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Expression of the chickpea CarNAC3 gene enhances salinity and drought tolerance in transgenic poplars

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CarNAC3 contains 285 amino acids and a conserved NAC domain. NAC genes, including NAM, ATAF1, ATAF2, and CUC2, are members of one of the largest transcription factor families in plants. CarNAC3, a member of the NAP group, plays an important role in plant development and responses to abiotic stresses. In this study, CarNAC3 was transformed into hybrid poplar plants (Populus deltoides × P. euramericana ‘Nanlin895’) using Agrobacterium tumefaciens. PCR analysis confirmed integration of the introduced T-DNA into the target genome. Reverse transcription PCR confirmed the transformation, and Southern and northern blotting verified the transgene copy number and gene expression, respectively. Fourteen lines of positive transformants were transplanted into a greenhouse to verify their drought and salt tolerances. Under normal conditions, transgenic poplar plants were shorter than the wild-type, but under drought and salt stresses, they maintained their normal rooting and stem growth rates, while those of the wild-type plants were suppressed. Under stress conditions, CarNAC3 expression caused increases in proline and photosynthetic pigment levels and in antioxidant enzyme activities. Furthermore, the expression of CarNAC3 lowered malondialdehyde concentrations compared with the wild-type control. Overall, our results indicated that the CarNAC3 transgene enhanced drought and salt tolerance in transgenic poplar plants.

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Neomycin phosphotransferase


Open reading frame


Guaiacol peroxidase


Reactive oxygen species


Superoxide dismutase




  1. Ábráham E, Horton C, Erdei L, Szabados L (2010) Methods for determination of proline in plants. Methods Mol Biol 639:317–331

  2. Ajithkumar IP, Panneerselvam R (2014) ROS scavenging system, osmotic maintenance, pigment and growth status of Panicum sumatrense roth. Under drought stress. Cell Biochem Biophys 68(3):587–595

  3. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399

  4. Asada K (1999) The water–water cycle in chloroplasts: scavenging of active oxygens and dissipation of excess photons. Annu Rev Plant Physiol 50:601–639

  5. Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207

  6. Boardman NK (1977) Comparative photosynthesis of sun and shade plants. Annu Rev Plant Physiol 28:355–377

  7. Bray A (1997) Plant responses to water deficit. Trends Plant Sci 2:48–54

  8. Doyle JJ, Dickson EE (1987) Preservation of plant samples for DNA restriction endonuclease analysis. Taxon 36:715–722

  9. Duval M, Hsieh TF, Kim SY, Thomas TL (2002) Molecular characterization of AtNAM: a member of the Arabidopsis NAC domain superfamily. Plant Mol Biol 50:237–248

  10. Foyer CH, Noctor G (2005) Oxidant and antioxidant signaling in plants: a reevaluation of the concept of oxidative stress in a physiological context. Plant Cell Environ 28:1056–1071

  11. Fridivich I (1989) Superoxide dismutase: an adaptation to a paramagnetic gas. J Biol Chem 264:7761–7764

  12. Fujita M, Fujita Y, Noutoshi Y et al (2006) Crosstalk between abiotic and biotic stress responses: a current view from the points of convergence in the stress signaling networks. Curr Opin Plant Biol 9:436–442

  13. Gara LD, Pinto MC, Tommasi F (2003) The antioxidant systems vis-à-vis reactive oxygen species during plant–pathogen interaction. Plant Physiol Biochem 41:863–870

  14. Guo YF, Gan SS (2006) AtNAP, a NAC family transcription factor, has an important role in leaf senescence. Plant J 46:601–612

  15. Holsters M, Waele D, Depicker A, Messens E, van Montagu M, Schell J (1978) Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet 163:181–187

  16. Horsch RB, Fry J, Hoffmann N, Neidermeyer J, Rogers SG, Fraley RT (1988) Leaf disc transformation: plant molecular biology manual. Kluwer Academic Publishers, Dordrecht, pp 1–9

  17. Hu HH, Dai MQ, Yao JL, Xiao BZ, Li XH (2006) Overexpressing a NAM, ATAF, and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice. Proc Natl Acad Sci USA 103:12987–12992

  18. Hui P, Cheng HY, Chen C, Yu XW, Yang JN, Gao WR, Shi QH, Zhang H, Li JG, Ma H (2009) A NAC transcription factor gene of chickpea (Cicer arietinum), CarNAC3, is involved in drought stress response and various developmental processes. J Plant Physiol 166:1934–1945

  19. Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382

  20. Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498

  21. Nakashima K, Ito Y, Yamaguchi-Shinozaki K (2009) Transcriptional regulatory networks in response to abiotic stresses in Arabidopsis and grasses. Plant Physiol 149:88–95

  22. Nakashima K, Takasaki H, Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K (2012) NAC transcription factors in plant abiotic stress responses. Biochim Biophys Acta 1819:97–103

  23. Niyogi KK (1999) Photoprotection revisited: genetic and molecular approaches. Annu Rev Plant Physiol Plant Mol Biol 50:333–359

  24. Ohtani M, Nishikubo N, Xu B, Yamaguchi M, Mitsuda N, Goue N, Shi F, Ohme-Takagi M, Demura T (2011) A NAC domain protein family contributing to the regulation of wood formation in poplar. Plant J 67:499–512

  25. Olsen AN, Ernst HA, Leggio LL, Skriver K (2005) NAC transcription factors: structurally distinct, functionally diverse. Trends Plant Sci 10:79–87

  26. 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

  27. Pagariya MC, Devarumath RM, Kawar PG (2012) Biochemical characterization and identification of differentially expressed candidate genes in salt-stressed sugarcane. Plant Sci 184:1–13

  28. Passioura B (1996) Drought, and drought tolerance. Plant Growth Reg 20:79–83

  29. Prochazkova D, Sairam RK, Srivastava GC, Singh DV (2001) Oxidative stress and antioxidant activity as the basis of senescence in maize leaves. Plant Sci 161:765–771

  30. Satchel S et al (1985) Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer in Agrobacterium tumefaciens. Nature 318:624–629

  31. Satoh K (1978) Serum lipid peroxide in cerebrovascular disorders determined by a new colorimetric method. Clin Chim Acta 90:37–43

  32. Singh B (1997) Climate changes in the greater and southern Caribbean. Int J Climatol 17:1093–1114

  33. Smirnoff N (1998) Plant resistance to environmental stress. Curr Opin Biotechnol 9:214–219

  34. Stewart RR, Bewley JD (1980) Lipid peroxidation associated with accelerated aging of soybean axes. Plant Physiol 65:245–248

  35. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739

  36. Torres MA, Dangl JL (2005) Functions of the respiratory burst oxidase in biotic interactions, abiotic stresses and development. Curr Opin Plant Biol 8:397–403

  37. Wang N, Hua H, Egrinya Eneji A, Li Z, Duan L, Tian X (2012) Genotypic variations in photosynthetic and physiological adjustment to potassium deficiency in cotton (Gossypium shirsutum). J Photochem Photobiol B 110:1–8

  38. 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

  39. Zhong R, Lee C, Ye Z (2010) Functional characterization of poplar wood-associated NAC domain transcription factors. Plant Physiol 152:1044–1055

  40. Zhou J, Wang J, Bi Y, Wang L, Tang L, Yu X, Ohtani M, Demura T, Zhuge Q (2014) Overexpression of PtSOS2 enhances salt tolerance in transgenic poplars. Plant Mol Biol Rep 32:185–197

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We thank Prof. Hao Ma (Nanjing Agricultural University, China) for providing the CarNAC3 plasmid and Prof. Mohammad Reza Mofid (Isfahan University of Medical Sciences, Iran) for his technical assistance. This work was supported by grants from the International Science and Technology Cooperation Program of China (2014DFG32440), the National 863 Program of China (No. 2013AA102703), the National Science Foundation of China (No. 31170561), the Priority Academic Program Development of Jiangsu Higher Education Institutions, and the Program for Innovative Research Team at the University of Educational Department and Jiangsu Province, China.

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Correspondence to Qiang Zhuge.

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Movahedi, A., Zhang, J., Gao, P. et al. Expression of the chickpea CarNAC3 gene enhances salinity and drought tolerance in transgenic poplars. Plant Cell Tiss Organ Cult 120, 141–154 (2015). https://doi.org/10.1007/s11240-014-0588-z

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  • CarNAC3
  • Populus
  • Agrobacterium tumefaciens
  • Abiotic stress
  • Transgenic