Gene Editing And Intercellular Migration Analysis Of MYB Transcription Factors Controlling Ear Development In Rice

Rice(Oryza sativa l.)is the most important food crop with the largest population in the world,and it is an important guarantee for China's food security to ensure that rice yield is higher than people's demand [1].China is a large traditional agricultural country,and the traditional hybrid breeding represented by yuan nongping is an important means to improve rice yield.In recent years,due to the development and improvement of molecular breeding technology,the ways to realize the growth of rice have become more diversified.Through the genetic improvement of the flower organs of rice,the yield and quality of rice can be directly increased,the breeding years can be greatly shortened,and the manpower,material resources and financial resources are saved.In this study,using bioinformatics methods and based on the previous study of flower pattern forming gene SFL(LOC_Os07g43580),the author further explored the regulation mechanism of SFL and its related genes on the development of ear in rice.We first obtained the four candidate genes of LOC_Os03g26130,LOC_Os07g43580,LOC_Os08g33940 and LOC_Os09g24180 by homologous comparison and pathway analysis.We used plant genome editing technology to knock out these genes for phenotypic identification.Then,the two candidate genes LOC_Os03g26130 and LOC_Os07g43580 were cloned from zhonghua 11 material to construct the relevant vector,and the subcellular localization was carried out.Finally,by designing different number of GFP fusion vectors,the intercellular migration pattern of ear development and gene ectopic expression was studied,and the following results were obtained:1.Through bioinformatics method and based on previous studies on tiller genes of SFL,the gene structure and physicochemical properties of the four candidate genes LOC_Os03g26130,LOC_Os07g43580,LOC_Os08g33940 and LOC_Os09g24180 were analyzed,and the subcellular localization of the proteins was performed by using ploc-mplant,a subcellular localization prediction tool.These genes have similar conserved domains for the protein,none are transmembrace proteins.Using chop-chop website online tool,the best gene editing target was obtained.2.Golden-gate-assemble technology was used to connect the sequence fragments to achieve multiple editing and simultaneously act on two target sites of a gene,so as to ensure the effectiveness of gene editing.Through the observation of the phenotype of the later experimental materials by transgenic technology,it was found that the plants were indeed small,weak or without spike.3.by PCR technique,we obtain the LOC_Os03g26130,LOC_Os07g43580 over the span of the two candidate genes sequence,the total length of 924 bp and 1856 bp respectively,through the homologous recombination technology to the GFP respectively with 1 X,2 X GFP,GFP 3 X and 4 X GFP gene construction of expression vector,using the method of rice pollen tube channel will build a carrier of the import of flowers of 11 material expression and combined with laser confocal microscope,rice seed regions have expressed,and its expression is associated with the number of GFP.It may not be affected by gated ion channels during Intercellular mobile.