绿原酸对油桃和苹果果实采后成熟衰老的调控作用

发布时间：2018-06-01 23:06

本文选题：绿原酸 + 苹果　；参考：《中国农业大学》2017年博士论文

【摘要】：酚类物质是植物体中重要的次级代谢产物,其在人体中的抗氧化、抗衰老、预防心血管疾病以及改善认知等作用已广泛获得研究者的关注,但是有关酚类物质对果实采后成熟衰老进程是否以及如何产生影响的研究还比较少。本文主要研究果实内源酚类物质如何对果实成熟衰老相关蛋白质的表达以及相应酶活性产生影响从而影响果实成熟衰老进程。研究内容与结果如下:实验发现相比于新绿原酸(油桃果实果肉组织中另一种主要酚类物质),绿原酸可以更显著地减少油桃果肉切片乙烯释放量、延缓切片硬度和可溶性固形物含量下降。绿原酸可以显著降低油桃果实在贮藏期间的乙烯释放量、呼吸速率、软化程度、腐烂率和丙二醛含量。同时,绿原酸处理也延缓了贮藏期间油桃果实果皮颜色的转红、可滴定酸含量下降和可溶性固形物上升。研究还发现,绿原酸处理显著增强了果实的抗氧化活性、自由基清除能力和还原能力。实验利用双向电泳以及基质辅助激光解析电离飞行二级质谱(MALDI-TOF-TOF/MS)技术成功分离鉴定了油桃果实中74个与成熟衰老相关并受到绿原酸影响的蛋白质。其中参与果实呼吸作用的蛋白质NADP-苹果酸酶(NADP-ME)、苹果酸脱氢酶和NAD-依赖型苹果酸脱氢酶,其蛋白表达量都由于绿原酸处理出现明显下调;绿原酸处理使超氧化物歧化酶,过敏蛋白Prup1和病程相关蛋白10等具有防御性功能的蛋白表达量出现上调,;碳水化合物代谢相关蛋白质,包括α-淀粉酶,蔗糖合成酶、UDP-葡萄糖焦磷酸化酶、UDP-糖基转移酶和α-半乳糖苷酶在油桃果实成熟过程中的表达量也明显受到绿原酸影响而有所下调。绿原酸还可以显著抑制1-氨基环丙烷-1-羧酸氧化酶(ACO)在油桃果实采后成熟过程的蛋白表达量上升。使用iTRAQ定量蛋白组学技术分析苹果果肉组织中与苹果内源多酚具有亲和作用的蛋白质。分析得到157个目标蛋白质,其中包括与果实成熟衰老相关的脂氧合酶、糖基转移酶、αα-淀粉酶、淀粉支链酶、纤维素合成酶、蔗糖磷酸酶、β-半乳糖苷酶、1-氨基环丙烷-1-羧酸合成酶(ACS)、ACO和乙烯受体蛋白等。实验以苹果果肉切片作为模型来研究绿原酸对苹果果实衰老的影响。与对照组相比,绿原酸处理显著降低了苹果果肉切片乙烯释放量和呼吸速率,减缓果肉切片硬度和可溶性固形物含量的下降。十二烷基苯磺酸钠-聚丙烯酰胺凝胶电泳和MALDI-TOF/TOF分析显示,绿原酸处理显著降低了苹果果肉切片中脂肪氧合酶、β-半乳糖苷酶、NADP-ME、UDP-葡萄糖焦磷酸化酶等蛋白的表达量,提高了类甜蛋白的蛋白表达量。在体外实验中发现,苹果多酚和绿原酸均可抑制ACS、ACO、α-淀粉酶、脂氧合酶、β-葡萄糖苷酶、β-半乳糖苷糖、聚半乳糖醛酸酶(PG)、果胶甲酯酶(PME)、NADP-ME、过氧化物酶(POD)、多酚氧化酶(PPO)的酶活性,并且绿原酸对这些酶活性的抑制效果与苹果多酚相比没有显著差异。进一步实验发现,当苹果果肉切片经过绿原酸处理后,其中ACO、ACS、α-淀粉酶、脂氧合酶、β-葡糖糖苷酶、β-半乳糖苷酶、NADP-苹果酸酶、POD、PPO的酶活性在切片孵育观察过程中会显著下降,而PG和PME酶活性没有受到明显的影响。
[Abstract]:Phenols are important secondary metabolites in plant body. They have been paid more and more attention to the antioxidation, anti aging, prevention of cardiovascular diseases and the improvement of cognition in human body. However, there are few studies on the effect of phenolic substances on the mature and aging process of postharvest fruits. The contents and results are as follows: compared with the new chlorogenic acid (another main phenolic substance in the fruit flesh of nectarine fruit), chlorogenic acid can be significantly reduced. The release of ethylene in Nectarine pulp sliced, and the content of microchip hardness and soluble solids decreased. Chlorogenic acid could significantly reduce the ethylene release, respiration rate, softening degree, decay rate and malondialdehyde content during storage. At the same time, chlorogenic acid treatment also postponed the red color of nectarine fruit peel during storage. The study also found that chlorogenic acid significantly enhanced the antioxidant activity, free radical scavenging ability and reduction ability of the fruit. The experiment was successfully separated and identified by two dimensional electrophoresis and matrix assisted laser analytical ionization flight two mass spectrometry (MALDI-TOF-TOF/MS). 74 of the fruits of Nectarine were successfully separated and identified. Protein NADP- malate enzyme (NADP-ME), malate dehydrogenase and NAD- dependent malate dehydrogenase, which are involved in fruit respiration, are all down regulated by chlorogenic acid treatment; chlorogenic acid treatment makes superoxide dismutase, allergy protein Prup1 The protein expression of defensive function, such as course related protein 10, was up-regulated, and carbohydrate metabolism related proteins, including alpha amylase, sucrose synthase, UDP- glucose pyrophosphorylase, UDP- glycosyltransferase and alpha galactosidase, were also affected by chlorogenic acid in the fruit ripening of nectarine fruit. The protein expression of 1- amino cyclopropane -1- carboxylic oxidase (ACO) in the Postharvest Ripening of nectarine fruit was also significantly inhibited. The protein content of Apple endogenous polyphenols in apple pulp tissues was analyzed by iTRAQ quantitative proteomics technology. 157 target proteins were obtained, including the analysis of the protein. Lipoxygenase, glycosyltransferase, alpha alpha amylase, amylopectin, cellulose synthetase, sucrose phosphatase, beta galactosidase, 1- aminopropane -1- carboxylic synthetase (ACS), ACO and ethylene receptor protein, etc. were used as a model to study the effect of chlorogenic acid on apple fruit senescence. Compared with the control group, chlorogenic acid treatment significantly reduced the ethylene release and respiration rate of the slices of apple pulp, and slowed down the content of the firmness and soluble solids in the slices of pulp. Twelve alkylbenzene sulfonates polyacrylamide gel electrophoresis and MALDI-TOF/TOF analysis showed that chlorogenic acid treatment significantly reduced the fat in the slices of apple pulp. The expression of lipoxygenase, beta galactosidase, NADP-ME, UDP- glucose pyrophosphorylase and other proteins increased the protein expression of sweet protein. In vitro, apple polyphenols and chlorogenic acids could inhibit ACS, ACO, alpha amylase, lipoxygenase, beta glucosidase, beta galactosidase, poly galacturonase (PG), pectin a, and pectin a The enzyme activity of esterase (PME), NADP-ME, peroxidase (POD) and polyphenol oxidase (PPO), and the inhibitory effect of chlorogenic acid on these enzymes was not significantly different from that of apple polyphenols. Further experiments showed that when the slices of apple pulp were treated with chlorogenic acid, ACO, ACS, alpha amylase, lipoxygenase, beta glucosidase, beta galactozyme, and beta galactosidase were found. The activities of glucosidase, NADP- malic enzyme, POD and PPO decreased significantly during the incubation process, while PG and PME enzyme activities were not significantly affected.
【学位授予单位】：中国农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：TS255.3

【参考文献】