热带作物学报2024，Vol.45Issue(8)：1651-1658,8.DOI:10.3969/j.issn.1000-2561.2024.08.014
酸碱滴定法优化香蕉磷脂酶A活性测定及炭疽病胁迫下其活性变化研究
Optimization on Activity Assay of Banana Phospholipase A by Acid-base Titration and Change of PLA Activity under Anthracnose Stress
摘要
Abstract
Plant phospholipase A(PLA)is a key enzyme in cell membrane phospholipid metabolism.It can catalyze the cleavage of multiple functional groups on phospholipids to generate signaling molecules,which are involved in regu-lating plant growth and development as well as environmental and biological stresses.PLA possesses important research value.In this study,the effects of various factors such as substrate concentration,reaction temperature,pH of buffer solution,buffer solution volume,and reaction time on catalytic activities of banana PLA for phospholipid hydrolysis were investigated,so as to optimize PLA activity assay method.According to the optimal assay conditions,the change of banana PLA activity under anthracnose stress was further studied.The results showed that the optimal conditions for the determination of banana PLA activity by acid-base titration method were as follows:pH 9,reaction temperature 55 ℃,substrate concentration 4 g/L,15 mL substrate,3 mL crude enzyme extract,the ratio of extraction buffer volume to peel weight(mL/g)3,and reaction time 7 h.According to the optimal conditions,the PLA activity of banana peel tissue under anthracnose stress was determined.It was found that the degree of disease in peel tissue was positively correlated with PLA activity,indicating that PLA activity increased with the deepening of disease.This suggested that the level of PLA activity was probably related to fruit resistance.As disease degree deepened,PLA activity increased,membrane phospholipids degraded,and cell membrane integrity disrupted,leading to fruit decay.In this article,a con-venient and feasible method for the determination of banana PLA activity was established,which laid a foundation for further study of the catalytic mechanism of banana phospholipase A.关键词
香蕉/磷脂酶A/酸碱滴定法/酶活性/炭疽病Key words
banana/phospholipase A/acid-base titration/enzyme activity/anthracnose分类
农业科技引用本文复制引用
陈瑜娴,李丽,李静,孙健,易萍,黄方,黄敏,甘婷..酸碱滴定法优化香蕉磷脂酶A活性测定及炭疽病胁迫下其活性变化研究[J].热带作物学报,2024,45(8):1651-1658,8.基金项目
国家自然科学基金项目(No.32272784,No.32160732) （No.32272784,No.32160732）
广西农业科学院基本科研业务专项(桂农科2024YP093). （桂农科2024YP093）
