首页> 中文期刊> 《河南农业大学学报》 >AtPHO2的CRISPR/Cas9靶向敲除及其在拟南芥中功能分析

AtPHO2的CRISPR/Cas9靶向敲除及其在拟南芥中功能分析

         

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.

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