转基因水稻中过表达OsHGGT基因提高种子三烯生育酚含量的研究

Increase of Tocotrienol Content by Overexpression of OsHGGT Gene in Transgenic Rice

旨在提高稻米中三烯生育酚的含量,将来源于日本晴尿黑酸牻牛儿基牛儿基牻转移酶(homogentisic acid gerany-lgeranyl transferase,HGGT)基因导入粳稻品种武育粳3号过量表达。经PCR和RT-PCR分析证明外源基因已导入水稻中并能够在水稻胚乳中表达。HPLC测定结果表明,过表达HGGT后,转基因水稻种子糠层及胚乳中γ-三烯生育酚和总三烯生育酚的含量分别是未转化对照的1.52和1.67倍,且三烯生育酚的积累并未导致总生育酚含量的降低,最终糠层及胚乳中总三烯生育酚与总生育酚的比值分别提高到0.82和1.82,极显著高于(P<0.01)未转化对照(分别为0.54和1.27)。

In present study, with the aim to improve tocotrienol content in rice seed, the eDNA encoding homogentisic acid gerany- lgeranyl transferase （ HGGT ） from Niponbare was over-expressed under the control of CaMV 35S promoter in an elite rice japonica cultivar Wuyujing 3 by Agrobacterium-mediated transformation. The presence and the expression of the input transgene were confirmed by PCR and RT- PCR analyses, respectively, as well as determination of tocopherol and tocotrienol contents by chromatographic analysis. HPLC results showed that tocotriennls in brans and endosperms of transgenie lines increased 1.52 and 1.67 folds compared with the wild type, respectively, and tocotrienols accumulation did not result in decreases of tocopherols content. As a resnh, the ratio of total tocotrienols to total tocopherols in brans or endosperms from transgenic lines were improved up to 0.82 and 1.82, respectively, which were significantly higher （ P〈0.01 ） than those of wild type （ 0.54 and 1.27, respectively ） .