基于广泛靶向代谢组学的桃芽抗寒代谢物的筛选与鉴定  

Screening and identification of several metabolites associated with cold resistance in peach buds based on widely targeted metabolomics

作　　者：刘苏宁 王力荣[1] 方伟超[1] 陈昌文[1] 王新卫[1] 李勇[1] 吴金龙 曹珂[1] LIU Suning;WANG Lirong;FANG Weichao;CHEN Changwen;WANG Xinwei;LI Yong;WU Jinlong;CAO Ke(Zhengzhou Fruit Research Institute,Chinese Academy of Agriculture Sciences,Zhengzhou 450009,Henan,China)

机构地区：[1]中国农业科学院郑州果树研究所,郑州450009

出　　处：《果树学报》2023年第1期1-12,共12页Journal of Fruit Science

基　　金：国家重点研发计划项目(2019YFD1000200)。

摘　　要：【目的】筛选与鉴定参与调控桃芽抗寒能力的代谢物质,以深入了解桃抗寒性形成的生理生化机制。【方法】采用超高效液相色谱串联质谱(UPLC-MS/MS)技术,-14℃低温处理中农金辉桃枝条,分别在胁迫处理后0、24和72 h时取芽,检测上述材料中相关代谢物质的变化,从中选取差异显著的代谢物质配制标准品溶液,外源喷施至-4℃低温处理的2年生盆栽桃苗,以喷施清水为对照,测定植株抗寒相关指标脯氨酸含量、脯氨酸合成关键基因PpP5CS和桃上已知的低温响应基因PpCBF2的表达量。【结果】经-14℃低温处理后,在桃芽中共检测到172种已知代谢物质,其中差异显著的物质达到32种,从中选择4种差异显著代谢物质——黄嘌呤核苷、腺苷、DL-同型半胱氨酸以及氨基嘌呤的标准品配制溶液用于体外验证试验。体外验证试验表明,与清水对照组相比,处理组的脯氨酸含量均呈上升趋势,其中又以黄嘌呤核苷处理后的样品上升最为明显。qPCR试验结果表明,低温处理后,低温响应基因PpCBF2受到诱导。同时,与喷施清水的对照组相比,经不同代谢物标准品溶液处理的材料中脯氨酸合成关键基因PpP5CS相对表达量上升,与脯氨酸含量的表型变化一致。【结论】利用广泛靶向代谢组学分析从-14℃低温处理的桃芽中鉴定出172种代谢物,体外验证试验表明,黄嘌呤核苷、腺苷、DL-同型半胱氨酸以及氨基嘌呤与桃芽的抗寒能力相关。【Objective】Peach has a long history of cultivation and wide distribution in China.Temperature is the main factor affecting the growth and development of peach trees as well as determining their natural distribution,so it is of great theoretical and practical significance to clarify its physiological and biochemical mechanisms in response to low temperature for breeding and screening peach cold-resistant varieties.Metabolomics,as important components of systems biology,can reveal differences in metabolite profiles among different species,between different tissues of the same species,and between the same tissues of the same species under different conditions of adversity stress.The aim of this study was to investigate the metabolites that would affect the cold resistance of peach buds and to facilitate a deeper understanding of the metabolic pathways of cold resistance in peach.【Methods】The material used for the cold treatment to identify the differential metabolites was 6-year-old Zhongnong Jinhui,and the annual branches were taken from the upper and middle periphery of the tree,cut into 30 cm segments,wrapped in plastic wrap and placed in the refrigerator for stress treatment.All the samples needed for the experiment were put in at once,and the time of starting the treatment was recorded.The branches were removed at 0 h,24 h and 72 h,and the leaves on the branches were removed before the branches were thawed,and the peach buds of the same size and without mechanical damage on the branches were selected and peeled out with forceps and dissecting needles and then were quickly put into liquid nitrogen.metabolite assay.Four significantly different metabolites,aminopurine,adenosine,xanthine nucleoside and DL-homocysteine were selected from the metabolome assay,and the standards were prepared in distilled water at different concentrations.Fifteen uniformly growing Zhongtao Kangzhen 1 rootstock plants were treated at-4℃in a light incubator for 16 h of light and 8 h of darkness.The seedlings were sprayed with d

关 键 词：桃芽 抗寒性 差异代谢物 代谢组 

分 类 号：S662.1[农业科学—果树学]