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Increased α-tocotrienol content in seeds of transgenic rice overexpressing Arabidopsis γ-tocopherol methyltransferase

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Abstract

Vitamin E comprises a group of eight lipid soluble antioxidant compounds that are an essential part of the human diet. The α-isomers of both tocopherol and tocotrienol are generally considered to have the highest antioxidant activities. γ-tocopherol methyltransferase (γ-TMT) catalyzes the final step in vitamin E biosynthesis, the methylation of γ- and δ-isomers to α- and β-isomers. In present study, the Arabidopsis γ-TMT (AtTMT) cDNA was overexpressed constitutively or in the endosperm of the elite japonica rice cultivar Wuyujing 3 (WY3) by Agrobacterium-mediated transformation. HPLC analysis showed that, in brown rice of the wild type or transgenic controls with empty vector, the α-/γ-tocotrienol ratio was only 0.7, much lower than that for tocopherol (~19.0). In transgenic rice overexpressing AtTMT driven by the constitutive Ubi promoter, most of the γ-isomers were converted to α-isomers, especially the γ- and δ-tocotrienol levels were dramatically decreased. As a result, the α-tocotrienol content was greatly increased in the transgenic seeds. Similarly, over-expression of AtTMT in the endosperm also resulted in an increase in the α-tocotrienol content. The results showed that the α-/γ-tocopherol ratio also increased in the transgenic seeds, but there was no significant effect on α-tocopherol level, which may reflect the fact that γ-tocopherol is present in very small amounts in wild type rice seeds. AtTMT overexpression had no effect on the absolute total content of either tocopherols or tocotrienols. Taken together, these results are the first demonstration that the overexpression of a foreign γ-TMT significantly shift the tocotrienol synthesis in rice, which is one of the world’s most important food crops.

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Acknowledgments

This study was supported by the National Key Basic Research Projects (2012CB944803), the National Special Program for Transgenic Research (2011ZX08001-006), and the Funds for Distinguished Young Scientists and Priority Academic Program Development from Jiangsu Government, China.

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Authors and Affiliations

  1. Key Laboratory of Crop Genetics and Physiology of Jiangsu Province, Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou, 225009, China

    Gui-Yun Zhang, Ru-Ru Liu, Geng Xu, Peng Zhang, Guo-Hua Liang & Qiao-Quan Liu

  2. School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China

    Yin Li & Ke-Xuan Tang

  3. National Center for Plant Gene Research (Shanghai), Shanghai, 200032, China

    Guo-Hua Liang & Qiao-Quan Liu

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  1. Gui-Yun Zhang
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Correspondence to Qiao-Quan Liu.

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11248_2012_9630_MOESM1_ESM.tif

Supplemental Figure 1. Vitamin E biosynthetic pathway in plants. HPPD, p-hydroxyphenyl-pyruvate dioxygenase; HPT, homogentisate phytyltransferase; TC, tocopherol cyclase; TMT, γ-tocopherol methyltransferase; HGGT, homogentisate geranylgeranyl transferase; MPBQ MT2, methylphytylbenzoquinone methyltransferase. (TIFF 240 kb)

11248_2012_9630_MOESM2_ESM.docx

Supplemental Table 1. Tocochromanol composition and content in AtTMT transgenic brown rice. Each value is the average ± SD of three independent experiments. WT, wild-type WY3; Lines C1 and C2 are transgenic control lines derived from empty vectors pUbi and pGt1, respectively; Lines 1-7 are pUbi-TMT-derived transgenic lines, and lines 8-13 are pGt1-TMT-derived transgenic lines. (DOCX 21 kb)

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Zhang, GY., Liu, RR., Xu, G. et al. Increased α-tocotrienol content in seeds of transgenic rice overexpressing Arabidopsis γ-tocopherol methyltransferase. Transgenic Res 22, 89–99 (2013). https://doi.org/10.1007/s11248-012-9630-2

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