枸杞果实发育期酶活性、基因表达与类黄酮积累的相关性分析  

Correlation of Enzyme Activity,Gene Expression and Flavonoid Accumulation in Lycium chinense Fruit Development

作　　者：张凯敏 耿贵工[2] 乔枫 Zhang Kaimin;Geng Guigong;Qiao Feng(Key Laboratory of Medicinal Animal and Plant Resources in Qinghai Tibet Plateau,College of Life Sciences,Qinghai Normal University,Xining,810008;Academy of Agricultural and Forestry Sciences,Qinghai University,Xining,810016)

机构地区：[1]青海师范大学生命科学学院,青藏高原药用动植物资源重点实验室,西宁810008 [2]青海大学农林科学院,西宁810016

出　　处：《分子植物育种》2024年第22期7515-7524,共10页Molecular Plant Breeding

基　　金：青海省科技厅项目(2021-ZJ-729);青海省青藏高原药用动植物资源重点实验室(2020-ZJ-Y40);农业农村部农业资源环境保护项目(125A0605)共同资助。

摘　　要：为了解枸杞果实发育期类黄酮合成途径中关键酶及基因表达的变化以及与类黄酮积累的相关性,本研究以枸杞发育期果实为试验材料,通过酶联免疫法测定了枸杞果实发育期类黄酮合成途径中10个关键酶活性:苯丙氨酸解氨酶(PAL)、肉桂酸-4-羟化酶(C4H)、4-香豆酸辅酶A连接酶(4CL)、查尔酮合成酶(CHS)、查尔酮异构酶(CHI)、黄酮3-羟化酶(F3H)、类黄酮3'-羟化酶(F3'H)、黄酮醇合酶(FLS)、类黄酮3'5'-羟化酶(F3'5'H)、花青素合成酶(ANS),通过紫外分光光度法测定了枸杞果实类黄酮含量;采用RT-qPCR法测定了其对应基因的相对表达量。结果显示:类黄酮含量呈先下降后上升的趋势,PAL活性呈先不变后下降的趋势,F3'H活性呈先不变后上升的趋势;CHS、CHI、F3H和F3'5'H活性呈现先上升后下降的趋势;4CL、ANS活性呈现先下降后上升的趋势;FLS呈一直上升的趋势。各对应基因的表达量与酶活性的趋势相似,其中4CL为类黄酮合成途径的关键枢纽,且4CL基因表达量与类黄酮的含量呈显著正相关(P<0.01)关系。本研究结果补充了枸杞果实发育期类黄酮积累研究的空白,为枸杞类黄酮代谢提供了理论基础。In order to understand the changes of key enzymes and gene expression, and the correlation with flavonoid accumulation in flavonoid synthesis pathway during Lycium chinense fruit development, in this study,the fruit of L. chinense in the development stage was used as the experimental material, the activities of 10 key enzymes in the flavonoids synthesis pathway of L. chinense fruit during the development period were determined by ELISA, including phenylalanine ammonia lyase(PAL), cinnamate 4-hydroxylase(C4H), 4-coumaric acid coenzyme A ligase(4CL), chalcone synthase(CHS), chalcone isomerase(CHI), flavonoids 3-hydroxylase(F3H), flavonoids3'-hydroxylase(F3'H), flavonol synthase(FLS), flavonoid 3'5'-hydroxylase(F3'5'H) and anthocyanin synthetase(ANS). The content of flavonoids in L. chinense fruit was determined by ultraviolet spectrophotometry. The relative expression of its corresponding genes was determined by RT-qPCR. The results showed that the content of flavonoids decreased first and then increased, PAL remained unchanged first and then decreased, and F3'H remained unchanged first and then increased;CHS, CHI, F3H and F3'5'H increased first and then decreased;4CL and ANS de creased first and then increased;FLS has been rising. The expression amount of each corresponding gene was similar to the trend of enzyme activity. Among them, 4CL was the key hub of flavonoid synthesis pathway, and the expression of 4CL gene had a significant positive correlation with flavonoid content(P<0.01). The results of this study supplement the blank of research on flavonoid accumulation during the development of Lycium barbarum fruit, and provide a theoretical basis for flavonoids metabolism of L. chinense.

关 键 词：枸杞果实 发育时期 类黄酮合成 RT-QPCR 代谢酶 

分 类 号：S567.19[农业科学—中草药栽培]