Abstract
Tomato is one of the major vegetable crops consumed worldwide. Tomato yellow leaf curl virus (TYLCV) and fungal Oidium sp. are devastating pathogens causing yellow leaf curl disease and powdery mildew. Such viral and fungal pathogens reduce tomato crop yields and cause substantial economic losses every year. Several commercial tomato varieties include Ty-5 (SlPelo) and Mildew resistance locus o 1 (SlMlo1) locus that carries the susceptibility (S-gene) factors for TYLCV and powdery mildew, respectively. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) is a valuable genome editing tool to develop disease-resistant crop varieties. In this regard, targeting susceptibility factors encoded by the host plant genome instead of the viral genome is a promising approach to achieve pathogen resistance without the need for stable inheritance of CRISPR components. In this study, the CRISPR/Cas9 system was employed to target the SlPelo and SlMlo1 for trait introgression in elite tomato cultivar BN-86 to confer host-mediated immunity against pathogens. SlPelo-knockout lines were successfully generated, carrying the biallelic indel mutations. The pathogen resistance assays in SlPelo mutant lines confirmed the suppressed accumulation of TYLCV and restricted the spread to non-inoculated plant parts. Generated knockout lines for the SlMlo1 showed complete resistance to powdery mildew fungus. Overall, our results demonstrate the efficiency of the CRISPR/Cas9 system to introduce targeted mutagenesis for the rapid development of pathogen-resistant varieties in tomato.
Highlights
Tomato (Solanum lycopersicum) is one of the agronomically important food crops consumed worldwide
We demonstrated the application of Streptococcus pyogenes Cas9 (SpCas9)-mediated genome-editing technology to confer Tomato yellow leaf curl virus (TYLCV) resistance by targeting the SlPelo gene in the Korean elite tomato line, BN-86
The present study demonstrates the potential of the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system to engineer elite tomato lines for multiple pathogen resistance
Summary
Tomato (Solanum lycopersicum) is one of the agronomically important food crops consumed worldwide. A recent analysis of the tomato market by Food and Agriculture. The tomato market is increasing every year. Tomato yellow leaf curl virus (TYLCV), a circular single-stranded. DNA (ssDNA) Begomovirus from the Geminiviridae family, is one of the most devastating viral pathogens affecting tomato cultivation, one that severely affects crop yield. TYLCV is transmitted by whiteflies (Bemisia tabaci) while feeding on plant phloem sap [3]. One recent report suggested that TYLCV can be seed-born because a viral particle can reside inside the seeds after infection and be transmitted to the generation [4]. Severe TYLCV infection can cause abnormal leaf morphology, including stunted growth, curling of margins, and yellowing
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