Abstract
The soybean Ep gene encodes an anionic peroxidase enzyme that accumulates in large amounts in seed coat tissues. We have isolated a second peroxidase gene, Prx2, that is also highly expressed in developing seed coat tissues. Sequence analysis of Prx2 cDNA indicates that this transcript encodes a cationic peroxidase isozyme that is far removed from Ep in peroxidase phylogeny. To determine the expression patterns for these two peroxidases in developing seeds, the abundance and localization of the Ep and Prx2 transcripts were compared by in situ hybridization. Results show the expression of Ep begins in a small number of cells flanking the vascular bundle in the seed coat, spreads to encircle the seed, and then migrates to the hourglass cells as they develop. Expression of Prx2 occurs throughout development in all cell layers of the seed coat, and is also evident in the pericarp and embryo. Nonetheless, the Ep-encoded enzyme accounts for virtually all of the peroxidase activity detected in mature seed coats. The Prx2 enzyme is either insoluble in a catalytically inactive form, or is subject to degradation during seed maturation.
Similar content being viewed by others
References
Buttery, B.R. and Buzzell, R.I. 1968. Peroxidase activity in seeds of soybean varieties. Crop Sci. 8: 722-725.
Buzzell, R.I. and Buttery, B.R. 1969. Inheritance of peroxidase activity in soybean seed coats. Crop Sci. 9: 387-388.
Campa, A. 1991. Biological roles of plant peroxidases: known and potential function. In: J. Everse, K.E. Everse and M.B. Grisham (Eds.), Peroxidases in Chemistry and Biology, vol. 2, CRC Press, Boca Raton, FL, pp. 25-50.
Carlson, J.B. and Lersten, N.R. 1987. Reproductive morphology. In: J.R. Wilcox (Ed.), Soybeans: Improvement, Production, and Uses, 2nd ed., Agronomy Society of America, Madison, WI, pp. 95-134.
Church, G.M. and Gilbert, W. 1994. Genomic sequencing. Proc. Natl. Acad. Sci. USA 81: 1991-1995.
Cox, K.H. and Goldberg, R.B. 1988. Analysis of plant gene expression. In: C.H. Shaw (Ed.), Plant Molecular Biology: A Practical Approach, IRL Press, Oxford, pp. 1-35.
Dowd, P.F. and Lagrimini, M.L. 1997. Examination of different tobacco (Nicotiana spp.) types under-and overproducing tobacco anionic peroxidase for their leaf resistance to Helicoverpa zea. J. Chem. Ecol. 23: 2357-2370.
El-Turk, J., Asemota, O., Leymarie, J., Sallaud, C., Mesnage, S., Breda, C., Buffard, D., Kondorosi, A. and Esnault, R. 1996. Nucleotide sequences of four pathogen-induced alfalfa peroxidase-encoding cDNAs. Gene 170: 213-216.
Fobert, P., Labbé, H., Cosmopoulos, J., Gottlob-McHugh, S., Ouellet, T., Hattori, J., Sunohara, G., Iyer, V.N. and Miki, B.L. 1994. T-DNA tagging of a seed coat-specific promoter in tobacco. Plant J. 6: 567-577.
Gerhartz, W. 1990. Enzymes in Industry. Production and Applications, VCH Verlagsgesellschaft, Weinheim, Germany.
Gijzen, M. 1997. A deletion mutation at the ep locus causes low seed coat peroxidase activity in soybean. Plant J. 12: 991-998.
Gijzen, M., van Huystee, R. and Buzzell, R.I. 1993. Soybean seed coat peroxidase. A comparison of high-activity and low-activity genotypes. Plant Physiol. 103: 1061-1066.
Gillikin, J.W. and Graham, J.S. 1991. Purification and developmental analysis of the major anionic peroxidase from the seed coat of Glycine max. Plant Physiol. 96: 214-220.
Harrison, S.J., Curtis, M.D., McIntyre, C.L., Maclean, D.J. and Manners, J.M. 1995. Differential expression of peroxidase isogenes during the early stages of infection of the tropical forage legume Stylosanthes humilis by Colletotrichum gloeosporioides. Mol. Plant-Microbe Interact. 8: 398-406.
Kjærsgård, I.V.H., Jespersen, H.M., Rasmussen, S.K. and Welinder, K.G. 1997. Sequence and RT-PCR expression analysis of two peroxidases from Arabidopsis thaliana belonging to a novel evolutionary branch of plant peroxidases. Plant Mol. Biol. 33: 699-708.
Klotz, K.L., Liu, T.T., Liu, L. and Lagrimini, L.M. 1998. Expression of tobacco anionic peroxidase is tissue-specific and developmentally regulated. Plant Mol. Biol. 36: 509-520.
Miller, S.S., Bowman, L.A., Gijzen, M. and Miki, B.L.A. 1999. Early development of the seed coat of soybean (Glycine max). Ann. Bot., in press.
Østergaard, L., Abelskov, A.K., Mattsson, O. and Welinder, K.G. 1996. Structure and organ specificity of an anionic peroxidase from Arabidopsis thaliana cell suspension culture. FEBS Lett. 398: 243-247.
Ouellet, T., Rutledge, R.G. and Miki, B.L. 1992. Members of the acetohydroxyacid synthase multigene family of Brassica napus have divergent patterns of expression. Plant J. 2: 321-330.
Peterson, C.M., Mosjidis, C.O.'H., Dute, R.R. and Westgate M.E. 1992. A flower and pod staging system for soybean. Ann. Bot. 69: 59-67.
Ritter, D., Allen, R.D., Trolinder, N., Hughes, W.D. and Galau, G.A. 1993. Cotton cotyledon cDNA encoding a peroxidase. Plant Physiol. 102: 1351.
Roberts, S., Kutchan, T. and Kulattukkudy, P.E. 1988. Cloning and sequencing of cDNA for a highly anionic peroxidase from potato and the induction of its mRNA in suberizing potato tubers and tomato fruits. Plant Mol. Biol. 11: 15-26.
Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Schmitz, N., Gijzen, M. and van Huystee, R. 1997. Characterization of anionic soybean (Glycine max) seed coat peroxidase. Can. J. Bot. 75: 1336-1341.
Wang, C.S. and Vodkin, L.O. 1994. Extraction of RNA from tissues containing high levels of procyanidins that bind RNA. PlantMol. Biol. Rep. 12: 132-145.
Welinder, K.G. 1992. Superfamily of plant, fungal and bacterial peroxidases. Curr. Opin. Struct. Biol. 2: 388-393.
Welinder, K.G., Jespersen, H.M., Kjærsgård, I.V.H., Østergaard, L., Abelskov, A.K., Hanseni, L.N. and Rasmussen, S.K. 1996. What can we learn from Arabidopsis peroxidases? In: C. Obinger, U. Burner, R. Ebermann, C. Penel and H. Greppin (Eds.), Plant Peroxidases: Biochemistry and Physiology, 4th International Symposium, University of Geneva, Switzerland, pp. 173-178.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gijzen, M., Miller, S.S., Bowman, LA. et al. Localization of peroxidase mRNAs in soybean seeds by in situ hybridization. Plant Mol Biol 41, 57–63 (1999). https://doi.org/10.1023/A:1006244500951
Issue Date:
DOI: https://doi.org/10.1023/A:1006244500951