Molecular Biology Reports

, Volume 40, Issue 2, pp 1407–1415 | Cite as

Cloning and evolutionary analysis of tobacco MAPK gene family

  • Xingtan Zhang
  • Tingcai Cheng
  • Genhong Wang
  • Yafei Yan
  • Qingyou XiaEmail author


The mitogen-activated protein (MAP) kinase cascade is an important signaling module which is involved in biotic and abiotic stress responses as well as plant growth and development. In this study, we identified 17 tobacco MAPKs including 11 novel tobacco MAPK genes that have not been identified before. Comparative analysis with MAPK gene families from other plants, such as Athaliana thaliana, rice and poplar, suggested that tobacco MAPKs (such as NtMPK1, NtMPK3 and NtMPK8) might play similar functions in response to abiotic and biotic stresses. QRT-PCR analysis revealed that a total of 14 NtMPKs were regulated by SA and/or MeJA, suggesting their potential roles involved in plant defense response. In addition, 6 NtMPKs were induced by drought treatment, implying their roles in response to drought stress. Our results indicated that most of tobacco MAPK might be involved in plant defense response, which provides the basis for further analysis on physiological functions of tobacco MAPKs.


Mitogen-activated protein (MAP) kinase Phylogenetic analysis QRT-PCR Tobacco 



This work was supported by grants from the National Basic Research Program of China (No. 2012CB114600), Fundamental Research Funds for the Central Universities (No. CDJZR10290003), the National Natural Science Foundation (No. 30901054, 31001034 and 31000563) and Key Project for National Tobacco Government (No. 110200902037).

Supplementary material

11033_2012_2184_MOESM1_ESM.gif (97 kb)
Supplementary Fig. 1 Full-length protein sequence alignment of tobacco MAPKs. The CDdomains were marked by a red box (GIF 97 kb)
11033_2012_2184_MOESM2_ESM.jpg (30 kb)
Supplementary Fig. 2 Expression of PR1a and ODC under treatments of SA and MeJA (JPG 31 kb)
11033_2012_2184_MOESM3_ESM.doc (24 kb)
Supplementary Table 1 The primer sequences used in RT-PCR for cloning of 11 novol NtMPKs. (DOC 24 kb)
11033_2012_2184_MOESM4_ESM.doc (38 kb)
Supplementary Table 2 The primer sequences used in qRT-PCR for detection of NtMPK expression. (DOC 37 kb)


  1. 1.
    Christian W et al (1999) Mitogen-activated protein kinase: conservation of a three-kinase module from yeast to human. Physiol Rev 79(1):143–180Google Scholar
  2. 2.
    Tsuneaki A et al (2002) MAP kinase signalling cascade in Arabidopsis innate immunity. Nature 415:977–983CrossRefGoogle Scholar
  3. 3.
    Ishihama N et al (2011) Phosphorylation of the Nicotiana benthamiana WRKY8 transcription factor by MAPK functions in the defense response. Plant Cell Online 23(3):1153–1170CrossRefGoogle Scholar
  4. 4.
    Hirt H (1997) Multiple roles of MAP kinases in plant signal transduction. Trends Plant Sci 2(1):131–144CrossRefGoogle Scholar
  5. 5.
    Shuqun Z, Daniel FK (2001) MAPK cascades in plant defense signaling. Trends Plant Sci 6(11):502–527CrossRefGoogle Scholar
  6. 6.
    Claudia J et al (2002) Complexity, cross talk and integration of plant MAP kinase signalling. Curr Opin Plant Biol 5:415–424CrossRefGoogle Scholar
  7. 7.
    Agrawal GK et al (2003) Novel rice MAP kinases OsMSRMK3 and OsWJUMK1 involved in encountering diverse environmental stresses and developmental regulation. Biochem Biophys Res Commun 300(3):775–783PubMedCrossRefGoogle Scholar
  8. 8.
    Nathan SR, Yinong Y (2006) Molecular analysis of the rice MAP kinase gene family in relation to Magnaporthe grisea infection. Mol Plant Microbe Interact 19:530–540CrossRefGoogle Scholar
  9. 9.
    Calderini O et al (1998) A cell cycle regulated MAP kinase with a possible role in cytokinesis in tobacco cells. J Cell Sci 111(Pt 20):3091–3100PubMedGoogle Scholar
  10. 10.
    László B et al (1999) A MAP kinase is activated late in plant mitosis and becomes localized to the plane of cell division. Plant Cell 11:101–113Google Scholar
  11. 11.
    Morten P, Peter B, Henrik N (2000) Arabidopsis MAP kinase 4 negatively regulates systemic aquired resistance. Cell 103:1111–1120CrossRefGoogle Scholar
  12. 12.
    Uwe C, Corné MJP, Brigitte M–M (2002) Priming in plant–pathogen interactions. Trends Plant Sci 7(5):210–216CrossRefGoogle Scholar
  13. 13.
    Beckers GJM, Conrath U (2007) Priming for stress resistance: from the lab to the field. Curr Opin Plant Biol 10(4):425–431PubMedCrossRefGoogle Scholar
  14. 14.
    Beckers GJM et al (2009) Mitogen-activated protein kinases 3 and 6 are required for full priming of stress responses in Arabidopsis thaliana. Plant Cell Online 21(3):944–953CrossRefGoogle Scholar
  15. 15.
    Guillaume T et al (2001) Plant mitogen-activated protein kinase signaling cascades. Curr Opin Plant Biol 4:392–400CrossRefGoogle Scholar
  16. 16.
    Ichimura Kazuya et al (2002) Mitogen-activated protein kinase cascades in plants: a new nomenclature. Trends Plant Sci 7:301–308CrossRefGoogle Scholar
  17. 17.
    Hamel L-P et al (2006) Ancient signals: comparative genomics of plant MAPK and MAPKK gene families. Trends Plant Sci 11(4):192–198PubMedCrossRefGoogle Scholar
  18. 18.
    Tae KH et al (2010) Comparative genomic analysis of mitogen activated protein kinase gene family in grapevine. Genes Genomics 32:275–281CrossRefGoogle Scholar
  19. 19.
    Krens S, Spaink H, Snaarjagalska B (2006) Functions of the MAPK family in vertebrate-development. FEBS Lett 580(21):4984–4990PubMedCrossRefGoogle Scholar
  20. 20.
    Krens SFG et al (2006) Characterization and expression patterns of the MAPK family in zebrafish. Gene Expr Patterns 6:1019–1026PubMedCrossRefGoogle Scholar
  21. 21.
    Shuqun Z, Klessig DF (1997) Salicylic acid activates a 48-kD MAP kinase in tobacco. Plant Cell 9:809–824Google Scholar
  22. 22.
    Shigemi S, Hiroshi S, Ohashi Y (1999) Jasmonate-based wound signal transduction requires activation of WIPK, a tobacco mitogen-activated protein kinase. Plant Cell 11:289–298Google Scholar
  23. 23.
    Shuqun Z, Yidong L, Daniel FK (2000) Multiple levels of tobacco WIPK activation during the induction of cell death by fungal elicitins. Plant J 23(3):339–347CrossRefGoogle Scholar
  24. 24.
    Dongtao R et al (2006) Activation of Ntf4, a tobacco mitogen-activated protein kinase, during plant defence response and its involvement in hypersensitive response-like cell death. Plant Physiol 141:1482–1493CrossRefGoogle Scholar
  25. 25.
    Yule L, Michael S, Dinesh-Kumar SP (2004) Involvement of MEK1 MAPKK, NTF6 MAPK, WRKY/MYB transcription factors, COL1 and CTR1 in N-mediated resistance to tobacco mosaic virus. Plant J 38:800–809CrossRefGoogle Scholar
  26. 26.
    Wilson C et al (1993) Isolation and characterization of a tobacco cDNA clone encoding a putative MAP kinase. Plant Mol Biol 23(3):543–551PubMedCrossRefGoogle Scholar
  27. 27.
    Gomi K et al (2005) A mitogen-activated protein kinase NtMPK4 activated by SIPKK is required for jasmonic acid signaling and involved in ozone tolerance via stomatal movement in tobacco. Plant Cell Physiol 46(12):1902–1914PubMedCrossRefGoogle Scholar
  28. 28.
    Dhirendra K, Daniel FK (1999) Differential induction of tobacco MAP kinases by the defense signals nitric oxide, salicylic acid, ethylene, and jasmonic acid. Mol Plant Microbe Interact 13:347–351Google Scholar
  29. 29.
    Kwang-Yeol Y, Yidong L, Shuqun Z (2000) Activation of a mitogen-activated protein kinase pathway is involved in disease resistance in tobacco. PNAS 98:741–746Google Scholar
  30. 30.
    Mary E, Hoyos, Shuqun Z (2000) calcium-independent activation of salicylic acid-induced protein kinase and a 40-kilodalton protein kinase by hyperosmotic stress. Plant Physiol 122:1355–1363CrossRefGoogle Scholar
  31. 31.
    Monika M, Ajczyk et al (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 12:165–178Google Scholar
  32. 32.
    Hailing J et al (2002) Function of a mitogen-activated protein kinase pathway in N gene-mediated resistance in tobacco. Plant J 33:719–731Google Scholar
  33. 33.
    Soyano T (2003) NQK1/NtMEK1 is a MAPKK that acts in the NPK1 MAPKKK-mediated MAPK cascade and is required for plant cytokinesis. Genes Dev 17(8):1055–1067PubMedCrossRefGoogle Scholar
  34. 34.
    Michiko S et al (2006) Phosphorylation of NtMAP65-1 by a MAP kinase down-regulates its activity of microtubule bundling and stimulates progression of cytokinesis of tobacco cells. Genes Dev 20:1004–1014CrossRefGoogle Scholar
  35. 35.
    Maria-Jose C et al (2007) In situ molecular identification of the Ntf4 MAPK expression sites in maturing and germinating pollen. Biol Cell 99:209–221CrossRefGoogle Scholar
  36. 36.
    Gaiyun Z et al (2009) Overexpression of the soybean GmERF3 gene, an AP2/ERF type transcription factor for increased tolerances to salt, drought, and diseases in transgenic tobacco. J Exp Bot 60(13):3781–3796CrossRefGoogle Scholar
  37. 37.
    Corpet F (1988) Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res 16(22):10881–10890PubMedCrossRefGoogle Scholar
  38. 38.
    Higgins DG, Thompson JD, Gibson TJ (1996) Using CLUSTAL for multiple sequence alignments. Comput Methods Macromol Seq Anal 266:383–402CrossRefGoogle Scholar
  39. 39.
    Tamura K et al (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24(8):1596–1599PubMedCrossRefGoogle Scholar
  40. 40.
    Jai SR, Yinong Y (2007) Rice Mitogen-activated Protein Kinase Gene Family and Its Role in Biotic and Abiotic Stress Response. J Integr Plant Biol 49(6):751–759CrossRefGoogle Scholar
  41. 41.
    Shen X, Yuan B, Liu H, Li X, Xu C, Wang S (2010) Opposite functions of a rice mitogen-activated protein kinase during the process of resistance against Xanthomonas oryzae. Plant J 64:86–99PubMedGoogle Scholar
  42. 42.
    Yang Y, Shah J, Klessig DF (1997) Signal perception and transduction in plant defense responses. Genes Dev 11(13):1621–1639PubMedCrossRefGoogle Scholar
  43. 43.
    Claudia J et al (1996) Stress signaling in plants: a mitogen-acticated protein kinase pathway is acticated by cold and drought. Proc Natl Acad Sci USA 93:11274–11279CrossRefGoogle Scholar
  44. 44.
    Shi J et al (2011) GhMPK16, a novel stress-responsive group D MAPK gene from cotton, is involved in disease resistance and drought sensitivity. BMC Mol Biol 12:22PubMedCrossRefGoogle Scholar
  45. 45.
    Bennetzen JL (2000) Transposable element contributions to plant gene and genome evolution. Plant Mol Biol 42:251–269PubMedCrossRefGoogle Scholar
  46. 46.
    Yuan B et al (2007) Mitogen-activated protein kinase OsMPK6 negatively regulates rice disease resistance to bacterial pathogens. Planta 226(4):953–960PubMedCrossRefGoogle Scholar
  47. 47.
    Ryals John et al (1995) Signal transduction in systemic acquired resistance. Proc Natl Acad Sci USA 92:4202–4205PubMedCrossRefGoogle Scholar
  48. 48.
    Heil Martin, Bostock RM (2002) Induced Systemic Resistance (ISR) Against Pathogens in the Context of Induced Plant Defences. Ann Bot 89:503–512PubMedCrossRefGoogle Scholar
  49. 49.
    Koo SC et al (2009) OsBWMK1 mediates SA-dependent defense responses by activating the transcription factor OsWRKY33. Biochem Biophys Res Commun 387(2):365–370PubMedCrossRefGoogle Scholar
  50. 50.
    Payne George et al (1989) Isolation and sequence of a genomic clone encoding the basic form of pathogenesis-related protein 1 from Nicotiana tabacum. Plant Mol Biol 12:595–596CrossRefGoogle Scholar
  51. 51.
    Xu Bingfang, Sheehan MoiraJ, Timko MP (2004) Differential induction of ornithine decarboxylase (ODC) gene family members in transgenic tobacco (Nicotiana tabacum L. cv. Bright Yellow 2)cell suspensions by methyl-jasmonate treatment. Plant Growth Regul 44:101–116CrossRefGoogle Scholar
  52. 52.
    Seo Shigemi et al (2007) The mitogen-activated protein kinases WIPK and SIPK regulate the levels of jasmonic and salicylic acids in wounded tobacco plants. Plant J 49:899–909PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Xingtan Zhang
    • 1
  • Tingcai Cheng
    • 2
  • Genhong Wang
    • 2
  • Yafei Yan
    • 1
  • Qingyou Xia
    • 2
    Email author
  1. 1.The Institute of Agricultural and Life ScienceChongqing UniversityChongqingChina
  2. 2.State Key Laboratory of Silkworm Genome BiologySouthwest UniversityChongqingChina

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