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Genetic and functional analysis of tocopherol biosynthesis pathway genes from rapeseed (Brassica napus L.)

机译:油菜生育酚生物合成途径基因的遗传和功能分析

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摘要

Brassica napus L. is one of the most important oilseed crops in the world. Improving oil quality by increasing vitamin E content is a major target of rapeseed breeding. Vitamin E compounds, or tocopherols, are lipid-soluble antioxidants that are essential nutrients for mammals. In this study, we report the characterization of rapeseed orthologs of the Arabidopsis tocopherol genes VTE1, VTE2 and PDS1. For each gene, at least two homologous sequences were found, and their expression was analysed in different tissues from three rapeseed genotypes. Genetic complementation experiments demonstrated that BnaX.VTE1.b and BnaA. VTE2.b homologs are capable of recovering seed gamma-tocopherol in Arabidopsis mutants. Overexpression of the genes in Arabidopsis Col-0 shifted the seed tocopherol composition towards higher alpha-tocopherol without increasing the total tocopherol content. To address the functionality of BnPDS1 sequences, we performed overexpression tests in Escherichia coli and enzymatic activity analyses. Overall, our results show that the identified sequences from rapeseed are functional orthologs of the Arabidopsis VTE genes and thus have considerable potential as molecular markers for selecting rapeseed with improved seed oil quality
机译:甘蓝型油菜是世界上最重要的油料作物之一。通过增加维生素E含量来改善油质是油菜育种的主要目标。维生素E化合物或生育酚是脂溶性抗氧化剂,是哺乳动物必不可少的营养素。在这项研究中,我们报告了拟南芥生育酚基因VTE1,VTE2和PDS1的油菜直向同源物的表征。对于每个基因,至少找到两个同源序列,并分析了来自三种油菜籽基因型的不同组织中它们的表达。遗传互补实验证明BnaX.VTE1.b和BnaA。 VTE2.b同源物能够回收拟南芥突变体中的种子γ-生育酚。拟南芥Col-0中基因的过表达使种子生育酚的组成向较高的α-生育酚转移,而没有增加总生育酚的含量。为了解决BnPDS1序列的功能,我们在大肠杆菌中进行了过表达测试,并进行了酶活性分析。总体而言,我们的结果表明,从油菜籽中鉴定出的序列是拟南芥VTE基因的功能直向同源物,因此具有作为分子标记物选择具有更高种子油品质的油菜籽的巨大潜力

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