DOI: 10.3724/SP.J.1249.2017.05464

Journal of Shenzhen University Science and Engineering (深圳大学学报理工版) 2017/34:5 PP.464-470

Identification of novel components of Arabidopsis miRNA pathway and mutant analysis

Micro ribonucleic acids (miRNAs) are 20-24 nucleotides non-coding RNAs that play key regulatory roles in developmental and physiological processes in plants. In order to screen the new components that are involved in miRNA biogenesis, turnover and movement, we establish a forward genetic screening system using Arabidopsis thaliana transgenic line (SUC2:amiR-SUL). After ethylmethylsulfone (EMS) mutagenesis on SUC2:amiR-SUL, one stable mutant line (SUP-E45) is isolated. The result of phenotype observation, quantitative real-time polymerase chain reaction (qRT-PCR) and whole genome sequencing on the SUP-E45 mutant plants indicates that the altered phenotype is caused by the mutation in Argonaute 1 (Ago1) gene. AGO1 protein encoded by ago1 gene is crucial in the miRNA pathways, which recruits mature miRNA to form miRISC silence complex (miRNAs-induced silencing complex, miRISC) to negatively regulate the expression of target genes. The screening system can be used to select other factors that participate in the processing such as miRNA synthesis, miRNA's activity or miRNA transport. The research can lay the foundation on the subsequent Arabidopsis miRNA pathway.

Key words:molecular biology,SUC2:amiR-SUL,Arabidopsis thaliana,forward genetic screening system,microRNA (miRNA),AGO1 protein,EMS mutagenesis,quantitative real-time PCR(qRT-PCR),whole genome sequencing

ReleaseDate:2017-10-20 02:07:29

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