Abstract
We sought to determine the genetic basis of expression of the ubiquitous (metabolic) urease of soybean. This isozyme is termed the metabolic urease because its loss, in eu4/eu4 mutants, leads to accumulation of urea, whereas loss of the embryo-specific urease isozyme does not. The eu4 lesion eliminated the expression of the ubiquitous urease in vegetative and embryonic tissues. RFLP analysis placed urease clone LC4 near, or within, the Eu4 locus. Sequence comparison of urease proteins (ubiquitous and embryo-specific) and clones (LC4 and LS1) indicated that LC4 and LS1 encode ubiquitous and embryo-specific ureases, respectively. That LC4 is transcribed into poly (A)+ RNA in all tissues was indicated by the amplification of its transcript by an LC4-specific PCR primer. (The LS1-specific primer, on the other hand, amplified poly(A)+ RNA only from developing embryos expressing the embryo-specific urease.) These observations are consistent with Eu4 being the ubiquitous urease structural gene contained in the LC4 clone. In agreement with this notion, the mutant phenotype of eu4/eu4 callus was partially corrected by the LC4 urease gene introduced by particle bombardment.
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References
Buttery BR, Buzzell RI (1971) Properties and inheritance of urease isozymes in soybean seeds. Can J Bot 49:1101–1105
Dellaporta SL, Woods J, Hicks JB (1983) A plant DNA minipreparation: Version II. Plant Mot Biol Rep 1:19–21
Devereaux J, Haeverli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
Eskew DL, Welch RM, Cary EE (1983) Nickel: an essential micronutrient for legumes and possibly all higher plants. Science 222:621–623
Finer JJ, Vain MW, Jones P, McMullen MD (1992) Development of the particle inflow gun for DNA delivery into plant cells. Plant Cell Rep 11:323–328
Griffin JD (1986) Genetic and germplasm studies with several isoenzyme loci in soybean (Glycine max [L] Merr.). PhD thesis, Iowa State University, Ames, USA
Herskovitz I (1987) Functional inactivation of genes by dominant negative mutations. Nature 219–222
Holland MA, Polacco JC (1992) Urease-null and hydrogenase-null phenotypes of a phylloplane bacterium reveal altered nickel metabolism in two soybean mutants. Plant Physiol 98:942–948
Holland MA, Griffin JD, Meyer-Bothling LE, Polacco JC (1987) Developmental genetics of the soybean urease isozymes. Dev Genet 8:375–387
Kerr PS, Blevins DG, Rapp B, Randall DD (1983) Soybean leaf urease: A comparison with seed urease. Physiol Plant 57:339–345
Klein TM, Wolf ED, Wu R, Sanford JC (1987) High-velocity microprojectiles for delivering nucleic acids into living cells. Nature 327:70–73
Kloth RH, Hymowitz T (1985) Re-evaluation of the inheritance of urease in soybean seed. Crop Sci 25:352–354
Krogmeier MJ, McCarty GW, Bremner JM (1989) Phytotoxicity of foliar-applied urea. Proc Natl Acad Sci USA 86:8189–8191
Krueger RW, Holland MA, Chisholm D, Polacco JC (1987) Recovery of a soybean urease genomic clone by sequential library screening with two synthetic oligonucleotides. Gene 54:41–50
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lütcke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA (1987) Selection of AUG initiation codons differs in plants and animals. EMBO J 6:43–48
Matsudaira P (1987) Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem 262:10035–10038
Mettler IJ (1987) A simple and rapid method for minipreparation of DNA from tissue cultured plant cells. Plant Mot Biol Rep 5:346–349
Meyer-Bothling LE, Polacco JC (1987) Mutational analysis of the embryo-specific urease locus of soybean. Mot Gen Genet 209:439–444
Meyer-Bothling LE, Polacco JC, Cianzio SR (1987) Pleiotropic soybean mutants defective in both urease isozymes. Mot Gen Genet 209:432–438
Polacco JC (1976) Nitrogen metabolism in soybean tissue culture. I. Assimilation of urea. Plant Physiol 58:350–357
Polacco JC, Havir EA (1979) Comparisons of soybean urease isolated from seed and tissue culture. J Biol Chem 254:1707–1715
Polacco JC, Holland MA (1993) Roles of ureases in plant cells. In: Jeon KW, Friedlander M (eds) International Review of Cytology, vol 145. Academic Press, San Diego, pp 65–103
Polacco JC, Sparks RB Jr (1982) Patterns of urease synthesis in developing soybeans. Plant Physiol 83:585–591
Polacco JC, Winkler RG (1984) Soybean leaf urease: a seed enzyme? Plant Physiol 74:800–803
Polacco JC, Thomas AL, Bledsoe PJ (1982) A soybean seed ureasenull produces urease in cell culture. Plant Physiol 69:1233–1240
Polacco JC, Krueger RW, Winkler RG (1985) Structure and possible ureide degrading function of the ubiquitous urease of soybean. Plant Physiol 79:794–800
Polacco JC, Judd AK, Dybing JK, Cianzio SR (1989) A new mutant class of soybean lacks urease in leaves but not in leafderived callus or in roots. Mol Gen Genet 217:257–262
Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81:8014–8019
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Stebbins NE, Holland MA, Cianzio SR, Polacco JC (1991) Genetic tests of the roles of the embryonic ureases of soybean. Plant Physiol 97:1004–1010
Takishima K, Suga T, Mamiya G (1988) The structure of jackbean urease. The complete amino acid sequence, limited proteolysis and reactive cysteine residues. Eur J Biochem 175:151–165
Torisky RS (1992) Evidence that a single gene encodes the metabolic urease of soybean. PhD thesis, University of Missouri, Columbia, USA
Torisky RS, Polacco JC (1990) Soybean roots retain the seed urease isozyme synthesized during embryo development. Plant Physiol 94:681–689
Towbin H, Staehelin T, Gordon G (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and applications. Proc Natl Acad Sci USA 76:4350–4354
Wadsworth GJ, Redinbaugh MG, Scandalios JG (1988) A procedure for the small-scale isolation of plant RNA suitable for RNA blot analysis. Anal Biochem 172:279–283
Yenofsky RL, Fine M, Liu C (1988) Isolation and characterization of a soybean (Glycine max) lipoxygenase-3 gene. Mol Gen Genet 211:215–222
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Communicated by E. Meyerowitz
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Torisky, R.S., Griffin, J.D., Yenofsky, R.L. et al. A single gene (Eu4) encodes the tissue-ubiquitous urease of soybean. Molec. Gen. Genet. 242, 404–414 (1994). https://doi.org/10.1007/BF00281790
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DOI: https://doi.org/10.1007/BF00281790