摘要
目的:克隆人参Panax ginseng独脚金内酯酶基因PgSL并进行生物信息学分析,确定PgSL基因在缺氮、缺磷及独脚金内酯类似物GR24处理下的表达水平。方法:根据高通量测序获得人参PgSL基因CDS序列,利用同源重组和golden gate无缝克隆的方法,扩增PgSL基因CDS序列,并将其编码的氨基酸序列提交至NCBI、Protparam、SOPMA等网站和软件进行生物信息学分析;利用荧光定量PCR(qRT-PCR)检测缺氮、缺磷及外源GR24处理下人参根部组织中PgSL基因的表达水平。结果:人参PgSL基因的CDS序列为816 bp,编码271个氨基酸;PgSL蛋白属于MhpC家族蛋白,该蛋白相对稳定,属于疏水性蛋白,α螺旋为主要组成部分;PgSL蛋白与琐氏独活Heracleum sosnowskyi、核桃树Juglans regia、木本棉Gossypium arboreum中的独脚金内酯酶具有较近的系统发育关系;与对照组相比,缺氮处理下PgSL基因的表达量升高,当施用外源GR24后,PgSL基因的表达量降低;缺磷处理下PgSL基因的表达量无显著变化,当施用外源GR24后,PgSL基因的表达量上升;同时缺氮缺磷处理下PgSL基因的表达量降低,当施用外源GR24后,PgSL基因的表达量升高。结论:克隆获得PgSL基因的CDS序列及表达信息,可为后续进行基因功能研究和人参栽培的氮磷营养调控提供参考。
Objective:To clone the strigolactone esterase gene PgSL from Panax ginseng,conduct bioinformatics analysis,and determine the expression level of PgSL gene under nitrogen deficiency,phosphorus deficiency,and treatment with the synthetic strigolactone analog GR24.Methods:Based on the CDS sequence of Panax ginseng PgSL gene obtained by high-throughput sequencing,the CDS sequence of PgSL gene was amplified by homologous recombination and golden gate seamless cloning method,and the encoded amino acid sequence was submitted to NCBI,Protparam,SOPMA and other websites or software for bioinformation analysis.Real-time fluorescent quantitative PCR(qRT-PCR)was used to detect the expression level of PgSL gene in root tissues of Panax ginseng treated with nitrogen deficiency,phosphorus deficiency and exogenous GR24.Results:The CDS sequence of Panax ginseng PgSL gene was 816 bp,encoding 271 amino acids.PgSL protein belonged to MhpC protein,which was relatively stable and belonged to hydrophobic protein,αhelix was the main component.There was a close phylogenetic relationship between PgSL protein and strigolactone esterase in Heracleum sosnowskyi,Juglans regia and Gossypium arboreum.In comparison to the control group,upregulation of the PgSL gene was observed in response to nitrogen deficiency treatment,whereas its expression decreased upon application of exogenous GR24.Conversely,no notable alteration was detected in the expression of the PgSL gene following phosphorus deficiency treatment,however,an increase in PgSL gene expression was noted following exogenous GR24 treatment.Moreover,under conditions of nitrogen and phosphorus deficiency treatment,a decrease in PgSL gene expression was observed,which was subsequently reversed to an increase upon application of exogenous GR24.Conclusion:The CDS sequence and expression information of PgSL gene are obtained by cloning,by cloning which can provide reference for the subsequent gene function study,and the regulation of nitrogen and phosphorus nutrition in Panax ginseng cultivation.
作者
梁浩
孙海
邵财
周季欣
吕柏辰
朱家鹏
张亚玉
LIANG Hao;SUN Hai;SHAO Cai;ZHOU Ji-xin;LYU Bo-chen;ZHU Jia-peng;ZHANG Ya-yu(Institute of Special Animal and Plant Sciences,Chinese Academy of Agricultural Sciences,Changchun 130112,China;College of Food and Biological Engineering,Chengdu University,Chengdu 610106,China)
出处
《中药材》
北大核心
2024年第6期1343-1350,共8页
Journal of Chinese Medicinal Materials
基金
国家现代农业产业技术体系项目(CARS-21)
中国农业科学院科技创新工程项目(CAAS-ASTIP-2021-ISAPS)。
关键词
人参
独脚金内酯
氮和磷
基因表达分析
Panax ginseng C.A.Mey.
Strigolactone
Nitrogen and phosphorus
Gene expression analysis
