BSMV-sg系统介导的大麦基因编辑体系的建立  

Heritable Genome Editing in Barley Using BSMV-sg System

作　　者：刘永菊 姜燕燕 薛瑞莹 刘光瑞 曹东[2] 刘宝龙[1,2,3] 李云[2] LIU Yongju;JIANG Yanyan;XUE Ruiying;LIU Guangrui;CAO Dong;LIU Baolong;LI Yun(School of Life Sciences,Qinghai Normal University,Xining,Qinghai 810008,China;Qinghai Provincial Key Laboratory of Crop Molecular Breeding,Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining,Qinghai 810008,China;Academy of Agriculture and Forestry Sciences,Qinghai University,Xining,Qinghai 810008,China)

机构地区：[1]青海师范大学生命科学学院,青海西宁810008 [2]中国科学院西北高原生物研究所青海省作物分子育种重点实验室,青海西宁810008 [3]青海大学农林科学院,青海西宁810008

出　　处：《麦类作物学报》2024年第3期281-288,共8页Journal of Triticeae Crops

基　　金：青海省科技厅基础研究计划-青年项目(2021-ZJ-958Q)。

摘　　要：基因编辑元件递送系统是基因编辑技术在大麦中广泛应用的制约因素之一,建立无需遗传转化的基因编辑元件递送系统,对于进一步提高大麦基因编辑技术的利用效率,加快大麦育种进程具有重要意义。本研究利用大麦条纹花叶病毒(BSMV)作为八氢番茄红素脱氢酶基因PDS的sgRNA的递送系统,侵染烟草并接种大麦Cas9过表达株系的叶片,分析了大麦叶片中BSMV的侵染效果和PDS基因的编辑效率。结果表明,接种大麦植株的主茎新生叶片和分蘖叶片均含有BSMV,BSMV-sg能够在大麦体内复制并移动。接种大麦植株的主茎新生叶片和分蘖叶片呈现不同程度的白化表型,15份叶片的PDS基因在靶点区域存在着1个或多个变异,包括碱基的插入、缺失或替换,为体细胞编辑,第3分蘖具有更高比例的PDS等位变异。在150个M1子代植株中,筛选到1株纯合编辑植株。本研究在大麦中建立了基于BSMV的可遗传的基因编辑系统,为大麦基因组编辑提供了一种方便、高效的方法。The delivery system for gene editing elements is a limiting factor in the widespread application of gene editing technology in barley.Developing a non-genetic transformation-based delivery system for gene editing elements holds great significance in enhancing the efficiency of barley gene editing technology and expediting the breeding process.In this study,barley striped mosaic virus(BSMV)was used as the sgRNA delivery system of the phytoene dehydrogenase PDS gene to infect tobacco and then inoculated the leaves of barley Cas9 overexpressed strains,and the infection effect of BSMV and the editing efficiency of PDS gene in barley leaves was analyzed.The presence of BSMV was detected in the young leaves of the main stem and tillering leaves of inoculated barley plants,indicating successful replication and movement of BSMV-sg within the barley plant.The newly emerged leaves on both the main stem and tillers exhibited varying degrees of albinism phenotype,with 15 PDS gene-containing leaves showing somatic editing through base insertion,deletion or replacement in the target region.Notably,a higher proportion of PDS allelic variation was observed in the third tiller.From a total of 150 M 1 progeny plants,one homozygously edited plant was selected as an outcome of this study s establishment of a heritable gene editing system based on BSMV in barley.This method offers a convenient and efficient approach for genome editing in barley.

关 键 词：基因编辑 PDS基因 大麦条纹花叶病毒 大麦遗传转化 

分 类 号：S512.3[农业科学—作物学] S330