摘要
[目的]本文旨在通过测定梅和杏果实不同发育时期有机酸含量、相关酶活性以及有机酸代谢酶基因表达量,分析2类果实有机酸生物合成与代谢的关键基因,进一步解析梅和杏果实有机酸含量差异形成机制。[方法]以梅品种‘养老’和‘丰后’以及杏品种‘金太阳’的果实为材料,首先利用高效液相色谱法(HPLC)测定不同生长期果实中有机酸组分和含量,然后用紫外分光光度计测定有机酸代谢相关酶的活性,最后用实时荧光定量PCR测定有机酸代谢酶关键基因在果实各个发育阶段的表达水平。[结果]梅和杏果实中积累的主要有机酸均为柠檬酸和苹果酸,且相关代谢关键酶活性存在显著差异。相关性分析表明:在梅和杏果实发育过程中,柠檬酸合成酶(CS)活性与柠檬酸含量极显著正相关;磷酸丙醇式丙酮酸羧化酶(PEPC)和NADP-苹果酸酶(NADP-ME)活性与苹果酸含量均具有显著相关性。荧光定量PCR分析结果表明:在梅果实中发现CS基因表达量与柠檬酸含量有显著的相关性;在梅和杏果实中同时发现ME基因表达量与苹果酸含量有显著相关性。[结论]梅和杏果实中有机酸含量差异主要是由CS、PEPC和ME活性不同引起的,在梅果实中发现CS和ME基因是有机酸合成与代谢的关键基因,在杏果实中发现ME基因是有机酸代谢的关键基因。
[Objectives]The aim of this research is to determine the content of organic acids,the activities of related enzymes and the gene expression of organic acid metabolizing enzymes in fruits of Prunus mume and Prunus armeniaca at different developmental stages.We analyze the key genes of organic acid biosynthesis and metabolism and further analyze the formation mechanism of organic acid content difference in fruits of P.mume and P.armeniaca.[Methods]The fruits of P.mume cultivar of‘Yourou’and‘Bungo’and P.armeniaca cultivar of‘Jintaiyang’were used as materials,we used high performance liquid chromatography(HPLC)to measure the components and contents of organic acids in different growth periods of fruit firstly and then use ultraviolet spectrophotometer to measure the related enzyme activity of the organic acid.Finally,we use real-time fluorescent quantitative PCR to find out the expression level of key genes of organic acid metabolizing enzymes in different developmental stages of fruit.[Results]The main organic acids accumulated in fruits of P.mume and P.armeniaca were citric acid and malic acid,and the activities of key enzymes related to metabolism were significantly different.We concluded from the correlation analysis that citrate synthase(CS)activity was positively correlated with citric acid content,phosphoenolpyruvate carboxylase(PEPC)and NADP-malic enzyme(NADP-ME)activities were significantly correlated with malic acid content in fruits of P.mume and P.armeniaca.Fluorescence quantitative PCR analysis results showed that the expression of CS gene was significantly correlated with citric acid content in P.mume fruit.The expression of ME gene was significantly correlated with malic acid content in fruits of P.mume and P.armeniaca.[Conclusions]The difference of organic acid content in fruits of P.mume and P.armeniaca is mainly caused by different enzyme activity of CS,PEPC and ME.We found that CS and ME are the key genes in the acid synthesis and metabolism in P.mume fruit,and ME is the key gene in the organic acid metabolism in P.armeniaca fruit.
作者
翁金洋
薛松
倪照君
阎依超
沈志军
高志红
WENG Jinyang;XUE Song;NI Zhaojun;YAN Yichao;SHEN Zhijun;GAO Zhihong(College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China;Institute of Pomology,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2018年第6期1009-1017,共9页
Journal of Nanjing Agricultural University
基金
农业部物种资源保护项目(1120162130135252032)
江苏省六大人才高峰项目(NY068)
关键词
梅
杏
有机酸
代谢
关键基因
Prunus mume
Prunus armeniaca
organic acid
metabolism
key gene
