Transformation of the US bread wheat 'Butte 86' and silencing of omega-5 gliadin genes.

GM crops Pub Date : 2011-01-01 DOI:10.4161/gmcr.2.1.15884
Susan B Altenbach, Paul V Allen
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引用次数: 59

Abstract

Complex groups of proteins determine the unique functional properties of wheat flour and are sometimes responsible for food intolerances and allergies in individuals that consume wheat products. Transgenic approaches can be used to explore the functions of different flour proteins, but are limited to the few wheat cultivars that can be transformed and also by the lack of detailed information about genes and proteins expressed in grain from those cultivars. The US bread wheat Butte 86 has been extensively characterized and a comprehensive proteome map was developed in which flour proteins were distinguished by mass spectrometry and associated with specific gene sequences. Here, this information has been used to design an RNA interference construct to silence the expression of genes encoding omega gliadins that trigger the food allergy wheat-dependent exercise-induced anaphylaxis (WDEIA). The construct was introduced into immature embryos from Butte 86 using biolistics and bialaphos-resistant plants were regenerated. Stable transformation and inheritance of the transgene were confirmed by PCR. Analysis of proteins in grain from transgenic plants demonstrated that the omega-5 gliadins were either absent or substantially reduced relative to non-transformed controls. The ability to genetically transform Butte 86 makes it possible to alter flour composition in a targeted manner in a commercial US wheat cultivar and should accelerate future research on flour quality and immunogenic potential.

美国面包小麦“Butte 86”的转化和ω -5麦胶蛋白基因的沉默。
复杂的蛋白质群决定了小麦粉的独特功能特性,有时会导致食用小麦产品的人产生食物不耐受和过敏。转基因方法可以用于探索不同面粉蛋白的功能,但仅限于少数几种可以转化的小麦品种,而且缺乏这些品种籽粒中表达的基因和蛋白质的详细信息。对美国面包小麦Butte 86进行了广泛的鉴定,并建立了一个全面的蛋白质组图谱,其中面粉蛋白通过质谱分析区分并与特定基因序列相关。在这里,这些信息被用于设计RNA干扰结构,以沉默编码omega麦胶蛋白的基因表达,这些基因会引发食物过敏小麦依赖运动诱导的过敏反应(WDEIA)。利用生物学方法将该构建体导入Butte 86的未成熟胚中,获得了抗双磷植株。PCR证实该转基因转化稳定,遗传良好。转基因植物谷物中的蛋白质分析表明,与未转化的对照相比,omega-5麦胶蛋白要么不存在,要么大幅减少。对Butte 86进行基因改造的能力使得有针对性地改变美国商业小麦品种的面粉成分成为可能,并将加速未来对面粉质量和免疫原性潜力的研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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