基于CRISPR/Cas9基因编辑的高油酸大豆品系创制  

Creation of high oleic acid soybean lines by CRISPR/Cas9

作　　者：张威 许文静[1,2] 许亚男 张红梅 刘晓庆[1] 崔晓艳[1] 陈新[1] 陈华涛[1] ZHANG Wei;XU Wen-jing;XU Ya-nan;ZHANG Hong-mei;LIU Xiao-qing;CUI Xiao-yan;CHEN Xin;CHEN Hua-tao(Institute of Industrial Crops,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China;College of Life Science,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]江苏省农业科学院经济作物研究所,江苏南京210014 [2]南京农业大学园艺学院,江苏南京210095 [3]南京农业大学生命科学学院,江苏南京210095

出　　处：《江苏农业学报》2023年第2期321-327,共7页Jiangsu Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2018YFE0112200);江苏省重点研发计划(现代农业)项目(BE2019376);江苏省种业振兴“揭榜挂帅”项目[JBGS(2021)060];江苏省自然科学基金项目(BK20210154)。

摘　　要：高油酸能够显著提升大豆食用油的稳定性和食用价值,是大豆品质改良的育种目标之一。为提高大豆籽粒油酸含量,本研究采用CRISPR/Cas9多基因编辑技术,同时敲除负调控大豆油酸含量的2个关键基因GmFAD2-1A和GmFAD2-1B。采用农杆菌介导的大豆子叶节转化方法获得20株T_(0)代阳性苗,但发生基因编辑的株系只有11个,其中GmFAD2-1A发生突变的有7株,GmFAD2-1B发生突变的有10株,这2个基因同时发生突变的有6株。经过加代纯合后,检测转基因株系(T_(3)代)大豆籽粒的脂肪酸组分,发现油酸含量显著提升,最高达81.38%,而亚油酸含量则显著降低,获得了高油酸新种质。本研究采取多基因敲除策略,成功突变大豆基因GmFAD2-1A和GmFAD2-1B,并获得高油酸大豆突变体株系,为高油酸大豆育种提供了新的种质资源。Improving the oleic acid is one of the important objectives for soybean breeding and improvement and will improve the stability and edible value of soybean oil.In the present study,we applied CRISPR/Cas9 gene editing to knock out GmFAD2-1A and GmFAD2-1B,which negatively regulate oleic acid content in soybean.By Agrobacterium-mediated transformation using mature cotyledonary node explants,twenty CRISPR/Cas9 T_(0) transgenic lines were obtained after stable transformation.Among of them,11 T 0 transgenic lines were detected mutations in the GmFAD2-1A or GmFAD2-1B,including seven GmFAD2-1A transgenic lines,10 GmFAD2-1B transgenic lines,and six GmFAD2-1A/GmFAD2-1B transgenic lines.The fatty acid profile analysis of T_(3) soybean seeds revealed that the oleic acid content improved obviously,and reached 81.38%,whereas linoleic acid decreased.In this study,the CRISPR/Cas9 gene editing can successfully edit GmFAD2-1A and GmFAD2-1B,and create high oleic acid soybean germplasm,which provides new germplasm resources for the breeding of high oleic acid.

关 键 词：CRISPR/Cas9基因编辑 大豆 油酸 GmFAD2-1A GmFAD2-1B 

分 类 号：S565.1[农业科学—作物学]