组蛋白去乙酰化酶抑制剂(HDACi)对小麦基因编辑效率的影响及转录组学分析  

Effect of Histone Deacetylase Inhibitor(HDACi)on CRISPR Editing Efficiency of Wheat and Transcriptomics Analysis

作　　者：代文双 刘会云[1] 杜庆国 邹枨[1] 王轲[1] DAI Wen-shuang;LIU Hui-yun;DU Qing-guo;ZOU Cheng;WANG Ke(Institute of Crop Science,Chinese Academy of Agricultural Sciences,Beijing 100081)

机构地区：[1]中国农业科学院作物科学研究所,北京100081

出　　处：《生物技术通报》2021年第1期2-14,共13页Biotechnology Bulletin

基　　金：国家自然科学基金面上项目(31871710)。

摘　　要：主要研究组蛋白去乙酰化酶抑制剂(HDACi)与染色质状态以及CRISPR/Cas9的编辑效率之间的关系。利用不同浓度的尼克酰胺(0,2.5和5 mmol/L)和丁酸钠(0,5和10 mmol/L)对小麦幼苗处理7 d和14 d,结果显示丁酸钠处理会抑制幼苗的生长,而尼克酰胺对幼苗影响较小。对尼克酰胺处理的小麦幼苗进行转录组测序,发现了一些有利于促进染色质状态开放的基因:6个甲基转移酶合成通路基因。此外对未发生编辑的TaAGO4a基因编辑转基因小麦材料的T2代进行尼克酰胺处理,结果显示5 mmol/L处理14 d时检测到1株3A和3B基因组均杂合编辑的植株,编辑效率从0提高到8.3%,其它处理组和对照组均没有检测到编辑。本研究证明尼克酰胺确实可以提高小麦基因编辑效率,为提高小麦基因编辑效率提供了一种新策略。This study focuses on the relationship between histone deacetylase inhibitors(HDACi)and chromatin status and CRISPR/Cas9 editing efficiency.Different concentrations of nicotinamide(0,2.5,and 5 mmol/L)and sodium butyrate(0,5,and 10 mmol/L)were used to treat wheat seedlings for 7 d and 14 d,and the results showed that sodium butyrate seriously inhibited wheat growth,however,nicotinamide had little effect on it.Moreover,transcriptome sequencing of wheat seedlings treated with nicotinamide revealed some genes which were conducive to promoting the opening of the chromatin state,including 6 methyltransferase synthesis pathway genes.In addition,the TaAGO4a transgenic wheat materials in T2 generation that had not been edited were treated with nicotinamide,and results showed that a heterozygous mutant plant with mutations in the homologous alleles on 3A and 3B chromosomes was detected in the 5 mmol/L treatment for 14 d,and the editing efficiency increased from 0 to 8.3%;there was no mutant found in other treatments or control group.This study proves that nicotinamide can indeed improve the editing efficiency of wheat genes as a new strategy.

关 键 词：小麦 组蛋白去乙酰化酶抑制剂 染色质状态 CRISPR/cas9 转录组 

分 类 号：Q943.2[生物学—植物学] S512.1[农业科学—作物学]