by polymerase chain reaction with primers designed to con- .... Additionally, none of the con- ...... Brunswick Scientific, Edison, NJ, U.S.A.) in 50 ml of LB con-.
MPMI Vol. 13, No. 10, 2000, pp. 1053–1070. Publication no. M-2000-0814-01R. © 2000 The American Phytopathological Society
Differential Expression of Two Soybean Apyrases, One of Which Is an Early Nodulin R. Bradley Day,1 Crystal B. McAlvin,1 John T. Loh,1 Roxanne L. Denny,2 Todd C. Wood,4 Nevin D. Young,2,3 and Gary Stacey1 1
Center for Legume Research, Department of Microbiology, University of Tennessee, Knoxville 37996, U.S.A.; 2Department of Plant Pathology and 3Department of Plant Biology, University of Minnesota, St. Paul 55108, U.S.A.; 4Clemson University Genomics Institute, Clemson University, Clemson, SC, 29634, U.S.A. Accepted 13 June 2000. Two cDNA clones were isolated from soybean (Glycine soja) by polymerase chain reaction with primers designed to conserved motifs found in apyrases (nucleotide phosphohydrolase). The two cDNAs are predicted to encode for two, distinct, apyrase proteins of approximately 50 kDa (i.e., GS50) and 52 kDa (i.e., GS52). Phylogenetic analysis indicated that GS52 is orthologous to a family of apyrases recently suggested to play a role in legume nodulation. GS50 is paralogous to this family and, therefore, likely plays a different physiological role. Consistent with this analysis, GS50 mRNA was detected in root, hypocotyls, flowers, and stems, while GS52 mRNA was found in root and flowers. Neither gene was expressed in leaves or cotyledons. Inoculation of roots with Bradyrhizobium japonicum, nitrogen-fixing symbiont of soybean, resulted in the rapid (
