Demonstration of CRISPR-cas9-mediated pds gene editing in a tomato hybrid parental line

Abstract

CRISPR-cas9 mediated gene editing is a powerful tool proven for crop improvement and has great potential for incorporating novel traits into important genetic resources. In this study, pds gene editing in a parent line of a tomato hybrid by using CRISPR-cas9 system was successfully conducted. Three small guide RNAs (sgRNAs) were designed to target three exons of a tomato pds gene and were transformed along with a cas9 gene into the line of interest. Observation of chimeric albino phenotypes in the regenerated tomato shoots indicated the disruption of pds gene. Furthermore, expression of cas9 gene was determined by demonstration of its transcript accumulation in the primary tomato transformants. Sequence analysis of targeted pds exons showed point as well as deletion mutations in the sgRNA target sites. The result confirmed functional nature of the CRISPR-cas9 assembly used in the present study, which will enable incorporation of novel traits directly into the parents of tomato hybrids.