Molecular and General Genetics MGG

, Volume 217, Issue 2–3, pp 257–262 | Cite as

A new mutant class of soybean lacks urease in leaves but not in leaf-derived callus or in roots

  • Joseph C. Polacco
  • Adam K. Judd
  • Jody K. Dybing
  • Silvia R. Cianzio


We reported earlier the recovery of two classes of soybean urease mutants in soybean (Glycine max L. Merr. cv. Williams). Class I mutants lack the embryo-specific urease while class II mutants lack the activities of both urease isozymes, the embryo-specific and the ubiquitous urease, the latter found in all tissues examined. We report here the recovery of a true-breeding mutant, aj3, which represents the third phenotypic class: normal levels of embryo-specific urease and little or no ubiquitous urease. Unlike class II mutant plants which lack urease in all tissue, aj3 lacks urease activity only in leaves (ca. 2% normal activity); its roots have near normal urease activity. Callus derived from leaves of aj3 has 14% to 40% the urease activity of Williams 82 callus. This partial reduction in urease activity in aj3 callus is sufficient to reduce growth with urea as sole nitrogen source and to confer resistance to 50 mM urea added to callus maintenance medium. Leaves of aj3 produce more than 40 times the urease antigen expected from their urease activity. The aj3 trait is due to a single recessive lesion which is not allelic with lesions at theEu2, Eu3 (class II) orEu1 (class I) loci. We designate the aj3 genotype aseu4/eu4.

Key words

Soybean mutant Ubiquitous urease Callus Leaf Root 



2,2′-azino-di[3-ethylbenzthiazolinesulfonic acid] diammonium salt


bovine serum albumin


enzyme-linked immunosorbent assay


ethyl methanesulfonate


Murashige and Skoog medium


N-methyl nitrosourea


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  1. Buttery BR, Buzzell RI (1971) Properties and inheritance of urease isozymes in soybean seeds. Can J Bot 49:1101–1105Google Scholar
  2. Hogan ME, Swift IE, Done J (1983) Urease assay and ammonia release from leaf tissue. Phytochemistry 22:663–667CrossRefGoogle Scholar
  3. Holland MA, Griffin JD, Meyer-Bothling LE, Polacco JC (1987) Developmental genetics of the soybean urease isozymes. Dev Genet 8:375–387CrossRefGoogle Scholar
  4. Kaplan A (1969) The determination of urea, ammonium and urease. Methods Biochem Anal 17:311–324PubMedGoogle Scholar
  5. Kloth RH, Hymowitz T (1985) Re-evaluation of the inheritance of urease in soybean seed. Crop Sci 25:352–354CrossRefGoogle Scholar
  6. Kloth RH, Polacco JC, Hymowitz T (1987) The inheritance of a urease null trait in soybean. Theor Appl Genet 73:410–418CrossRefGoogle Scholar
  7. Krueger RW, Holland MA, Chisholm D, Polacco JC (1987) Recovery of a soybean urease genomic clone by sequential library screening with two synthetic oligodeoxynucleotides. Gene 54:41–50PubMedCrossRefGoogle Scholar
  8. Meyer-Bothling LE, Polacco JC (1987) Mutational analysis of the embryo-specific urease locus of soybean. Mol Gen Genet 209:439–444CrossRefPubMedGoogle Scholar
  9. Meyer-Bothling LE, Polacco JC, Cianzio SR (1987) Pleiotropic soybean mutants defective in both urease isozymes. Mol Gen Genet 209:432–438CrossRefPubMedGoogle Scholar
  10. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  11. Polacco JC (1976) Nitrogen metabolism in soybean tissue culture. I. Assimilation of urea. Plant Physiol 58:350–357PubMedGoogle Scholar
  12. Polacco JC, Havir EA (1979) Comparison of soybean urease isolated from seed and tissue culture. J Biol Chem 254:1707–1715PubMedGoogle Scholar
  13. Polacco JC, Winkler RG (1984) Soybean leaf urease: A seed enzyme? Plant Physiol 74:800–803PubMedGoogle Scholar
  14. Polacco JC, Thomas AL, Bledsoe PJ (1982) A soybean seed ureasenull produces urease in cell culture. Plant Physiol 69:1233–1240PubMedGoogle Scholar
  15. Polacco JC, Krueger RW, Winkler RG (1985) Structure and possible ureide degrading function of the ubiquitous urease of soybean. Plant Physiol 79:794–800PubMedGoogle Scholar
  16. Ryan SA, Harper JE (1983) Mutagenesis of soybeans. Soybean Genet Newslett 10:29–32Google Scholar
  17. Sack DA, Huda S, Neogi PKB, Daniel RR, Spira W (1980) Microtiter ganglioside enzyme-linked immunosorbent assay forVibrio andEscherichia coli heat-labile enterotoxins and antitoxin. J Clin Microbiol 11:35–40PubMedGoogle Scholar
  18. Triplett EW, Heitholt JJ, Evensen KB, Blevins DG (1981) Increase in internode length ofPhaseolus lunatus L. caused by inoculation with a nitrate reductase-deficient strain ofRhizobium sp. Plant Physiol 67:1–4PubMedGoogle Scholar
  19. Winkler RG, Polacco JC, Eskew DL, Welch RM (1983) Nickel is not required for apourease synthesis in soybean seeds. Plant Physiol 72:262–263PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Joseph C. Polacco
    • 1
  • Adam K. Judd
    • 1
  • Jody K. Dybing
    • 1
  • Silvia R. Cianzio
    • 2
  1. 1.Biochemistry DepartmentUniversity of MissouriColumbiaUSA
  2. 2.Agronomy DepartmentIowa State UniversityAmesUSA

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