生物技术通报2024，Vol.40Issue(9)：11-19,9.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0520
CRISPR/Cas9编辑MeHNL基因创制低生氰糖苷木薯
CRISPR/Cas9 Editing MeHNL Gene to Generate Cassava Plants with Low Cyanogenic Glycoside
摘要
Abstract
[Objective]Because cassava(Manihot esculenta Crantz)contains potentially toxic cyanogenic glucosides,its food safety is affected,which leads to the increase of processing costs.It is of great significance to cultivate cassava with low cyanogenic glycosides by biotechnology.[Method]CRISPR/Cas9 technology was used to edit the α-hydroxynitrile lyase gene MeHNL.This gene encodes α-hydroxynitrile lyase that catalyzes the decomposition of cyanogenic glycosides.The editing target is located on its first exon,and 27 positive plants were obtained through Agrobacterium-mediated stable transformation.[Result]Sequencing analysis indicated successful editing in 26 out of 27 transgenic plants,and a high editing efficiency was achieved by 96.3%.The editing types predominantly comprised base insertions and deletions,alongside minor base substitutions and large fragment deletions.HCN colorimetric kit and HPLC analysis confirmed a significant reduction in hydrocyanic acid and cyanogenic glycosides in the mutant lines.Additionally,transgenic plants demonstrated slender leaves compared to non-transgenic counterparts,implicating MeHNL's impact on plant growth and development.[Conclusion]The cassava germplasm with low cyanide is obtained by CRISPR/Cas9 technology,which provides materials for exploring the study of cyanogenic glycoside metabolism on cassava growth and development.关键词
木薯/基因编辑/醇氰酶/低氰化物/生氰糖苷Key words
cassava/gene editing/HNL/low cyanide/cyanogenic glycosides引用本文复制引用
童玮婧,罗数,陆新露,沈建福,陆柏益,李开绵,马秋香,张鹏..CRISPR/Cas9编辑MeHNL基因创制低生氰糖苷木薯[J].生物技术通报,2024,40(9):11-19,9.基金项目
国家重点研发计划(2023YFD1600605-4),热带作物生物育种全国重点实验室 2023 年度开放课题项目(1630052024001),农业农村部国家现代农业产业技术体系项目(CARS-11) （2023YFD1600605-4）
