Transformation of the US bread wheat 'Butte 86' and silencing of omega-5 gliadin genes.
Transformation of the US bread wheat 'Butte 86' and silencing of omega-5 gliadin genes.
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DOI：
10.4161/gmcr.2.1.15884
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发表时间：
2011-01-01
期刊：
影响因子：
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通讯作者：
Allen, Paul V
中科院分区：
文献类型：
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作者：
Altenbach, Susan B;Allen, Paul V
复杂的蛋白质组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个体的食物不耐受和过敏。转基因方法可用于探索不同面粉蛋白质的功能，但限于少数可转化的小麦品种，并且缺乏关于这些品种的基因和蛋白质在谷物中表达的详细信息。美国面包小麦Butte 86已被广泛表征，并开发了一个全面的蛋白质组图谱，其中面粉蛋白质通过质谱法区分并与特定基因序列相关。在这里，这些信息已被用来设计一个RNA干扰构建沉默的基因编码的ω麦醇溶蛋白，触发食物过敏小麦依赖性运动诱导的过敏反应（WDEIA）的表达。使用生物基因枪将该构建体导入Butte 86的未成熟胚中，并再生出双丙氨磷抗性植物。通过PCR证实了转基因的稳定转化和遗传。对转基因植物谷粒中蛋白质的分析表明，相对于非转化对照，ω-5醇溶蛋白要么不存在，要么显著减少。转基因Butte 86的能力使得有可能在美国商业小麦品种中以有针对性的方式改变面粉成分，并应加速未来对面粉质量和免疫原性潜力的研究。
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.