Metabolite Profiling of Soybean Seed Extracts from Near-Isogenic Low and Normal Phytate Lines Using Orthogonal Separation Strategies.
Untargeted metabolomic profiling using liquid chromatography-mass spectrometry (LC-MS) was applied to lipid-depleted methanolic extracts of soybean seeds utilizing orthogonal chromatographic separations (reversed-phase and hydrophilic interaction) in both positive and negative ionization modes. Four near-isogenic lines (NILs) differing in mutations for two genes encoding highly homologous multidrug resistant proteins (MRPs) were evaluated. The double mutant exhibited a low phytate phenotype, whereas the other three NILs, the two single mutants and the wild type, did not. Principal component analysis (PCA) of the four LC-MS data sets fully separated the low phytate line from the other three. While the levels of neutral oligosaccharides were the same for all lines, there were significant metabolite differences residing in the levels of malonyl isoflavones, soyasaponins, and arginine. Two methanol-soluble polypeptides were also found as differing in abundance levels, one of which was identified as the allergen Gly m 1.
Departments of Biochemistry, ‡Crop and Soil Environmental Sciences, §Biological Sciences, and Plant Pathology, and ∥Physiology and Weed Science, Virginia Tech , Blacksburg, Virginia 24061, United States.
Judith
J
Jervis
Departments of Biochemistry, ‡Crop and Soil Environmental Sciences, §Biological Sciences, and Plant Pathology, and ∥Physiology and Weed Science, Virginia Tech , Blacksburg, Virginia 24061, United States.
Christin
C
Kastl
Departments of Biochemistry, ‡Crop and Soil Environmental Sciences, §Biological Sciences, and Plant Pathology, and ∥Physiology and Weed Science, Virginia Tech , Blacksburg, Virginia 24061, United States.
Sherry B
SB
Hildreth
Departments of Biochemistry, ‡Crop and Soil Environmental Sciences, §Biological Sciences, and Plant Pathology, and ∥Physiology and Weed Science, Virginia Tech , Blacksburg, Virginia 24061, United States.
Ruslan
R
Biyashev
Departments of Biochemistry, ‡Crop and Soil Environmental Sciences, §Biological Sciences, and Plant Pathology, and ∥Physiology and Weed Science, Virginia Tech , Blacksburg, Virginia 24061, United States.
Elizabeth A
EA
Grabau
Departments of Biochemistry, ‡Crop and Soil Environmental Sciences, §Biological Sciences, and Plant Pathology, and ∥Physiology and Weed Science, Virginia Tech , Blacksburg, Virginia 24061, United States.
Mohammad A
MA
Saghai-Maroof
Departments of Biochemistry, ‡Crop and Soil Environmental Sciences, §Biological Sciences, and Plant Pathology, and ∥Physiology and Weed Science, Virginia Tech , Blacksburg, Virginia 24061, United States.
Richard F
RF
Helm
D002853:Chromatography, Liquid; D007529:Isoflavones / Q000737:chemistry / Q000378:metabolism; D055432:Metabolomics; D009154:Mutation; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D010833:Phytic Acid / Q000737:chemistry* / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D012503:Saponins / Q000737:chemistry / Q000378:metabolism; D012639:Seeds / Q000737:chemistry / Q000378:metabolism; D013025:Glycine max / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry
D007529:Isoflavones; D009844:Oligosaccharides; D010936:Plant Extracts; D010940:Plant Proteins; D012503:Saponins; D010833:Phytic Acid
Glycine max; hydrophobic seed protein; metabolomics; oligosaccharides; phytate; seed; soyasaponin