作物学报2024，Vol.50Issue(1)：100-109,10.DOI:10.3724/SP.J.1006.2024.34056
利用CRISPR/Cas9技术编辑GmBADH1基因改变大豆耐盐性
Knockout of GmBADH1 gene using CRISPR/Cas9 technique to reduce salt tolerance in soybean
摘要
Abstract
Functional identification of salt-tolerant genes is crucial for the improvement of soybean varieties and the development and utilization of saline-alkali land.Under salt stress,crops reduce the effect of salt damage on crop yield by synthesizing and accumulating betaine as osmotic protective agent.BADH gene is been proved to regulate plant response to stress in many plants,but the regulation mechanism in soybean is not clear.In this study,GmBADH1 gene was cloned,and qRT-PCR showed that the gene was expressed stems and leaves,especially in roots.The expression vector constructed by CRISPR/Cas9 system was trans-ferred into soybean variety JACK by Agrobacterium-mediated soybean genetic transformation technology,and three targeted mu-tations that could regulate the salt tolerance of soybean were produced.They all occurred in the coding region,which had three difference of deletion 19 bp,insertion 1 bp,and insertion 9 bp to replace 2 bp.The first two,premature termination codons,re-sulting in truncated BADH1 protein,the latter occurred frameshift mutation.The mutant homozygous plants were treated with salt at the emergence stage and seedling stage,respectively.The results showed that the salt tolerance of the mutant homozygous plants was significantly lower than that of the wild type at seedling stage,and there was no significant difference compared with the wild type at seeding stage,which indicating that GmBADH1 gene may mainly regulate the salt tolerance of seedling stage.This study provides a basis for further excavation of soybean salt-tolerant genes and cultivation of soybean salt-tolerant varieties.关键词
大豆/GmBADH1/CRISPR/Cas9/耐盐性Key words
soybean/GmBADH1/CRISPR/Cas9/salt tolerance引用本文复制引用
石宇欣,刘欣玥,孙建强,李晓菲,郭潇阳,周雅,邱丽娟..利用CRISPR/Cas9技术编辑GmBADH1基因改变大豆耐盐性[J].作物学报,2024,50(1):100-109,10.基金项目
本研究由国家重点研发计划项目:"北方大豆高产优质耐密新种质创制与应用项目"(2021YFD1201104)资助. This study was supported by the National Key Research and Development Program of China:"the Development and Application of High Yield,High Quality and Dense Tolerance New Germplasm of Soybean in Northern China"(2021YFD1201104). （2021YFD1201104）
