鲜食橄榄发育成熟过程中多酚及相关酶活性的动态变化
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摘要
【目的】酚类物质由于本身的涩味而影响果实的鲜食品质,探究不同品种(系)橄榄果实中多酚及与酚类代谢相关酶的变化规律,以期探究影响果实鲜食的酚类物质及关键酶。【方法】选用涩味明显的普通橄榄‘檀头23’及适宜鲜食的实生变异株系——涩味较淡、回甘明显的清橄榄‘马坑22’为试验材料,研究其果实发育成熟过程中酚类物质的动态变化和苯丙烷类3个代谢相关酶(PAL、C4H、4CL)和多酚分解相关酶(PPO、POD)的活性变化。【结果】在果实发育成熟过程中‘,马坑22’总酚、鞣花酸、没食子酸甲酯与金丝桃苷的含量显著低于‘檀头23’(p <0.05),且含量变化趋势基本相似‘;马坑22’酚类物质合成酶(C4H、4CL)活性显著低于‘檀头23’,酚类物质分解酶PPO的活性在成熟后期出现活性高峰且显著高于‘檀头23’,使得两个品种(系)中酚类物质含量出现明显差异。【结论】总酚及多酚组分间的含量差异导致普通橄榄和清橄榄苦涩味不同,其中鞣花酸、没食子酸甲酯与金丝桃苷之间的差异可能是导致鲜食橄榄口感风味不同的重要影响因子;PPO、C4H、4CL活性的协调差异调控与橄榄酚类物质含量的差异形成有关。
【Objective】The astringency of phenols affects the quality of fresh fruits. The change of polyphenols in different lines of Chinese olive(Canarium album(Lour.) Raeusch.) fruits and the relationship between polyphenols and enzymes related to phenol metabolism were explored, and the phenol components and key enzymes affecting the fresh eating of fruits were revealed.【Methods】The contents of polyphenols(gallic acid, rutin, ellagic acid, methyl gallate, hyperoside) of common line C. album‘Tantou23'with obvious astringency and C. album‘Makeng 22', a selection for fresh eating from its seedlings with mild astringency, obvious aftertaste, were analyzed by high performance liquid chromatography(HPLC) during fruit development and maturation for studying the dynamic changes of phenolic substances and three phenylpropane metabolic enzymes(PAL, C4 H, 4 CL) and polyphenol decomposition related enzymes(PPO, POD) of phenylpropane.【Results】The contents of total phenols and polyphenols in C. album‘Makeng 22'were lower than those in C. album‘Tantou 23'in ripe fruits. The contents of total phenols, ellagic acid, rutin, hyperoside, methyl gallate and gallic acid in C. album‘Tantou 23'were 1.44,1.32, 1.57, 1.53, 3.08 and 1.24 times of C. album‘Makeng 22', respectively. Except for gallic acid, total phenol, ellagic acid, rutin, methyl gallate and hyperoside were significantly different(p < 0.01); The content of rutin was the highest in C. album‘Makeng 22', followed by ellagic acid, which was slightly lower than rutin, and the content of ellagic acid was the highest in C. album‘Tantou 23'. The total phenol contents of the two lines reached the peak at 102 days after anthesis, and then decreased gradually with fruit maturation. The total phenol content of C. album‘Makeng 22'was significantly lower than that of C. album‘Tantou 23'(p < 0.01) at every stage. Except for gallic acid and rutin, the contents of ellagic acid,methyl gallate and hyperoside in C. album‘Makeng 22'were also significantly lower than those in C. album‘Tantou 23'(p < 0.01). The change trends of PAL activity in two lines were basically similar, and there was no significant difference in PAL activity at maturity(p > 0.05). It was suggested that the accumulation of phenolic substances during olive fruit development and maturity might not be related to PAL,which might not be the main cause of the difference in the content of phenolic substances between two lines. The trends of C4 H activity in two lines were also similar. The C4 H activity in C. album‘Makeng22'was significantly lower than that of C. album‘Tantou 23'(p < 0.01) during the whole fruit development and maturity period. Although the change trends of 4 CL activity were different between the two lines, the activity of 4 CL in C. album‘Makeng 22'was lower than that of C. album‘Tantou 23'during olive fruit maturity except for the later stage of fruit development. Therefore, it was suggested that C4 H and 4 CL might be related to the difference of the content of polyphenols between two the lines. There was no significant difference in POD activity between the two lines(p > 0.05). Although POD activity peaked in C. album‘Makeng 22', which was significantly higher than that of C. album‘Tantou 23', at the later stage of fruit development. But there was no significant difference in POD activity between the two lines during fruit ripening. Therefore, POD might not be the main reason for the difference of fruit flavor between the two lines during fruit ripening, but the difference and influence of POD during fruit development need further to study. The PPO activity in C. album‘Makeng 22'peaked at the later stage of maturation and was significantly higher than that in C. album‘Tantou 23'(p < 0.05). The PPO activity also decreased as the decrease of polyphenol content. During the olive fruit development and maturation,the pericarp is green, and chlorophyll in the pericarp can be photosynthesized to accumulate nutrients.Therefore, PPO is not only related to polyphenol decomposition, but also may promote polyphenol accumulation by promoting metabolic activity.【Conclusion】The difference of total phenols and polyphenol components result in different bitterness and astringency of common olive and the new selection, and then affect the quality of fresh eating. The difference of ellagic acid, methyl gallate and hyperoside might lead to different taste of fresh-eating Chinese olive. And the coordinated regulation of PPO, C4 H and4 CL results in the difference of the formation of phenols in the two lines of Chinese olive.
【Objective】The astringency of phenols affects the quality of fresh fruits. The change of polyphenols in different lines of Chinese olive(Canarium album(Lour.) Raeusch.) fruits and the relationship between polyphenols and enzymes related to phenol metabolism were explored, and the phenol components and key enzymes affecting the fresh eating of fruits were revealed.【Methods】The contents of polyphenols(gallic acid, rutin, ellagic acid, methyl gallate, hyperoside) of common line C. album‘Tantou23'with obvious astringency and C. album‘Makeng 22', a selection for fresh eating from its seedlings with mild astringency, obvious aftertaste, were analyzed by high performance liquid chromatography(HPLC) during fruit development and maturation for studying the dynamic changes of phenolic substances and three phenylpropane metabolic enzymes(PAL, C4 H, 4 CL) and polyphenol decomposition related enzymes(PPO, POD) of phenylpropane.【Results】The contents of total phenols and polyphenols in C. album‘Makeng 22'were lower than those in C. album‘Tantou 23'in ripe fruits. The contents of total phenols, ellagic acid, rutin, hyperoside, methyl gallate and gallic acid in C. album‘Tantou 23'were 1.44,1.32, 1.57, 1.53, 3.08 and 1.24 times of C. album‘Makeng 22', respectively. Except for gallic acid, total phenol, ellagic acid, rutin, methyl gallate and hyperoside were significantly different(p < 0.01); The content of rutin was the highest in C. album‘Makeng 22', followed by ellagic acid, which was slightly lower than rutin, and the content of ellagic acid was the highest in C. album‘Tantou 23'. The total phenol contents of the two lines reached the peak at 102 days after anthesis, and then decreased gradually with fruit maturation. The total phenol content of C. album‘Makeng 22'was significantly lower than that of C. album‘Tantou 23'(p < 0.01) at every stage. Except for gallic acid and rutin, the contents of ellagic acid,methyl gallate and hyperoside in C. album‘Makeng 22'were also significantly lower than those in C. album‘Tantou 23'(p < 0.01). The change trends of PAL activity in two lines were basically similar, and there was no significant difference in PAL activity at maturity(p > 0.05). It was suggested that the accumulation of phenolic substances during olive fruit development and maturity might not be related to PAL,which might not be the main cause of the difference in the content of phenolic substances between two lines. The trends of C4 H activity in two lines were also similar. The C4 H activity in C. album‘Makeng22'was significantly lower than that of C. album‘Tantou 23'(p < 0.01) during the whole fruit development and maturity period. Although the change trends of 4 CL activity were different between the two lines, the activity of 4 CL in C. album‘Makeng 22'was lower than that of C. album‘Tantou 23'during olive fruit maturity except for the later stage of fruit development. Therefore, it was suggested that C4 H and 4 CL might be related to the difference of the content of polyphenols between two the lines. There was no significant difference in POD activity between the two lines(p > 0.05). Although POD activity peaked in C. album‘Makeng 22', which was significantly higher than that of C. album‘Tantou 23', at the later stage of fruit development. But there was no significant difference in POD activity between the two lines during fruit ripening. Therefore, POD might not be the main reason for the difference of fruit flavor between the two lines during fruit ripening, but the difference and influence of POD during fruit development need further to study. The PPO activity in C. album‘Makeng 22'peaked at the later stage of maturation and was significantly higher than that in C. album‘Tantou 23'(p < 0.05). The PPO activity also decreased as the decrease of polyphenol content. During the olive fruit development and maturation,the pericarp is green, and chlorophyll in the pericarp can be photosynthesized to accumulate nutrients.Therefore, PPO is not only related to polyphenol decomposition, but also may promote polyphenol accumulation by promoting metabolic activity.【Conclusion】The difference of total phenols and polyphenol components result in different bitterness and astringency of common olive and the new selection, and then affect the quality of fresh eating. The difference of ellagic acid, methyl gallate and hyperoside might lead to different taste of fresh-eating Chinese olive. And the coordinated regulation of PPO, C4 H and4 CL results in the difference of the formation of phenols in the two lines of Chinese olive.
引文
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[2] CHANG Q,SU M H,CHEN Q X,ZENG B Y,LI H H,WANG W. Physicochemical properties and antioxidant capacity of Chinese olive(Canarium album L.)cultivars[J]. Journal of Food Science,2017,82(6):1369-1377.
[3] HE Z Y,XIA W S,CHEN J. Isolation and structure elucidation of phenolic compounds in Chinese olive(Canarium album L.)fruit[J]. European Food Research and Technology,2008,226(5):1191-1196.
[4]林玉芳,杜正花,陈清西.橄榄果实品质评价因子的筛选及指标确定[J].热带作物学报,2014,35(4):805-810.LIN Yufang,DU Zhenghua,CHEN Qingxi. Selection of quality evaluation indices for Chinese olive[J]. Chinese Journal of Tropical Crops,2014,35(4):805-810.
[5]池毓斌,朱丽娟,黄敏杰,彭真汾,叶清华,张静芳,陈清西,许长同.鲜食橄榄品质综合评价模型的建立与验证[J].果树学报,2017,34(8):1051-1060.CHI Yubin,ZHU Lijuan,HUANG Minjie,PENG Zhenfen,YE Qinghua,ZHANG Jingfang,CHEN Qingxi,XU Changtong. Establishment and verification of a comprehensive evaluation model for quality of fresh Chinese olive[J]. Journal of Fruit Science,2017,34(8):1051-1060.
[6]林玉芳,谢倩,陈清西.橄榄多酚类化合物组分分析[J].热带作物学报,2014,35(3):460-465.LIN Yufang,XIE Qian,CHEN Qingxi. Polyphenols of Chinese olive fruits[J]. Chinese Journal of Tropical Crops,2014,35(3):460-465.
[7]何志勇.橄榄酚类化合物的分离纯化和结构研究[D].无锡:江南大学,2007.HE Zhiyong. Separation,purification and structure elucidation of phenolic compounds from Canarium Album L.[D]. Wuxi:Jiangnan University,2007.
[8]陈岗.橄榄中多酚类物质的分离纯化及功能性研究[D].重庆:西南大学,2009.CHEN Gang. Studies on separation,purification and functionalities of polyphenols from Canarium Album[D]. Chongqing:Southwest University,2009.
[9]林玉芳,陈清西,关夏玉,陈明贤,欧高政.橄榄总多酚提取工艺优化研究[J].中国农学通报,2011,27(5):396-400.LIN Yufang,CHEN Qingxi,GUAN Xiayu,CHEN Mingxian,OU Gaozheng. Extraction of total polyphenol from Chinese olive(Canarium ablum L.)[J]. Chinese Agricultural Science Bulletin,2011,27(5):396-400.
[10]谢倩,王威,陈清西.橄榄多酚含量测定方法的比较[J].食品科学,2014,35(8):204-207.XIE Qian,WANG Wei,CHEN Qingxi. Comparative study on three different methods for the determination of total phenolics in Chinese olive[J]. Food Science,2014,35(8):204-207.
[11]陈建业.葡萄酒中酚酸及葡萄果实苯丙烷类代谢途径研究[D].北京:中国农业大学,2005.CHEN Jianye. Study on the phenolic acids in wines and phenylpropanoid metabolism in grape berries[D]. Beijing:China Agricultural University,2005.
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