Faculty of Environment and Information Sciences, Fukui University of Technology, Fukui 910-8505, Japan.
Department of Medical Technology, Yokkaichi Nursing and Medical Care University, Yokkaichi 512-8045, Japan.
Int J Mol Sci. 2021 Aug 22;22(16):9046. doi: 10.3390/ijms22169046.
To investigate the mechanism of flooding tolerance of soybean, flooding-tolerant mutants derived from gamma-ray irradiated soybean were crossed with parent cultivar Enrei for removal of other factors besides the genes related to flooding tolerance in primary generated mutant soybean. Although the growth of the wild type was significantly suppressed by flooding compared with the non-flooding condition, that of the mutant lines was better than that of the wild type even if it was treated with flooding. A two-day-old mutant line was subjected to flooding for 2 days and proteins were analyzed using a gel-free/label-free proteomic technique. Oppositely changed proteins in abundance between the wild type and mutant line under flooding stress were associated in endoplasmic reticulum according to gene-ontology categorization. Immunoblot analysis confirmed that calnexin accumulation increased in both the wild type and mutant line; however, calreticulin accumulated in only the mutant line under flooding stress. Furthermore, although glycoproteins in the wild type decreased by flooding compared with the non-flooding condition, those in the mutant line increased even if it was under flooding stress. Alcohol dehydrogenase accumulated in the wild type and mutant line; however, this enzyme activity significantly increased and mildly increased in the wild type and mutant line, respectively, under flooding stress compared with the non-flooding condition. Cell death increased and decreased in the wild type and mutant line, respectively, by flooding stress. These results suggest that the regulation of cell death through the fermentation system and glycoprotein folding might be an important factor for the acquisition of flooding tolerance in mutant soybean.
为了研究大豆耐淹水的机制，利用γ射线辐照大豆获得的耐淹水突变体与亲本品种恩瑞进行杂交，以去除初级生成的突变体大豆中除了与耐淹水相关的基因以外的其他因素。虽然与非淹水条件相比，野生型的生长在淹水条件下受到显著抑制，但与野生型相比，突变体系的生长更好，即使在淹水条件下也是如此。将两天大的突变体系在淹水条件下处理 2 天，然后使用无胶/无标记蛋白质组学技术分析蛋白质。根据基因本体论分类，丰度在淹水胁迫下在野生型和突变体系之间发生相反变化的蛋白质与内质网相关。免疫印迹分析证实，钙连蛋白在野生型和突变体系中均有积累；然而，在淹水胁迫下，只有突变体系中钙网蛋白积累。此外，尽管与非淹水条件相比，野生型中的糖蛋白在淹水条件下减少，但在突变体系中即使在淹水条件下也增加。醇脱氢酶在野生型和突变体系中积累；然而，与非淹水条件相比，该酶活性在野生型和突变体系中分别显著增加和轻度增加。在野生型和突变体系中，细胞死亡分别因淹水胁迫而增加和减少。这些结果表明，通过发酵系统和糖蛋白折叠调节细胞死亡可能是突变体大豆获得耐淹水能力的一个重要因素。
