Abstract
A cDNA encoding γ-tocopherol methyltransferase from Brassica napus (BnTMT) was overexpressed in soybean [Glycine max (L.) Merr.] under the control of seed-specific promoter of Arabidopsis fatty acid elongase 1 (FAE1) or soybean glycinin G1. Two and three transgenic plants were selected, respectively, after Agrobacterium-mediated transformation. Polymerase chain reaction (PCR) and Southern blots confirmed that BnTMT was single-copy integrated into the genome of transgenic plants. RT-PCR analysis showed that the expression of BnTMT was higher in the immature cotyledons than in the mature cotyledons, while no expression was detected in the leaves. Moreover, the expression level under the control of FAE1 was higher than that of G1. HPLC analysis indicated that the seed-specific expression of BnTMT resulted in 11.1-fold and 18.9-fold increase in α- and β-tocopherol content, respectively, in T2 seed. These results suggested that introducing BnTMT into soybean can be used to increase the vitamin E composition in seeds.
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Abbreviations
- BnTMT:
-
γ-tocopherol methyltransferase gene from Brassica napus
- FAE1:
-
fatty acid elongase 1
- PCR:
-
polymerase chain reaction
- RT-PCR:
-
real time PCR
References
Cho, E.A., Lee, C.A., Kim, Y.S., Back, S.H., de los Reyes, B.G., Yun, S.J.: Expression of γ-tocopherol methyltransferase transgene improves tocopherol composition in lettuce (Latuca sativa L.). — Mol. Cells 19: 16–22, 2005.
Demurin, Y., Skoric, D., Karlovic, D.: Genetic variability of tocopherol composition in sunflower seeds as a basis of breeding for improved oil quality. — Plant Breed. 115: 33–36, 1996.
Ding, S.H., Huang, L.Y., Wang, Y.D., Sun, H.C., Xiang, Z.H.: High-level expression of basic fibroblast growth factor in transgenic soybean seeds and characterization of its biological activity. — Biotechnol Lett. 28: 869–875, 2006.
Grusak, M.A., Della Penna, D.: Improving the nutrient composition of plants to enhance human nutrition and health. — Annu. Rev. Plant Physiol. Plant mol. Biol. 50: 133–161, 1999.
Hofius, D., Sonnewald, U.: Vitamin E biosynthesis: biochemistry meets cell biology. — Trends Plant Sci. 8: 6–8, 2003.
Koch, M., Lemke, R., Heise, K.P., Mock, H.P.: Characterization of gamma-tocopherol methyltransferases from Capsicum annuum L and Arabidopsis thaliana. — Eur. J. Biochem. 270: 84–92, 2003.
Lee, B.K., Kim, S.L., Kim, K.H., Yu, S.H., Lee, S.C., Zhang, Z.Y., Kim, M.S., Park, H.M., Lee, J.Y.: Seed specific expression of perilla γ-tocopherol methyltransferase gene increases α-tocopherol content in transgenic perilla (Perilla frutescens). — Plant Cell Tissue Organ Cult. 92: 47–54, 2008.
Li, Y., Wang, G., Hou, R., Zhou, Y., Gong, R., Sun, X., Tang, K.: Engineering tocopherol biosynthesis pathway in lettuce. — Biol. Plant. 55: 453–460, 2011.
Liu, H.K., Yang, C., Wei, Z.M.: Efficient Agrobacterium tumefaciens— mediated transformation of soybeans using an embryonic tip regeneration system. — Planta 219: 1042–1049, 2004.
Maren, R., Mark, S., Ljerka, K.: Expression of the FAE1 gene and FAE1 promoter activity in developing seeds of Arabidopsis thaliana. — Plant mol. Biol. 46: 717–725, 2001.
Qian, W.C., Chen, D.F., Wang, H.Z., Wang, Y.Q., Chen, X.W.: [Gene cloning and function confirmation of γ-tocopherol methyltransferase from Brassica napus L.] — Plant Physiol. Commun. 43: 68–688, 2007. [In Chin.]
Sambrook, J., Russel, D.W. (ed.): Molecular Cloning: a Laboratory Manual. 3rd Ed. — Cold Spring Harbor Laboratory Press, Cold Spring Harbor 2001.
Savidge, B., Weiss, J.D., Wong, Y.H., Lassner, M.W., Mitsky, T.A., Shewmaker, C.K., Post-Beittenmiller, D., Valentin, H.E.: Isolation and characterization of homogentisate phytyltransferase genes from Synechocystis sp. PCC 6803 and Arabidopsis. — Plant Physiol. 129: 321–332, 2002.
Shinatani, D., Della Penna, D.: Elevating the vitamin E content of plants through metabolic engineering. — Science 282: 2098–2100, 1998.
Seong, E.S., Ghimire, B.K., Goh, E.J., Lim, J.D., Kim, M.J., Chung, I.M., Yu, C.Y.: Overexpression of the γ-TMT gene in Codonopsis lanceolata. — Biol. Plant. 53: 631–636, 2009.
Tan, B.: Palm carotenoids, tocopherols and tocotrienols. — J.amer. oil. chem. Soc. 66: 770–776, 1989.
Tavva, V.S., Kim, Y.H., Kagan, I.A., Dinkins, R.D., Kim, K.H., Collins, G.B.: Increased α-tocopherol content in soybean seed overexpressing the Perilla frutescens γ-tocopherol methyltransferase gene. — Plant Cell Rep. 26: 61–70, 2007.
Van Eenennaam, A.L., Lincoln, K., Durrett, T.P., Valentin, H.E., Shewmaker, C.K., Thorne, G.M., Jiang, J., Baszis, S.R., Levering, C.K., Aasen E.D., Hao, M., Stein, J.C., Norris, S.R., Last, R.L.: Engineering vitamin E content: from Arabidopsis mutant to soy oil. — Plant Cell 15: 3007–3019, 2003.
Yusuf, M.A., Sarin, N.B.: Antioxidant value addition in human diets: genetic transformation of Brassica juncea with γ-TMT gene for increased α-tocopherol content. — Transgenic Res. 16: 109–113, 2007.
Zeng, J., Wang, Y.C., Shen, G., Zheng X.B.: [A Phytophthora sojae gene of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) induced in host infection and its anti-oxidative function in yeast.] — Chin. Sci. Bull. 51: 1316–1323, 2006. [In Chin.]
Acknowledgements
The research was supported by the key program of the Natural Science Foundation of Tianjin (grant No. 07JCZDJC03800), the grant of the National Natural Science Foundation of China (No. 31070273), and the R & D Special Projects for Public Welfare Industry of State Oceanic Administration People’s Republic of China (No. 200805044).
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Chen, D.F., Zhang, M., Wang, Y.Q. et al. Expression of γ-tocopherol methyltransferase gene from Brassica napus increased α-tocopherol content in soybean seed. Biol Plant 56, 131–134 (2012). https://doi.org/10.1007/s10535-012-0028-z
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DOI: https://doi.org/10.1007/s10535-012-0028-z