利用基因编辑技术研究BnaFZP基因调控油菜分枝的功能  

Regulation of shoot branching by BnaFZP in Brassica napus based on gene editing technology

作　　者：刘倩倩 申潇潇 李淮琳 余凯迪 范楚川[1,2] LIU Qian-qian;SHEN Xiao-xiao;LI Huai-lin;YU Kai-di;FAN Chu-chuan(College of Plant Science&Technology,Huazhong Agricultural University,Wuhan 430070,China;Hubei Hongshan Laboratory,Wuhan 430070,China;Xiangyang Academy of Agricultural Sciences,Xiangyang 441057,China)

机构地区：[1]华中农业大学植物科学技术学院,湖北武汉430070 [2]湖北洪山实验室,湖北武汉430070 [3]襄阳市农业科学院,湖北襄阳441057

出　　处：《中国油料作物学报》2024年第5期985-992,共8页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金(32172021,31971976)。

摘　　要：FRIZZY PANICLE(FZP)基因编码AP2/ERF类型的转录因子,在模式植物拟南芥中参与株型(分枝)调控,是植物株型改良的重要靶基因。本研究首次针对油菜中的同源基因BnaFZP开展了功能研究。通过基因克隆和生物信息学分析,发现BnaFZP在甘蓝型油菜基因组中有六个同源拷贝,它们均只有一个外显子,且在N端含有一个AP2保守结构域。组织表达特异性分析发现,BnaFZP在各个组织中的表达量都较低,其中根、花瓣和角果中的表达量相对较高。利用CRISPR/Cas9基因编辑技术技术和农杆菌介导的遗传转化方法,成功创制出BnaFZP基因不同拷贝发生编辑的48个T0植株。这些定点突变可以稳定遗传,并在后代筛选得到非转基因的编辑单株。对T1突变体进行表型观察发现,BnaFZP的BnaA02g35090D/BnaC02g08170D纯合双突变体和BnaC03g09100D/BnaA02g35090D/BnaC02g08170D纯合三突变体均表现出明显的多分枝表型,表明BnaFZP参与油菜分枝和株型调控。该研究为甘蓝型油菜株型与分枝调控的分子机理研究与株型改良提供了重要研究材料。The FRIZZYPANICLE(FZP)gene encodes the AP2/ERF transcription factor and is involved in plant architecture(shoot branching)regulation in the model plant Arabidopsis thaliana,which is an important target gene for the improvement of plant architecture in rapeseed.In the present study,we assessed the functions of rape-seed homologues of FZP gene in rapeseed at first time.According to the rapeseed bioinformatics analysis and gene cloning,we identified six copies of BnaFZP in Brassica napus genome.There is only an exon for all copies,which contains a conserved AP2-domain at N-terminal of their proteins.Gene expression analysis revealed that the BnaFZP expression levels were very low in different tissues,with a relative higher expression in roots,petals and si-liques.A total of 48 targeted mutants with loss-of-function alleles at different copies of BnaFZP gene were ob-tained at T0 generation using the CRISPR/Cas9 system and Agrobacterium-mediated transformation.The induced mutations were stably transmitted to successive generations,and a variety of BnaFZP homozygous T-DNA-free mu-tants were obtained in the T1 generation.Phenotypic observation of the obtained mutants showed that the homozy-gous BnaA02g35090D/BnaC02g08170D double mutants and homozygous BnaC03g09100D/BnaA02g35090D/BnaC02g08170D triple mutants presented significant much shoot branching,which indicted that BnaFZP is in-volved in the regulation of plant architecture.Collectively,the mutants generated in this study would provide valu-able resources for both basic studies and breeding programs.

关 键 词：甘蓝型油菜 BnaFZP AP2/ERF CRISPR/Cas9 株型 

分 类 号：S565.4[农业科学—作物学]