[1]张威,许文静,许亚男,等.基于CRISPR/Cas9基因编辑的高油酸大豆品系创制[J].江苏农业学报,2023,(02):321-327.[doi:doi:10.3969/j.issn.1000-4440.2023.02.003]
　ZHANG Wei,XU Wen-jing,XU Ya-nan,et al.Creation of high oleic acid soybean lines by CRISPR/Cas9[J].,2023,(02):321-327.[doi:doi:10.3969/j.issn.1000-4440.2023.02.003]
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基于CRISPR/Cas9基因编辑的高油酸大豆品系创制()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2023年02期
页码:
321-327
栏目:
遗传育种·生理生化
出版日期:
2023-04-30

文章信息/Info

Title:
Creation of high oleic acid soybean lines by CRISPR/Cas9
作者:
张威1; 许文静1; 2; 许亚男1; 3; 张红梅1; 刘晓庆1; 崔晓艳1; 陈新1; 陈华涛1
(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)
Author(s):
ZHANG Wei1; XU Wen-jing1; 2; XU Ya-nan1; 3; ZHANG Hong-mei1; LIU Xiao-qing1; CUI Xiao-yan1; CHEN Xin1; CHEN Hua-tao1
(1.Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2.College of Horticulture， Nanjing Agricultural University, Nanjing 210095, China;3.College of Life Science, Nanjing Agricultural University, Nanjing 210095, China)
关键词:
CRISPR/Cas9基因编辑; 大豆; 油酸; GmFAD2-1A; GmFAD2-1B
Keywords:
CRISPR/Cas9 gene editing; soybean; oleic acid; GmFAD2-1A; GmFAD2-1B
分类号:
S565.1
DOI:
doi:10.3969/j.issn.1000-4440.2023.02.003
文献标志码:
A
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T0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（T3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。
Abstract:
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 T0 transgenic lines were obtained after stable transformation. Among of them, 11 T0 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 T3 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.

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备注/Memo

备注/Memo:
收稿日期：2022-10-11 基金项目：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) 作者简介：张威（1990-），男，江苏徐州人，博士，助理研究员 主要从事大豆品质育种质研究。（E-mail）20220015@jaas.ac.cn 通讯作者：陈华涛, （E-mail） cht@jaas.ac.cn
更新日期/Last Update: 2023-05-12