CRISPR/Cas9 gene editing and natural variation analysis demonstrate the potential for HvARE1 in improvement of nitrogen use efficiency in barley

作     者：Sakura D.Karunarathne Yong Han Xiao-Qi Zhang Chengdao Li Sakura D.Karunarathne;Yong Han;Xiao-Qi Zhang;Chengdao Li

作者机构：Western Crop Genetics AllianceCollege of ScienceHealthEngineering and EducationMurdoch UniversityPerthWA6150 Australia Western Australian State Agricultural Biotechnology CentreMurdoch UniversityPerthWA6150 Australia Department of Primary Industries and Regional DevelopmentPerthWA6151 Australia 

出 版 物：《Journal of Integrative Plant Biology》 (植物学报（英文版）)

年 卷 期：2022年第64卷第3期

页      面：756-770页

核心收录：

学科分类：0710[理学-生物学] 09[农学] 0901[农学-作物学] 

基　　金：generous support of Western Crop Genetics Alliance,Murdoch University,Western Australia awarded a Murdoch International Postgraduate Scholarship。 

主　　题：ARE1 barley CRISPR/Cas9 gene editing leaf senescence NUE 

摘      要：Nitrogen is a major determinant of grain yield and quality.As excessive use of nitrogen fertilizer leads to environmental pollution and high production costs,improving nitrogen use efficiency(NUE)is fundamental for a sustainable agriculture.Here,we dissected the role of the barley abnormal cytokinin response1 repressor 1(Hv ARE1)gene,a candidate for involvement in NUE previously identified in a genome-wide association study,through natural variation analysis and clustered regularly interspacedshort palindromic repeats(CRISPR)/CRISPRassociated protein 9(Cas9)-mediated gene editing.Hv ARE1 was predominantly expressed in leaves and shoots,with very low expression in roots under low nitrogen conditions.Agrobacterium-mediated genetic transformation of immature embryos(cv.Golden Promise)with single guide RNAs targeting Hv ARE1 generated 22 T0 plants,from which four T1 lines harbored missense and/or frameshift mutations based on genotyping.Mutant are1 lines exhibited an increase in plant height,tiller number,grain protein content,and yield.Moreover,we observed a 1.5-to2.8-fold increase in total chlorophyll content in the flag leaf at the grain filling stage.Delayed senescence by 10–14 d was also observed in mutant lines.Barley are1 mutants had high nitrogen content in shoots under low nitrogen conditions.These findings demonstrate the potential of ARE1 in NUE improvement in barley.