利用CRISPR/Cas9技术编辑AFP1基因提高水稻耐逆性  

CRISPR/Cas9-mediated Editing of AFP1 Improves Rice Stress Tolerance

作　　者：周天顺 余东[1,3] 刘玲 欧阳宁 袁贵龙[1,3] 段美娟[4] 袁定阳[1,2,3] ZHOU Tianshun;YU Dong;LIU Ling;OUYANG Ning;YUAN Guilong;DUAN Meijuan;YUAN Dingyang(Hunan Hybrid Rice Research Center/State Key Laboratory of Hybrid Rice,Changsha 410125,China;Longping Branch,Graduate School of Hunan University,Changsha 410125,China;Hunan Academy of Agricultural Sciences,Changsha 410125,China;College of Agronomy,Hunan Agricultural University,Changsha 410126,China)

机构地区：[1]湖南杂交水稻研究中心/杂交水稻国家重点实验室,长沙410125 [2]湖南大学研究生院隆平分院,长沙410125 [3]湖南省农业科学院,长沙410125 [4]湖南农业大学农学院,长沙410126

出　　处：《中国水稻科学》2021年第1期11-18,共8页Chinese Journal of Rice Science

基　　金：国家重点研发计划资助项目(2016YFD0101100);湖南省自然科学基金资助项目(2019JJ40208);湖南省科技人才托举工程资助项目(2019TJ-Q08)。

摘　　要：【目的】为鉴定水稻AFP1在非生物胁迫响应中的作用,创制非生物胁迫抗性的水稻新材料。【方法】以优异籼稻恢复系华占为转化受体,利用CRISPR/Cas9技术创制afp1突变体,并对afp1突变体的耐逆性进行初步鉴定。【结果】AFP1靶点1和靶点2的编辑效率分别为66.67%和75.00%。所有突变株系中,突变类型仅有插入和缺失突变,90%突变株系的突变长度为小片段突变(<5 bp)。获得了6种无转基因成分的afp1纯合突变体。正常条件下,afp1突变体株高和结实率降低,有效分蘖增加,穗长显著升高,单株产量在-4.06%和11.75%之间变化。和野生型相比,afp1突变体的ABA敏感性和叶片水分散失率降低,耐干旱、热和渗透胁迫能力提高。【结论】编辑AFP1基因可提高水稻多种非生物胁迫抗性。【Objective】It is important to reveal the functions of rice AFP1 in response to abiotic stress,and create new stress-resistant rice germplasm in innovative ways.【Method】Excellent indica restoring line Huazhan was used as transgenic receptor material to generate afp1 mutants using CRISPR/Cas9 system,and abiotic stress tolerances of afp1 knockout lines were preliminary analyzed.【Results】The editing efficiency of AFP1 in two targets were 66.67%and 75.00%,respectively.In all mutant plants,only insertions and deletions occurred.About 90%of mutant bases were small fragments,with a length less than 5 bp.Six homozygous mutants without genetically modified ingredients were generated.Under normal conditions,plant height and seed setting rate of afp1 mutants were reduced,but effective tiller number per plant and panicle length were significantly increased.Interestingly,the yield per plant of mutants changed between-4.06%and+11.75%.Compared with the wild type,the ABA sensitivity and water loss rate of afp1 mutants were obviously decreased.Drought,heat and osmotic stress resistances were significantly improved.【Conclusion】Mutation of AFP1 gene could improve multiple abiotic stress resistances in rice.

关 键 词：水稻 CRISPR/Cas9基因编辑技术 AFP1 耐逆性 

分 类 号：Q943.2[生物学—植物学] S511[农业科学—作物学]