生物技术通报2024，Vol.40Issue(2)：245-252,8.DOI:10.13560/j.cnki.biotech.bull.1985.2022-1508
日本落叶松LkF3H2基因克隆及调控类黄酮代谢功能研究
Cloning of the LkF3H2 Gene in Larix kaempferi and Its Function in Regulating Flavonoid Metabolism
摘要
Abstract
[Objective]The Larix kaempferi flavanone 3-hydroxylase 2(LkF3H2)gene in Larix kaempferi plays a crucial role in flavonoid biosynthesis.This study is aimed to investigate the function of the LkF3H2 in L.kaempferi and plant flavonoid biometabolism.[Method]The LkF3H2 gene of L.kaempferi was cloned based on preliminary transcriptome data,and bioinformatics analysis and tissue expression analysis were performed.The gene was then stably transformed into tobacco to examine the relationship between gene expression and flavonoid content using tissue expression analysis and flavonoid content determination.[Result]The cDNA sequence length of LkF3H2 gene is 1074 bp,and its protein encodes 358 amino acids with a molecular formula of C1785H2807N481O541S18 and a molecular weight of 40.24 kD,indicating that it is an unstable hydrophilic protein lacking a signal peptide.Moreover,phylogenetic analysis revealed that LkF3H2 is closely related to the F3H genes of Pinus taeda,Pinus radiata,Picea abies,and Picea glauca.In addition,the LkF3H2 gene had the highest expression in the leaves of one-month-old L.kaempferi seedlings and the stems of one-year L.kaempferi seedlings,as determined by tissue expression analysis.Moreover,it had the highest expression in the transgenic tobacco leaves;notably,transgenic tobacco leaves had the highest flavonoid content,followed by stems and roots,indicating a positive relationship between flavonoid content and LkF3H2 gene expression in transgenic tobacco.[Conclusion]The LkF3H2 gene of L.kaempfer belongs to the 2-oxoglutarate-dependent dioxygenase family and is pivotal in flavonoid biosynthesis.关键词
黄烷酮3-羟化酶基因/转基因烟草/组织表达/类黄酮Key words
flavanone 3-hydroxylase gene/transgenic tobacco/tissue expression/flavonoids引用本文复制引用
李灿,蒋湘宁,盖颖..日本落叶松LkF3H2基因克隆及调控类黄酮代谢功能研究[J].生物技术通报,2024,40(2):245-252,8.基金项目
国家重点研发计划(2022YFD2200100) （2022YFD2200100）
