Abstract
The NAC (NAM, ATAF1, -2, and CUC2) gene family encodes a large family of plant-specific transcription factors that play diverse roles in plant development and stress regulation. In this study, we performed a survey of citrus NAC transcription factors in the HarvEST: Citrus database, in which 45 NAC domain-containing proteins were identified and phylogenetically classified into 13 different subfamilies. The results suggest the existence of a structurally diversified family of NAC transcription factors in citrus, which has not been previously characterized. One of these NAC genes, CsNAC1 was found to be a member of the stress-NAC subfamily, whose homologs from other plant species function in pathways of environmental stress response and tolerance, and was further characterized. The CsNAC1 deduced protein was shown to contain the five N-terminal A through E NAC subdomains, a C-terminal region containing three transcriptional activation motifs, and a predicted NAC nuclear localization signal, consistent with its putative role as a NAC transcription factor. In silico analysis indicated that CsNAC1 was primarily expressed in leaves and shoot meristems, and was involved in general stress responses. Quantitative real-time reverse transcription PCR analysis revealed that CsNAC1 was strongly induced by drought stress in leaves of Citrus reshni and Citrus limonia, and also by salt stress, cold, and ABA in leaves and roots of C. reshni. Collectively, these results suggest that CsNAC1 encodes a novel stress-responsive NAC transcription factor that is potentially useful for engineering tolerance to multiple abiotic stresses in citrus.
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References
Boyer JS (1982) Plant productivity and environment. Science 218:443–448
Chaves MM, Oliveira MM (2004) Mechanisms underlying plant resilience to water deficits: prospects for water-saving agriculture. J Exp Bot 55:2365–2384
Chen C, Costa MGC, Yu Q, Moore GA, Gmitter FG Jr (2010) Identification of novel members in sweet orange carotenoid biosynthesis gene families. Tree Genet Genome. 6:905–914
Collinge M, Boller T (2001) Differential induction of two potato genes, Stprx2 and StNAC, in response to infection by Phytophthora infestans and to wounding. Plant Mol Biol 46:521–529
Delessert C, Wilson IW, Van Der Straeten D, Dennis ES, Dolferus R (2004) Spatial and temporal analysis of the local response to wounding in Arabidopsis leaves. Plant Mol Biol 55:165–181
Duval M, Hsieh T, Kim S, Thomas T (2002) Molecular characterization of AtNAM: a member of the Arabidopsis NAC domain superfamily. Plant Mol Biol 50:237–248
Fan J, Gao X, Yang Y-W, Deng W, Li Z-G (2007) Molecular cloning and characterization of a NAC-like gene in “navel” orange fruit response to postharvest stresses. Plant Mol Biol Report 25:145–153
Fang Y, You J, Xie K, Xie W, Xiong L (2008) Systematic sequence analysis and identification of tissue-specific or stress-responsive genes of NAC transcription factor family in rice. Mol Genet Genomics 280:535–546
Fujita M, Fujita Y, Maruyama K, Seki M, Hiratsu K, Ohme-Takagi M, Tran L-SP, Yamaguchi-Shinozaki K, Shinozaki K (2004) A dehydration-induced NAC protein, RD26, is involved in a novel ABA-dependent stress-signaling pathway. Plant J 39:863–876
Greve K, La Cour T, Jensen MK, Poulsen FM, Skriver K (2003) Interactions between plant RING-H2 and plant-specific NAC (NAM/ATAF1/2/CUC2) proteins: RING-H2 molecular specificity and cellular localization. Biochem J 371:97–108
Guo Y, Gan S (2006) AtNAP, a NAC family transcription factor, has an important role in leaf senescence. Plant J 46:601–612
Hauck P, Thilmony R, He SY (2003) A Pseudomonas syringae type III effector suppresses cell wall-based extracellular defense in susceptible Arabidopsis plants. Proc Natl Acad Sci USA 100:8577–8582
He XJ, Mu RL, Cao WH, Zhang ZG, Zhang JS, Chen SY (2005) AtNAC2, a transcription factor downstream of ethylene and auxin signaling pathways, is involved in salt stress response and lateral root development. Plant J 44:903–916
Hegedus D, Yu M, Baldwin D, Gruber M, Sharpe A, Parkin I, Whitwill S, Lydiate D (2003) Molecular characterization of Brassica napus NAC domain transcriptional activators induced in response to biotic and abiotic stress. Plant Mol Biol 53:383–397
Hu H, You J, Fang Y, Zhu X, Qi Z, Xiong L (2008) Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice. Plant Mol Biol 67:169–181
Liu Y-Z, Baig MNR, Fan R, Ye J-L, Cao Y-C, Deng X-X (2009) Identification and expression pattern of a novel NAM, ATAF, and CUC-like gene from Citrus sinensis Osbeck. Plant Mol Biol Report 27:292–297
Mahajan S, Tuteja N (2005) Cold, salinity and drought stresses: an overview. Arch Biochem Biophys 444:139–158
Meng C, Cai C, Zhang T, Guo W (2009) Characterization of six novel NAC genes and their responses to abiotic stresses in Gossypium hirsutum L. Plant Sci 176:352–359
Mitsuda N, Iwase A, Yamamoto H, Yoshida M, Seki M, Shinozaki K, Ohme-Takagi M (2007) NAC transcription factors, NST1 and NST3 are key regulators of the formation of secondary walls in woody tissues of Arabidopsis. Plant Cell 19:270–280
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497
Nakashima K, Tran LS, Van Nguyen D, Fujita M, Maruyama K, Todaka D, Ito Y, Hayashi N, Shinozaki K, Yamaguchi-Shinozaki K (2007) Functional analysis of a NAC-type transcription factor OsNAC6 involved in abiotic and biotic stress-responsive gene expression in rice. Plant J 51:617–630
Nogueira FTS, Schlogl PS, Camargo SR, Fernandez JH, De Rosa VE, Jr PP, Arruda P (2005) SsNAC23, a member of the NAC domain protein family, is associated with cold, herbivory and water stress in sugarcane. Plant Sci 169:93–106
Ohnishi T, Sugahara S, Yamada T, Kikuchi K, Yoshiba Y, Hirano HY, Tsutsumi N (2005) OsNAC6, a member of the NAC gene family, is induced by various stresses in rice. Genes Genet Syst 80:135–139
Oliveira MLP, Febres VJ, Costa MGC, Moore GA, Otoni WC (2009) High-efficiency Agrobacterium-mediated transformation of citrus via sonication and vacuum infiltration. Plant Cell Rep 28:387–395
Olsen AN, Ernst HA, Leggio LL, Skriver K (2005) NAC transcription factors: structurally distinct, functionally diverse. Trends Plant Sci 10:79–87
Ooka H, Satoh K, Doi K et al (2003) Comprehensive analysis of NAC family genes in Oryza sativa and Arabidopsis thaliana. DNA Res 10:239–247
Pinheiro GL, Marques CS, Costa MDBL, Reis PAB, Alves MS, Carvalho CM, Fietto LG, Fontes EPB (2009) Complete inventory of soybean NAC transcription factors: sequence conservation and expression analysis uncover their distinct roles in stress response. Gene 444:10–23
Ren T, Qu F, Morris TJ (2000) HRT gene function requires interaction between a NAC protein and viral capsid protein to confer resistance to turnip crinkle virus. Plant Cell 12:1917–1926
Riechmann JL, Heard J, Martin G et al (2000) Arabidopsis transcription factors: genome-wide comparative analysis among eukaryotes. Science 290:2105–2110
Sablowski RW, Meyerowitz EM (1998) A homolog of NO APICAL MERISTEM is an immediate target of the floral homeotic genes APETALA3/PISTILLATA. Cell 92:93–103
Saibo NJM, Lourenço T, Oliveira MM (2009) Transcription factors and regulation of photosynthetic and related metabolism under environmental stresses. Ann Bot 103:609–623
Saitou N, Nei M (1987) The Neighbor-Joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Sharp RE, Poroyko V, Hejlek LG, Spollen WG, Springer GK, Bohnert HJ, Nguyen HT (2004) Root growth maintenance during water deficits: physiology to functional genomics. J Exp Bot 55:2343–2351
Sinclair TR, Ludlow MM (1986) Influence of soil water supply on the plant water balance of four tropical grain legumes. Aust J Plant Physiol 13:329–341
Souer E, van Houwelingen A, Kloos D, Mol J, Koes R (1996) The no apical meristem gene of Petunia is required for pattern formation in embryos and flowers and is expressed at meristem and primordial boundaries. Cell 85:159–170
Storey R, Walker RR (1999) Citrus and salinity. Sci Hortic 78:39–81
Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599
Thompson JD, Higgins DG, Gibson TJ (1994) Clustal-W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680
Tran LS, Nakashima K, Sakuma Y, Simpson SD, Fujita Y, Maruyama K, Fujita M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2004) Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter. Plant Cell 16:2481–2498
Tran L-SP, Quach TN, Guttikonda SK, Aldrich DL, Kumar R, Neelakandan A, Valliyodan B, Nguyen HT (2009) Molecular characterization of stress-inducible GmNAC genes in soybean. Mol Genet Genomics 28:647–664
Uauy C, Distelfeld A, Fahima T, Blechl A, Dubcovsky J (2006) A NAC gene regulating senescence improves grain protein, zinc, and iron content in wheat. Science 314:1298–1301
Wahbi A, Sinclair TR (2007) Transpiration response of Arabidopsis, maize, and soybean to drying of artificial and mineral soil. Environ Exp Bot 59:188–192
Xie Q, Frugis G, Colgan D, Chua NH (2000) Arabidopsis NAC1 transduces auxin signal downstream of TIR1 to promote lateral root development. Genes Dev 14:3024–3036
Yamaguchi-Shinozaki K, Shinozaki K (2006) Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Annu Rev Plant Biol 57:781–803
Yoo YS, Kim Y, Kim YS, Lee JS, Ahn HJ (2007) Control of flowering time and cold response by a NAC-domain protein in Arabidopsis. PLoS ONE 2:e642
Zheng X, Chen B, Lu G, Han B (2009) Overexpression of a NAC transcription factor enhances rice drought and salt tolerance. Biochem Biophys Res Commun 379:985–989
Zhong R, Demura T, Ye ZH (2006) SND1, a NAC domain transcription factor, is a key regulator of secondary wall synthesis in fibers of Arabidopsis. Plant Cell 18:3158–3170
Acknowledgments
This research was supported by research grants from Embrapa (Macroprograma 2), CNPq (Brasília, Brazil), and FAPESP (São Paulo, Brazil). We gratefully acknowledge the Masters Degree scholarships to T.M. de Oliveira and L.C. Cidade by FAPESB (Salvador, Brazil) and CAPES (Brasília, Brazil), respectively. We thank Dr. Claudia Fortes Ferreira (Embrapa, Brazil) for helpful advices and the critical reading of the manuscript.
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Deduced amino acid sequences of the CsNAC proteins identified in the HarvEST: Citrus database (DOC 35 kb)
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de Oliveira, T.M., Cidade, L.C., Gesteira, A.S. et al. Analysis of the NAC transcription factor gene family in citrus reveals a novel member involved in multiple abiotic stress responses. Tree Genetics & Genomes 7, 1123–1134 (2011). https://doi.org/10.1007/s11295-011-0400-8
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DOI: https://doi.org/10.1007/s11295-011-0400-8