吉林农业大学学报2025，Vol.47Issue(6)：977-986,10.DOI:10.13327/j.jjlau.2022.1858
大豆GmFAD2基因家族鉴定及基于CRISPR/Cas9编辑载体的构建
Identification of Soybean GmFAD2 Gene Family and Construction of CRISPR/Cas9-based Editing Vector
摘要
Abstract
The GmFAD2 gene is a key gene that controls the conversion of oleic acid to linoleic acid and determines the content and proportion of polyunsaturated fatty acids in soybean seeds.In recent years,research on the GmFAD2 gene family has focused on preventing the conversion of oleic acid to linoleic acid catalyzed by soybean fatty acid dehydrogenase through gene editing techniques or muta-genesis breeding,thereby increasing the oleic acid content of soybean.In this study,we performed bioinformatics analysis of the soybean GmFAD2 gene family.Additionally,using CRISPR/Cas9 edit-ing technology,based on gene sequences retrieved from NCBI,we designed sgRNA sequences and performed target site analysis for five key enzyme genes with the CRISPR-P online tool:GmFAD2-1A(Glyma10g42470.1),GmFAD2-1B(Glyma20g24530.5),GmFAD2-2A(Glyma19g32930.1),GmFAD2-2B(Glyma19g32940.1)and GmFAD2-2C(Glyma03g30070.1).Using pBGK015 as the vector back-bone and GmU6 as the specific promoter,the five single gene editing vectors were constructed sepa-rately and the efficiency of the editing vectors was effectively evaluated with the help of Agrobacte-rium tumefaciens transformation technology.Bioinformatics analysis showed that the five key enzyme genes of the soybean GmFAD2 gene family were unevenly distributed on three different chromo-somes,encoding proteins predicted to be positioned in the endoplasmic reticulum and interacting with several related proteins involved in oil synthesis.The mutations included base substitutions,de-letions and insertions.This study has laid foundation for the subsequent CRISPR/Cas9-mediated ge-netic transformation of soybean and the acquisition of high oleic acid mutant germplasm resources.关键词
大豆/GmFAD2基因/CRISPR/Cas9/编辑载体/生物信息学分析Key words
soybean/GmFAD2 gene/CRISPR/Cas9/editing vector/bioinformatics analysis分类
农业科技引用本文复制引用
ZHOU Junming,LI Zeyuan,LI Mihan,LUAN Xinchao,WANG Lixue,LIU Yixuan,YAO Dan..大豆GmFAD2基因家族鉴定及基于CRISPR/Cas9编辑载体的构建[J].吉林农业大学学报,2025,47(6):977-986,10.基金项目
吉林省重点研发项目(20210202006NC) （20210202006NC）
