Preliminary Establishment Of System Editing Potato StCRK1/2 Gene Using CRISPR/Cas9

The potato is the fourth most important food crop in the world after wheat,maize and rice.China has listed potatoes as a staple food and proposed a 21 st century potato staple food strategy.Potato production is also increasing year by year,but there are still various diseases that hinder its growth and development during the potato cultivation process,which seriously affects Yield and tuber quality.Plants have developed their own defense mechanisms against various diseases during the long-term evolution.Receptor-like kinase is a signaling molecule that can regulate plant growth and development and adapt to stress.Among them,cysteine-rich receptor-like kinase(CRK)plays an important role in the process of plant disease resistance and programmed cell death.The current research on CRK gene function is mainly concentrated in rice,wheat,Arabidopsis,tomato and other plants,and there are relatively few reports on potato.In this experiment,the pMDI35S:GFP plasmid was transformed into Agrobacterium,and the potato bottom Ciri explants were infected by Agrobacterium-mediated method to obtain 4 GFP transgenic potato lines,and the lines with the highest GFP expression were screened by RT-qPCR.Using CRISPR/Cas9 technology,two vectors for editing GFP gene were constructed.The CDS sequences of StCRKl and StCRK2 in potato were analyzed to predict the target site,two vectors for editing StCRKl gene and one vector for StCRK2 gene were constructed,which laid the foundation for the subsequent study of potato StCRK1/2 gene function.