Watanabe Daisuke, Adányi Nóra, Takács Krisztina, Maczó Anita, Nagy András, Gelencsér Éva, Pachner Martin, Lauter Kathrin, Baumgartner Sabine, Vollmann Johann
Department of Crop Sciences, Division of Plant Breeding, University of Natural Resources and Life Sciences Vienna (BOKU), 3430, Tulln an der Donau, Austria.
National Agricultural Research and Innovation Centre, Food Science Research Institute (NARIC-FSRI), 1022, Budapest, Hungary.
J Sci Food Agric. 2017 Feb;97(3):1010-1017. doi: 10.1002/jsfa.7827. Epub 2016 Jul 4.
In soybean, at least 16 seed proteins have been identified as causing allergenic reactions in sensitive individuals. As a soybean genebank accession low in the immunodominant protein P34 (Gly m Bd 30K) has recently been found, introgression of the low-P34 trait into adapted soybean germplasm has been attempted in order to improve the safety of food products containing soybean protein. Therefore, marker-assisted selection and proteomics were applied to identify and characterize low-P34 soybeans.
In low-P34 lines selected from a cross-population, concentrations of the P34 protein as identified with a polyclonal antibody were reduced by 50-70% as compared to P34-containing controls. Using 2D electrophoresis and immunoblotting, the reduction of P34 protein was verified in low-P34 lines. This result was confirmed by liquid chromatographic-tandem mass spectrometric analysis, which revealed either a reduction or complete absence of the authentic P34 protein as suggested from presence or absence of a unique peptide useful for discriminating between conventional and low-P34 lines.
Marker-assisted selection proved useful for identifying low-P34 soybean lines for the development of hypoallergenic soy foods. The status of the P34 protein in low-P34 lines needs further characterization. In addition, the food safety relevance of low-P34 soybeans should be tested in clinical studies. © 2016 Society of Chemical Industry.
在大豆中,至少有16种种子蛋白已被确定会在敏感个体中引发过敏反应。由于最近发现了一种免疫显性蛋白P34(Gly m Bd 30K)含量低的大豆基因库种质,人们尝试将低P34性状导入适应性强的大豆种质中,以提高含大豆蛋白食品的安全性。因此,应用了标记辅助选择和蛋白质组学技术来鉴定和表征低P34大豆。
在从杂交群体中选出的低P34品系中,用多克隆抗体鉴定的P34蛋白浓度与含P34的对照相比降低了50 - 70%。使用二维电泳和免疫印迹法,在低P34品系中证实了P34蛋白的减少。液相色谱 - 串联质谱分析证实了这一结果,该分析表明,根据用于区分常规品系和低P34品系的独特肽段的存在与否,真实的P34蛋白要么减少,要么完全不存在。
标记辅助选择被证明有助于鉴定低P34大豆品系,用于开发低过敏性大豆食品。低P34品系中P34蛋白的状态需要进一步表征。此外,低P34大豆的食品安全相关性应在临床研究中进行测试。© 2016化学工业协会。