The role of 5′-adenylylsulfate reductase in the sulfur assimilation pathway of soybean: Molecular cloning, kinetic characterization, and gene expression

Soybean seeds are a major source of protein, but contain low levels of sulfur-containing amino acids. With the objective of studying the sulfur assimilation pathway of soybean, a full-length cDNA clone for 5′-adenylylsulfate reductase (APS reductase) was isolated and characterized. The cDNA clone contained an open reading frame of 1414 bp encoding a 52 kDa protein with a N-terminal chloroplast/plastid transit peptide. Southern blot analysis of genomic DNA indicated that the APS reductase in soybean is encoded by a small multigene family. Biochemical characterization of the heterologously expressed and purified protein shows that the clone encoded a functional APS reductase. Although expressed in tissues throughout the plant, these analyses established an abundant expression of the gene and activity of the encoded protein in the early developmental stages of soybean seed, which declined with seed maturity. Sulfur and phosphorus deprivation increased this expression level, while nitrogen starvation repressed APS reductase mRNA transcript and protein levels. Cold-treatment increased expression and the total activity of APS reductase in root tissues. This study provides insight into the sulfur assimilation pathway of this nutritionally important legume.