Abstract
We report the identification and biotechnological utility of a plant gene encoding the tocopherol (vitamin E) biosynthetic enzyme 2-methyl-6-phytylbenzoquinol methyltransferase. This gene was identified by map-based cloning of the Arabidopsis mutation vitamin E pathway gene3-1 (vte3-1), which causes increased accumulation of delta-tocopherol and decreased gamma-tocopherol in the seed. Enzyme assays of recombinant protein supported the hypothesis that At-VTE3 encodes a 2-methyl-6-phytylbenzoquinol methyltransferase. Seed-specific expression of At-VTE3 in transgenic soybean reduced seed delta-tocopherol from 20 to 2%. These results confirm that At-VTE3 protein catalyzes the methylation of 2-methyl-6-phytylbenzoquinol in planta and show the utility of this gene in altering soybean tocopherol composition. When At-VTE3 was coexpressed with At-VTE4 (gamma-tocopherol methyltransferase) in soybean, the seed accumulated to >95% alpha-tocopherol, a dramatic change from the normal 10%, resulting in a greater than eightfold increase of alpha-tocopherol and an up to fivefold increase in seed vitamin E activity. These findings demonstrate the utility of a gene identified in Arabidopsis to alter the tocopherol composition of commercial seed oils, a result with both nutritional and food quality implications.
MeSH Terms
Alleles
Amino Acid Sequence
Arabidopsis/chemistry,genetics,metabolism
Arabidopsis Proteins/genetics,metabolism
Cloning, Molecular
DNA, Complementary/chemistry,genetics
Escherichia coli/genetics,metabolism
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Plant
Methyltransferases/genetics,metabolism
Molecular Sequence Data
Mutation
Plants, Genetically Modified
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Soybean Oil/metabolism
Soybeans/enzymology,genetics
Tocopherols/metabolism
Vitamin E/biosynthesis
Chemicals
Arabidopsis Proteins
DNA, Complementary
Vitamin E
Soybean Oil
2-methyl-6-phytylbenzoquinol methyltransferase, Arabidopsis
2-methyl-6-phytylbenzoquinone methyltransferase
Methyltransferases
delta-tocopherol
Tocopherols
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Van Eenennaam Alison L
Monsanto Company, Calgene Campus, Davis, California 95616, USA.
Lincoln Kim
Durrett Timothy P
Valentin Henry E
Shewmaker Christine K
Thorne Greg M
Jiang Jian
Baszis Susan R
Levering Charlene K
Aasen Eric D
Hao Ming
Stein Joshua C
Norris Susan R
Last Robert L
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