水稻ABERRANT PANICLE ORGANIZATION 1 （APO1）/ ABERRANT PANICLE ORGANIZATION 2（APO2）互作基因
关键词：水稻；APO1；APO2/RFL；iTRAQ；酵母双杂；CRISPR/Cas9
The construct of the mutants interact with ABERRANT PANICLE ORGANIZATION 1 （APO1）/ABERRANT PANICLE ORGANIZATION 2 （APO2/RFL）of rice and the analyze of the function
Abstract: [Objectives] rice is an important crop. The shape of spike has a tight connection to the production. With which, do research about the genes of rice spike and dig out new function gene so that improve the production has a good meaning. The proteins of APO1 and APO2/RFL could interact with each other and cooperate to control the dedifferentiation of young spike of rice. APO1 is F-box protein of rice, mediate substrate protein ubiquitination degradation. The research analyzes the function of APO1 and APO2/RFL, try to make it clear about the substrate protein which APO1 degradation. At the same time, construct the mutants of the genes interact with APO1 and APO2/RFL. [Methods] make use of apo1 and apo2/rfl mutants have gained. First, observe the phenotypes compared with wild type. And connect the gene of mchery protein with APO1,connect the gene of GFP with APO2/RFL,turned them into apo1 and apo2/rfl mutant respectively. Observe the phenotype whether it recovers. Then, extract the proteins of young spikes of apo1 mutant and wild type, analyze with the technology of iTRAQ. Then filter by the technology of yeast two hybrid, so that gain the proteins interact with each other. Then we use the technology of CRISPR/Cas9 to construct the mutants of the genes interact with APO1 and APO2/RFL. [Results] Compared with wild type, the shape of spike of apo1 mutant was obviously smaller, the amount of secondary branches was also less. When gRFL:GFP turned into apo2/rfl mutant, the phenotype of spike was recovered to a certain extent. When gAPO1:mchery turned into apo1 mutant, the APO1 gene was overexpressed, with the phenotype of spike become bigger. The analyze of iTRAQ found that there are 279 proteins express with obvious difference, including cell pision, cell dedifferentiation, and metabolism, and so on. We also did Yeast two hybrid, to find the proteins which gene APO1 and APO2/RFL controls directly. The technology of CRISPR/Cas9 has constructed some mutants. [Conclusions] We demonstrated how APO1 and APO2/RFL have an influence at rice spike dedifferentiation at the angle of protein, which makes a base on the research on how APO1 and APO2/RFL interaction.
Keywords: rice; APO1; APO2/RFL; iTRAQ; Yeast two hybrid;CRISPR/Cas9