Plant Molecular Biology Reporter

, Volume 36, Issue 4, pp 543–552 | Cite as

Overexpression of the Transcription Factor NtNAC2 Confers Drought Tolerance in Tobacco

  • Xiaoyan Xu
  • Xinzhuan Yao
  • Litang LuEmail author
  • Degang ZhaoEmail author
Original Paper


NAC proteins constitute one of the largest families of plant-specific transcription factors and play an important role in biological processes, including plant development and phytohormone homeostasis, and in responses to various environmental stresses. In this study, we isolated an NAC group A gene (named NtNAC2) from Nicotiana tabacum L. Quantitative RT-PCR (qRT-PCR) analysis indicated that NtNAC2 was significantly upregulated under drought stress, which implied NtNAC2 was important in tobacco under such conditions. Overexpression of NtNAC2 in tobacco plants exhibited enhanced drought tolerance by means of improved seedling growth. Under drought stress, organic osmoprotectants were significantly accumulated in these plants. Additionally, the activities of antioxidant defense enzymes, like superoxide dismutase (SOD) and peroxidase (POD), which could effectively scavenge accumulated reactive oxygen species (ROS), increased in NtNAC2-overexpression transgenic tobacco plants compared with wild-type plants. The net photosynthetic rate was also significantly increased in NtNAC2-overexpression transgenic lines compared with wild-type plants, and the content of malondialdehyde (MDA) and proline was lower in NtNAC2-overexpression transgenic lines than that in wild-type plants (P < 0.01). Furthermore, the expression of NtWRKY28, a drought resistance gene, was significantly increased and the δ-OAT gene was downregulated in NtNAC2-overexpression plants relative to wild-type plants. Taken together, these results indicated that NtNAC2 functions as a positive regulator of drought stress tolerance. This study provides a basis for further study of drought resistance conferred by the NtNAC2 gene.


NtNAC2 Antioxidant defense Drought resistance Nicotiana tabacum 



This work was funded by the National Natural Science Foundation (No. 31160149) and by the Major Projects of National New Varieties of Genetically Modified Organisms.

(No. 2014ZX08010-003-2016ZX08010-003). The authors thank Litang Lu for the technical assistance.

Author Contribution Statement

XX and YX designed and conducted the experiments. XX wrote the manuscript. LL contributed by helping with some experiments presented in the manuscript. LL helped to edit the manuscript. LL and ZD supervised the studies and revised the manuscript. All authors read and approved the manuscript.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. Agarwal PK, Agarwal P, Reddy MK, Sopory S (2006) Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Rep 25(12):1263–1274CrossRefPubMedGoogle Scholar
  2. Ashraf M, Foolad MR (2007) Roles of glycine betaine and proline in improving plant biotic stress resistance. Environ Exp Bot 59(2):206–216CrossRefGoogle Scholar
  3. Benjamin JG, Nielsen DC (2006) Water deficit effects on root distribution of soybean, field pea and chickpea. Field Crop Res 97(2):248–253CrossRefGoogle Scholar
  4. Carvalho MHC (2008) Drought stress and reactive oxygen species. Plant Signal Behav 3:156–165CrossRefGoogle Scholar
  5. Choudhury S, Panda P, Sahoo L, Panda SK (2013) Reactive oxygen species signaling in plants under abiotic stress. Plant Signal Behav 8(4)CrossRefGoogle Scholar
  6. Die HU, Liu H, Chen DM, Tian Y (2011) Cloning and sequence analysis of the NAC transcription factor NtNAC8 from tobacco. Journal of Huaihua University 30(05):33–37Google Scholar
  7. Erpen L, Devi HS, Grosser JW, Dutt M(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):1–25CrossRefGoogle Scholar
  8. Fujita M, Fujita Y, Maruyama K, Seki M, Hiratsu K, Ohme-Takagi M,Tran LS, Yamaguchi-Shinozaki K, Shinozaki K(2004) A dehydration-induced nac protein, RD26, is involved in a novel aba-dependent stresssignaling pathway. Plant Journal 39(6):863–876CrossRefPubMedGoogle Scholar
  9. Guo GH, Liu HY, Li GH, Liu M, Li Y, Wang SH, Liu ZH, Tang S, Ding YF (2014) Analysis of physiological characteristics about ABA alleviating rice booting stage drought stress. Chinese Journal of Agricultural Sciences 47(22):4380–4391Google Scholar
  10. Han QQ, Qiao P, Song YZ, Zhang JY (2014) Structural analysis and tissue-specific expression patterns of a novel salt-inducible NAC transcription factor gene from Nicotiana tabacum cv. Xanthi. Journal of Pomology & Horticultural Science 89(6):700–706Google Scholar
  11. Hanin M, Brini F, Ebel C, Toda Y, Takeda S, Masmoudi K (2011) Plant dehydrins and stress tolerance: versatile proteins for complex mechanisms. Plant Signal Behav 6(10):1503–1509CrossRefPubMedPubMedCentralGoogle Scholar
  12. Hifzur R, Valarmathi R, Jagedeeshselvam N, Sudhakar D, Raveendran M (2016) Over-expression of a NAC 67 transcription factor from finger millet (Eleusine coracana L.) confers tolerance against salinity and drought stress in rice. BMC Biotechnol 1(1):35Google Scholar
  13. Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231CrossRefGoogle Scholar
  14. Huang QJ, Wang Y, Li B, Chang JL, Chen MJ, Li KX, Yang GX, He GY (2015) TaNAC29, a NAC transcription factor from wheat, enhances salt and drought tolerance in transgenic Arabidopsis. BMC Plant Biol 15(1):268CrossRefPubMedPubMedCentralGoogle Scholar
  15. Li Y, Chen Q, Nan H, Li X, Lu S, Zhao X, Liu B, Guo C, Kong F, Cao D (2017) Overexpression of GmFDL19 enhances tolerance to drought and salt stresses in soybean. PLoS One 12(6):e0179554CrossRefPubMedPubMedCentralGoogle Scholar
  16. Liu Y, Yao XZ, Lu LT, Lei YT, Dai TT, Zhao DG (2016a) Cloning of Sb SKIP gene from sorghum (Sorghum bicolor) and analysis of drought-resistant function in tobacco (Nicotiana tabacum). J Agric Biotechnol 24(10):1500–1511Google Scholar
  17. Liu YM, Yu XW, Liu SS, Peng H, Mijiti A, Wang Z, Zhang H, Ma H (2016b) A chickpea NAC-type transcription factor, CarNAC6, confers enhanced dehydration tolerance in Arabidopsis. Plant Mol Biol Report 35(1):1–14Google Scholar
  18. Liu H, Zhou Y, Li H, Wang T, Zhang J, Ouyang B (2018) Molecular and functional characterization of shnac1, an NAC transcription factor from solanum habrochaites. Plant Sci 271:9–19CrossRefPubMedGoogle Scholar
  19. Long L, Gao W, Xu L, Liu M, Luo X, He X, Yang X, Zhang X, Zhu L (2013) GbMPK3, a mitogen-activated protein kinase from cotton, enhances drought and oxidative stress tolerance in tobacco. Plant Cell Tissue Organ Cult 116(2):153–162CrossRefGoogle Scholar
  20. Molinari HBC, Marur CJ, Daros E, Campos MKF, Carvalho JFRP, Bespalhok-Filho JC, Pereira LFP (2007) Evaluation of the stress-inducible production of proline in transgenic sugarcane (Saccharum spp.): osmotic adjustment, chlorophyll fluorescence and oxidative stress. Physiol Plant 130(2):218–229CrossRefGoogle Scholar
  21. Nakashima K (2006) Regulons involved in osmotic stress-responsive and cold stress-responsive gene expression in plants. Physiol Plant 126(1):62–71CrossRefGoogle Scholar
  22. Nakashima K, Takasaki H, Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K (2012) NAC transcription factors in plant abiotic stress responses. Biochim Biophys Acta 1819(2):97–103CrossRefPubMedGoogle Scholar
  23. Ning W, Zhai H, Yu J, Liang S, Yang X, Xing X (2017) Overexpression of glycine soja WRKY20, enhances drought tolerance and improves plant yields under drought stress in transgenic soybean. Mol Breed 37(2):19CrossRefGoogle Scholar
  24. Olsen AN, Ernst HA, Leggio LL, Skriver K (2005) NAC transcription factors: structurally distinct, functionally diverse. Trends Plant Sci 10(2):79–87CrossRefPubMedGoogle Scholar
  25. Puranik S, Sahu PP, Srivastava PS, Prasad M (2012) NAC proteins: regulation and role in stress tolerance. Trends Plant Sci 17(6):369–381CrossRefPubMedGoogle Scholar
  26. Shao H, Wang H, Tang X (2015) NAC transcription factors in plant multiple abiotic stress responses: progress and prospects. Front Plant Sci 6:902CrossRefPubMedPubMedCentralGoogle Scholar
  27. Sheng SY, Wu YX, Zheng YS (2017) Review on drought response in plants from phenotype to molecular. Curr Biotechnol 7(3):169–176Google Scholar
  28. Sun CH, Du W, Cheng XL, Xu XN, Zhang YH, Sun D, Shi JJ (2010) The effects of drought stress on the activity of acid phosphatase and its protective enzymes in pigweed leaves. Afr J Biotechnol 9(6):825–833CrossRefGoogle Scholar
  29. Thirumalaikumar VP, Devkar V, Mehterov N, Ali S, Ozgur R, and Turkan I et al. (2017) Nac transcription factor JUNGBRUNNEN1 enhances drought tolerance in tomato. Plant Biotechnol J 16(2):354–366CrossRefPubMedPubMedCentralGoogle Scholar
  30. Tian Y (2016) The responding mechanism of physiology and proteomics in Prunus mira root under drought. Northeast Forestry University, Haerbin ShiGoogle Scholar
  31. Tian Y, Fang J, Xiang-Yang LU (2009) Cloning and bioinformatics analysis of nac transcription factor ntnac1 in tobacco. Acta Tabacaria Sinica 15(5):67–72Google Scholar
  32. Tran L-SP, 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 zinc finger homeodomain transcription factor ZFHD1: role of the ZFHD1 and NAC transcription factors that bind to a drought-responsive ciselement in the early responsive to dehy-dration stress 1 promoter. Plant Cell Physiol Suppl 16:2481–2498Google Scholar
  33. Umezawa T, Fujita M, Fujita Y, Yamaguchi-Shinozaki K, Shinozaki K (2006) Engineering drought tolerance in plants: discovering and tailoring genes to unlock the future. Curr Opin Biotechnol 17(2):113–122CrossRefPubMedGoogle Scholar
  34. Wang WB, Kim YH, Deng XP, Kwak SS, Wang ZH, Zhao ZP (2009) Physiological and biological responses of alfalfa shoots and roots to salt stress. J Northwest A & F Univ 37(5):217–223Google Scholar
  35. Wang H, Zhao Q, Chen F, Wang M, Dixon RA (2011) NAC domain function and transcriptional control of a secondary cell wall master switch. Plant J 68(6):1104–1114CrossRefPubMedGoogle Scholar
  36. Wang Y, Wang Q, Liu ML, Bo C, Wang X, Ma Q (2017) Overexpression of a maize MYB48 gene confers drought tolerance in transgenic arabidopsis plants. Journal of Plant Biology 60(6):612–621CrossRefGoogle Scholar
  37. Yang ZQ, Qiu YX, Liu CX, Chen YQ, Tan W (2016) The effects of soil moisture stress on the growth of root and above-ground parts of greenhouse tomato crops. Chinese Journal of Ecology 36(3):748–757Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.College of Life Sciences and The Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Institute of Agro-BioengineeringGuizhou UniversityGuiyangChina
  2. 2.College of Tea ScienceGuizhou UniversityGuiyangChina

Personalised recommendations