Molecular cloning and heterologous expression analysis of JrVTE1 gene from walnut (Juglans regia)

文献类型： 外文期刊

第一作者： Wang, Cancan

作者： Wang, Cancan;Li, Chuanrong;Guo, Xianfeng;Yang, Keqiang;Wang, Cancan;Li, Chuanrong;Yang, Keqiang;Leslie, Charles A.;Sun, Qingrong

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关键词： Juglans regia;JrVTE1;Genetic transformation;Zizyphus jujuba var. spinosa;Pyrus communis;Tocopherols

期刊名称：MOLECULAR BREEDING （ 影响因子：2.589； 五年影响因子：2.75 ）

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收录情况： SCI

摘要： Tocopherol cyclase (VTE1) plays a key role in promoting the production of gamma-tocopherol and improving total tocopherol content in photosynthetic organisms. Walnut is an important source of tocopherols in the human diet, and gamma-tocopherol is the major tocopherol compound in walnut kernels. In this study, a full-length cDNA of the VTE1 gene was isolated from walnut using RT-PCR and RACE, and designated as JrVTE1. The full-length cDNA of the JrVTE1 gene contained a 1353-bp open-reading frame encoding a 451-amino-acid protein with a calculated molecular weight of 49.5 kDa. The deduced JrVTE1 protein had a considerable homology with other plant VTE1s and belonged to the tocopherol cyclase family. Functional characterization of JrVTE1 by heterologous expression was carried out in E. coli BL21 (DE3) and microshoot lines of the fruit trees jujube (Zizyphus jujuba var. spinosa) and pear (Pyrus communis) cultivar 'Old Home'. JrVTE1 in E. coli expressed as a 50 kDa protein, as expected. One or two copies of the transferred JrVTE1 gene were detected in the genomes of representative transgenic lines (from the initial transgenic plants) of jujube and pear by gel blots analysis. Over-expression of JrVTE1 in jujube and pear resulted in an accumulation of tocopherol and a shift in tocopherol composition in leaf, root and stem tissues. In the transgenic jujube, the total tocopherol content increased by 29.8 mu g/g in the stems of line J3, 43.7 and 22.5 mu g/g in the roots and leaves of line J1, respectively, whereas in the transgenic pear it increased by 47.3 mu g/g in the leaf of line P3, and 16.7 and 10.4 mu g/g in roots and stems of line P9, respectively. In the examined tissues of transgenic plants, the highest accumulation rate was the gamma-tocopherol. These results indicate that JrVTE1 is one of the rate-limiting enzymes for tocopherol production and could be used to improve the tocopherol content of tree crops through genetic engineering.

分类号： Q94

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