Citation:[Copy]
[Copy]
【Print page】 【Online reading】【PDF Full text】 查看/发表评论下载PDF阅读器关闭

←Previous page|Page Next →

Back Issue    Advanced search

This Paper:Browse 129   Download 154 本文二维码信息
码上扫一扫!
利用CRISPR/Cas9技术创制高油酸甘蓝型油菜新种质
高谢旺, 谭安琪, 胡信畅, 祝孟洋, 阮颖, 刘春林
0
(植物表观遗传和分子生物学湖南省重点实验室)
摘要:
在低硫苷低芥酸的双低甘蓝型油菜基础上,培育稳定的高油酸油菜是当前重要的育种目标之一,本研究通过利用 CRISPR/Cas9 基因编辑对湖南农业大学选育的品种 XY15 进行品种改良。BnaFAD2 和 BnaFAE1 是油酸合成不饱和脂肪酸和超长 链脂肪酸的两个关键基因。利用 CRISPR/Cas9 基因编辑系统与高效油菜转化技术相结合,对甘蓝型油菜湘油 15 号(XY15) BnaFAD2.A05、BnaFAD2.C05 、BnaFAE1.A08 和 BnaFAE1.C03 同时进行基因编辑,获得了 7 株 T0 代转基因植株,通过靶基因 克隆测序,发现均未发生基因编辑。然而,在 T1 代植株中发现并检测到了基因编辑的发生,BnaFAD2.A05、BnaFAD2.C05 、 BnaFAE1.A08 和 BnaFAE1.C03 的总基因编辑效率分别为 38.89%,27.78%,22 .22%,30.56%。在 T2 代植株中,筛选到了 5 株 4 个靶基因均发生了突变,且无外源基因的突变体材料 Cas9-1-12-4、Cas9-1-12-5、Cas9-2-7-11、Cas9-5-5-3、Cas9-6-6-1,它们的 相对油酸含量为 68.69%、80.16%、75.82%、75.97%、74.37%,相比于野生型 XY15(相对油酸含量为 62.04%)均有显著提高。
关键词:  甘蓝型油菜  BnaFAD2  BnaFAE1  CRISPR/Cas9  基因编辑  高油酸种质
DOI:10.13430/j.cnki.jpgr.20191211001
投稿时间:2019-12-11修订日期:2020-03-07
基金项目:湖南省教育厅重点资助科研项目
Creation of New Germplasm of High-Oleic Rapeseed Using CRISPR/Cas9
GAO Xie-wang, TAN An-qi, HU Xin-chang, ZHU Meng-yang, RUAN Ying, LIU Chun-lin
(Hunan Provincial Key Laboratory of Crop Epigenetic Regulation and Development)
Abstract:
On the basis of low-glucosinolate and low-erucic rapeseed (Brassica napus), stable high-oleic rapeseed is one of the most important breeding objectives. In this study, CRISPR/cas9 system was used to improve the variety XY15 selected by Hunan Agricultural University. BnaFAD2 and BnaFAE1 are two key genes for the conversion of oleic acid to unsaturated fatty acids and ultra-long-chain fatty acids. The CRISPR/Cas9 system was introduced into XY15 by using highly efficient rapeseed transformation technology. The results of target gene cloning and sequencing showed that none of the seven T0 transgenic plants obtained were edited. However, gene editing was found and detected in T1 plants. The total gene editing efficiency of BnaFAD2.A05, BnaFAD2.C05, BnaFAE1.A08 and BnaFAE1.C03 were 38.89%, 27.78%, 22.22% and 30.56%, respectively. Among the T2 plants, 5 plants and 4 target genes were all mutated, and the relative oleic acid content of the mutants Cas9-1-12-4, Cas9-1-12-5, Cas9-2-7-11, Cas9-5-5-3 and Cas9-6-6-1 without foreign genes was 68.69%, 80.16%, 75.82%, 75.97% and 74.37%, respectively, which were significantly higher than that of the wild type XY15 (62.04%).
Key words:  Brassica napus  BnaFAD2  BnaFAE1  CRISPR/Cas9  gene editing  high-oleic germplasm