Abstract
Stylosanthes (stylo) species are economically important tropical and subtropical forage legumes. They are vulnerable to chilling and frost, and little is known about the genetic and molecular mechanisms of their responses or adaptations to low temperature stress. Two cold-responsive NAC genes, SgNAC1 and SgNAC2, were selected from a whole transcriptome profiling study of fine-stem stylo (S. guianensis var. intermedia) and further investigated for their roles in cold stress tolerance. Bioinformatic analysis indicated that SgNAC1 and SgNAC2 belonged to ATAF subgroup of NAC family, and shared highly conserved N-terminal A–E NAC subdomains and a EVQS[E/x]PK[W/I] motif with ATAF-like NAC transcription factors. Expression profiling, subcellular location and transactivation assay revealed that SgNAC1 and SgNAC2 encode nucleus-localized polypeptides with transactivation activities and responds promptly to cold stress. Phenotypic and physiological changes indicated that the transgenic tobacco plants overexpressing SgNAC1 and SgNAC2 were more tolerant to cold stress than the wild-type plants. Meanwhile, the expression of SgNAC1 and SgNAC2 were significantly enhanced at the early stages of cold treatment, indicating that overexpression of SgNAC1 and SgNAC2 is sufficient to confer cold tolerance to tobacco plants. These results suggest that SgNAC1 and SgNAC2 are promising candidate genes for cold tolerance improvement strategies in stylo.
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Abbreviations
- NAC:
-
[No apical meristem (NAM), Arabidopsis transcription activation factor (ATAF), cup-shaped cotyledon (CUC)]
- MS medium:
-
Murashige and Skoog medium
- BA:
-
Benzylaminopurine
- NAA:
-
1-Naphthylacetic acid
- WT:
-
Wild-type
- GFP:
-
Green fluorescent protein
- cDNA:
-
Complementary DNA
- His:
-
Histidine
- Trp:
-
Tryptophan
- Leu:
-
Leucine
- YPDA medium:
-
Yeast, peptone, dextrose and adenine medium
References
Ahmad M (2011) Polyamines: natural and engineered abiotic and biotic stress tolerance in plants. Biotechnol Adv 29:300–311
Ariel FD, Manavella PA, Dezar CA, Chan RL (2007) The true story of the HD-Zip family. Trends Plant Sci 12:419–426. https://doi.org/10.1016/j.tplants.2007.08.003
Aslam M, Grover A, Sinha VB et al (2012) Isolation and characterization of cold responsive NAC gene from Lepidium latifolium. Mol Biol Rep 39:9629
Bais HP, Ravishankar GA (2002) Role of polyamines in the ontogeny of plants and their biotechnological applications. Plant Cell Tissue Organ Cult 69:1–34
Bao G, Zhuo C, Qian C et al (2015) Co-expression of NCED and ALO improves vitamin C level and tolerance to drought and chilling in transgenic tobacco and stylo plants. Plant Biotechnol J 1–9. https://doi.org/10.1111/pbi.12374
Bart R, Chern M, Park C-J et al (2006) A novel system for gene silencing using siRNAs in rice leaf and stem-derived protoplasts. Plant Methods 2:13. https://doi.org/10.1186/1746-4811-2-13
Castilhos G, Lazzarotto F, Spagnolo-Fonini L et al (2014) Possible roles of basic helix-loop-helix transcription factors in adaptation to drought. Plant Sci 223:1–7. https://doi.org/10.1016/j.plantsci.2014.02.010
Chen Q, Quan W, Xiong L, Lou Z (2011) A structural view of the conserved domain of rice stress-responsive NAC1. Protein Cell 2:55–63
Chen L, Song Y, Li S et al (2012) The role of WRKY transcription factors in plant abiotic stresses. Biochim Biophys Acta Gene Regul Mech 1819:120–128. https://doi.org/10.1016/j.bbagrm.2011.09.002
Christianson JA, Dennis ES, Llewellyn DJ, Wilson IW (2010) ATAF NAC transcription factors: regulators of plant stress signaling. Plant Signal Behav 5:428–432
Cui Y, Wang Q (2006) Physiological responses of maize to elemental sulphur and cadmium stress. Plant Soil Environ 52:523–529
Delessert C, Wilson IW, Van Der Straeten D et al (2004) Spatial and temporal analysis of the local response to wounding in Arabidopsis leaves. Plant Mol Biol 55:165–181. https://doi.org/10.1007/s11103-004-0112-7
Delessert C, Kazan K, Wilson IW et al (2005) The transcription factor ATAF2 represses the expression of pathogenesis-related genes in Arabidopsis. Plant J 43:745–757. https://doi.org/10.1111/j.1365-313X.2005.02488.x
Ernst HA, Olsen AN, Larsen S, Lo LL (2004) Structure of the conserved domain of ANAC, a member of the NAC family of transcription factors. EMBO Rep 5:297–303
Fang Y, You J, Xie K et al (2008) Systematic sequence analysis and identification of tissue-specific or stress-responsive genes of NAC transcription factor family in rice. Mol Genet Genom 280:547–563
Farrant JM (2010) Mechanisms of desiccation tolerance in resurrection plants: a review from the molecular to whole plant physiological level. S Afr J Bot 76:389
Gill SS, Tuteja N (2010) Polyamines and abiotic stress tolerance in plants. Plant Signal Behav 5:26–33
Goel D, Singh AK, Yadav V et al (2010) Overexpression of osmotin gene confers tolerance to salt and drought stresses in transgenic tomato (Solanum lycopersicum L.). Protoplasma 245:133–141. https://doi.org/10.1007/s00709-010-0158-0
Grigg A, Shelton M, Mullen B (2000) The nature and management of rehabilitated pastures on open-cut coal mines in central Queensland. Trop Grassl 34:241–250
Guo W-L, Wang S-B, Chen R-G et al (2015) Characterization and expression profile of CaNAC2 pepper gene. Front Plant Sci 6:1–9. https://doi.org/10.3389/fpls.2015.00755
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. https://doi.org/10.1073/pnas.1431173100
He XJ, Mu RL, Cao WH et al (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
Hong Y, Zhang H, Huang L et al (2016) Overexpression of a stress-responsive NAC transcription factor gene ONAC022 improves drought and salt tolerance in rice. Front Plant Sci 7:1–19. https://doi.org/10.3389/fpls.2016.00004
Hu H, You J, Fang Y et al (2010) Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice. Plant Mol Biol 72:567–568
Huang G-T, Ma S-L, Bai L-P et al (2012a) Signal transduction during cold, salt, and drought stresses in plants. Mol Biol Rep 39:969–987. https://doi.org/10.1007/s11033-011-0823-1
Huang H, Wang Y, Wang S et al (2012b) Transcriptome-wide survey and expression analysis of stress-responsive NAC genes in Chrysanthemum lavandulifolium. Plant Sci 193–194:18–27
Humphrey LR (1980) A guide to better pastures for the tropics and sub-tropics, 4th edn. Wright Stephenson and Co. (Australia) Pty. Ltd, Melbourne
Jensen MK, Rung JH, Gregersen PL et al (2007) The HvNAC6 transcription factor: a positive regulator of penetration resistance in barley and Arabidopsis. Plant Mol Biol 65:137–150. https://doi.org/10.1007/s11103-007-9204-5
Jensen MK, Kjaersgaard T, Nielsen MM et al (2010) The Arabidopsis thaliana NAC transcription factor family: structure-function relationships and determinants of ANAC019 stress signalling. Biochem J 426:183–196
Jiang Y, Deyholos MK (2006) Comprehensive transcriptional profiling of NaCl-stressed Arabidopsis roots reveals novel classes of responsive genes. BMC Plant Biol 6:25. https://doi.org/10.1186/1471-2229-6-25
Jin H, Huang F, Cheng H et al (2013) Overexpression of the GmNAC2 Gene, an NAC transcription factor, reduces abiotic stress tolerance in tobacco. Plant Mol Biol Rep 31:435–442. https://doi.org/10.1007/s11105-012-0514-7
Kato H, Motomura T, Komeda Y et al (2010) Overexpression of the NAC transcription factor family gene ANAC036 results in a dwarf phenotype in Arabidopsis thaliana. J Plant Physiol 167:571–577. https://doi.org/10.1016/j.jplph.2009.11.004
Kjaersgaard T, Jensen MK, Christiansen MW et al (2011) Senescence-associated barley NAC (NAM, ATAF1,2, CUC) transcription factor interacts with radical-induced cell death 1 through a disordered regulatory domain. J Biol Chem 286:35418–35429
Le DT, Nishiyama R, Watanabe Y et al (2011) Genome-wide survey and expression analysis of the plant-specific NAC transcription factor family in soybean during development and dehydration stress. DNA Res 18:263–276. https://doi.org/10.1093/dnares/dsr015
Li X, Yang X, Hu YX et al (2014) A novel NAC transcription factor from Suaeda liaotungensis K. enhanced transgenic Arabidopsis drought, salt, and cold stress tolerance. Plant Cell Rep 33:767–778. https://doi.org/10.1007/s00299-014-1602-y
Licausi F, Ohme-Takagi M, Perata P (2013) APETALA2/ethylene responsive factor (AP2/ERF) transcription factors: mediators of stress responses and developmental programs. New Phytol. https://doi.org/10.1111/nph.12291
Liu YG, Chen Y (2007) High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences. Biotechniques 43:649–650, 652, 654 passim
Liu X, Zhang BY, Hong L et al (2010) Molecular characterization of Arachis hypogaea NAC 2 (AhNAC2) reveals it as a NAC-like protein in peanut. Biotechnol Biotechnol Equip 24:2066–2070. https://doi.org/10.2478/V10133-010-0085-4
Lu S, Wang X, Guo Z (2013) Differential responses to chilling in Stylosanthes guianensis (Aublet) Sw. and Its mutants. Agron J 105:377–382
Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K (2012) AP2/ERF family transcription factors in plant abiotic stress responses. Biochim Biophys Acta Gene Regul Mech 1819:86–96. https://doi.org/10.1016/j.bbagrm.2011.08.004
Nakashima K, Tran LSP, Van Nguyen D et al (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. https://doi.org/10.1111/j.1365-313X.2007.03168.x
Nakashima K, Takasaki H, Mizoi J et al (2012) NAC transcription factors in plant abiotic stress responses. Biochim Biophys Acta 1819:97–103
Noble AD, Orr DM, Middleton CH, Rogers LG (2000) Legumes in native pasture - asset or liability? a case history with stylo. Trop Grassl 34:199–206
Nogueira FTS, Schlögl PS, Camargo SR et al (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
Nuruzzaman M, Sharoni AM, Kikuchi S (2013) Roles of NAC transcription factors in the regulation of biotic and abiotic stress responses in plants. Front Microbiol 4:1–16. https://doi.org/10.3389/fmicb.2013.00248
Oh SK, Lee S, Yu SH, Choi D (2005) Expression of a novel NAC domain-containing transcription factor (CaNAC1) is preferentially associated with incompatible interactions between chili pepper and pathogens. Planta 222:876–887. https://doi.org/10.1007/s00425-005-0030-1
Ohnishi T, Sugahara S, Yamada T et al (2005) OsNAC6, a member of the NAC gene family, is induced by various stresses in rice. Genes Genet Syst 80:135–139. https://doi.org/10.1266/ggs.80.135
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
Peng H, Yu X, Cheng H et al (2010) Cloning and characterization of a novel NAC family gene CarNAC1 from chickpea (Cicer arietinum L.). Mol Biotechnol 44:30–40
Puranik S, Sahu PP, Srivastava PS, Prasad M (2012) NAC proteins: regulation and role in stress tolerance. Trends Plant Sci 17:369–381. https://doi.org/10.1016/j.tplants.2012.02.004
Qu Y, Duan M, Zhang Z et al (2016) Overexpression of the Medicago falcata NAC transcription factor MfNAC3 enhances cold tolerance in Medicago truncatula. Environ Exp Bot 129:67–76
Rushton DL, Tripathi P, Rabara RC et al (2012) WRKY transcription factors: key components in abscisic acid signalling. Plant Biotechnol J 10:2–11
Seki M, Urano K (2007) Regulatory metabolic networks in drought stress responses. Curr Opin Plant Biol 10:296–302
Shan W, Kuang JF, Lu WJ, Chen JY (2014) Banana fruit NAC transcription factor MaNAC1 is a direct target of MaICE1 and involved in cold stress through interacting with MaCBF1. Plant Cell Environ 37:2116–2127. https://doi.org/10.1111/pce.12303
Shao H, Wang H, Tang X (2015) NAC transcription factors in plant multiple abiotic stress responses: progress and prospects. Front Plant Sci 6:902. https://doi.org/10.3389/fpls.2015.00902
Shen H, Yin Y, Chen F et al (2009) A bioinformatic analysis of NAC genes for plant cell wall development in relation to lignocellulosic bioenergy production. BioEnergy Res 2:217–232
Shen C, Wang S, Bai Y et al (2010) Functional analysis of the structural domain of ARF proteins in rice (Oryza sativa L.). J Exp Bot 61:3971–3981. https://doi.org/10.1093/jxb/erq208
Tran LSP, Quach TN, Guttikonda SK et al (2009) Molecular characterization of stress-inducible GmNAC genes in soybean. Mol Genet Genom 281:647–664. https://doi.org/10.1007/s00438-009-0436-8
Tran LS, Nishiyama R, Yamaguchi-Shinozaki K, Shinozaki K (2010) Potential utilization of NAC transcription factors to enhance abiotic stress tolerance in plants by biotechnological approach. GM Crops 1:32–39
Urano K, Maruyama K, Ogata Y et al (2009) Characterization of the ABA-regulated global responses to dehydration in Arabidopsis by metabolomics. Plant J 57:1065–1078
Wang Z, Rashotte AM, Moss AG, Dane F (2014) Two NAC transcription factors from Citrullus colocynthis, CcNAC1, CcNAC2 implicated in multiple stress responses. Acta Physiol Plant 36:621–634
Wang L, Hu Z, Zhu M et al (2017) The abiotic stress-responsive NAC transcription factor SlNAC11 is involved in drought and salt response in tomato (Solanum lycopersicum L.). Plant Cell Tissue Organ Cult 129:161–174. https://doi.org/10.1007/s11240-017-1167-x
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. https://doi.org/10.1101/gad.852200
Yang J, Guo Z (2007) Cloning of a 9-cis-epoxycarotenoid dioxygenase gene (SgNCED1) from Stylosanthes guianensis and its expression in response to abiotic stresses. Plant Cell Rep 26:1383–1390. https://doi.org/10.1007/s00299-007-0325-8
Yang SD, Seo PJ, Yoon HK, Park CM (2011) The Arabidopsis NAC transcription factor VNI2 integrates abscisic acid signals into leaf senescence via the COR/RD genes. Plant Cell 23:2155–2168
Yu X, Liu Y, Wang S et al (2016) CarNAC4, a NAC-type chickpea transcription factor conferring enhanced drought and salt stress tolerances in Arabidopsis. Plant Cell Rep 35:613–627. https://doi.org/10.1007/s00299-015-1907-5
Yu D, Zhang L, Zhao K et al (2017) VaERD15, a transcription factor gene associated with cold-tolerance in Chinese Wild Vitis amurensis. Front Plant Sci 8:1–13. https://doi.org/10.3389/fpls.2017.00297
Zhai C, Zhang Y, Yao N et al (2014) Function and interaction of the coupled genes responsible for Pik-h encoded rice blast resistance. PLoS ONE. https://doi.org/10.1371/journal.pone.0098067
Zhang L, Zhao G, Jia J et al (2012) Molecular characterization of 60 isolated wheat MYB genes and analysis of their expression during abiotic stress. J Exp Bot 63:203–214
Zhong R, Lee C, Ye ZH (2010) Global analysis of direct targets of secondary wall NAC master switches in Arabidopsis. Mol Plant 3:1087–1103
Zhou B, Guo Z, Liu Z (2005a) Effects of abscisic acid on antioxidant systems of Stylosanthes guianensis (Aublet) Sw. under chilling stress. Crop Sci 45:599–605
Zhou B, Guo Z, Xing J, Huang B (2005b) Nitric oxide is involved in abscisic acid-induced antioxidant activities in Stylosanthes guianensis. J Exp Bot 56:3223–3228. https://doi.org/10.1093/jxb/eri319
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Financial support from the National Natural Science Foundation of China (Nos. 31601990, 31272491 and 31802116) is gratefully acknowledged.
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XMX conceived and initiated the project. PLZ and SWK designed and performed most of the experiments and data analysis. PLZ and SC wrote the article; XQZ and BLF participated in plasmid construction, plant transformation and physiological determination; SC, TXZ, LL and PYZ conducted the data collection and bioinformatic analysis.
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Communicated by Sergio J. Ochatt.
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Zhan, PL., Ke, SW., Zhang, PY. et al. Overexpression of two cold-responsive ATAF-like NAC transcription factors from fine-stem stylo (Stylosanthes guianensis var. intermedia) enhances cold tolerance in tobacco plants. Plant Cell Tiss Organ Cult 135, 545–558 (2018). https://doi.org/10.1007/s11240-018-1486-6
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DOI: https://doi.org/10.1007/s11240-018-1486-6