Plant Molecular Biology

, Volume 60, Issue 5, pp 747–758 | Cite as

Rejuvenation by Shoot Apex Culture Recapitulates the Developmental Increase of Methylation at the Maize Gene Pl-Blotched

  • Erin E. Irish
  • Douglas McMurray


Cytosine methylation provides an attractive epigenetic modification for the global maintenance of phases in plant development; however, there are few known examples of specific genes whose methylation status changes in a developmentally regulated manner. Pl-Blotched, an allele of purple plant1 (pl1), which encodes a myb-like transcription factor that regulates anthocyanin production in maize, is one such gene: certain cytosines at the 3′ end of this allele are hypomethylated in seedlings, become hypermethylated in organs formed in the adult phase, and are hypomethylated again in the next generation. We tested whether this developmental pattern of low juvenile cytosine methylation followed by higher methylation in adult tissues could also be observed in plants “rejuvenated” via shoot apex culture. We found that cytosine methylation patterns at Pl-Blotched were indeed recapitulated in culture-rejuvenated plants, showing hypomethylation in leaves with juvenile patterns of differentiation (even though they were made by an old meristem) followed by hypermethylation in later-formed leaves. Our results show that methylation status at that locus is determined by the developmental phase of the shoot, rather than by the age of the meristem forming it. These results support the hypothesis that DNA methylation is employed by the plant to maintain an epigenetic state.


cytosine methylation epigenetic phase change plant development 


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  1. Alleman, M., Doctor, J. 2000Genomic imprinting in plants: observations and evolutionary implicationsPlant Mol. Biol.43147161CrossRefPubMedGoogle Scholar
  2. Banks, J.A., Federoff, N.V. 1989Patterns of developmental and heritable change in methylation of the Suppressor-mutator transposable elementDevel. Genet.10425437CrossRefGoogle Scholar
  3. Banks, J.A., Masson, P., Federoff, N. 1988Molecular mechanisms in the developmental regulation of the maize Suppressor-mutator transposable elementGenes Dev.213641380PubMedGoogle Scholar
  4. Bao, N., Lye, K.W., Barton, M.K. 2004MicroRNA binding sites in Arabidopsis class III HD-ZIP mRNAs are required for methylation of the template chromosomeDev. Cell.5653662CrossRefGoogle Scholar
  5. Barkan, A., Martienssen, R.A. 1991Inactivation of maize transposon Mu suppresses a mutant phenotype by activating an outward-reading promoter near the end of Mu1Proc. Natl. Acad. Sci. USA8835023506PubMedGoogle Scholar
  6. Bennetzen, J.L. 1987Covalent DNA modification and the regulation of Mutator element transposition in maizeMol. Gen. Genet.2085762CrossRefGoogle Scholar
  7. Bitonti, M.B., Cozza, R., Chiappetta, A., Giannino, D., Castiglione, M., Dewitte, W., Mariotti, D., Onokelen, H., Innocenti, A.M. 2002Distinct nuclear organization, DNA methylation pattern and cytokinin distribution mark juvenile, juvenile-like and adult vegetative apical meristems in peach (Prunus persica L. Batsch)J. Exp. Bot.5310471054CrossRefPubMedGoogle Scholar
  8. Bongard-Pierce, D.K., Evans, M.M.S., Poethig, R.S. 1996Heteroblastic features of leaf anatomy in maize and their genetic regulationInt. J. Plant. Sci.157331340CrossRefGoogle Scholar
  9. Bowman, J.L., Sakai, H., Jack, T., Weigel, D., Mayer, U., Meyerowitz, E.M. 1992SUPERMAN, a regulator of floral homeotic genes in ArabidopsisDevelopment114599615PubMedGoogle Scholar
  10. Brink, R.A. 1962Phase change in higher plants and somatic cell heredityQuart. Rev. Biol.37122CrossRefGoogle Scholar
  11. Brown, W.E., Springer, P.S., Bennetzen, J.L. 1994Progressive modification of Mu transposable elements during developmentMaydica39119126Google Scholar
  12. Brutnell, T.P., Dellaporta, S.L. 1994Somatic inactivation and reactivation of Ac associated with changes in cytosine methylation and transposase expressionGenetics138213225PubMedGoogle Scholar
  13. Chandler, V.L., Walbot, V. 1986DNA modification of a maize transposable element correlates with loss of activityProc. Natl. Acad. Sci. USA8317671771PubMedGoogle Scholar
  14. Chen, J., Dellaporta, S. 1993Urea-based plant DNA miniprepFreeling, M.Walbot, V. eds. The Maize HandbookSpringer-VerlagNew York526527Google Scholar
  15. Chomet, P.S., Wessler, S., Dellaporta, S.L. 1987Inactivation of the maize transposable element Activator (Ac) is associated with its DNA modificationEMBO J.6295302PubMedGoogle Scholar
  16. Cocciolone, S.M., Cone, K.C. 1993Pl-Bh, an anthocyanin regulatory gene of maize that leads to variegated pigmentationGenetics135575588PubMedGoogle Scholar
  17. Cocciolone, S.M., Chopra, S., Flint-Garcia, S.A., McMullen, M.D., Peterson, T. 2001Tissue-specific patterns of a maize Myb transcription factor are epigenetically regulatedPlant J.27467478CrossRefPubMedGoogle Scholar
  18. Cone, K.C., Cocciolone, S.M., Burr, F.A., Burr, B. 1993Maize anthocyanin regulatory gene pl is a duplicate of c1 that functions in the plantPlant Cell517951805CrossRefPubMedGoogle Scholar
  19. Federoff, N., Masson, P., Banks, J.A. 1989Mutations, epimutations, and the developmental programming of the maize Suppressor-mutator transposable elementBioEssays10139144CrossRefGoogle Scholar
  20. Federoff, N.V., Banks, J.A. 1988Is the Suppressor-mutator element controlled by a basic developmental regulatory mechanism? Genetics120559577Google Scholar
  21. Federoff, N. 1989The heritable activation of cryptic Suppressor-mutator elements by an active elementGenetics121591608Google Scholar
  22. Finnegan, E.J., Genger, R.K., Kovac, K., Peacock, W.J., Dennis, E.S. 1996DNA methylation and the promotion of flowering by vernalizationProc. Natl. Acad. Sci. USA9558245829CrossRefGoogle Scholar
  23. Fraga, M., Canal, J., Rodriguez, R. 2002Phase-change related epigenetic and physiological changes in Pinus radiata D.DonPlanta215672678CrossRefPubMedGoogle Scholar
  24. Goodrich, J., Tweedie, S. 2002Remembrance of things past: chromatin remodeling in plant developmentAnn. Rev. Cell Dev. Biol.18707746CrossRefGoogle Scholar
  25. Henikoff, S., Matzke, M.A. 1997Exploring and explaining epigenetic effectsTrends Genet.13293295CrossRefPubMedGoogle Scholar
  26. Hoekenga, O.A., Muszynski, M.G., Cone, K.C. 2000Developmental patterns of chromatin structure and DNA methylation responsible for epigenetic expression of a maize regulatory geneGenetics15518891902PubMedGoogle Scholar
  27. Hollick, J.B., Dorweiler, J.E., Chandler, V.L. 1997Paramutation and related allelic interactionsTrends Genet.13302308CrossRefPubMedGoogle Scholar
  28. Hsieh, T.-F., Fischer, R.L. 2005Biology of chromatin dynamicsAnn. Rev. Plant Biol.56327351CrossRefGoogle Scholar
  29. Irish, E., Jegla, D. 1997Regulation of extent of vegetative development of the maize shoot meristemPlant J.116371CrossRefGoogle Scholar
  30. Irish, E.E., Karlen, S. 1998Restoration of juvenility in maize shoots by meristem cultureInt. J. Plant Sci.159695701CrossRefGoogle Scholar
  31. Irish, E.E., Nelson, T. 1988Development of maize plants from cultured shoot apicesPlanta175912CrossRefGoogle Scholar
  32. Jacobsen, S.W., Meyerowitz, E.M. 1997Hypermethylated SUPERMAN epigenetic alleles in ArabidopsisScience27711001103CrossRefPubMedGoogle Scholar
  33. Jones, P.A. 1985Altering gene expression with 5-azacytidineCell40485486CrossRefPubMedGoogle Scholar
  34. Kakutani, T., Jeddeloh, J.A., Flowers, S.K., Munakata, K., Richard, E.J. 1996Developmental abnormalities and epimutations associated with DNA hypomethylation mutationsProc. Natl. Acad. Sci. USA931240612411CrossRefPubMedGoogle Scholar
  35. Lawson, E.J.R., Poethig, R.S. 1995Shoot development in plants: time for a changeTrends Genet.11263268CrossRefPubMedGoogle Scholar
  36. Li, G., Hall, T.C., Holmes-Davis, R. 2002Plant chromatin: development and gene controlBioEssays24234243CrossRefPubMedGoogle Scholar
  37. Lisch, D., Carey, C.C., Dorweiler, J.E., Chandler, V.L. 2002A mutation that prevents paramutation in maize also reverses Mutator transposon methylation and silencingProc. Natl. Acad. Sci. USA9961306135CrossRefPubMedGoogle Scholar
  38. Lund, G., Ciceri, P., Viotti, A. 1995aMaternal-specific demethylation and expression of specific alleles of zein genes in the endosperm of Zea mays LPlant J.8571581CrossRefGoogle Scholar
  39. Lund, G., Messing, J., Viotti, A. 1995bEndosperm-specific demethylation and activation of specific alleles of alpha-tubulin genes of Zea mays L MoleGen. Genet.246716722CrossRefGoogle Scholar
  40. Martienssen, R., Baron, A. 1994Coordinate suppression of mutations caused by ROBERTSON’S Mutator transposons in maizeGenetics13611571170PubMedGoogle Scholar
  41. Martienssen, R.A., Colot, V. 2001DNA methylation and epigenetic inheritance in plants and filamentous fungiScience29310701073CrossRefPubMedGoogle Scholar
  42. Martienssen, R., Barkan, A., Taylor, W.C., Freeling, M. 1990Somatically heritable switches in the DNA modification of Mu transposable elements monitored with a suppressible mutant in maizeGenes Dev.4331343PubMedGoogle Scholar
  43. Matzke, M.A., Mette, M.F., Matzke, A.J. 2000Transgene silencing by the host genome defense: implications for the evolution of epigenetic control mechanisms in plants and vertebratesPlant Mol. Biol.43401415CrossRefPubMedGoogle Scholar
  44. Poethig, R.S. 1990Phase change and regulation of shoot morphogenesis in plantsScience250923930Google Scholar
  45. Richards, E.J. 1997DNA methylation and plant developmentTrends Genet.13319323CrossRefPubMedGoogle Scholar
  46. Ronemus, M.J., Galbiati, M., Ticknor, C., Chen, J., Dellaporta, S.L. 1996Demethylation-induced developmental pleiotropy in ArabidopsisScience273654657PubMedGoogle Scholar
  47. Soppe, W.J., Jacobsen, S.E., Alonso-Blanco, C., Jackson, J.P., Kakutani, T., Koornneef, M., Peeters, A.J.M. 2000The late flowering phenotype of fwa mutants is caused by gain-of-function epigenetic alleles of a homeodomain geneMol. Cell6791802CrossRefPubMedGoogle Scholar
  48. Stam, M., Belele, C., Dorweiler, J.E., Chandler, V.L. 2002Differential chromatin structure within a tandem array 100 kb upstream of the maize b1 locus is associated with paramutationGenes Dev.1619061918CrossRefPubMedGoogle Scholar
  49. Sussex, I.M. 1976Phase change: physiological and genetic aspectsActa Hort.56275280Google Scholar
  50. Vongs, A., Kakutani, T., Martienssen, R.A., Richards, E. 1993Arabidopsis thaliana DNA methylation mutantsScience26019261928PubMedGoogle Scholar
  51. Wagner, D. 2003Chromatin regulation of plant developmentCurr. Opin. Plant Biol.62028CrossRefPubMedGoogle Scholar
  52. Walbot, V. 2001Imprinting of R-r, paramutation of B-I and Pl, and epigenetic silencing of MuDR/Mu transposons in Zea mays L. are coordinately affected by inbred backgroundGenet. Res. Camb.77219226Google Scholar
  53. Walker, E. 1998Paramutation of the r1 locus of maize is associated with increased cytosine methylationGenetics14819731981PubMedGoogle Scholar
  54. Zluvova, J., Janousek, B., Vyskot, B. 2001Immunohistochemical study of DNA methylation dynamics during plant developmentJ. Exp. Bot.5222652273CrossRefPubMedGoogle Scholar

Copyright information

© Springer 2006

Authors and Affiliations

  1. 1.Department of Biological SciencesThe University of IowaIowa CityUSA

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