基于系统生物学方法研究五味子鲜果干燥过程中抑制采后生理对药材品质的影响

Fruits of Schisandra chinensis(Turcz.)Baill belongs to berries, having the property that thin fruit peel, high moisture, high sugar content. For the convenience of storage and transportation, fresh fruits should be dried after picking. It is well known that the tissues of fresh fruits are still undertaking active metabolism after picking，called Postharvest Physiology, assuming that the immature fruits can continue to ripe and the mature fruits can continue to decompose. Schisandra chinensis matures later in Heilongjiang province. And the traditional drying method(TDM) has a long period. Therefore the postharvest physiological activities will continue to consume the contents of the fruits in the drying cycle. The early experiments showed that if the enzyme activity in the fruits were destroyed by microwave, it can not only make the contents of schizandrin, deoxyschizandrin and schisandrin B more than control group(TDM), but also inhibit the enzymatic Browning, make the color of dried herbs more bright, and reducing the damage of the anthocyanin. This project aims to reserch the effects of post-harvest physiology on the metabolites and protein using plant metabolomics and differential proteomics techniques by suppressing the post-harvest physiological activities of fresh fruits through some means (steaming, ironing, and microwave irradiation), and analyze the correlation between characteristic components and key-proteins by bioinformatics. Finally, to confirm what is the effect of suppress post-harvest physiology on the quality of Schisandra chinensis at the molecular level. It will provide new ideas and explorations for the scientization and rationalization of the processing of Chinese medicinal materials.

五味子果实属于浆果，鲜果果皮薄，含水率、含糖率高，为便于储运鲜果采摘后多需要干制。众所周知，鲜果采摘后组织仍进行着活跃的新陈代谢，称之为采后生理，表现为未成熟的果实可以继续成熟，已成熟的会老化腐烂。黑龙江省的五味子采收期较晚，传统干燥方法周期长，干燥周期内采后生理活动会继续消耗果实中的内含物质。前期实验发现利用微波破坏果实中的内源酶活性，不仅能使五味子醇甲、甲素和乙素的含量高于传统晒干对照品，还能抑制酶促褐变，使干燥药材颜色更加鲜艳，减少了五味子色素的破坏。本课题拟通过一定方法（蒸、烫漂、微波照射）抑制五味子鲜果的采后生理活动，运用植物代谢组学和差异蛋白质组学技术，研究抑制采后生理对五味子代谢产物和蛋白表达的影响，再利用生物信息学技术分析特征成分与关键蛋白之间的相关性，最终从分子水平论证抑制五味子鲜果采后生理对其品质的影响，为中药材产地加工的科学化、合理化提供新思路和新探索。

五味子果实属于浆果，鲜果果皮薄，含水率、含糖率高，为便于储运鲜果采摘后多需要干制。众所周知，鲜果采摘后组织仍进行着活跃的新陈代谢，称之为采后生理，表现为未成熟的果实可以继续成熟，已成熟的会老化腐烂。黑龙江省的五味子采收期较晚，传统干燥方法周期长，干燥周期内采后生理活动会继续消耗果实中的内含物质。本课题采用50 ℃烘干、晒干、微波“杀青”后50 ℃烘干、烫漂“杀青”后50 ℃烘干、蒸制“杀青”后50 ℃烘干及冷冻干燥6种方法对五味子鲜果进行初加工，运用LC-MS、GC-MS、NRM等植物代谢组学技术和蛋白质组学技术，以50 ℃烘干和晒干样品为对照，对五味子鲜果采后“杀青”样品的代谢产物和蛋白表达影响进行研究。从总木脂素、总多糖和挥发油三者含量的总和来看，烫漂和蒸制样品最高，微波350 W样品次之；测定五味子醇甲、五味子醇乙、五味子酯甲、五味子酯乙、五味子甲素、五味子乙素和五味子丙素7种木脂素类成分的含量证实了先采用烫漂、微波350 W等方法“杀青”处理后再干燥处理的五味子样品的品质要优于直接晒干和烘干的样品；利用LC-MS对五味子醇提物进行分析，进一步证实了先采用烫漂和微波350 W等方法“杀青”处理后再干燥处理的五味子样品的品质要优于直接晒干和烘干的样品；LC-MS对五味子水提物的分析表明不同初加工方法的影响并不显著；GC-MS代谢组学技术研究还发现微波“杀青”后50 ℃烘干、烫漂“杀青”后50 ℃烘干、蒸制“杀青”后50 ℃烘干和冻干对五味子挥发油总体成分谱影响显著；NMR代谢组学技术鉴定出的27个代谢产物绝大多数为初级代谢产物，主要为氨基酸、有机酸和糖类，而这些初级代谢产物是果实采后生理能量代谢的主要参与者，说明“杀青”可能干扰了五味子鲜果干燥过程中的能量代谢活动；蛋白质组学研究证实“杀青”确实能干扰五味子鲜果干燥过程中的能量代谢活动。本研究从分子水平论证抑制五味子鲜果采后生理对其品质的影响，为中药材产地加工的科学化、合理化提供新思路和新探索。