AtPHO2的CRISPR/Cas9靶向敲除及其在拟南芥中功能分析  

Gene knock out and functional analysis of AtPHO2 by CRISPR/Cas9-induced mutagenesis

作　　者：王钊辉 石超男 杨天啸[1] 薛亚东[1] 孙虎威[1] 唐贵良[1] 张战辉[1] 

机构地区：[1]河南农业大学农学院,河南郑州450002

出　　处：《河南农业大学学报》2017年第4期554-560,565,共8页Journal of Henan Agricultural University

基　　金：省部共建小麦玉米作物学国家重点实验室项目(39990002)

摘　　要：利用CRISPR/Cas9基因编辑技术对AtPHO2进行精确编辑,在拟南芥转基因后代中筛选到3个Atpho2阳性突变体。突变体中AtPHO2的表达量下降了10倍。对获得突变体的表型及无机磷含量变化进行分析,结果表明,突变体叶片衰老速度明显快于野生型,突变体叶片无机磷含量是野生型的2~4倍。随着衰老进程的推移,地上部新叶中无机磷含量逐渐增加,突变体中由于磷代谢途径受阻而大量积累。本研究也表明,AtPHO2不仅可能通过磷代谢调控来响应非生物胁迫,而且与拟南芥的衰老进程存在紧密联系。We constructed CRISPR/Cas9-mediated AtPHO2 knock-out vector,then transformed in Arabidopsis.In the transgenic descendants,three Atpho2 positive mutants were screened.In the transgenic mutant,the gene expression of AtPHO2 was significantly down-regulated,only one in ten of the wild type.The phosphorus contents in mutants and in the wild types were tested,the results of which revealed that Pi gradually accumulated in the new leaves with the aging process.The leaf phosphorus contents of the mutant Atpho2 were 2 ~ 4 times for the wild type.The results implied that more phosphorus accumulated in Atpho2 mutants for the phosphorus metabolism blocking.Additionally,AtPHO2 is probably associated with abiotic stress through regulating phosphorus metabolism and Arabidopsis senescence.

关 键 词：拟南芥 基因组编辑技术 规律成簇短回文重复序列 泛素结合酶24 磷代谢 

分 类 号：Q754[生物学—分子生物学]