Plant Cell Reports

, Volume 23, Issue 6, pp 397–403 | Cite as

The shooty callus induced by suppression of tobacco CHRK1 receptor-like kinase is a phenocopy of the tobacco genetic tumor

  • J. H. Lee
  • D.-M. Kim
  • Y. P. Lim
  • H.-S. Pai
Genetics and Genomics


CHRK1 encodes a tobacco receptor-like kinase that contains a chitinase-like sequence in the extracellular domain. In a previous study, CHRK1-suppressed transgenic tobacco plants exhibited pleiotropic developmental abnormalities including spontaneous growth of shooty callus from emerging embryos in the absence of any exogenous hormones. In this study, we show that the CHRK1 shooty callus mimics tobacco genetic tumors in its morphology, physiology, and gene expression profiles. Similar to CHRK1 shooty callus, tobacco genetic tumors exhibit shooty callus morphology and hormone-independent shoot organogenesis. Both the CHRK1 callus and genetic tumors constitutively expressed KNOTTED1-type homeobox genes at the high levels, consistent with their vigorous shoot formation. These two types of calli exhibited cell death phenotypes, accompanied by high H2O2 production, increased ion leakage, and callose accumulation. Consistently, both types of calli constitutively expressed high levels of defense genes induced during pathogen-mediated HR cell death. These results, together with previous reports that both the CHRK1 shooty callus and tobacco genetic tumor contained high levels of cytokinin, indicate that CHRK1 shooty callus is a phenocopy of tobacco genetic tumor. CHRK1-mediated signal transduction may play a role in the formation of the genetic tumor in tobacco.


Cell death Cytokinin Defense-related genes Homeobox genes Shoot proliferation 



The authors wish to thank Dr. Myeong-Hyeon Wang (Kang-Won University, Korea) for providing tobacco genetic tumor lines. This research was supported by a grant from Plant Diversity Research Center of 21st Century Frontier Research Program funded by Ministry of Science and Technology of Korean government.


  1. Ames IH (1972) The influence of cytokinins on genetic tumor formation. Can J Bot 50:2235–2238Google Scholar
  2. Bayer MH (1982) Genetic tumors: physiological aspects of tumor formation in interspecies hybrids. In: Kahl G, Schell J (eds) Molecular biology of plant tumors. Academic, New York, pp 33–67Google Scholar
  3. Brodersen P, Petersen M, Pike HM, Olszak B, Skov S, Odum N, Jorgensen LB, Brown RE, Mundy J (2002) Knockout of Arabidopsis accelerated-cell-death11 encoding a sphingosine transfer protein causes activation of programmed cell death and defense. Genes Dev 16:490–502CrossRefPubMedGoogle Scholar
  4. Feng XH, Dube SK, Bottino PJ, Kung SD (1990) Restoration of shooty morphology of a nontumorous mutant of Nicotiana glauca × N. langsdorffii by cytokinin and the isopentenyltransferase gene. Plant Mol Biol 15:407–420PubMedGoogle Scholar
  5. Frugis G, Giannino D, Mele G, Nicolodi C, Chiappetta A, Bitonti MB, Innocenti AM, Dewitte W, van Onckelen H, Mariotti D (2001) Overexpression of KNAT1 in lettuce shifts leaf determinate growth to a shoot-like indeterminate growth associated with an accumulation of isopentenyl-type cytokinins. Plant Physiol 126:1370–1380CrossRefPubMedGoogle Scholar
  6. Fujita T, Kouchi H, Ichikawa T, Syono K (1994) Cloning of cDNAs for genes that are specifically or preferentially expressed during the development of tobacco genetic tumors. Plant J 5:645–654PubMedGoogle Scholar
  7. Ichikawa T, Syono K (1991) Tobacco genetic tumors. Plant Cell Physiol 32:1123–1128Google Scholar
  8. Ito Y, Eiguchi M, Kurata N (2001) KNOX homeobox genes are sufficient in maintaining cultured cells in an undifferentiated state of rice. Genesis 30:231–238CrossRefPubMedGoogle Scholar
  9. Kim YS, Lee JH, Yoon GM, Cho HS, Park SW, Suh MC, Choi D, Ha HJ, Liu JR, Pai H-S (2000) CHRK1, a chitinase-related receptor-like kinase in tobacco. Plant Physiol 123:905–915CrossRefPubMedGoogle Scholar
  10. Kim M, Ahn J-W, Jin U-H, Paek K-H, Pai H-S (2003) Activation of the programmed cell death pathway by inihibition of proteasome function in plants. J Biol Chem 278:19406–19415CrossRefPubMedGoogle Scholar
  11. Lee JH, Takei K, Sakakibara H, Cho HS, Kim DM, Kim YS, Min SR, Kim WT, Sohn DY, Lim YP, Pai H-S (2003) CHRK1, a chitinase-related receptor-like kinase, plays a role in plant development and cytokinin homeostasis in tobacco. Plant Mol Biol 53:877–890CrossRefPubMedGoogle Scholar
  12. Nandi SK, de Klerk GJM, Parker CW, Palni LMS (1990) Endogenous cytokinin levels and metabolism of zeatin riboside in genetic tumor tissues and non-tumorous tissues of tobacco. Physiol Plant 78:197–204CrossRefGoogle Scholar
  13. Näf U (1958) Studies on tumor formation in Nicotiana hybrids. Growth 22:167–180PubMedGoogle Scholar
  14. Nishimura A, Tamaoki M, Sakamoto T, Matsuoka M (2000) Over-expression of tobacco knotted1-type class1 homeobox genes alters various leaf morphology. Plant Cell Physiol 41:583–590PubMedGoogle Scholar
  15. Osakabe Y, Miyata S, Urao T, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2002) Overexpression of Arabidopsis response regulators, ARR4/ATRR1/IBC7 and ARR8/ATRR3, alters cytokinin responses differentially in the shoot and in callus formation. Biochem Biophys Res Commun 293:806–815CrossRefPubMedGoogle Scholar
  16. Pontier D, Tronchet M, Rogowsky P, Lam E, Roby D (1998) Activation of hsr203, a plant gene expressed during incompatible plant-pathogen interactions, is correlated with programmed cell death. Mol Plant Microbe Interact 11:544–554PubMedGoogle Scholar
  17. Riou-Khamlichi C, Huntley R, Jacqmard A, Murray JAH (1999) Cytokinin activation of Arabidopsis cell division through a D-type cyclin. Science 283:1541–1544CrossRefPubMedGoogle Scholar
  18. Rupp HM, Frank M, Werner T, Strnad M, Schmülling T (1999) Increased steady state mRNA levels of the STM and KNAT1 homeobox genes in cytokinin overexpressing Arabidopsis thaliana indicate a role for cytokinins in the shoot apical meristem. Plant J 18:557–563CrossRefPubMedGoogle Scholar
  19. Sakamoto T, Nishimura A, Tamaoki M, Kuba M, Tanaka H, Iwahori S, Matsuoka M (1999) The conserved KNOX domain mediates specificity of tobacco KNOTTED1-type homeodomain proteins. Plant Cell 11:1419–1431CrossRefPubMedGoogle Scholar
  20. Sharma VJ, Fletcher JC (2002) Maintenance of shoot and floral meristem cell proliferation and fate. Plant Physiol 129:31–39CrossRefPubMedGoogle Scholar
  21. Sinha NR, Williams RE, Hake S (1993) Overexpression of the maize homeobox gene, KNOTTED-1, causes a switch from determinate to indeterminate cell fates. Genes Dev 7:787–795PubMedGoogle Scholar
  22. Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp Soc Exp Biol 11:118–131Google Scholar
  23. Smith HH (1972) Plant genetic tumors. Prog Exp Tumor Res 15:138–164PubMedGoogle Scholar
  24. Tamaoki M, Kusaba S, Kano-Murakami Y, Matsuoka M (1997) Ectopic expression of a tobacco homeobox gene, NTH15, dramatically alters leaf morphology and hormone levels in transgenic tobacco. Plant Cell Physiol 38:917–927PubMedGoogle Scholar
  25. Thomas JC, Smigocki AC, Bohnert HJ (1995) Light-induced expression of ipt gene from Agrobacterium tumefaciens results in cytokinnin accumulation and osmotic stress symptoms in transgenic tobacco. Plant Mol Biol 27:225–235PubMedGoogle Scholar
  26. Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localization of H2O2 in plants: H2O2 accumulation in papillae and hypersensitive response during the barley–powdery mildew interaction. Plant J 11:1187–1194CrossRefGoogle Scholar
  27. Wang MH, Doonan JH, Sastry GRK (1999) Cloning and characterization of the unusual cyclin gene from an amphidiploid of Nicotiana glauca-Nicotiana langsdorffii hybrid. Biochim Biophys Acta 1489:399–404CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Laboratory of Plant GenomicsKorea Research Institute of Bioscience and BiotechnologyTaejonSouth Korea
  2. 2.Department of HorticultureChungnam UniversityTaejonSouth Korea
  3. 3.Division of Biosciences and BioinformaticsMyongji UniversityKyonggi-doSouth Korea

Personalised recommendations