不同春生叶龄期追施氮肥对小麦粉蛋白致敏性的影响

Effects of Nitrogen Topdressing on Allergenicity of Wheat Flour Protein at Different Spring Leaf Stages

为研究不同春生叶龄期施氮对小麦粉蛋白致敏性的影响,以河北省主产区3种强筋小麦科农2009(KN2009)、石4366(SH4366)、藁优2018(GY2018)为原材料,在大田中进行不同叶龄时期追施氮肥的实验,采用微量凯氏定氮法测定蛋白质含量,不连续聚丙烯酰胺凝胶电泳(SDS-PAGE)测定分析蛋白质亚基组分,双抗体夹芯式酶联免疫法(ELISA)测定过敏原含量,免疫印迹技术分析蛋白质组分致敏性。结果表明,随着施氮时间的延迟,科农2009和藁优2018小麦粉蛋白致敏性增强,石4366小麦致敏蛋白含量逐渐降低。KN2009和SH4366小麦品种在N3时期施氮肥GY2018小麦品种在N5时期施氮肥较适宜,施氮时间的延迟主要影响小麦粉的致敏蛋白,包括高分子质量麦谷蛋白、ω-麦醇溶蛋白以及26、29、38、43 ku的蛋白亚基。

The effects of nitrogen application on protein allergenicity of wheat flour at different spring leaf ages were studied.Three kinds of strong gluten wheat(KN2009,SH4366 and GY2018)were the raw materials in the main producing areas of Hebei Province.The experiment of topdressing nitrogen fertilizer at different leaf ages was carried out in the field.Protein content was determined by the micro Kjeldahl method;protein subunit components were determined by discontinuous polyacrylamide gel electrophoresis(SDS-PAGE);allergen content was determined by double antibody sandwich ELISA;allergenicity of protein components was analyzed by western blotting.The results indicated that with the delay of nitrogen application time,the allergenicity expression of wheat flour protein of KN2009 and GY2018 increased,and the content of wheat allergenic protein of SH4366 gradually decreased.N top-dressing at third spring leaf instar was significantly effective and safety fertilizer period Application of KN2009 and SH4366.N top-dressing at fifth spring leaf instar was significantly effective and safety fertilizer period Application of GY2018.In addition,with the delay of nitrogen application time,high molecular weight glutenin,ω-gliadin and protein subunits with molecular weight of 26,29,38,and 43 ku were the principal allergens of wheat flour.