Abstract
The NAC transcription factor (TF) plays a pivotal role in resisting various abiotic stresses, and it has been proved that the overexpression of NAC transcription factors (TFs) in Arabidopsis is beneficial to increase drought tolerance. However, the function of NAC genes in chrysanthemum remains unclear. Here, we conducted physiological and molecular experiments to assess the role of DgNAC1 in response to drought stress of chrysanthemum. DgNAC1-overexpressed chrysanthemum was significantly more drought-resistant than wild type (WT). Especially, the transgenic chrysanthemum presented a greater survival rates (86.42, 88.89 and 87.65%, respectively) than WT (41.98%) under drought conditions. It also showed higher leaf water potential and relative water content; lower relative electrolyte conductivity; fewer accumulations of malondialdehyde, hydrogen peroxide (H2O2), superoxide anion (O2−); higher activities of superoxide dismutase, peroxidase, catalase; and more content of proline, chlorophyll, soluble protein, soluble sugar, glutathione (GSH), glutathione disulphide (GSSG), as well as higher ratio of GSH/GSSG than those of WT during drought stress. Moreover, stress-responsive genes in transgenic chrysanthemum showed a significant up-regulation than in WT under drought stress. Therefore, all the above results suggested that DgNAC1 served as an active regulator in chrysanthemum’s responses to drought stress, and it may have a significant impact on the molecular breeding of drought-resistant plants as well.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Agarwal PK, Agarwal P, Reddy MK, Sopory SK (2006) Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Rep 25(12):1263–1274. https://doi.org/10.1007/s00299-006-0204-8
Aida M, Ishida T, Fukaki H, Fujisawa H, Tasaka M (1997) Genes involved in organ separation in Arabidopsis: an analysis of the cup-shaped cotyledon mutant. Plant Cell 9(6):841–857. https://doi.org/10.1105/tpc.9.6.841
Anderson ME (1985) Determination of glutathione and glutathione disulfide in biological samples. Methods Enzymol 113(4):548–555. https://doi.org/10.1016/s0076-6879(85)13073-9
Ashraf M, Foolad M (2007) Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environ Exp Bot 59(2):206–216. https://doi.org/10.1016/j.envexpbot.2005.12.006
Barrs HD, Weatherley PE (1962) A re-examination of the relative turgidity technique for estimating water deficits in leaves. Aust J Biol Sci 15(3):413–428. https://doi.org/10.1071/bi9620413
Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39(1):205–207. https://doi.org/10.1007/bf00018060
Boase MR, Bradley JM, Borst NK (1998) Genetic transformation mediated by Agrobacterium tumefaciens of florists’ chrysanthemum (Dendranthema × grandiflorum) cultivar ‘Peach Margaret’. In Vitro Cell Dev Biol Plant 34(1):46–51. https://doi.org/10.1007/bf02823122
Borochov A, Tirosh T, Mayak S (1986) The fate of membrane proteins during flower senescence. Acta Hort 181:75–80. https://doi.org/10.17660/actahortic.1986.181.7
Chen KM, Gong HJ, Chen GC, Wang SM, Zhang CL (2004) Gradual drought under field conditions influences the glutathione metabolism, redox balance and energy supply in spring wheat. J Plant Growth Regul 23(1):20–28. https://doi.org/10.1007/s00344-003-0053-4
Chen S, Cui X, Chen Y, Gu C, Miao H, Gao H et al (2011) CgDREBa transgenic chrysanthemum confers drought and salinity tolerance. Environ Exp Bot 74:255–260. https://doi.org/10.1016/j.envexpbot.2011.06.007
Chen L, Chen Y, Jiang J, Chen S, Chen F, Guan Z et al (2012) The constitutive expression of Chrysanthemum dichrum ICE1 in Chrysanthemum grandiflorum improves the level of low temperature, salinity and drought tolerance. Plant Cell Rep 31(9):1747–1758. https://doi.org/10.1007/s00299-012-1288-y
Da Silva JAT (2003) Chrysanthemum: advances in tissue culture, cryopreservation, postharvest technology, genetics and transgenic biotechnology. Biotechnol Adv 21(8):715–766. https://doi.org/10.1016/s0734-9750(03)00117-4
Duval M, Hsieh TF, Kim SY, Thomas TL (2002) Molecular characterization of AtNAM: a member of the Arabidopsis NAC domain superfamily. Plant Mol Biol 50(2):237–248. https://doi.org/10.1023/A:1016028530943
Erpen L, Devi HS, Grosser JW et al (2018) Potential use of the DREB/ERF, MYB, NAC and WRKY transcription factors to improve abiotic and biotic stress in transgenic plants. Plant Cell Tissue Organ Cult 132(1):1–25. https://doi.org/10.1007/s11240-017-1320-6
Fan Q, Song A, Xin J, Chen S, Jiang J, Wang Y et al (2015) CmWRKY15 facilitates Alternaria tenuissima infection of chrysanthemum. PLoS ONE 10(11):e0143349. https://doi.org/10.1371/journal.pone.0143349
Fan Q, Song A, Jiang J, Zhang T, Sun H, Wang Y et al (2016) CmWRKY1 enhances the dehydration tolerance of chrysanthemum through the regulation of ABA-associated genes. PLoS ONE 11(3):e0150572. https://doi.org/10.1371/journal.pone.0150572
Farmer EE, Mueller MJ (2013) ROS-mediated lipid peroxidation and RES-activated signaling. Annu Rev Plant Biol 64:429–450. https://doi.org/10.1146/annurev-arplant-050312-120132
Foyer CH, Shigeoka S (2011) Understanding oxidative stress and antioxidant functions to enhance photosynthesis. Plant Physiol 155(1):93–100. https://doi.org/10.1104/pp.110.166181
Gao F, Xiong A, Peng R, Jin X, Xu J, Zhu B et al (2010) OsNAC52, a rice NAC transcription factor, potentially responds to ABA and confers drought tolerance in transgenic plants. Plant Cell Tissue Organ Cult 100(3):255–262. https://doi.org/10.1007/s11240-009-9640-9
Giannopolitis CN, Ries SK (1977) Superoxide dismutases I. Occurrence in higher plants. Plant physiol 59(2):309–314. https://doi.org/10.1104/pp.59.2.309
He C, He Y, Liu Q, Liu T, Liu C, Wang L et al (2013) Co-expression of genes ApGSMT2 and ApDMT2 for glycinebetaine synthesis in maize enhances the drought tolerance of plants. Mol Breed 31(3):559–573. https://doi.org/10.1007/s11032-012-9815-7
Hong B, Tong Z, Ma N, Li J, Kasuga M, Yamaguchi-Shinozaki K et al (2006) Heterologous expression of the AtDREB1A gene in chrysanthemum increases drought and salt stress tolerance. Sci China Ser C 49(5):436–445. https://doi.org/10.1007/s11427-006-2014-1
Hu L, Wang Z, Huang B (2013) Effects of cytokinin and potassium on stomatal and photosynthetic recovery of kentucky bluegrass from drought stress. Crop Sci 53(1):221–231. https://doi.org/10.2135/cropsci2012.05.0284
Hussain SS, Kayani MA, Amjad M (2011) Transcription factors as tools to engineer enhanced drought stress tolerance in plants. Biotechnol Prog 27(2):297–306. https://doi.org/10.1002/btpr.514
Irigoyen JJ, Einerich DW, Sánchez-Díaz M (1992) Water stress induced changes in concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativa) plants. Physiol Plant 84(1):55–60. https://doi.org/10.1111/j.1399-3054.1992.tb08764.x
Jaffar AM, Song A, Muhammad F, Chen S, Jiang J, Liu C et al (2016) Involvement of CmWRKY10 in drought tolerance of chrysanthemum through the ABA-signaling pathway. Int J Mol Sci 17(5):693. https://doi.org/10.3390/ijms17050693
Jensen MK, Lindemose S, De Masi F, Reimer JJ, Nielsen M, Perera V et al (2013) ATAF1 transcription factor directly regulates abscisic acid biosynthetic gene NCED3 in Arabidopsis thaliana. FEBS Open Biol 3(1):321–327. https://doi.org/10.1016/j.fob.2013.07.006
Jia JJ, Meng XP, Liu R, Xia M, Wang XP (2008) Overexpression of AtNCED3 gene in rice exhibits improved drought tolerance in transgenic plants. J Fudan Univ Nat Sci 47(3):288–294. https://doi.org/10.3724/sp.j.1005.2008.011051
Kalir A, Poljakoffmayber A (1981) Changes in activity of malate dehydrogenase, catalase, peroxidase and superoxide dismutase in leaves of halimione portulacoides (l.) aellen exposed to high sodium chloride concentrations. Ann Bot 47(1):75–85. https://doi.org/10.1093/oxfordjournals.aob.a086002
Kjaersgaard T, Jensen MK, Christiansen MW, Gregersen P, Kragelund BB, Skriver K (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(41):35418–35429. https://doi.org/10.1074/jbc.m111.247221
Li Z, Zhang J, Li J, Li H, Zhang G (2016) The functional and regulatory mechanisms of the thellungiella salsuginea ascorbate peroxidase 6 (TsAPX6) in response to salinity and water deficit stresses. PLoS ONE 11(4):e0154042. https://doi.org/10.1371/journal.pone.0154042
Liang QY, Wu YH, Wang K, Bai ZY, Liu QL, Pan YZ et al (2017) Chrysanthemum WRKY gene DgWRKY5 enhances tolerance to salt stress in transgenic chrysanthemum. Sci Rep 7(1):4799. https://doi.org/10.1038/s41598-017-05170-x
Lissarre M, Ohta M, Sato A, Miura K (2010) Cold-responsive gene regulation during cold acclimation in plants. Plant Signal Behav 5(8):948–952. https://doi.org/10.4161/psb.5.8.12135
Liu QL, Xu KD, Zhao LJ, Pan YZ, Jiang BB, Zhang HQ et al (2011) Overexpression of a novel chrysanthemum NAC transcription factor gene enhances salt tolerance in tobacco. Biotechnol Lett 33(10):2073–2082. https://doi.org/10.1007/s10529-011-0659-8
Liu QL, Xu KD, Zhong M, Pan YZ, Jiang BB, Liu GL et al (2013) Overexpression of a novel chrysanthemum Cys2/His2-type zinc finger protein gene DgZFP3 confers drought tolerance in tobacco. Biotechnol Lett 35(11):1953–1959. https://doi.org/10.1007/s10529-013-1289-0
Liu QL, Xu KD, Pan YZ, Jiang BB, Liu GL, Jia Y et al (2014) Functional analysis of a novel chrysanthemum WRKY transcription factor gene involved in salt tolerance. Plant Mol Biol Rep 32(1):282–289. https://doi.org/10.1007/s11105-013-0639-3
Lu PL, Chen NZ, An R, Su Z, Qi BS, Ren F et al (2007) A novel drought-inducible gene, ATAF1, encodes a NAC family protein that negatively regulates the expression of stress-responsive genes in Arabidopsis. Plant Mol Biol 63(2):289–305. https://doi.org/10.1007/s11103-006-9089-8
Lu M, Zhang DF, Shi YS et al (2013) Expression of SbSNAC1, a NAC transcription factor from sorghum, confers drought tolerance to transgenic Arabidopsis. Plant Cell Tissue Organ Cult 115(3):443–455. https://doi.org/10.1007/s11240-013-0375-2
Mao X, Zhang H, Qian X, Li A, Zhao G, Jing R (2012) TaNAC2, a NAC-type wheat transcription factor conferring enhanced multiple abiotic stress tolerances in Arabidopsis. J Exp Bot 63(8):2933–2946. https://doi.org/10.1093/jxb/err462
Miller GAD, Suzuki N, Ciftci-Yilmaz S, Mittler RO (2010) Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant Cell Environ 33(4):453–467. https://doi.org/10.1111/j.1365-3040.2009.02041.x
Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9(10):490–498. https://doi.org/10.1016/j.tplants.2004.08.009
Morran S, Eini O, Pyvovarenko T, Parent B, Singh R, Ismagul A et al (2011) Improvement of stress tolerance of wheat and barley by modulation of expression of DREB/CBF factors. Plant Biotechnol J 9(2):230–249. https://doi.org/10.1111/j.1467-7652.2010.00547.x
Nakashima K, Ito Y, Yamaguchi-Shinozaki K (2009) Transcriptional regulatory networks in response to abiotic stresses in Arabidopsis and grasses. Plant Physiol 149(1):88–95. https://doi.org/10.1104/pp.108.129791
Nakashima K, Takasaki H, Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K (2012) NAC transcription factors in plant abiotic stress responses. BBA-Gene Regul Mech 1819(2):97–103. https://doi.org/10.3724/sp.j.1005.2012.00993
Negi NP, Shrivastava DC, Sharma V, Sarin NB (2015) Overexpression of CuZnSOD from Arachis hypogaea alleviates salinity and drought stress in tobacco. Plant Cell Rep 34(7):1109–1126. https://doi.org/10.1007/s00299-015-1770-4
Nuruzzaman M, Manimekalai R, Sharoni AM, Satoh K, Kondoh H, Ooka H et al (2010) Genome-wide analysis of NAC transcription factor family in rice. Gene 465(1):30–44. https://doi.org/10.1016/j.gene.2010.06.008
Olsen AN, Ernst HA, Leggio LL, Skriver K (2005) NAC transcription factors: structurally distinct, functionally diverse. Trends Plant Sci 10(2):79–87. https://doi.org/10.1016/j.tplants.2004.12.010
Ooka H, Satoh K, Doi K, Nagata T, Otomo Y, Murakami K et al (2003) Comprehensive analysis of NAC family genes in Oryza sativa and Arabidopsis thaliana. DNA Res 10(6):239–247. https://doi.org/10.1093/dnares/10.6.239
Pan Y, Wu LJ, Yu ZL (2006) Effect of salt and drought stress on antioxidant enzymes activities and SOD isoenzymes of liquorice (Glycyrrhiza uralensis Fisch). Plant Growth Regul 49(2–3):157–165. https://doi.org/10.1007/s10725-006-9101-y
Ranieri A, Petacco F, Castagna A, Soldatini GF (2000) Redox state and peroxidase system in sunflower plants exposed to ozone. Plant Sci 159(1):159–167. https://doi.org/10.1016/s0168-9452(00)00352-6
Scholander PF, Bradstreet ED, Hemmingsen EA, Hammel HT (1965) Sap pressure in vascular plants: negative hydrostatic pressure can be measured in plants. Science 148(3668):339–346. https://doi.org/10.1126/science.148.3668.339
Shan H, Chen S, Jiang J, Chen F, Chen Y, Gu C et al (2011) Heterologous expression of the chrysanthemum R2R3-MYB transcription factor CmMYB2 enhances drought and salinity tolerance, increases hypersensitivity to ABA and delays flowering in Arabidopsis thaliana. Mol Biotechnol 51(2):160–173. https://doi.org/10.1007/s12033-011-9451-1
Shi J, Fu XZ, Peng T, Huang XS, Fan QJ, Liu JH (2010) Spermine pretreatment confers dehydration tolerance of citrus in vitro plants via modulation of antioxidative capacity and stomatal response. Tree Physiol 30(7):914–922. https://doi.org/10.1093/treephys/tpq030
Shiriga K, Sharma R, Kumar K, Yadav SK, Hossain F, Thirunavukkarasu N (2014) Genome-wide identification and expression pattern of drought-responsive members of the NAC family in maize. Meta Gene 2:407–417. https://doi.org/10.1016/j.mgene.2014.05.001
Singh TN, Aspinall D, Paleg LG (1972) Proline accumulation and varietal adaptability to drought in barley: a potential metabolic measure of drought resistance. Nat New Biol 236(67):188–190. https://doi.org/10.1038/newbio236188a0
Singh M, Srivastava JP, Kumar A (1992) Cell membrane stability in relation to drought tolerance in wheat genotypes. J Agron Crop Sci 168(3):186–190. https://doi.org/10.1111/j.1439-037x.1992.tb00997.x
Song A, An J, Guan Z, Jiang J, Chen F, Lou W et al (2014) The constitutive expression of a two transgene construct enhances the abiotic stress tolerance of chrysanthemum. Plant Physiol Biochem 80:114–120. https://doi.org/10.1016/j.plaphy.2014.03.030
Talaat NB, Shawky BT, Ibrahim AS (2015) Alleviation of drought-induced oxidative stress in maize (Zea mays L.) plants by dual application of 24-epibrassinolide and spermine. Environ Exp Bot 113:47–58. https://doi.org/10.1016/j.envexpbot.2015.01.006
Tian Z, Wang F, Zhang W, Liu C, Zhao X (2012) Antioxidant mechanism and lipid peroxidation patterns in leaves and petals of marigold in response to drought stress. Hortic Environ Biotechnol 53(3):183–192. https://doi.org/10.1007/s13580-012-0069-4
Tran LSP, 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(1):32–39. https://doi.org/10.4161/gmcr.1.1.10569
Turner NC (1975) Concurrent comparisons of stomatal behavior, water status, and evaporation of maize in soil at high or low water potential. Plant Physiol 55(5):932–936. https://doi.org/10.1104/pp.55.5.932
Van der Mescht A, De Ronde JA (1999) Chlorophyll fluorescence and chlorophyll content as a measure of drought tolerance in potato. S Afr J Sci 95(9):407–412
Verma S, Mishra SN (2005) Putrescine alleviation of growth in salt stressed Brassica juncea by inducing antioxidative defense system. J Plant Physiol 162(6):669–677. https://doi.org/10.1016/j.jplph.2004.08.008
Wang G, Zhang S, Ma X, Wang Y, Kong F, Meng Q (2016) A stress-associated NAC transcription factor (SlNAC35) from tomato plays a positive role in biotic and abiotic stresses. Physiol Plant 158(1):45–64. https://doi.org/10.1111/ppl.12444
Wang K, Wu YH, Tian XQ, Bai ZY, Liang QY, Liu QL et al (2017a) Overexpression of DgWRKY4 enhances salt tolerance in chrysanthemum seedlings. Front Plant Sci 8:1592. https://doi.org/10.3389/fpls.2017.01592
Wang K, Zhong M, Wu YH, Bai ZY, Liang QY, Liu QL et al (2017b) Overexpression of a chrysanthemum transcription factor gene DgNAC1 improves the salinity tolerance in chrysanthemum. Plant Cell Rep 36(4):571–581. https://doi.org/10.1007/s00299-017-2103-6
Wang L, Hu Z, Zhu M, Zhu Z, Hu J, Qanmber G, Chen G (2017c) The abiotic stress-responsive NAC transcription factor SlNAC11 is involved in drought and salt tolerance response in tomato (Solanum lycopersicum L.). Plant Cell Tissue Organ Cult 129:161–174. https://doi.org/10.1007/s11240-017-1167-x
Xing X, Liu Y, Kong X, Liu Y, Li D (2011) Overexpression of a maize dehydrin gene, ZmDHN2b, in tobacco enhances tolerance to low temperature. Plant Growth Regul 65(1):109–118. https://doi.org/10.1007/s10725-011-9580-3
Xiong L, Schumaker KS, Zhu JK (2002) Cell signaling during cold, drought, and salt stress. Plant Cell 14(1):165–183. https://doi.org/10.1105/tpc.000596
Xuan N, Jin Y, Zhang H, Xie Y, Liu Y, Wang G (2011) A putative maize zinc-finger protein gene, ZmAN13, participates in abiotic stress response. Plant Cell Tissue Organ Cult 107(1):101–112. https://doi.org/10.1007/s11240-011-9962-2
Yang Y, Zhu K, Wu J, Liu L, Sun G, He Y et al (2016) Identification and characterization of a novel NAC. Plant Cell Rep 35(8):1783–1798. https://doi.org/10.1007/s00299-016-1996-9
Yokotani N, Ichikawa T, Kondou Y, Matsui M, Hirochika H, Iwabuchi M et al (2009) Tolerance to various environmental stresses conferred by the salt-responsive rice gene ONAC063 in transgenic Arabidopsis. Planta 229(5):1065–1075. https://doi.org/10.1007/s00425-009-0895-5
Zhang L, Tian LH, Zhao JF, Song Y, Zhang CJ, Guo Y (2009) Identification of an apoplastic protein involved in the initial phase of salt stress response in rice root by two-dimensional electrophoresis. Plant Physiol 149:916–928. https://doi.org/10.1104/pp.108.131144
Acknowledgements
This research was supported by National Natural Science Foundation of China (31770742).
Author information
Authors and Affiliations
Contributions
QZ, MZ and QLL conceived and designed the experiments. QZ, QLL, LH and BW performed the experiments. YP, BJ and LZ analyzed the data. QZ wrote the paper. All authors read and approved the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Additional information
Communicated by Sergio J. Ochatt.
Rights and permissions
About this article
Cite this article
Zhao, Q., Zhong, M., He, L. et al. Overexpression of a chrysanthemum transcription factor gene DgNAC1 improves drought tolerance in chrysanthemum. Plant Cell Tiss Organ Cult 135, 119–132 (2018). https://doi.org/10.1007/s11240-018-1449-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-018-1449-y