期刊论文

Using CRISPR/Cas9-Mediated Targeted Mutagenesis of qSH1 Reduces the Seed Shattering in Rice

中文

中国农业科学

0578-1752

2018

51

14

2631-2641

10.3864/j.issn.0578-1752.2018.14.001

31771767:国家自然科学基金 2016ZX08001-001:抗虫转基因水稻新品种培育项目 2014ZX0801001B-002:转基因生物新品种培育科技重大专项 TK1711793:香港中文大学合作项目 2017JJ3166:湖南省自然科学基金青年项目

本校为第一机构

【目的】易落粒既不利于稻谷收获,也不适宜于水稻机械化生产。利用现阶段前沿的分子育种手段— —CRISPR/Cas9技术对水稻落粒性主效基因qSH1进行定点编辑,并调查分析易落粒性状的改良效果,为创制稳产和适合机械化生产的水稻新种质奠定材料基础和探索新途径。【方法】以qSH1为靶标基因,根据CRISPR/Cas9技术原理设计靶标位点。将所设计的靶点序列在水稻参考基因组中比对分析以排除非特异性靶位点,最终筛选出qSH1-T1和qSH1-T5靶标位点。化学合成靶位点寡核苷酸序列,退火后分别与pYLgRNA-U3、pYLgRNA-U6a载体连接构建U3-qSH1T5-gRNA、U6a-qSH1T1-gRNA表达盒,最后将2个gRNA表达盒同时连接至pYLCRISPR...

【Objective】Seed shattering is not good for seeds harvest,nor suitable for mechanized production of rice.In this study,using the latest molecular breeding method-CRISPR/Cas9 system to site-directed edit of the rice seed shattering gene qSH1 and evaluated the improvement effect,in order to lay materials foundation and explore new method for creating stable-yielding rice germplasm suitable for mechanized seed production.【Method】The sgRNA was designed based on the principle of CRISPR/Cas9 technology.After excluding non-specific target sites by ...