通过CRISPR/Cas9建立豌豆基因组大片段敲除体系  

Establishment of large fragment knockout in pea genome by CRISPR/Cas9 technology

作　　者：黄淑贤 刘荣[1] 李冠 疏琴 徐斐 宗绪晓[1] 杨涛[1] HUANG Shu-Xian;LIU Rong;LI Guan;SHU Qin;XU Fei;ZONG Xu-Xiao;YANG Tao(Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Institute of Crop Germplasm Resources,Shandong Academy of Agricultural Sciences,Jinan 250100,Shandong,China)

机构地区：[1]中国农业科学院作物科学研究所,北京100081 [2]山东省农业科学院农作物种质资源研究所,山东济南250100

出　　处：《作物学报》2024年第7期1658-1668,共11页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(32241042);财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-08-G11);作物基因资源与育种全国重点实验项目资助。

摘　　要：豌豆是世界上主要食用豆类作物之一,因其丰富的营养价值和良好的生态价值而受到越来越多的重视。CRISPR/Cas9作为一种生物育种的新型技术手段,已广泛应用于各种作物的品种改良中,但在豌豆中的应用报道非常少见。本研究以“中豌6号”品种为试验材料,采用CRISPR/Cas9技术手段成功地实现了豌豆基因组中Psat01G0240600-T1、Psat03G0303300-T1和Psat03G0304700-T1基因的大片段缺失,其大片段敲除效率分别是24.1%、13.0%和3.6%。进一步对结果分析发现,不同靶点的编辑效率相差较大,并且大片段缺失的效率取决于2个靶点中较低编辑效率的靶点。本研究利用发状根体系探索了CRISPR/Cas9在豌豆中的应用,首次在豌豆中实现了大片段敲除,对于豌豆的研究具有重要意义。Pea is one of the main food legumes in the world and has been paid more and more attention due to its rich nutritional value and good ecological value.CRISPR/Cas9 as a new biology breeding tool has been widely used in many crops,however,its usage in pea is very limited.In this study,we successfully achieved large fragment deletion of the Psat01G0240600-T1,Psat03G0303300-T1,and Psat03G0304700-T1 genes in“Zhongwan 6”variety by CRISPR/Cas9 technology.The large fragment knockout ratio was 24.1%,13.0%,and 3.6%,respectively.Further analysis revealed that there were significant differences in editing efficiency among different target sites and large fragment deletion depend on the specific target site with the lower editing efficiency between the two target sites.In this study,we explored the application of CRISPR/Cas9 by using the hairy root system in pea and achieved large fragment knockout for the first time in pea,which is of great significant for the research of peas.

关 键 词：豌豆 CRISPR/Cas9 发状根 大片段敲除 

分 类 号：S643.3[农业科学—蔬菜学]