中国农业科学 ›› 2020, Vol. 53 ›› Issue (16): 3356-3371.doi: 10.3864/j.issn.0578-1752.2020.16.013
马宁1(),王鹤潼1,方东路2,赵立艳2,杨文建1,裴斐1,胡秋辉1
收稿日期:
2020-03-21
接受日期:
2020-05-27
出版日期:
2020-08-16
发布日期:
2020-08-27
作者简介:
马宁,Tel:13951729396;E-mail:基金资助:
MA Ning1(),WANG HeTong1,FANG DongLu2,ZHAO LiYan2,YANG WenJian1,PEI Fei1,HU QiuHui1
Received:
2020-03-21
Accepted:
2020-05-27
Online:
2020-08-16
Published:
2020-08-27
摘要:
【目的】从线粒体水平上揭示不同包装对金针菇能量代谢的影响,为进一步揭示纳米包装保鲜机制提供新思路。【方法】以金针菇为材料,对比不连续密度梯度离心法(DDGCM)、酵母线粒体提取试剂盒方法(YMEKM)和改进普利莱法(IPKM)3种提取方法对金针菇子实体与菌丝体中线粒体功能活性的影响,确定最佳提取方法。通过测定不同包装及不同贮藏期金针菇线粒体中ATP代谢系统物质含量(ATP、ADP、AMP及能荷)和线粒体主要复合体活性,揭示不同包装金针菇采后能量代谢规律。【结果】通过标志性污染物酶和线粒体呼吸速率指标测定,3种方法以IPKM提取的线粒体内乙醇脱氢酶活性及胞液内细胞色素C氧化酶活性最低,呼吸速率最高,证明IPKM所提的线粒体结构完整性最好;IPKM提取的线粒体超氧化物歧化酶活性最高,达到17.82 U·mg-1 pro,比DDGCM-1和YMEKM分别高7.31%和25.59%。结合透射电镜和健那绿染色结果可知,IPKM提取的线粒体结构相对完整,活性线粒体数量最多。在金针菇冷藏期间,纳米包装金针菇的ATP水平、能荷含量以及线粒体主要复合体活性都显著高于普通PE包装。纳米包装材料通过维持高水平的ATP含量,延缓能荷值的下降,促进了线粒体的氧化磷酸化过程。纳米包装组在第21天的ATP含量仍然比普通PE包装组高113.83 μg·g-1FW,从而避免金针菇线粒体复合体I、Ⅲ活性降低,减缓线粒体复合体Ⅳ的活性降低。普通PE包装金针菇的线粒体复合体Ⅳ活性在第9天达到第一个峰值,而纳米包装金针菇线粒体复合体IV活性在第15天达到第一个峰值(5.14 U·mg-1 pro),显著高于普通PE包装(1.12 U·mg-1 pro),相差达到78.23%。证明纳米包装能有效控制线粒体呼吸链中的电子传递不受阻碍,保证细胞能量供应,从而更好地保持金针菇的能量状态。【结论】改进普利莱法对线粒体损伤最小,并维持线粒体活性,适合金针菇采后线粒体能量代谢研究。同时,纳米包装通过维持较高水平的ATP含量与线粒体复合体I、Ⅲ活性并延缓能荷与线粒体复合体Ⅳ活性的下降,进而保持金针菇的能量状态并延缓衰老,延长贮藏期。
马宁,王鹤潼,方东路,赵立艳,杨文建,裴斐,胡秋辉. 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[J]. 中国农业科学, 2020, 53(16): 3356-3371.
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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