Overexpression of ATP sulfurylase improves the sulfur amino acid content, enhances the accumulation of Bowman-Birk protease inhibitor and suppresses the accumulation of the β-subunit of β-conglycinin in soybean seeds
| Autor: | Won-Seok Kim, Nathan W Oehrle, Hari B. Krishnan, Joseph M. Jez, Jeong Sun-Hyung |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine Transgene Sulfur metabolism lcsh:Medicine 01 natural sciences Article 03 medical and health sciences chemistry.chemical_compound Gene Expression Regulation Plant Plastid Amino Acids lcsh:Science Trypsin Inhibitor Bowman-Birk Soybean Gel electrophoresis chemistry.chemical_classification Multidisciplinary Methionine lcsh:R Seed Storage Proteins food and beverages Globulins Antigens Plant Plants Genetically Modified Sulfate Adenylyltransferase Amino Acids Sulfur 030104 developmental biology Enzyme Biochemistry chemistry Seeds Soybean Proteins lcsh:Q Soybeans Cell fractionation Plant sciences Biotechnology 010606 plant biology & botany Cysteine |
| Zdroj: | Scientific Reports, Vol 10, Iss 1, Pp 1-13 (2020) Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | ATP sulfurylase, an enzyme which catalyzes the conversion of sulfate to adenosine 5′-phosphosulfate (APS), plays a significant role in controlling sulfur metabolism in plants. In this study, we have expressed soybean plastid ATP sulfurylase isoform 1 in transgenic soybean without its transit peptide under the control of the 35S CaMV promoter. Subcellular fractionation and immunoblot analysis revealed that ATP sulfurylase isoform 1 was predominantly expressed in the cell cytoplasm. Compared with that of untransformed plants, the ATP sulfurylase activity was about 2.5-fold higher in developing seeds. High-resolution 2-D gel electrophoresis and immunoblot analyses revealed that transgenic soybean seeds overexpressing ATP sulfurylase accumulated very low levels of the β-subunit of β-conglycinin. In contrast, the accumulation of the cysteine-rich Bowman–Birk protease inhibitor was several fold higher in transgenic soybean plants when compared to the non-transgenic wild-type seeds. The overall protein content of the transgenic seeds was lowered by about 3% when compared to the wild-type seeds. Metabolite profiling by LC–MS and GC–MS quantified 124 seed metabolites out of which 84 were present in higher amounts and 40 were present in lower amounts in ATP sulfurylase overexpressing seeds compared to the wild-type seeds. Sulfate, cysteine, and some sulfur-containing secondary metabolites accumulated in higher amounts in ATP sulfurylase transgenic seeds. Additionally, ATP sulfurylase overexpressing seeds contained significantly higher amounts of phospholipids, lysophospholipids, diacylglycerols, sterols, and sulfolipids. Importantly, over expression of ATP sulfurylase resulted in 37–52% and 15–19% increases in the protein-bound cysteine and methionine content of transgenic seeds, respectively. Our results demonstrate that manipulating the expression levels of key sulfur assimilatory enzymes could be exploited to improve the nutritive value of soybean seeds. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|