Plant Molecular Biology

, Volume 77, Issue 1–2, pp 17–31 | Cite as

A cotton group C MAP kinase gene, GhMPK2, positively regulates salt and drought tolerance in tobacco

  • Liang Zhang
  • Dongmei Xi
  • Shanwei Li
  • Zheng Gao
  • Shuoli Zhao
  • Jing Shi
  • Changai Wu
  • Xingqi GuoEmail author


Mitogen-activated protein kinase (MAPK) cascades play important roles in mediating biotic and abiotic stress responses. In plants, MAPKs are classified into four major groups (A–D) according to their sequence homology and conserved phosphorylation motifs. Compared with well-studied MAPKs in groups A and B, little is known about group C. In this study, we functionally characterised a stress-responsive group C MAPK gene (GhMPK2) from cotton (Gossypium hirsutum). Northern blot analysis indicated that GhMPK2 was induced by abscisic acid (ABA) and abiotic stresses, such as NaCl, PEG, and dehydration. Subcellular localization analysis suggested that GhMPK2 may activate its specific targets in the nucleus. Constitutive overexpression of GhMPK2 in tobacco (Nicotiana tabacum) conferred reduced sensitivity to ABA during both seed germination and vegetative growth. Interestingly, transgenic plants had a decreased rate of water loss and exhibited enhanced drought and salt tolerance. Additionally, transgenic plants showed improved osmotic adjustment capacity, elevated proline accumulation and up-regulated expression of several stress-related genes, including DIN1, Osmotin and NtLEA5. β-glucuronidase (GUS) expression driven by the GhMPK2 promoter was clearly enhanced by treatment with NaCl, PEG, and ABA. These results strongly suggest that GhMPK2 positively regulates salt and drought tolerance in transgenic plants.


Cotton (GossypiumhirsutumGhMPK2 Salt Drought Osmotic adjustment Transgenic tobacco 



Abscisic acid


Green fluorescence protein




Inverse polymerase chain reaction


Mitogen-activated protein kinase


Osmotic adjustment


Open reading frame


Polymerase chain reaction


Polyethylene glycol



This work was financially supported by China National Transgenic Plant Research and Commercialization Projects (2009ZX08009-092B; 2009ZX08009-113B), and the National Natural Science Foundation of China (30970225).

Supplementary material

11103_2011_9788_MOESM1_ESM.tif (549 kb)
Supplementary material 1 (TIFF 497 kb)
11103_2011_9788_MOESM2_ESM.doc (32 kb)
Supplementary material 2 (DOC 33 kb)


  1. Abe H, Urao T, Ito T, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling. Plant Cell 15:63–78PubMedCrossRefGoogle Scholar
  2. Baumann K, De Paolis A, Costantino P, Gualberti G (1999) The DNA binding site of the Dof protein NtBBF1 is essential for tissue-specific and auxin-regulated expression of the rolB oncogene in plants. Plant Cell 11:323–334PubMedCrossRefGoogle Scholar
  3. Beckers GJ, Jaskiewicz M, Liu Y, Underwood WR, He SY, Zhang S, Conrath U (2009) Mitogen-activated protein kinases 3 and 6 are required for full priming of stress responses in Arabidopsis thaliana. Plant Cell 21:944–953PubMedCrossRefGoogle Scholar
  4. Blum A, Munns R, Passioura JB, Turner NC, Sharp RE, Boyer JS, Nguyen HT, Hsiao TC, Verma D, Hong Z (1996) Genetically engineered plants resistant to soil drying and salt stress: how to interpret osmotic relations? Plant Physiol 110:1051–1053PubMedGoogle Scholar
  5. Burnett EC, Desikan R, Moser RC, Neill SJ (2000) ABA activation of an MBP kinase in Pisum sativum epidermal peels correlates with stomatal responses to ABA. J Exp Bot 51:197–205PubMedCrossRefGoogle Scholar
  6. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743PubMedCrossRefGoogle Scholar
  7. Deuschle K, Funck D, Forlani G, Stransky H, Biehl A, Leister D, van der Graaff E, Kunze R, Frommer WB (2004) The role of Δ1-pyrroline-5-carboxylate dehydrogenase in proline degradation. Plant Cell 16:3413–3425PubMedCrossRefGoogle Scholar
  8. Droillard MJ, Boudsocq M, Barbier-Brygoo H, Lauriere C (2004) Involvement of MPK4 in osmotic stress response pathways in cell suspensions and plantlets of Arabidopsis thaliana: activation by hypoosmolarity and negative role in hyperosmolarity tolerance. FEBS Lett 574:42–48PubMedCrossRefGoogle Scholar
  9. Fiil BK, Petersen K, Petersen M, Mundy J (2009) Gene regulation by MAP kinase cascades. Curr Opin Plant Biol 12:615–621PubMedCrossRefGoogle Scholar
  10. Gao L, Xiang CB (2008) The genetic locus At1g73660 encodes a putative MAPKKK and negatively regulates salt tolerance in Arabidopsis. Plant Mol Biol 67:125–134PubMedCrossRefGoogle Scholar
  11. Gu L, Liu Y, Zong X, Liu L, Li DP, Li DQ (2010) Overexpression of maize mitogen-activated protein kinase gene, ZmSIMK1 in Arabidopsis increases tolerance to salt stress. Mol Biol Rep 37:4067–4073PubMedCrossRefGoogle Scholar
  12. Hamel LP, Nicole MC, Sritubtim S, Morency MJ, Ellis M, Ehlting J, Beaudoin N, Barbazuk B, Klessig D, Lee J et al (2006) Ancient signals: comparative genomics of plant MAPK and MAPKK gene families. Trends Plant Sci 11:192–198PubMedCrossRefGoogle Scholar
  13. He C, Fong SH, Yang D, Wang GL (1999) BWMK1, a novel MAP kinase induced by fungal infection and mechanical wounding in rice. Mol Plant Microbe Interact 12:1064–1073PubMedCrossRefGoogle Scholar
  14. Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database: 1999. Nucleic Acids Res 27:297–300PubMedCrossRefGoogle Scholar
  15. Horsch RB, Rogers SG, Fraley RT (1985) Transgenic plants. Cold Spring Harb Symp Quant Biol 50:433–437PubMedGoogle Scholar
  16. Jammes F, Song C, Shin D, Munemasa S, Takeda K, Gu D, Cho D, Lee S, Giordo R, Sritubtim S et al (2009) MAP kinases MPK9 and MPK12 are preferentially expressed in guard cells and positively regulate ROS-mediated ABA signaling. Proc Natl Acad Sci USA 106:20520–20525PubMedCrossRefGoogle Scholar
  17. Jonak C, Kiegerl S, Ligterink W, Barker PJ, Huskisson NS, Hirt H (1996) Stress signaling in plants: a mitogen-activated protein kinase pathway is activated by cold and drought. Proc Natl Acad Sci USA 93:11274–11279PubMedCrossRefGoogle Scholar
  18. Knetsch M, Wang M, Snaar-Jagalska BE, Heimovaara-Dijkstra S (1996) Abscisic acid induces mitogen-activated protein kinase activation in barley aleurone protoplasts. Plant Cell 8:1061–1067PubMedCrossRefGoogle Scholar
  19. Lescot M, Dehais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouze P, Rombauts S (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res 30:325–327PubMedCrossRefGoogle Scholar
  20. Leung J, Giraudat J (1998) Abscisic acid signal transduction. Annu Rev Plant Physiol Plant Mol Biol 49:199–222PubMedCrossRefGoogle Scholar
  21. Liu Q, Xue Q (2007) Computational identification and phylogenetic analysis of the MAPK gene family in Oryza sativa. Plant Physiol Biochem 45:6–14PubMedCrossRefGoogle Scholar
  22. Luo Y, Liu YB, Dong YX, Gao XQ, Zhang XS (2009) Expression of a putative alfalfa helicase increases tolerance to abiotic stress in Arabidopsis by enhancing the capacities for ROS scavenging and osmotic adjustment. J Plant Physiol 166:385–394PubMedCrossRefGoogle Scholar
  23. MacRobbie EA, Kurup S (2007) Signalling mechanisms in the regulation of vacuolar ion release in guard cells. New Phytol 175:630–640PubMedCrossRefGoogle Scholar
  24. MAPK Group (2002) Mitogen-activated protein kinase cascades in plants: a new nomenclature. Trends Plant Sci 7:301–308CrossRefGoogle Scholar
  25. Mikolajczyk M, Awotunde OS, Muszynska G, Klessig DF, Dobrowolska G (2000) Osmotic stress induces rapid activation of a salicylic acid-induced protein kinase and a homolog of protein kinase ASK1 in tobacco cells. Plant Cell 12:165–178PubMedCrossRefGoogle Scholar
  26. Mori IC, Muto S (1997) Abscisic acid activates a 48-kilodalton protein kinase in guard cell protoplasts. Plant Physiol 113:833–839PubMedGoogle Scholar
  27. Munnik T, Ligterink W, Meskiene II, Calderini O, Beyerly J, Musgrave A, Hirt H (1999) Distinct osmo-sensing protein kinase pathways are involved in signalling moderate and severe hyper-osmotic stress. Plant J 20:381–388PubMedCrossRefGoogle Scholar
  28. Nakashima K, Fujita Y, Katsura K, Maruyama K, Narusaka Y, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2006) Transcriptional regulation of ABI3- and ABA-responsive genes including RD29B and RD29A in seeds, germinating embryos, and seedlings of Arabidopsis. Plant Mol Biol 60:51–68PubMedCrossRefGoogle Scholar
  29. Nakashima K, Ito Y, Yamaguchi-Shinozaki K (2009) Transcriptional regulatory networks in response to abiotic stresses in Arabidopsis and grasses. Plant Physiol 149:88–95PubMedCrossRefGoogle Scholar
  30. Nicole MC, Hamel LP, Morency MJ, Beaudoin N, Ellis BE, Seguin A (2006) MAP-ping genomic organization and organ-specific expression profiles of poplar MAP kinases and MAP kinase kinases. BMC Genomics 7:223–245PubMedCrossRefGoogle Scholar
  31. Ning J, Li X, Hicks LM, Xiong L (2010) A Raf-like MAPKKK gene DSM1 mediates drought resistance through reactive oxygen species scavenging in rice. Plant Physiol 152:876–890PubMedCrossRefGoogle Scholar
  32. Ortiz-Masia D, Perez-Amador MA, Carbonell J, Marcote MJ (2007) Diverse stress signals activate the C1 subgroup MAP kinases of Arabidopsis. FEBS Lett 581:1834–1840PubMedCrossRefGoogle Scholar
  33. Ortiz-Masia D, Perez-Amador MA, Carbonell P, Aniento F, Carbonell J, Marcote MJ (2008) Characterization of PsMPK2, the first C1 subgroup MAP kinase from pea (Pisum sativum L.). Planta 227:1333–1342PubMedCrossRefGoogle Scholar
  34. Park JM, Park CJ, Lee SB, Ham BK, Shin R, Paek KH (2001) Overexpression of the tobacco Tsi1 gene encoding an EREBP/AP2-type transcription factor enhances resistance against pathogen attack and osmotic stress in tobacco. Plant Cell 13:1035–1046PubMedCrossRefGoogle Scholar
  35. Pitzschke A, Hirt H (2009) Disentangling the complexity of mitogen-activated protein kinases and reactive oxygen species signaling. Plant Physiol 149:606–615PubMedCrossRefGoogle Scholar
  36. Reyna NS, Yang Y (2006) Molecular analysis of the rice MAP kinase gene family in relation to Magnaporthe grisea infection. Mol Plant Microbe Interact 19:530–540PubMedCrossRefGoogle Scholar
  37. Rodriguez MC, Petersen M, Mundy J (2010) Mitogen-activated protein kinase signaling in plants. Annu Rev Plant Biol 61:621–649PubMedCrossRefGoogle Scholar
  38. Shan DP, Huang JG, Yang YT, Guo YH, Wu CA, Yang GD, Gao Z, Zheng CC (2007) Cotton GhDREB1 increases plant tolerance to low temperature and is negatively regulated by gibberellic acid. New Phytol 176:70–81PubMedCrossRefGoogle Scholar
  39. Shi J, An HL, Zhang L, Gao Z, Guo XQ (2010) GhMPK7, a novel multiple stress-responsive cotton group C MAPK gene, has a role in broad spectrum disease resistance and plant development. Plant Mol Biol 74:1–17PubMedCrossRefGoogle Scholar
  40. Shinozaki K, Yamaguchi-Shinozaki K, Seki M (2003) Regulatory network of gene expression in the drought and cold stress responses. Curr Opin Plant Biol 6:410–417PubMedCrossRefGoogle Scholar
  41. Takahashi W, Oishi H, Ikeda S, Takamizo T, Komatsu T (2006) Molecular cloning and expression analysis of the replacement histone H3 gene of Italian ryegrass (Lolium multiflorum). J Plant Physiol 163:58–68PubMedCrossRefGoogle Scholar
  42. Tamura T, Hara K, Yamaguchi Y, Koizumi N, Sano H (2003) Osmotic stress tolerance of transgenic tobacco expressing a gene encoding a membrane-located receptor-like protein from tobacco plants. Plant Physiol 131:454–462PubMedCrossRefGoogle Scholar
  43. Tuteja N (2007) Abscisic acid and abiotic stress signaling. Plant Signal Behav 2:135–138PubMedCrossRefGoogle Scholar
  44. Varagona MJ, Schmidt RJ, Raikhel NV (1992) Nuclear localization signal(s) required for nuclear targeting of the maize regulatory protein Opaque-2. Plant Cell 4:1213–1227PubMedCrossRefGoogle Scholar
  45. Verbruggen N, Hermans C (2008) Proline accumulation in plants: a review. Amino Acids 35:753–759PubMedCrossRefGoogle Scholar
  46. Wang W, Vinocur B, Altman A (2003) Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218:1–14PubMedCrossRefGoogle Scholar
  47. Wang M, Zhang Y, Wang J, Wu X, Guo X (2007) A novel MAP kinase gene in cotton (Gossypium hirsutum L.), GhMAPK, is involved in response to diverse environmental stresses. J Biochem Mol Biol 40:325–332PubMedCrossRefGoogle Scholar
  48. Wang XJ, Zhu SY, Lu YF, Zhao R, Xin Q, Wang XF, Zhang DP (2010) Two coupled components of the mitogen-activated protein kinase cascade MdMPK1 and MdMKK1 from apple function in ABA signal transduction. Plant Cell Physiol 51:754–766PubMedCrossRefGoogle Scholar
  49. Xiao B, Huang Y, Tang N, Xiong L (2007) Over-expression of a LEA gene in rice improves drought resistance under the field conditions. Theor Appl Genet 115:35–46PubMedCrossRefGoogle Scholar
  50. Xing Y, Jia W, Zhang J (2009) AtMKK1 and AtMPK6 are involved in abscisic acid and sugar signaling in Arabidopsis seed germination. Plant Mol Biol 70:725–736PubMedCrossRefGoogle Scholar
  51. Xiong L, Yang Y (2003) Disease resistance and abiotic stress tolerance in rice are inversely modulated by an abscisic acid-inducible mitogen-activated protein kinase. Plant Cell 15:745–759PubMedCrossRefGoogle Scholar
  52. Xiong L, Schumaker KS, Zhu JK (2002) Cell signaling during cold, drought, and salt stress. Plant Cell 14(Suppl):S165–S183Google Scholar
  53. Xiong L, Wang RG, Mao G, Koczan JM (2006) Identification of drought tolerance determinants by genetic analysis of root response to drought stress and abscisic Acid. Plant Physiol 142:1065–1074PubMedCrossRefGoogle Scholar
  54. Zhang S, Klessig DF (1998) Resistance gene N-mediated de novo synthesis and activation of a tobacco mitogen-activated protein kinase by tobacco mosaic virus infection. Proc Natl Acad Sci USA 95:7433–7438PubMedCrossRefGoogle Scholar
  55. Zhu JK (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53:247–273PubMedCrossRefGoogle Scholar
  56. Zong XJ, Li DP, Gu LK, Li DQ, Liu LX, Hu XL (2009) Abscisic acid and hydrogen peroxide induce a novel maize group C MAP kinase gene, ZmMPK7, which is responsible for the removal of reactive oxygen species. Planta 229:485–495PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Liang Zhang
    • 1
  • Dongmei Xi
    • 2
  • Shanwei Li
    • 1
  • Zheng Gao
    • 1
  • Shuoli Zhao
    • 3
  • Jing Shi
    • 1
  • Changai Wu
    • 1
  • Xingqi Guo
    • 1
    Email author
  1. 1.State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop BiologyShandong Agricultural UniversityTaianPeople’s Republic of China
  2. 2.Experimental CenterLinyi UniversityLinyiPeople’s Republic of China
  3. 3.College of Food Science and EngineeringShandong Agricultural UniversityTaianPeople’s Republic of China

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