水稻种胚脂肪酶酶活的精确定量分析技术研究

Study on accurate analysis technique of rice seed lipase

建立了一种新的适合在微乳液体系中采用脂肪酸比色法定量分析水稻种子脂肪酶活力的方法,并通过酶动力学实验优化了该方法的最适条件。优化的结果是：底物浓度30mg/mL,酶反应时间20h,各种品种只用10-50个种胚即可精确测出种子脂肪酶的总活力,方法的重现性好。用该方法检测了皖鉴2090及其后代材料的脂肪酶活并利用人工老化实验进行了耐储藏性评价,实验结果显示,该酶活性与储藏特性关系十分密切。该方法可用于大规模选育耐储藏水稻品种,并为全面揭示耐储藏机理打下坚实基础。

A new fatty acid colorimetry method for accurate analysis of activities of rice seed lipase was established based on the microemulsion with the optimum conditions investigated. Based on the enzyme kinetic study, the activities of lipase in different rice varieties could be precisely determined in 10-50 rice embryos, with the substrate concentration of 30 mg/mL and the reaction period of 20 hours. The experimental results show that activities of lipase in Wanjian2090 and its offspring mutants, determined by this method, affect rice storability significantly during accelerating aging experiments. This method could benefit cultivating rice varieties with good storability and studying the mechanism of rice aging.