Research On Rose Chinensis RcAP2 Gene Editing Based On CRISPR/Cas9

In this study,in order to explore the effect of Rose chinensis RcAP2 gene on plant growth and environmental stress,a plant binary expression vector based on CRISPR / Cas9 gene editing technology was constructed.Using Arabidopsis thaliana as the receptor material,Agrobacterium-mediated method was used to infect and transform A.thaliana inflorescence,and A.thaliana transformed plants expressing eGFP were obtained;using R.chinensis as the receptor material,the regeneration system of R.chinensis plants was optimized;The Agrobacterium-mediated method was used to infect and transform the R.chinensis embryogenic callus and R.chinensis stem buds,and R.chinensis axillary buds and leaves expressing eGFP were obtained.Conducted a preliminary study on the function of R.chinensis RcAP2 gene and got the following main conclusions.i)This study is based on CRISPR/Cas9 gene editing technology,based on the conserved sequence of R.chinensis RcAP2 gene,a pair of target site genes were screened,and the target fragment was ligated into pKSE402 vector by PCR technology.The plant expression vector of RcAP2 gene was successfully constructed.ii)The axillary buds of R.chinensis stems were used as test materials,and 2,4-D was used as the main growth regulator to optimized the R.chinensis regeneration system.iii)Using A.thaliana as the recipient material,Agrobacterium-mediated method was used to infect and transform A.thaliana inflorescence,and A.thaliana transformed plants expressing eGFP were obtained.iv)Using R.chinensis embryonic callus and R.chinensis stem buds as recipient materials,the wound pairs were infected by Agrobacterium-mediated method,R.chinensis transformed plants were obtained,and green fluorescence was detected on axillary buds and leaves of R.chinensis.