McMYB4 improves temperature adaptation by regulating phenylpropanoid metabolism and hormone signaling in apple

Abstract

Temperature changes affect apple development and production. Phenylpropanoid metabolism and hormone signaling play a crucial role in regulating apple growth and development in response to temperature changes. Here, we found that McMYB4 is induced by treatment at 28 °C and 18 °C, and McMYB4 overexpression results in flavonol and lignin accumulation in apple leaves. Yeast one-hybrid (Y1H) assays and electrophoretic mobility shift assays (EMSAs) further revealed that McMYB4 targets the promoters of the flavonol biosynthesis genes CHS and FLS and the lignin biosynthesis genes CAD and F5H. McMYB4 expression resulted in higher levels of flavonol and lignin biosynthesis in apple during growth at 28 °C and 18 °C than during growth at 23 °C. At 28 °C and 18 °C, McMYB4 also binds to the AUX/ARF and BRI/BIN promoters to activate gene expression, resulting in acceleration of the auxin and brassinolide signaling pathways. Taken together, our results demonstrate that McMYB4 promotes flavonol biosynthesis and brassinolide signaling, which decreases ROS contents to improve plant resistance and promotes lignin biosynthesis and auxin signaling to regulate plant growth. This study suggests that McMYB4 participates in the abiotic resistance and growth of apple in response to temperature changes by regulating phenylpropanoid metabolism and hormone signaling.

References

[1] Ding, Y. L., Shi, Y. T. & Yang, S. H. Molecular regulation of plant responses to environmental temperatures. Mol. Plant. 13, 544-564 (2010).

[2] Li, H. et al. MPK3- and MPK6-mediated ICE1 phosphorylation negatively regulates ICE1 stability and freezing tolerance in Arabidopsis. Dev. Cell. 43, 630-642 (2017).

[3] Xu, Y. et al. Natural variations of SLG1 confer high-temperature tolerance in indica rice. Nat. Commun. 11, 5441 (2020).

[4] Prinzenberg, A. E., Campos-Dominguez, L., Kruijer, W., Harbinson, J. & Aarts, M. G. M. Natural variation of photosynthetic efficiency in Arabidopsis thaliana accessions under low temperature conditions. Plant Cell Environ. 43, 2000-2013 (2020).

[5] Huo, L. Q. et al. MdATG18a overexpression improves basal thermotolerance in transgenic apple by decreasing damage to chloroplasts. Hortic. Res. 7, 1-15 (2020).

[6] Tian, J. et al. McMYB10 regulates coloration via activating McF3’H and later structural genes in ever-red leaf crabapple. Plant Biotechnol. J. 13, 948-961 (2015).

[7] Wang, Y. C. et al. Ethylene increases the cold tolerance of apple via the MdERF1B-MdCIbHLH1 regulatory module. Plant J. 105, 1-15 (2021).

[8] Ma, D. W. & Constabel, C. P. MYB repressors as regulators of phenylpropanoid metabolism in plants. Trends Plant Sci. 24, 275-289 (2019).

[9] Korn, M., Peterek, S., Mock, H. P., Heyer, A. G. & Hincha, D. K. Heterosis in the freezing tolerance, and sugar and flavonoid contents of crosses between Arabidopsis thaliana accessions of widely varying freezing tolerance. Plant Cell Environ. 31, 813-827 (2008).

[10] Yin, R. et al. Kaempferol 3-O-rhamnoside-7-O-rhamnoside is an endogenous flavonol inhibitor of polar auxin transport in Arabidopsis shoots. N. Phytol. 201, 466-475 (2014).

[11] Yang, C. et al. Lignin metabolism involves Botrytis cinerea BcGs1-induced defense response in tomato. BMC Plant Biol. 18, 103 (2018).

[12] Jin, K. et al. Imaging the dynamic deposition of cell wall polymer in xylem and phloem in Populus × euramericana. Planta 248, 849-858 (2018).

[13] Takeda, Y. et al. Down regulation of p-COUMAROYL ESTER 3-HYDROXYLASE in rice leads to altered cell wall structures and improves biomass saccharification. Plant J. 95, 796-811 (2018).

[14] Šípošová, K., Kollárová, K., Lišková, D. & Vivodová, Z. The effects of IBA on the composition of maize root cell walls. J. Plant Physiol. 239, 10-17 (2019).

[15] Ployet, R. et al. Long cold exposure induces transcriptional and biochemical remodelling of xylem secondary cell wall in Eucalyptus. Tree Physiol. 38, 409-422 (2018).

[16] Artlip, T., McDermaid, A., Ma, Q. & Wisniewski, M. Differential gene expression in nontransgenic and transgenic “M.26” apple overexpressing a peach CBF gene during the transition from eco-dormancy to bud break. Hortic. Res. 6, 86 (2019).

[17] Song, C. et al. Genome-wide identification and expression profiling of the YUCCA gene family in Malus domestica. Sci. Rep. 10, 10866 (2020).

[18] Bawa, G. et al. Gibberellins and auxin regulate soybean hypocotyl elongation under low light and high-temperature interaction. Physiol. Plant. 170, 345-356 (2020).

[19] Chen, T. et al. TRANSTHYRETIN-LIKE and BYPASS1-LIKE co-regulate growth and cold tolerance in Arabidopsis. BMC Plant Biol. 20, 332 (2020).

[20] Zhang, F. et al. Effects of low-temperature stress and brassinolide application on the photosynthesis and leaf structure of tung tree seedlings. Front Plant Sci. 10, 1767 (2020).

[21] Song, T. T. et al. Identification of new regulators through transcriptome analysis that regulate anthocyanin biosynthesis in apple leaves at low temperatures. PLoS ONE 14, e0210672 (2019).

[22] Bhatia, C., Pandey, A., Gaddam, S. R., Hoecker, U. & Trivedi, P. K. Low temperature-enhanced flavonol synthesis requires light-associated regulatory components in Arabidopsis thaliana. Plant Cell Physiol. 59, 2099-2112 (2018).

[23] An, J. P. et al. An apple MYB transcription factor regulates cold tolerance and anthocyanin accumulation and undergoes MIEL1-mediated degradation. Plant Biotechnol. J. 18, 337-353 (2020).

[24] Rao, X. & Dixon, R. A. Current models for transcriptional regulation of secondary cell wall biosynthesis in Grasses. Front Plant Sci. 9, 399 (2018).

[25] Chezem, W. R., Memon, A., Li, F. S., Weng, J. K. & Clay, N. K. SG2-type R2R3-MYB transcription factor MYB15 controls defense-induced lignification and basal immunity in Arabidopsis. Plant Cell 29, 1907-1926 (2017).

[26] Moser, M. et al. MdDAM1 The MADS-Box gene controls growth cessation and bud dormancy in Apple. Front Plant Sci. 11, 1003 (2020).

[27] Fang, H. et al. MdCOL4 interaction mediates crosstalk between UV-B and high temperature to control fruit coloration in Apple. Plant Cell Physiol. 60, 1055-1066 (2019).

[28] Pastore, C. et al. Whole plant temperature manipulation affects flavonoid metabolism and the transcriptome of grapevine berries. Front Plant Sci. 8, 929 (2017).

[29] Sun, C. H., Yang, C. Y. & Tzen, J. T. C. Camellia sinensis molecular identification and characterization of hydroxycinnamoyl transferase in tea plants (L.). Int J. Mol. Sci. 19, 3938 (2018).

[30] Yu, X. et al. Accumulation of flavonoid glycosides and UFGT gene expression in mulberry leaves (Morus alba L.) before and after Frost. Chem. Biodivers. 14, 1-8 (2017).

[31] Xu, Q. et al. 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 (2014).

[32] Zeng, J. K. et al. 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 (2015).

[33] Fornalé, S. et al. ZmMYB31 directly represses maize lignin genes and redirects the phenylpropanoid metabolic flux. Plant J. 64, 633-644 (2010).

[34] Liu, W. et al. Switchgrass (Panicum virgatum L.) promoters for green tissue-specific expression of the MYB4 transcription factor for reduced-recalcitrance transgenic switchgrass. Biotechnol. Biofuels 11, 122 (2018).

[35] Zhou, K. et al. MdUGT88F1-mediated phloridzin biosynthesis regulates Apple development and Valsa canker resistance. Plant Physiol. 180, 2290-2305 (2019).

[36] Wang, F. et al. SlHY5 integrates temperature, light, and hormone signaling to balance plant growth and cold tolerance. Plant Physiol. 179, 749-760 (2019).

[37] van der Woude, L. C. et al. Arabidopsis thaliana HISTONE DEACETYLASE 9 stimulates auxin-dependent thermomorphogenesis in by mediating H2A.Z depletion. PNAS 116, 25343-25354 (2019).

[38] Lucie, C. et al. Root-derived GA12 contributes to temperature-induced shoot growth in Arabidopsis. Nat. Plants 5, 1216-1221 (2019).

[39] Nieto, C., Luengo, L. M. & Prat, S. Regulation of COP1 function by Brassinosteroid signaling. Front Plant Sci. 11, 1151 (2020).

[40] Zhao, M. et al. Transcriptome analysis reveals a positive effect of brassinosteroids on the photosynthetic capacity of wucai under low temperature. BMC Genomics 20, 810 (2019).

[41] Carole, L. G. et al. A conditional mutation in SCD1 reveals linkage between PIN protein trafficking, auxin transport, gravitropism, and lateral root initiation. Front Plant Sci. 11, 910 (2020).

[42] Huot, B., Yao, J., Montgomery, B. L. & He, S. Y. Growth-defense tradeoffs in plants: a balancing act to optimize fitness. Mol. Plant. 7, 1267-1287 (2014).

[43] Maloney, G. S., DiNapoli, K. T. & Muday, G. K. The anthocyanin reduced tomato mutant demonstrates the role of flavonols in tomato lateral root and root hair development. Plant Physiol. 166, 614-631 (2014).

[44] Muhlemann, J. K., Younts, T. L. B. & Muday, G. K. Flavonols control pollen tube growth and integrity by regulating ROS homeostasis during high-temperature stress. PNAS 115, E11188-E11197 (2018).

[45] Silva-Navas, J. et al. Flavonols mediate root phototropism and growth through regulation of proliferation-to-differentiation transition. Plant Cell 28, 1372-1387 (2016).

[46] Sudheeran, P. K. et al. Induced defense response in red mango fruit against Colletotrichum gloeosporioides. Hortic. Res. 8, 17 (2021).

[47] Dong, N. N. et al. Regulation of Brassinosteroid signaling and salt resistance by SERK2 and potential utilization for crop improvement in rice. Front Plant Sci. 11, 621859 (2020).

[48] He, H. M. et al. The Arabidopsis mediator complex subunit 8 regulates oxidative stress responses. Plant Cell koab079, 1-64 (2021).

[49] Li, H. et al. Methyl jasmonate mediates melatonin-induced cold tolerance of grafted watermelon plants. Hortic. Res. 8, 57 (2021).

[50] Han, M. H. et al. Exogenous melatonin positively regulates lignin biosynthesis in Camellia sinensis. Int J. Biol. Macromol. 179, 485-499 (2021).

[51] Xi, Y. L. et al. IAA3-mediated repression of PIF proteins coordinates light and auxin signaling in Arabidopsis. PLoS Genet. 17, e1009384 (2021).

[52] Zhao, C. D. et al. Apple TIME FOR COFFEE contributes to freezing tolerance by promoting unsaturation of fatty acids. Plant Sci. 302, 110695 (2021).

[53] Ma, W. Y. et al. Melatonin alters the secondary metabolite profile of grape berry skin by promoting VvMYB14-mediated ethylene biosynthesis. Hortic. Res. 8, 43 (2021).