基于线粒体能量代谢途径的金针菇采后纳米包装保鲜机制

doi:10.3864/j.issn.0578-1752.2020.16.013

Abstract

Abstract:

【Objective】 The purposes of this study were to illuminate the effects of different packaging on energy metabolism of Flammulina filiformis from mitochondrial level, providing a new idea for further revealing the preservation mechanism of nano-package. 【Method】 In this experiment, F. filiformis was used as the research object. By using three common methods, including discontinuous density gradient centrifugation method (DDGCM), yeast mitochondrial extraction kit method (YMEKM) and improved Pulilai kit method (IPKM), the optimal way to extract F. filiformis mitochondria was determined. The regularity of post-harvest energy metabolism in packaged F. filiformis was expounded via measuring the content of ATP metabolism system substances (ATP, ADP, AMP and energy charge) and mitochondrial complex activity. 【Result】 The alcohol dehydrogenase activity in mitochondria and the cytochrome C oxidase activity in cytosol were the lowest, and mitochondrial respiration rates were the highest in IPKM extracted samples compared to other two methods, demonstrating the IPKM obtained mitochondria had the best membrane integrity. The mitochondrial superoxide dismutase activity extracted by IPKM was the highest, reaching 17.82 U·mg^-1 pro, which was 7.31% and 25.59% higher than that extracted by DDGCM-1 and YMEKM, respectively. Combined with the results of transmission electron microscopy and Janus green B staining test showed that the IPKM treatment had the largest number of active mitochondria, and their mitochondrial structure was relatively complete. During the cold storage period of F. filiformis, the sample in nano-packaging (NP) groups showed that not only the ATP and energy content but also the activity of the main mitochondrial complex was significantly higher than those of polyethylene (PE) packaged bags (control). NP materials promoted the process of oxidative phosphorylation of mitochondria by maintaining a high level of ATP content and delaying the decline of energy charge. At the 21st day, the ATP content of NP groups was still 113.83 μg·g^-1FW, which was higher than that of control, thus avoiding the reduction of the activity of F. filiformis mitochondrial complex I and Ⅲ, and slowing down the activity of mitochondrial complex Ⅳ. The mitochondrial complex Ⅳ activity of F. filiformis in the PE packaging groups reached the first peak at the 9th day, while NP groups reached the first peak at the 15th day, and the activity of the mitochondrial complex IV (5.14 U·mg^-1 pro) in the NP materials reached the first peak on the 15th day, which was significantly higher than that in the PE packaging groups (1.12 U·mg^-1 pro), with a difference of 78.23%. It proved that NP could effectively control the electron transfer in the mitochondrial respiratory chain without hindrance, ensure the energy supply of cells, thus better maintain the energy state of F. filiformis . 【Conclusion】 Compared with other methods, the mitochondria proposed by IPKM was the best approach for leading to the minimal damage on mitochondria structure and bioactivity, which laid the foundation of mitochondrial energy metabolism research on F. filiformis in the future. Meanwhile, the energy state of F. filiformis was maintained and its shelf-life was prolonged by NP treatment via improving ATP content and mitochondrial complex I, Ⅲ activities, and delaying the decline of energy charge and mitochondrial complex Ⅳ activity.

Key words: Flammulina filiformis, mitochondria, improved Pulilai method, storage, energy metabolism

MA Ning,WANG HeTong,FANG DongLu,ZHAO LiYan,YANG WenJian,PEI Fei,HU QiuHui. Nano-Packaging Preservative Mechanism of Flammulina filiformis After Harvest Based on Mitochondrial Energy Status Pathways[J].Scientia Agricultura Sinica, 2020, 53(16): 3356-3371.

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能量组分 Component of energy charge	回归方程 Regression equation	相关系数 Correlation coefficient
ATP	y=29.933x-4.275	R²=0.9998
ADP	y=27.48x+28.358	R²=1
AMP	y=36.92x+12.892	R²=1

Nano-Packaging Preservative Mechanism of Flammulina filiformis After Harvest Based on Mitochondrial Energy Status Pathways

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