZH (2010) The poplar MYB transcription factors, PtrMYB3 and PtrMYB20, are involved in the regulation of secondary wall biosynthesis. Plant Cell Physiol 51:1084–1090

    Article  CAS  Google Scholar 

  • Misra A, Mcknight TD, Mandadi KK (2018) Bromodomain proteins GTE9 and GTE11 are essential for specific BT2-mediated sugar and ABA responses in Arabidopsis thaliana. Plant Mol Bio 96:393–402

    Article  CAS  Google Scholar 

  • Nakano Y, Yamaguchi M, Endo H, Rejab NA, Ohtani M (2015) NAC-MYB-based transcriptional regulation of secondary cell wall biosynthesis in land plants. Front Plant Sci 6:288

    Article  Google Scholar 

  • Patzlaff A, McInnis S, Courtenay A, Surman C, Newman LJ, Smith C, Bevan MW, Mansfield S, Whetten RW, Sederoff RR, Campbell MM (2003) Characterization of a pine MYB that regulates lignification. Plant J 36:743–754

    Article  CAS  Google Scholar 

  • Preston J, Wheeler J, Heazlewood J, Li SF, Parish RW (2004) AtMYB32 is required for normal pollen development in Arabidopsis thaliana. Plant J 40:979–995

    Article  CAS  Google Scholar 

  • Shan LL, Li X, Wang P, Cai C, Zhang B, Sun CD, Zhang WS, Xu CJ, Ferguson IB, Chen KS (2008) Characterization of cDNAs associated with lignification and their expression profiles in loquat fruit with different lignin accumulation. Planta 227:1243–1254

    Article  CAS  Google Scholar 

  • Sonbol FM, Fornale S, Capellades M, Encina A, Tourino S, Torres JL, Rovira P, Ruel K, Puigdomenech P, Rigau J, Caparrós-Ruiz D (2009) The maize ZmMYB42 represses the phenylpropanoid pathway and affects the cell wall structure, composition and degradability in Arabidopsis thaliana. Plant Mol Biol 70:283–296

    Article  CAS  Google Scholar 

  • Tucker G, Yin XR, Zhang AD, Wang MM, Zhu QG, Liu XF, Xie XL, Chen KS, Grierson D (2017) Ethylene and fruit softening. Food Qual Saf 1:253–267

    Article  Google Scholar 

  • Vanholme R, Demedts B, Morreel K, Ralph J, Boerjan W (2010) Lignin biosynthesis and structure. Plant Physiol 153:895–905

    Article  CAS  Google Scholar 

  • Wang P, Zhang B, Li X, Xu CJ, Yin XR, Shan LL, Ferguson IB, Chen KS (2010) Ethylene signal transduction elements involved in chilling injury in non-climacteric loquat fruit. J Exp Bot 61:179–190

    Article  CAS  Google Scholar 

  • Wang WQ, Zhang J, Ge H, Li SJ, Li X, Yin XR, Donald G, Chen KS (2016) EjMYB8 transcriptionally regulates flesh lignification in loquat fruit. PLoS ONE 11:e0154399

    Article  Google Scholar 

  • Wei QH, Zhang F, Sun FS, Luo QC, Wang RB, Hu R, Chen MJ, Chang JL, Yang GX, He GY (2017) A wheat MYB transcriptional repressor TaMyb1D regulates phenylpropanoid metabolism and enhances tolerance to drought and oxidative stresses in transgenic tobacco plants. Plant Sci 265:112–123

    Article  CAS  Google Scholar 

  • Xu Q, Yin XR, Zeng JK, Ge H, Song M, Xu CJ, Li X, Ferguson IB (2014) Activator- and repressor-type MYB transcription factors are involved in chilling injury induced flesh lignification in loquat via their interactions with the phenylpropanoid pathway. J Exp Bot 65:4349–4359

    Article  CAS  Google Scholar 

  • Xu Q, Wang WW, Zeng JK, Zhang J, Grierson D, Li X, Xin XR, Chen KS (2015) A NAC transcription factor, EjNAC1, affects lignification of loquat fruit by regulating lignin. Postharvest Biol Technol 102:25–31

    Article  CAS  Google Scholar 

  • Xu JJ, Zhang XF, Xue HW (2016) Rice aleurone layer specific OsNF-YB1 regulates grain filling and endosperm development by interacting with an ERF transcription factor. J Exp Bot 67:6399–6411

    Article  CAS  Google Scholar 

  • Xue C, Yao JL, Qin MF, Zhang MY, Allan AC, Wang DF, Wu J (2018) PbrmiR397a regulates lignification during stone cell development in pear fruit. Plant Biotechnol J. https://doi.org/10.1111/pbi.12950

    Article  PubMed  PubMed Central  Google Scholar 

  • Yin XR, Allan AC, Chen KS, Ferguson IB (2010) Kiwifruit EIL and ERF genes involved in regulating fruit ripening. Plant Physiol 153:1280–1292

    Article  CAS  Google Scholar 

  • Zeng JK, Li X, Xu Q, Chen JY, Yin XR, Ferguson IB, Chen KS (2015) EjAP2-1, an AP2/ERF gene, is a novel regulator of fruit lignification induced by chilling injury, via interaction with EjMYB transcription factors. Plant Biotechnol J 13:1325–1334

    Article  CAS  Google Scholar 

  • Zeng JK, Li X, Zhang J, Ge H, Yin XR, Chen KS (2016) Regulation of loquat fruit low temperature response and lignification involves interaction of heat shock factors and genes associated with lignin biosynthesis. Plant Cell Environ 39:1780–1789

    Article  CAS  Google Scholar 

  • Zhang J, Ge H, Zang C, Li X, Donald G, Chen KS, Yin XR (2016) EjODO1, a MYB transcription factor, regulating lignin biosynthesis in developing loquat (Eriobotrya japonica) fruit. Front Plant Sci 7:1360

    PubMed  PubMed Central  Google Scholar 

  • Zhang AD, Wang WQ, Tong Y, Li MJ, Grierson D, Ferguson IB, Chen KS, Yin XR (2018) Transcriptome analysis identifies a zinc finger protein regulating starch degradation in kiwifruit. Plant Physiol. https://doi.org/10.1104/pp.18.00427

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhao Q (2016) Lignification: flexibility, biosynthesis and regulation. Trends Plant Sci 21:713–721

    Article  CAS  Google Scholar 

  • Zhong R, Lee C, Zhou J, McCarthy RL, Ye ZH (2008) A battery of transcription factors involved in the regulation of secondary cell wall biosynthesis in Arabidopsis. Plant Cell 20:2763–2782

    Article  CAS  Google Scholar 

  • Zhong R, McCarthy RL, Haghighat M, Ye ZH (2013) The poplar MYB master switches bind to the SMRE site and activate the secondary wall biosynthetic program during wood formation. PLoS ONE 8:e69219

    Article  CAS  Google Scholar 

  • Zhou J, Lee C, Zhong R, Ye ZH (2009) MYB58 and MYB63 are transcriptional activators of the lignin biosynthetic pathway during secondary cell wall formation in Arabidopsis. Plant Cell 21:248–266

    Article  CAS  Google Scholar 

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Acknowledgements

This research was supported by the National Natural Science Foundation of China (31630067), the National Natural Science Foundation of China (31722042), the Natural Science Foundation of Zhejiang Province, China (LR16C150001), and the 111 Project (B17039).

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Authors and Affiliations

  1. Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology, Zhejiang University, Zijingang Campus, Hangzhou, People’s Republic of China

    Jing Zhang, Meng-xue Zhang, Meng Xu, Xue-ren Yin, Donald Grierson & Kun-song Chen

  2. The State Agriculture Ministry Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Zhejiang University, Zijingang Campus, Hangzhou, People’s Republic of China

    Xue-ren Yin & Kun-song Chen

  3. Plant & Crop Sciences Division, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, UK

    Donald Grierson

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  1. Jing Zhang
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  2. Meng-xue Zhang
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Zhang, J., Zhang, Mx., Xu, M. et al. EjMYB4 is a transcriptional activator of 4-Coumarate:coenzyme A ligase involved in lignin biosynthesis in loquat (Eriobotrya japonica). Plant Growth Regul 86, 413–421 (2018). https://doi.org/10.1007/s10725-018-0439-8

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