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. 1991 Nov;97(3):1004–1010. doi: 10.1104/pp.97.3.1004

Genetic Tests of the Roles of the Embryonic Ureases of Soybean 1

Norm Stebbins 1,2, Mark A Holland 1,2, Silvia R Cianzio 1,2, Joseph C Polacco 1,2
PMCID: PMC1081116  PMID: 16668483

Abstract

We assayed the in vivo activity of the ureases of soybean (Glycine max) embryos by genetically eliminating the abundant embryo-specific urease, the ubiquitous urease, or a background urease. Mutant embryos accumulated urea (250-fold over progenitor) only when lacking all three ureases and only when developed on plants lacking the ubiquitous urease. Thus, embryo urea is generated in maternal tissue where its accumulation is not mitigated by the background urease. However, the background urease can hydrolyze virtually all urea delivered to the developing embryo. Radicles of 2-day-old germinants accumulated urea in the presence or absence of the embryo-specific urease (2 micromoles per gram dry weight radicle). However, mutants lacking the ubiquitous urease exhibited increased accumulation of urea (to 4-5 micromoles urea per gram dry weight radicle). Thus, the ubiquitous and not the embryo-specific urease hydrolyzes urea generated during germination. In the absence of both of these ureases, the background urease activity (4% of ubiquitous urease) may hydrolyze most of the urea generated. A pleiotropic mutant lacking all urease accumulated 34 micromoles urea per gram dry weight radicle (increasing 2.5-fold at 3 days after germination). Urea (20 millimolar) was toxic to in vitro-cultured cotyledons which contained active embryo-specific urease. Cotyledons lacking the embryo-specific urease accumulated more protein when grown with urea than with no nitrogen source. Among cotyledons lacking the embryo-specific urease, fresh weight increases were virtually unchanged whether grown on urea or on no nitrogen and whether in the presence or absence of the ubiquitous urease. However, elimination of the ubiquitous urease reduced protein deposition on urea-N, and elimination of both the ubiquitous and background ureases further reduced urea-derived protein. The evidence is consistent with the lack of a role in urea hydrolysis for the embryo-specific urease in developing embryos or germinating seeds. Because the embryo-specific urease is deleterious to cotyledons cultured in vitro on urea-N, its role may be to hydrolyze urea in wounded or infected embryos, creating a hostile environment for pest or pathogen. While the ubiquitous urease is operative in leaves and in seedlings, all or most of its function can be assumed by the background urease in embryos and in seedlings.

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Selected References

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  1. Dizengremel P., Chauveau M., Roussaux J. Inhibition by adenine derivatives of the cyanide-insensitive electron transport pathway of plant mitochondria. Plant Physiol. 1982 Aug;70(2):585–589. doi: 10.1104/pp.70.2.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Eskew D. L., Welch R. M., Cary E. E. Nickel: an essential micronutrient for legumes and possibly all higher plants. Science. 1983 Nov 11;222(4624):621–623. doi: 10.1126/science.222.4624.621. [DOI] [PubMed] [Google Scholar]
  3. Krueger R. W., Holland M. A., Chisholm D., Polacco J. C. Recovery of a soybean urease genomic clone by sequential library screening with two synthetic oligodeoxynucleotides. Gene. 1987;54(1):41–50. doi: 10.1016/0378-1119(87)90345-3. [DOI] [PubMed] [Google Scholar]
  4. Meyer-Bothling L E, Polacco J C, Cianzio S R. Pleiotropic soybean mutants defective in both urease isozymes. Mol Gen Genet. 1987 Oct;209(3):432–438. doi: 10.1007/BF00331146. [DOI] [PubMed] [Google Scholar]
  5. Meyer-Bothling L E, Polacco J C. Mutational analysis of the embryo-specific urease locus of soybean. Mol Gen Genet. 1987 Oct;209(3):439–444. doi: 10.1007/BF00331147. [DOI] [PubMed] [Google Scholar]
  6. Micallef B. J., Shelp B. J. Arginine metabolism in developing soybean cotyledons : I. Relationship to nitrogen nutrition. Plant Physiol. 1989 Jun;90(2):624–630. doi: 10.1104/pp.90.2.624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Polacco J. C., Havir E. A. Comparisons of soybean urease isolated from seed and tissue culture. J Biol Chem. 1979 Mar 10;254(5):1707–1715. [PubMed] [Google Scholar]
  8. Polacco J. C., Krueger R. W., Winkler R. G. Structure and possible ureide degrading function of the ubiquitous urease of soybean. Plant Physiol. 1985 Nov;79(3):794–800. doi: 10.1104/pp.79.3.794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Polacco J. C., Thomas A. L., Bledsoe P. J. A soybean seed urease-null produces urease in cell culture. Plant Physiol. 1982 May;69(5):1233–1240. doi: 10.1104/pp.69.5.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Polacco J. C., Winkler R. G. Soybean leaf urease: a seed enzyme? Plant Physiol. 1984 Apr;74(4):800–803. doi: 10.1104/pp.74.4.800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Torisky R. S., Polacco J. C. Soybean Roots Retain the Seed Urease Isozyme Synthesized during Embryo Development. Plant Physiol. 1990 Oct;94(2):681–689. doi: 10.1104/pp.94.2.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Winkler R. G., Polacco J. C., Eskew D. L., Welch R. M. Nickel is not required for apourease synthesis in soybean seeds. Plant Physiol. 1983 May;72(1):262–263. doi: 10.1104/pp.72.1.262. [DOI] [PMC free article] [PubMed] [Google Scholar]

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