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Characterization of Citrus WRKY transcription factors and their responses to phytohormones and abiotic stresses

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Biologia Plantarum

Abstract

WRKY transcription factors (TF) family is involved in a huge variety of plant processes, including seed germination, plant development, phytohormone signalling, and defence against both biotic or abiotic stresses. In this work, WRKY TF family has been characterized in citrus. In a first experiment, the relative expression of CsWRKYs was analyzed in shoots and roots of plants treated with abscisic acid (ABA), salicylic acid (SA), and methyl jasmonate (MeJA) under in vitro conditions. Expression of CsWRKYs was also determined in roots of commercial citrus rootstocks subjected to osmotic and salt stresses. A total amount of 50 CsWRKYs has been found and devided into different groups of WRKY family according to the WRKY domain sequences. In response to the applications of phytohormones, the highest differences were observed in roots, and it was found that ABA and SA generally repressed CsWRKYs expressions, but MeJA induced their overexpression. Osmotic stress repressed the expression of most of the CsWRKYs, whereas salt stress induced their expression. Moreover, salt stress induced higher increase in CsWRKY expressions in the tolerant rootstock Citrus macrophylla than in the sensitive rootstock Carrizo citrange, suggesting that these TFs may play an important role in response to this stress.

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Abbreviations

ABA:

abscisic acid

JA:

jasmonic acid

MeJA:

methyl jasmonate

PEG:

polyethylene glycol

RT-qPCR:

reverse transcriptase quantitative polymerase chain reaction

SA:

salicylic acid

TF:

transcription factor

References

  • Agarwal, P., Reddy, M.P., Chikara, J.: WRKY: its structure, evolutionary relationship, DNA-binding selectivity, role in stress tolerance and development of plants. — Mol. Biol. Rep. 38: 3883–3896, 2011.

    Article  CAS  PubMed  Google Scholar 

  • Bakshi, M., Oelmüller, R.: WRKY transcription factors: jack of many trades in plants. — Plant Signal. Behav. 9: 1–18, 2014.

    Article  Google Scholar 

  • Bansal, S., Harrington, C.A., St. Clair, J.B.: Tolerance to multiple climate stressors: a case study of Douglas-fir drought and cold hardiness. — Ecol. Evol. 6: 2074–2083, 2016.

    Article  PubMed  PubMed Central  Google Scholar 

  • Chen, H., Lai, Z., Shi, J., Xiao, Y., Chen, Z., Xu, X.: Roles of Arabidopsis WRKY18, WRKY40 and WRKY60 transcription factors in plant responses to abscisic acid and abiotic stress. — BMC Plant Biol. 10: 281, 2010.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen, L., Liu, Y., Liu, H., Kang, L., Geng, J., Gai, Y., Ding, Y., Sun, H., Li, Y.: Identification and expression analysis of MATE genes involved in flavonoid transport in blueberry plants. — PLoS ONE 10: e0118578, 2015.

    Google Scholar 

  • Chen, L., Song, Y., Li, S., Zhang, L., Zou, C., Yu, D.: The role of WRKY transcription factors in plant abiotic stresses. — Biochim. biophys. Acta 1819: 120–128, 2012.

    Article  CAS  PubMed  Google Scholar 

  • Czarnecki, O., Yang, J., Wang, X., Wang, S., Muchero, W., Tuskan, G.A., Chen, J.-G.: Characterization of MORE AXILLARY GROWTH genes in Populus. — PLoS ONE 9: e102757, 2014.

    Article  Google Scholar 

  • De Ollas, C., Hernando, B., Arbona, V., Gómez-Cadenas, A.: Jasmonic acid transient accumulation is needed for abscisic acid increase in citrus roots under drought stress conditions. — Physiol. Plant. 147: 296–306, 2013.

    Article  PubMed  Google Scholar 

  • Durgbanshi, A., Arbona, V., Pozo, O., Miersch, O., Sancho, J. V., Gómez-Cadenas, A.: Simultaneous determination of multiple phytohormones in plant extracts by liquid chromatography-electrospray tandem mass spectrometry. — J. Agr. Food Chem. 53: 8437–8442, 2005.

    Article  CAS  Google Scholar 

  • Eulgem, T., Rushton, P.J., Robatzek, S., Somssich, I.E.: The WRKY superfamily of plant transcription factors. — Trends Plant Sci. 5: 199–206, 2000.

    Article  CAS  PubMed  Google Scholar 

  • Gómez-Cadenas, A., De Ollas, C., Manzi, M., Arbona, V.: Phytohormonal crosstalk under abiotic stress. - In: Phan Tran, L.S., Pal, S. (ed.): Phytohormones: A Window to Metabolism, Signaling and Biotechnological Applications. Vol. 1. Pp. 289–321. Springer, New York - Heidelberg - Dordrecht - London 2014.

    Chapter  Google Scholar 

  • Gómez-Cadenas, A., Tadeo, F.R., Primo-Millo, E., Talon, M.: Involvement of abscisic acid and ethylene in the responses of citrus seedlings to salt shock. — Physiol. Plant. 103: 475–484, 1998.

    Article  Google Scholar 

  • Gómez-Cadenas, A., Vives, V., Zandalinas, S.I., Manzi, M., Sánchez-Pérez, A.M., Pérez-Clemente, R.M., Arbona, V.: Abscisic acid: a versatile phytohormone in plant signaling and beyond. — Curr. Protein Pept. Sci. 16: 413–434, 2015.

    Article  PubMed  Google Scholar 

  • Gong, X., Zhang, J., Hu, J., Wang, W., Wu, H., Zhang, Q., Liu, J.-H.: FcWRKY70, a WRKY protein of Fortunella crassifolia, functions in drought tolerance and modulates putrescine synthesis by regulating arginine decarboxylase gene. — Plant Cell Environ. 38: 2248–2262, 2015.

    Article  CAS  PubMed  Google Scholar 

  • Guo, C., Guo, R., Xu, X., Gao, M., Li, X., Song, J., Zheng, Y., Wang, X.: Evolution and expression analysis of the grape (Vitis vinifera L.) WRKY gene family. — J. exp. Bot. 65: 1513–1528, 2014.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Huang, G.T., Ma, S.L., Bai, L.P., Zhang, L., Ma, H., Jia, P., Liu, J., Zhong, M., Guo, Z.F.: Signal transduction during cold, salt, and drought stresses in plants. — Mol. Biol. Rep. 39: 969–987, 2012.

    Article  PubMed  Google Scholar 

  • Iglesias, D.J., Levy, Y., Gómez-Cadenas, A., Tadeo, F.R., Primo-Millo, E., Talon, M.: Nitrate improves growth in saltstressed citrus seedlings through effects on photosynthetic activity and chloride accumulation. — Tree Physiol. 24: 1027–1034, 2004.

    Article  CAS  PubMed  Google Scholar 

  • Jiang, Y., Duan, Y., Yin, J., Ye, S., Zhu, J., Zhang, F., Lu, W., Fan, D., Luo, K.: Genome-wide identification and characterization of the Populus WRKY transcription factor family and analysis of their expression in response to biotic and abiotic stresses. — J. exp. Bot. 65: 6629–6644, 2014.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Johnson, C.S., Kolevski, B., Smyth, D.R.: TRANSPARENT TESTA GLABRA2, a trichome and seed coat development gene of Arabidopsis, encodes a WRKY transcription factor. — Plant Cell 14: 1359–1375, 2002.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lamesch, P., Berardini, T.Z., Li, D., Swarbreck, D., Wilks, C., Sasidharan, R., Muller, R., Dreher, K., Alexander, D.L., Garcia-Hernandez, M., Karthikeyan, A.S., Lee, C.H., Nelson, W.D., Ploetz, L., Singh, S., Wensel, A., Huala, E.: The Arabidopsis Information Resource (TAIR): improved gene annotation and new tools. — Nucl. Acids Res. 40: D1202–D1210, 2012.

    Article  CAS  PubMed  Google Scholar 

  • Li, J., Brader, G., Palva, E.T.: The WRKY70 transcription factor: a node of convergence for jasmonate-mediated and salicylate-mediated signals in plant defense. — Plant Cell 16: 319–331, 2004.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu, J.-H., Peng, T., Dai, W.: Critical cis-acting elements and interacting transcription factors: key players associated with abiotic stress responses in plants. — Plant mol. Biol. Rep. 32: 303–317, 2014.

    Article  CAS  Google Scholar 

  • Llorca, C.M., Potschin, M., Zentgraf, U.: bZIPs and WRKYs: two large transcription factor families executing two different functional strategies. — Front. Plant Sci. 5: 169, 2014.

    Article  PubMed  PubMed Central  Google Scholar 

  • Mangelsen, E., Kilian, J., Berendzen, K.W., Kolukisaoglu, U.H., Harter, K., Jansson, C., Wanke, D.: Phylogenetic and comparative gene expression analysis of barley (Hordeum vulgare) WRKY transcription factor family reveals putatively retained functions between monocots and dicots. — BMC Genomics 9: 194, 2008.

    Article  PubMed  PubMed Central  Google Scholar 

  • Michel, B.E., Kaufmann, M.R.: The osmotic potential of polyethylene glycol 6000. — Plant Physiol. 51: 914–916, 1973.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Montoliu, A., Gómez-Cadenas, A., Pérez-Clemente, R.M.: In vitro adventitious rooting of Carrizo citrange microshoots. — HortScience 45: 988–990, 2010.

    Google Scholar 

  • Montoliu, A., López-Climent, M.F., Arbona, V., Pérez-Clemente, R.M., Gómez-Cadenas, A.: A novel in vitro tissue culture approach to study salt stress responses in citrus. — Plant Growth Regul. 59: 179–187, 2009.

    Article  CAS  Google Scholar 

  • Moya, J.L., Gómez-Cadenas, A., Primo-Millo, E., Talon, M.: Chloride absorption in salt-sensitive Carrizo citrange and salt-tolerant Cleopatra mandarin citrus rootstocks is linked to water use. — J. exp. Bot. 54: 825–833, 2003.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nei, M., Kumar, S.: Molecular Evolution and Phylogenetics. - Oxford University Press, New York 2000.

    Google Scholar 

  • Pan, L.-J., Jiang, L.: Identification and expression of the WRKY transcription factors of Carica papaya in response to abiotic and biotic stresses. — Mol. Biol. Rep. 41: 1215–1225, 2014.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pfaffl, M.W.: A new mathematical model for relative quantification in real-time RT-PCR. — Nucl. Acids Res. 29: 2002–2007, 2001.

    Article  Google Scholar 

  • Pfaffl, M.W., Horgan, G.W., Dempfle, L.: Relative expression software tool ( REST © ) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. — Nucl. Acids Res. 30: 1–10, 2002.

    Article  Google Scholar 

  • Ramamoorthy, R., Jiang, S.Y., Kumar, N., Venkatesh, P.N., Ramachandran, S.: A comprehensive transcriptional profiling of the WRKY gene family in rice under various abiotic and phytohormone treatments. — Plant Cell Physiol. 49: 865–879, 2008.

    Article  CAS  PubMed  Google Scholar 

  • Robatzek, S., Somssich, I.E.: Targets of AtWRKY6 regulation during plant senescence and pathogen defense. — Genes Dev. 16: 1139–1149, 2002.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Roy Choudhury, A., Gupta, B., Sengupta, D.N.: Trans-acting factor designated OSBZ8 interacts with both typical abscisic acid responsive elements as well as abscisic acid responsive element-like sequences in the vegetative tissues of indica rice cultivars. — Plant Cell Rep. 27: 779–794, 2008.

    Article  CAS  Google Scholar 

  • Rushton, P.J., Somssich, I.E., Ringler, P., Shen, Q.J.: WRKY transcription factors. — Trends Plant Sci. 15: 247–258, 2010.

    Article  CAS  PubMed  Google Scholar 

  • Saeed, A.I., Bhagabati, N.K., Braisted, J.C., Liang, W., Sharov, V., Howe, E.A., Li, J., Thiagarajan, M., White, J.A., Quackenbush, J.: TM4 microarray software suite. — Methods Enzymol. 411: 134–193, 2006.

    Article  CAS  PubMed  Google Scholar 

  • Sah, S.K., Reddy, K.R., Li, J.: Abscisic acid and abiotic stress tolerance in crop plants. — Front. Plant Sci. 7: 1–26, 2016.

    Article  Google Scholar 

  • Şahin-Çevik, M.: A WRKY transcription factor gene isolated from Poncirus trifoliata shows differential responses to cold and drought stresses. — Plant Omics J. 5: 438–445, 2012.

    Google Scholar 

  • Şahin-Çevik, M., Moore, G.A.: Identification of a drought-and cold-stress inducible WRKY gene in the cold-hardy Citrus relative Poncirus trifoliata. — New Zeal. J. Crops hort. Sci. 41: 57–68, 2013.

    Article  Google Scholar 

  • Saitou, N., Nei, M.: The neighbor-joining nethod: a new method for reconstructing phylogenetic trees. — Mol. Biol. Evol. 4: 406–425, 1987.

    CAS  PubMed  Google Scholar 

  • Shakirova, F.M., Allagulova, C.R., Maslennikova, D.R., Klyuchnikova, E.O., Avalbaev, A.M., Bezrukova, M.V.: Salicylic acid-induced protection against cadmium toxicity in wheat plants. — Environ. exp. Bot. 122: 19–28, 2016.

    Article  CAS  Google Scholar 

  • Skibbe, M., Qu, N., Galis, I., Baldwin, I.T.: Induced plant defenses in the natural environment: Nicotiana attenuata WRKY3 and WRKY6 coordinate responses to herbivory. — Plant Cell 20: 1984–2000, 2008.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Song, H., Wang, P., Hou, L., Zhao, S., Zhao, C., Xia, H., Li, P.: Global analysis of WRKY genes and their response to dehydration and salt stress in soybean. — Front. Plant Sci. 7: 9, 2016.

    PubMed  PubMed Central  Google Scholar 

  • Tamura, K., Stecher, G., Peterson, D., Filipski, A., Kumar, S.: MEGA6: molecular evolutionary genetics analysis version 6.0. — Mol. Biol. Evol. 30: 2725–2729, 2013.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Van Verk, M.C., Pappaioannou, D., Neeleman, L., Bol, J.F., Linthorst, H.J.M.: A Novel WRKY transcription factor is required for induction of PR-1a gene expression by salicylic acid and bacterial elicitors. - Plant Physiol. 146, 1983–1995, 2008.

    Article  PubMed  PubMed Central  Google Scholar 

  • Wei, W., Zhang, Y., Han, L., Guan, Z., Chai, T.: A novel WRKY transcriptional factor from Thlaspi caerulescens negatively regulates the osmotic stress tolerance of transgenic tobacco. — Plant Cell Rep. 27: 795–803, 2008.

    Article  CAS  PubMed  Google Scholar 

  • Woo, H.R., Kim, H.J., Nam, H.G., Lim, P.O.: Plant leaf senescence and death–regulation by multiple layers of control and implications for aging in general. — J. cell. Sci. 126: 4823–4833, 2013.

    Article  CAS  PubMed  Google Scholar 

  • Xie, Z., Zhang, Z.L., Zou, X., Yang, G., Komatsu, S., Shen, Q.J.: Interactions of two abscisic-acid induced WRKY genes in repressing gibberellin signaling in aleurone cells. — Plant J. 46: 231–242, 2006.

    Article  CAS  PubMed  Google Scholar 

  • Yang, B., Jiang, Y., Rahman, M.H., Deyholos, M.K., Kav, N.N. V: Identification and expression analysis of WRKY transcription factor genes in canola (Brassica napus L.) in response to fungal pathogens and hormone treatments. — BMC Plant Biol. 9: 1–19, 2009.

    Article  Google Scholar 

  • Zandalinas, S.I., Rivero, R.M., Martínez, V., Gómez-Cadenas, A., Arbona, V.: Tolerance of citrus plants to the combination of high temperatures and drought is associated to the increase in transpiration modulated by a reduction in abscisic acid levels. — BMC Plant Biol. 16: 105, 2016.

    Article  PubMed  PubMed Central  Google Scholar 

  • Zheng, Z., Qamar, S.A., Chen, Z., Mengiste, T.: Arabidopsis WRKY33 transcription factor is required for resistance to necrotrophic fungal pathogens. — Plant J. 48: 592–605, 2006.

    Article  CAS  PubMed  Google Scholar 

  • Zhou, L., Wang, N.-N., Gong, S.-Y., Lu, R., Li, Y., Li, X.-B.: Overexpression of a cotton (Gossypium hirsutum) WRKY gene, GhWRKY34, in Arabidopsis enhances salt-tolerance of the transgenic plants. — Plant Physiol. Biochem. 96: 311–320, 2015.

    Article  CAS  PubMed  Google Scholar 

  • Zhou, Q.Y., Tian, A.G., Zou, H.F., Xie, Z.M., Lei, G., Huang, J., Wang, C.M., Wang, H.W., Zhang, J.S., Chen, S.Y.: Soybean WRKY-type transcription factor genes, GmWRKY13, GmWRKY21, and GmWRKY54, confer differential tolerance to abiotic stresses in transgenic Arabidopsis plants. — Plant Biotechnol. J. 6: 486–503, 2008.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Departament de Ciències Agràries i del Medi Natural, Universitat Jaume I, E-12071, Castellón de la Plana, Spain

    V. Vives-Peris, D. Marmaneu, A. Gómez-Cadenas & R. M. Pérez-Clemente

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  1. V. Vives-Peris
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  2. D. Marmaneu
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  3. A. Gómez-Cadenas
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  4. R. M. Pérez-Clemente
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Correspondence to R. M. Pérez-Clemente.

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Acknowledgements: This work was supported by the Spanish Ministerio de Economia y Competitividad (MINECO) and the Universitat Jaume I through grants AGL2016-76574-R and UJI-B2016-23, respectively. V.V.-P. was a recipient of a predoctoral contract from the Universitat Jaume I (PREDOC/2013/31).

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Vives-Peris, V., Marmaneu, D., Gómez-Cadenas, A. et al. Characterization of Citrus WRKY transcription factors and their responses to phytohormones and abiotic stresses. Biol Plant 62, 33–44 (2018). https://doi.org/10.1007/s10535-017-0737-4

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  • DOI: https://doi.org/10.1007/s10535-017-0737-4

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