Skip to main content
Log in

Root cap specific expression of an endo-β-1,4--glucanase (cellulase): a new marker to study root development in Arabidopsis

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

The sloughing of root cap cells from the root tip is important because it assists the growing root in penetrating the soil. Using a promoter–reporter (GUS) and RT-PCR analysis, we identified an endo-β-1,4-glucanase (AtCel5) of Arabidopsis thaliana that is expressed exclusively in root cap cells of both primary and secondary roots. Expression is inhibited by high concentrations of IAA, both exogenous and internal, as well as by ABA. AtCel5 expression begins once the mature tissue pattern is established and continues for 3 weeks. GUS staining is observed in both root cap cells that are still attached and cells that have already been shed. Using AtCel5-GUS as a marker, we observed that the root cap cells begin to separate at the sides of the tip while the cells of the central region of the tip separate last. Separation involves sequential tiers of intact cells that separate from the periphery of the root tip. A homozygous T-DNA insertion mutant that does not express AtCel5 forms the root cap and sheds root cap cells but sloughing is less efficient compared to wild type. The reduction in sloughing in the mutant does not affect the overall growth performance of the plant in loose media. The modest effect of abolishing AtCel5 expression suggests that there are multiple redundant genes regulating the process of sloughing of the root cap, including AtCel3/At1g71380, the paralog of the AtCel5 gene that is also expressed in the root cap cells. Thus, these two endo-1,4-β-<sc>d</sc>-glucanases may have a role in the sloughing of border cells from the root tip. We propose that AtCel5, provides a new molecular marker to further analyze the process of root cap cell separation and a root cap specific promoter for targeting to the environment genes with beneficial properties for plant growth.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Alonso, J. M., Stepanova, A. N., Leisse, T. J., Kim, C. J., Chen, H., Shinn, P., Denise K. S., Zimmerman, J., Barajas, P., Cheuk, R., Gadrinab, C., Heller, C., Jeske, A., Koesema, E., Meyers, C. C., Parker, H., Prednis, L., Ansari, Y., Choy, N., Deen, H., Geralt, M., Hazari, N., Hom, E., Karnes, M., Mulholland, C., Ndubaku, R., Schmidt, I., Guzman, P., Aguilar-Henonin, L., Schmid, M., Weige, D., Carter, D. E., Marchand, T., Risseeuw, E., Brogden, D., Zeko, A., Crosby, W. L., Berry, C. C. and Ecker, J. R. 2003. Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301: 653-657.

    PubMed  Google Scholar 

  • Bengough, A. and Mckenzie, B. 1997. Sloughing of root-cap cells decreases the frictional resistance to maize root growth. J. Exp. Bot. 48: 885-893.

    Google Scholar 

  • Bent, A. F. 2000. Arabidopsis in planta transformation: uses, mechanisms, and prospects for transformation of other species. Plant Physiol. 124: 1540-1547.

    PubMed  Google Scholar 

  • Birnbaum, K., Shasha, D. E., Wang, J. Y., Jung, J. W., Lambert, G. M., Galbraith, D. W. and Benfey, P. N. 2003. A gene expression map of the Arabidopsis root. Science 302: 1956-dy1960.

    Google Scholar 

  • Borderies, G., Jamet, E., La tte, C., Rossignol, M., Jauneau, A., Boudart, G., Monserrat, B., Esquarreed-Tugaye, M.-T., Bouder, A. and Pont-Lezica, R. 2003. Proteomics of loosely bound cell wall proteins of Arabidopsis thaliana cell suspension cultures: a critical analysis. Electrophoresis 24: 3421-3432.

    PubMed  Google Scholar 

  • Brigham, L., Woo, H., Wen, F. and Hawes, M. 1998. Meristem-specific suppression of mitosis and a global switch in gene expression in the root cap of pea by endogenous signals. Plant Physiol. 118: 1223-1231.

    PubMed  Google Scholar 

  • Casimiro, I., Marchant, A., Bhalerao, R. P., Beeckman, T., Dhooge, S., Swarup, R., Graham, N., Inze ´, D., Sandberg, G., Pedro J. Casero and Bennett, M. 2001. Auxin transport promotes Arabidopsis lateral root initiation. Plant Cell 13: 843-852.

    PubMed  Google Scholar 

  • Corpet, F. 1988. Multiple sequence alignment with hierarchical clustering. Nucl. Acids Res. 16: 10881-10890.

    PubMed  Google Scholar 

  • del Campillo, E. 1999. Multiple endo-1, 4-b D D-glucanase (cellulase)genes in Arabidopsis. Curr. Top. Dev. Biol. 46: 39-61.

    PubMed  Google Scholar 

  • del Campillo, E. and Bennett, A. B. 1996. Pedicel breakstrength and cellulase gene expression during tomato. ower abscission. Plant Physiol. 111: 813-820.

    PubMed  Google Scholar 

  • del Campillo, E. and Lewis, L. 1992. Occurrence of 9. 5 cellulase and other hydrolases in. ower reproductive organs undergoing major cell wall disruption. Plant Physiol. 99: 1015-dy1020.

    Google Scholar 

  • DiDonato, R. J., Arbuckle, E., Buker, S., Sheets, J., Tobar, J., Totong, R., Grisa, P., Fink, G. R. and Celenza, J. L. 2004. Arabidopsis ALF4 encodes a nuclear localized protein required for lateral root formation. Plant J. 37: 340-353.

    PubMed  Google Scholar 

  • Gonzalez-Bosch, C., Campillo, E. D. and Bennett, A. B. 1997. Immunodetection and characterization of tomato endo-[beta ]-1, 4-glucanase cel1 protein in. ower abscission zones. Plant Physiol. 114: 1541-1546.

    PubMed  Google Scholar 

  • Hawes, M. 1990. Living plant cells released from the root cap: a regulator of microbial populations in the rhizosphere? Plant Soil 129: 19-27.

    Google Scholar 

  • Hawes, M. C., Brigham, L. A., Wen, F., Woo, H. H. and Zhu, Y. 1998. Function of root border cells in plant health: pioneers in the rhizosphere. Annu. Rev. Phytopathol. 36: 311-327.

    PubMed  Google Scholar 

  • Henrissat, B. 1991. A classification of glycosyl hydrolases based on amino-acid sequence similarities. Biochem. J. 280: 309-316.

    Google Scholar 

  • Henrissat, B. and Bairoch, A. 1993. New families in the classi cation of glycosyl hydrolases based on amino acid sequence similarities. Biochem. J. 293: 781-788.

    PubMed  Google Scholar 

  • Henrissat, B. and Bairoch, A. 1996. Updating the sequence-based classification of glycosyl hydrolases. Biochem. J. 316: 695-696.

    PubMed  Google Scholar 

  • Higo, K., Ugawa, Y., Iwamoto, M. and Korenaga, T. 1999. Plant cis-acting regulatory DNA elements (PLACE)data-base. Nucleic Acids Res. 27: 297-300.

    PubMed  Google Scholar 

  • Iijima, M., Higuchi, T., Barlow, P. and Bengough, A. 2003. Root cap removal increases root penetration resistance in maize (Zea mays L). J. Exp. Bot. 54: 2105-2109.

    PubMed  Google Scholar 

  • Iwasaki, T., Yamaguchi-Shinozaki, K. and Shinozaki, K. 1995. Identification of a cis-regulatory region of a gene in Arabidopsis thaliana whose induction by dehydration is mediated by abscisic acid and requires protein synthesis. Mol. Gen. Genet. 247: 391-398.

    PubMed  Google Scholar 

  • Jaglo-Ottosen, K. R., Gilmour, S. J., Zarka, D. G., Schabenber-ger, O. and Thomashow, M. F. 1998. Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance. Science 3: 104-106.

    Google Scholar 

  • Je. erson, R. A., Kavanagh, T. A. and Bevan, M. W. 1987. GUS fusions: βglucuronidase is a sensitive and versatile fusion marker in higher plants. EMBO J. 6: 3901-3907.

    PubMed  Google Scholar 

  • Jensen, P. J., Hangarter, R. P. and Estelle, M. 1998. Auxin transport is required for hypocotyl elongation in light-grown but not dark-grown Arabidopsis. Plant Physiol. 116: 455-462.

    PubMed  Google Scholar 

  • Kalaitzis, P., Hong, S. B., Solomos, T. and Tucker, M. L. 1999. Molecular characterization of a tomato [Lycopersicon esculentum ]endo-beta-1, 4-glucanase gene expressed in mature pistils, abscission zones and fruit. Plant Cell Physiol.: 905-908.

  • Lashbrook, C. C., Gonzalez-Bosch, C. and Bennett, A. B. 1994. Two divergent Endo-[beta ]-1, 4-glucanase genes exhibit over-lapping expression in ripening fruit and abscising flowers. Plant Cell 6: 1485-1493.

    PubMed  Google Scholar 

  • Laskowski, M., Williams, M., Nusbaum, C. and Sussex, I. M. 1995. Formation of lateral root meristems is a two-stage process. Development 121: 3303-3310.

    PubMed  Google Scholar 

  • Lu, G. and Ferl, R. 1995. An Arabidopsis cDNA encoding beta-glucanase. Plant Mol. Biol. 29: 883.

    Google Scholar 

  • Milioni, D., Sado, P.-E., Stacey, N. J., Domingo, C., Roberts, K. and McCann, M. 2001. Di. erential expression of cell-wall-related genes during the formation of tracheary elements in the Zinnia mesophyll cell system. Plant Mol. Biol. 47: 221-238.

    PubMed  Google Scholar 

  • Milioni, D., Sado, P.-E., Stacey, N. J., Roberts, K. and McCann, M. 2002. Early gene expression associated with the commitment and differentiation of a plant tracheary element is revealed by cDNA-ampli ed fragment length polymorphism analysis. Plant Cell 14: 2813-2824.

    PubMed  Google Scholar 

  • Miyasaka, S. and Hawes, M. 2001. Possible role of root border cells in detection and avoidance of aluminum toxicity. Plant Physiol. 125: 1978-1987.

    PubMed  Google Scholar 

  • Moseyko, N., Zhu, T., Chang, H.-S., Wang, X. and Feldman, L. J. 2002. Transcription pro ling of the early gravitropic response in Arabidopsis using high-density oligonucleotide probe microarrays. Plant Physiol 130: 720-728.

    PubMed  Google Scholar 

  • Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15: 473-497.

    Google Scholar 

  • Murphy, A. and Taiz, L. 1995. A new vertical mesh transfer technique for metal-tolerance studies in Arabidopsis. Ecotypic variation and copper-sensitive mutants. Plant Physiol. 108: 29-38.

    PubMed  Google Scholar 

  • Nicol, F., His, I., Jauneau, A., Vernhettes, S., Canut, H. and Ho ¨fte, H. 1998. A plasma membrane-bound putative endo-1, 4-b D D-glucanase is required for normal wall assembly and cell elongation in Arabidopsis. EMBO 17: 5563-5576.

    Google Scholar 

  • Thoma, S. L., Glass, T., Most, A. and Patterson, S. E. 2003. Analysis of an abscission-associated cellulase in Arabidopsis. 14th International Conference on Arabidopsis Research, Madison, WI.

  • Tucker, M., Sexton, R., del Campillo, E. and Lewis, L. 1988. Bean abscission cellulase: characterization of a cDNA clone and regulation of gene expression by ethylene and auxin. Plant Physiol. 88: 1257-1262.

    Google Scholar 

  • Ulmasov, T., Murfett, J., Hagen, G. and Guilfoyle, T. J. 1997. Aux/IAA proteins repress expression of reporter genes containing natural and highly active synthetic auxin response elements. Plant Cell 9: 1963-1971.

    PubMed  Google Scholar 

  • Wen, F., Zhu, Y. and Hawes, M. 1999. Effect of pectin methyl-esterase gene expression on pea root development. Plant Cell 11: 1129-1140.

    PubMed  Google Scholar 

  • Willats, W. G. T., Willats W. G. T., McCartney L., Steele-King C. G., Marcus S. E., Mort A., Huisman M., van Alebeek G.-J., Schols H. A., Voragen A. G. J., Le Goff A., Bonnin E., Thibault J.-F. and J. P., K. 2004. A xylogalacturonan epitope is specifically associated with plant cell detachment. Planta 218: 673-681.

    PubMed  Google Scholar 

  • Zeevaart, J. A. D. and Creelman, R. A. 1988. Metabolism and physiology of abscisic acid. Annu. Rev. Plant. Physiol. 39: 438-473

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Campillo, E.d., Abdel-Aziz, A., Crawford, D. et al. Root cap specific expression of an endo-β-1,4--glucanase (cellulase): a new marker to study root development in Arabidopsis . Plant Mol Biol 56, 309–323 (2004). https://doi.org/10.1007/s11103-004-3380-3

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11103-004-3380-3

Navigation