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Three non-allelic epistatically interacting methyltransferase mutations produce novel tocopherol (vitamin E) profiles in sunflower

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Abstract

Wildtype sunflower (Helianthus annuus L.) seeds are a rich source of α-tocopherol (vitamin E). The g = Tph 2 mutation disrupts the synthesis of α-tocopherol, enhances the synthesis of γ-tocopherol, and was predicted to knock out a γ-tocopherol methyltransferase (γ-TMT) necessary for the synthesis of α-tocopherol in sunflower seeds—wildtype (g + g +) lines accumulated > 90% α-tocopherol, whereas mutant (g g) lines accumulated > 90% γ-tocopherol. We identified and isolated two γ-TMT paralogs (γ-TMT-1 and γ-TMT-2). Both mapped to linkage group 8, cosegregated with the g locus, and were transcribed in developing seeds of wildtype lines. The g mutation greatly decreased γ-TMT-1 transcription, caused alternative splicing of γ-TMT-1, disrupted γ-TMT-2 transcription, and knocked out one of two transcription initiation sites identified in the wildtype; γ-TMT transcription was 36 to 51-fold greater in developing seeds of wildtype (g + g +) than mutant (g g) lines. F2 populations (B109 × LG24 and R112 × LG24) developed for mapping the g locus segregated for a previously unidentified locus (d). B109, R112, and LG24 were homozygous for a null mutation (m = Tph 1) in MT-1, one of two 2-methyl-6-phytyl-1,4-benzoquinone/2-methyl-6-solanyl-1,4-benzoquinone methyltransferase (MPBQ/MSBQ-MT) paralogs identified in sunflower. The d mutations segregating in B109 × LG24 and R112 × LG24 were allelic to a cryptic mutation identified in the other MPBQ/MSBQ-MT paralog (MT-2) and disrupted the synthesis of α- and γ-tocopherol in F2 progeny carrying m or g mutations—m m g + g + d d homozygotes accumulated 41.5% α- and 58.5% β-T, whereas m m g g d d homozygotes accumulated 58.1% γ- and 41.9% δ-T. MT-2 cosegregated with d and mapped to linkage group 4. Hence, novel tocopherol profiles are produced in sunflower seed oil by three non-allelic epistatically interacting methyltransferase mutations.

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Acknowledgements

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 the University of Georgia Research Foundation.

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Authors and Affiliations

  1. Department of Crop and Soil Science, Oregon State University, Corvallis, OR, 97331, USA

    Catherine G. Hass

  2. Center for Applied Genetic Technologies, The University of Georgia, 111 Riverbend Road, Athens, GA, 30602, USA

    Shunxue Tang & Steven J. Knapp

  3. Linus Pauling Institute, Oregon State University, Corvallis, OR, 97331, USA

    Scott Leonard & Maret G. Traber

  4. USDA-ARS, Northern Crop Science Laboratory, Fargo, ND, 58105, USA

    Jerry F. Miller

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  1. Catherine G. Hass
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  2. Shunxue Tang
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  4. Maret G. Traber
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  5. Jerry F. Miller
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  6. Steven J. Knapp
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Correspondence to Steven J. Knapp.

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Communicated by C. Gebhardt

Catherine G. Hass and Shunxue Tang contributed equally to this work.

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Hass, C.G., Tang, S., Leonard, S. et al. Three non-allelic epistatically interacting methyltransferase mutations produce novel tocopherol (vitamin E) profiles in sunflower. Theor Appl Genet 113, 767–782 (2006). https://doi.org/10.1007/s00122-006-0320-4

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