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Characterization of a γ-tocopherol methyltransferase mutant gene in wild (Carthamus oxyacanthus M. Bieb.) and cultivated safflower (C. tinctorius L.)

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Abstract

Safflower (Carthamus tinctorius L.) seeds contain a high proportion of tocopherols (>90 %) in the α-tocopherol form. A mutant with a high concentration of γ-tocopherol (>85 %) was identified in germplasm of wild safflower (Carthamus oxyacanthus M. Bieb.) that showed strong introgression of C. tinctorius, which allowed selection of individuals of both species with high concentrations of either α- or γ-tocopherol. The trait is controlled by a γ-tocopherol methyltransferase (γ-TMT) locus. The objective of this research was to identify γ-TMT sequence mutations associated with the high γ-tocopherol trait. Full length genomic and cDNA sequences of the γ-TMT gene were obtained from plants of C. tinctorius and C. oxyacanthus with both tocopherol profiles. Sequences from high γ-tocopherol plants showed an 11 bp deletion in exon 6 of the γ-TMT gene that disrupted the reading frame and created a premature stop codon, resulting in a predicted protein with a drastically altered amino acid sequence downstream the frameshift site. The data suggested that the frameshift mutation was underlying the γ-TMT loss of function mutant allele that determines the high γ-tocopherol phenotype. The characterized sequence change of 11 bp deletion could be used directly as a functional marker for introgression of the high γ-tocopherol trait into elite safflower cultivars.

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Acknowledgments

The research was funded by the Spanish Ministry of Science and Innovation and the European Union FEDER funds (research project AGL2007-62834). The authors are grateful to Dr. María J. Giménez and Dr. Fernando Piston (Instituto de Agricultura Sostenible, CSIC) for their advice on experimental procedures.

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Authors and Affiliations

  1. Instituto de Agricultura Sostenible (IAS-CSIC), Alameda del Obispo s/n, 14080, Córdoba, Spain

    María J. García-Moreno, José M. Fernández-Martínez, Leonardo Velasco & Begoña Pérez-Vich

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  1. María J. García-Moreno
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  2. José M. Fernández-Martínez
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  3. Leonardo Velasco
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  4. Begoña Pérez-Vich
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Corresponding author

Correspondence to Leonardo Velasco.

Electronic supplementary material

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10681_2014_1149_MOESM1_ESM.pdf

Fig. S1. γ-tocopherol methyltransferase genomic DNA sequence alignment for high α-tocopherol Carthamus tinctorius lines CL-1 and Rancho, high α-tocopherol C. tinctorius line IASC-1, high α-tocopherol C. oxyacanthus selections PI426472-A5 and PI426472-A12, and high γ-tocopherol C. oxyacanthus selection PI426472-A10. The cDNA sequence of the safflower line CL-1 (CL-1_cDNA) is also included. (PDF 160 kb)

10681_2014_1149_MOESM2_ESM.pdf

Fig. S2. γ-tocopherol methyltransferase cDNA sequence alignment for Carthamus tinctorius lines CL-1 and Rancho, with high α-tocopherol content, and IASC-1, with high γ-tocopherol content. (PDF 35 kb)

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García-Moreno, M.J., Fernández-Martínez, J.M., Velasco, L. et al. Characterization of a γ-tocopherol methyltransferase mutant gene in wild (Carthamus oxyacanthus M. Bieb.) and cultivated safflower (C. tinctorius L.). Euphytica 200, 231–238 (2014). https://doi.org/10.1007/s10681-014-1149-6

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  • DOI: https://doi.org/10.1007/s10681-014-1149-6

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