Abstract
The m (Tph 1) mutation partially disrupts the synthesis of α-tocopherol (vitamin E) in sunflower (Helianthus annuus L.) seeds and was predicted to disrupt a methyltransferase activity necessary for the synthesis of α- and γ-tocopherol. We identified and isolated two 2-methyl-6-phytyl-1,4-benzoquinone/2-methyl-6-solanyl-1,4-benzoquinone methyltransferase (MPBQ/MSBQ-MT) paralogs from sunflower (MT-1 and MT-2), resequenced MT-1 and MT-2 alleles from wildtype (m + m +) and mutant (m m) inbred lines, identified m as a non-lethal knockout mutation of MT-1 caused by the insertion of a 5.2 kb Ty3/gypsy-like retrotransposon in exon 1, and uncovered a cryptic codominant mutation (d) in a wildtype × mutant F2 population predicted to be segregating for the m mutation only. MT-1 and m cosegregated and mapped to linkage group 1 and MT-1 was not transcribed in mutant homozygotes (m m). The m locus was epistatic to the d locus—the d locus had no effect in m + m + and m + m individuals, but significantly increased β-tocopherol percentages in m m individuals. MT-2 and d cosegregated, MT-2 alleles isolated from mutant homozygotes (d d) carried a 30 bp insertion at the start of the 5′-UTR, and MT-2 was more strongly transcribed in seeds and leaves of wildtype (d + d +) than mutant (d d) homozygotes (transcripts were 2.2- to 5.0-fold more abundant in the former than the latter). The double mutant (m m d d) was non-lethal and produced 24–45% α- and 55–74% β-tocopherol (the wildtype produced 96% α- and 4% β-tocopherol). MT-2 compensated for the loss of the MT-1 function, and the MT-2 mutation profoundly affected the synthesis of tocopherols without adversely affecting the synthesis of plastoquinone crucial for normal plant growth and development.
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Acknowledgments
This work was supported by funding to S.J.K. from the National Research Initiative of the United States Department of Agriculture Cooperative State Research, Education, and Extension Service Plant Genome Program (Grant No. 2003-35300-15184), the Paul C. Berger Endowment at Oregon State University, the Georgia Research Alliance, and University of Georgia Research Foundation.
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Communicated by C. Gebhardt
Shunxue Tang and Catherine G. Hass contributed equally to this work.
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Tang, S., Hass, C.G. & Knapp, S.J. Ty3/gypsy-like retrotransposon knockout of a 2-methyl-6-phytyl-1,4-benzoquinone methyltransferase is non-lethal, uncovers a cryptic paralogous mutation, and produces novel tocopherol (vitamin E) profiles in sunflower. Theor Appl Genet 113, 783–799 (2006). https://doi.org/10.1007/s00122-006-0321-3
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DOI: https://doi.org/10.1007/s00122-006-0321-3