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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 regulating 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 convenient 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.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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