Plant Growth Regulation

, Volume 76, Issue 3, pp 269–279 | Cite as

LcMKK, a novel group A mitogen-activated protein kinase kinase gene in Lycium chinense, confers dehydration and drought tolerance in transgenic tobacco via scavenging ROS and modulating expression of stress-responsive genes

  • Dianyun Wu
  • Jing Ji
  • Gang Wang
  • Wenzhu Guan
  • Chunfeng Guan
  • Chao Jin
  • Xiaowei Tian
Original Paper


The mitogen-activated protein kinase (MAPK) cascades have been previously implicated in signal transduction during plant responses to various environmental stresses. As the convergent point of the MAPK cascades, MAPKKs play paramount roles in amplifying, integrating, and channeling information between the extracellular stimuli and intracellular responses. However, the functional role of MAPKKs in Lycium chinense has never been explored. In this study, a novel MAPKK gene, LcMKK, in L. chinense belonging to group A MAPKKs was isolated and functionally characterized. The transcript level of LcMKK rapidly increased in L. chinense after drought treatments. Overexpression of LcMKK in tobacco conferred dehydration and drought tolerance. Under dehydration and drought conditions, the transgenic tobacco lines exhibited better water status, less accumulation of reactive oxygen species (ROS), higher levels of germination rate and antioxidant enzyme activity than the wild type. In addition, overexpression of LcMKK enhanced the expression of ROS-related and stress-responsive genes under drought conditions. Taken together, these data demonstrate that LcMKK acts as a positive regulator in dehydration/drought stress responses by either regulating ROS homeostasis through the activation of the cellular antioxidant defense system or modulating transcriptional levels of a variety of stress-associated genes.


Antioxidant system Dehydration/drought stress tolerance Lycium chinense LcMKK Reactive oxygen species Stress-responsive genes 



This work was supported financially by the National Science and Technology Key Project of China on GMO cultivation for new varieties (No. 2014ZX08003-002B) and the National Natural Science Foundation of China (No. 31271419, No. 31271793 and No. 31401391).

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10725_2014_9998_MOESM1_ESM.docx (1.3 mb)
Supplementary material 1 (DOCX 1337 kb)


  1. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126PubMedGoogle Scholar
  2. Agarwal PK, Gupta K, Jha B (2010) Molecular characterization of the Salicornia brachiata SbMAPKK gene and its expression by abiotic stress. Mol Biol Rep 37:981–986PubMedCrossRefGoogle Scholar
  3. Andreasson E, Ellis B (2010) Convergence and specificity in the Arabidopsis MAPK nexus. Trends Plant Sci 15:106–113PubMedCrossRefGoogle Scholar
  4. Cai G, Wang G, Wang L, Pan J, Liu Y, Li D (2014) ZmMKK1, a novel group A mitogen-activated protein kinase kinase gene in maize, conferred chilling stress tolerance and was involved in pathogen defense in transgenic tobacco. Plant Sci 214:57–73PubMedCrossRefGoogle Scholar
  5. Chen J-Q, Meng X-P, Zhang Y, Xia M, Wang X-P (2008) Over-expression of OsDREB genes lead to enhanced drought tolerance in rice. Biotechnol Lett 30:2191–2198PubMedCrossRefGoogle Scholar
  6. Chen L et al (2012) Genome-wide identification and analysis of MAPK and MAPKK gene families in Brachypodium distachyon. PLoS One 7:e46744PubMedCentralPubMedCrossRefGoogle Scholar
  7. Dong JZ et al (2013) Selenium increases chlorogenic acid, chlorophyll and carotenoids of Lycium chinense leaves. J Sci Food Agric 93:310–315PubMedCrossRefGoogle Scholar
  8. Foyer CH, Shigeoka S (2011) Understanding oxidative stress and antioxidant functions to enhance photosynthesis. Plant Physiol 155:93–100PubMedCentralPubMedCrossRefGoogle Scholar
  9. Giannopolitis CN, Ries SK (1977) Superoxide dismutases I. Occurrence in higher plants. Plant Physiol 59:309–314PubMedCentralPubMedCrossRefGoogle Scholar
  10. Hadiarto T et al (2006) Activation of Arabidopsis MAPK kinase kinase (AtMEKK1) and induction of AtMEKK1–AtMEK1 pathway by wounding. Planta 223:708–713PubMedCrossRefGoogle Scholar
  11. Horsch R, Fry J, Hoffmann N, Eichholtz D, Rogers SA, Fraley R (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231CrossRefGoogle Scholar
  12. Huang X-S, Liu J-H, Chen X-J (2010) Overexpression of PtrABF gene, a bZIP transcription factor isolated from Poncirus trifoliata, enhances dehydration and drought tolerance in tobacco via scavenging ROS and modulating expression of stress-responsive genes. BMC Plant Biol 10:230PubMedCentralPubMedCrossRefGoogle Scholar
  13. Hundertmark M, Hincha DK (2008) LEA (late embryogenesis abundant) proteins and their encoding genes in Arabidopsis thaliana. BMC Genomics 9:118PubMedCentralPubMedCrossRefGoogle Scholar
  14. Kiegerl S et al (2000) SIMKK, a mitogen-activated protein kinase (MAPK) kinase, is a specific activator of the salt stress–induced MAPK, SIMK. Plant Cell 12:2247–2258PubMedCentralPubMedCrossRefGoogle Scholar
  15. Kim HP et al (2002) Zeaxanthin dipalmitate from Lycium chinense fruit reduces experimentally induced hepatic fibrosis in rats. Biol Pharm Bull 25:390–392PubMedCrossRefGoogle Scholar
  16. Kong X et al (2011a) ZmMKK4, a novel group C mitogen-activated protein kinase kinase in maize (Zea mays), confers salt and cold tolerance in transgenic Arabidopsis. Plant Cell Environ 34:1291–1303PubMedCrossRefGoogle Scholar
  17. Kong X, Sun L, Zhou Y, Zhang M, Liu Y, Pan J, Li D (2011b) ZmMKK4 regulates osmotic stress through reactive oxygen species scavenging in transgenic tobacco. Plant Cell Rep 30:2097–2104PubMedCrossRefGoogle Scholar
  18. Kong F, Wang J, Cheng L, Liu S, Wu J, Peng Z, Lu G (2012) Genome-wide analysis of the mitogen-activated protein kinase gene family in Solanum lycopersicum. Gene 499:108–120PubMedCrossRefGoogle Scholar
  19. Lu W, Chu X, Li Y, Wang C, Guo X (2013) Cotton GhMKK1 Induces the tolerance of salt and drought stress, and mediates defence responses to pathogen infection in transgenic Nicotiana benthamiana. PLoS One 8:e68503PubMedCentralPubMedCrossRefGoogle Scholar
  20. Mikołajczyk M, Awotunde OS, Muszyńska 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 12:165–178PubMedCentralPubMedCrossRefGoogle Scholar
  21. Miller G, Suzuki N, CIFTCI-YILMAZ S, Mittler R (2010) Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant Cell Environ 33:453–467PubMedCrossRefGoogle Scholar
  22. Mizoguchi T, Ichimura K, Irie K, Morris P, Giraudat J, Matsumoto K, Shinozaki K (1998) Identification of a possible MAP kinase cascade in Arabidopsis thaliana based on pairwise yeast two-hybrid analysis and functional complementation tests of yeast mutants. FEBS Lett 437:56–60PubMedCrossRefGoogle Scholar
  23. Nadarajah K, Sidek HM (2010) The green MAPKS. Asian J Plant Sci 9:1CrossRefGoogle Scholar
  24. Nicole M-C, Hamel L-P, Morency M-J, Beaudoin N, Ellis B, Séguin A (2006) MAP-ping genomic organization and organ-specific expression profiles of poplar MAP kinases and MAP kinase kinases. BMC Genomics 7:223PubMedCentralPubMedCrossRefGoogle Scholar
  25. Pitzschke A, Djamei A, Bitton F, Hirt H (2009) A major role of the MEKK1–MKK1/2–MPK4 pathway in ROS signalling. Mol Plant 2:120–137PubMedCentralPubMedCrossRefGoogle Scholar
  26. Rao KP, Richa T, Kumar K, Raghuram B, Sinha AK (2010) In silico analysis reveals 75 members of mitogen-activated protein kinase kinase kinase gene family in rice. DNA Res 17:139–153PubMedCentralPubMedCrossRefGoogle Scholar
  27. Ravikumar G et al (2014) Stress-inducible expression of AtDREB1A transcription factor greatly improves drought stress tolerance in transgenic indica rice. Transgenic Res 23:421–439PubMedCentralPubMedCrossRefGoogle Scholar
  28. Samuel MA, Ellis BE (2002) Double jeopardy both overexpression and suppression of a redox-activated plant mitogen-activated protein kinase render tobacco plants ozone sensitive. Plant Cell 14:2059–2069PubMedCentralPubMedCrossRefGoogle Scholar
  29. Stulemeijer IJ, Stratmann JW, Joosten MH (2007) Tomato mitogen-activated protein kinases LeMPK1, LeMPK2, and LeMPK3 are activated during the Cf-4/Avr4-induced hypersensitive response and have distinct phosphorylation specificities. Plant Physiol 144:1481–1494PubMedCentralPubMedCrossRefGoogle Scholar
  30. Tena G, Asai T, Chiu W-L, Sheen J (2001) Plant mitogen-activated protein kinase signaling cascades. Curr Opin Plant Biol 4:392–400PubMedCrossRefGoogle Scholar
  31. 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 17:113–122PubMedCrossRefGoogle Scholar
  32. Wintermans J, De Mots A (1965) Spectrophotometric characteristics of chlorophylls a and b and their phenophytins in ethanol. BBA 109:448–453PubMedGoogle Scholar
  33. Xing Y, Jia W, Zhang J (2008) AtMKK1 mediates ABA-induced CAT1 expression and H2O2 production via AtMPK6-coupled signaling in Arabidopsis. Plant J 54:440–451PubMedCrossRefGoogle Scholar
  34. Zhang A, Jiang M, Zhang J, Tan M, Hu X (2006) Mitogen-activated protein kinase is involved in abscisic acid-induced antioxidant defense and acts downstream of reactive oxygen species production in leaves of maize plants. Plant Physiol 141:475–487PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Dianyun Wu
    • 1
  • Jing Ji
    • 2
  • Gang Wang
    • 2
  • Wenzhu Guan
    • 2
  • Chunfeng Guan
    • 2
  • Chao Jin
    • 2
  • Xiaowei Tian
    • 1
  1. 1.School of Chemical Engineering and TechnologyTianjin UniversityTianjinPeople’s Republic of China
  2. 2.School of Environmental Science and EngineeringTianjin UniversityTianjinPeople’s Republic of China

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