Executive Summary : | The integration of genome editing technologies has significantly improved crop enhancement through genomic investigation. Strategies involving simultaneous modification of multiple genomic targets offer a remarkable avenue for the concurrent manipulation of multiple genes. The progression of genomics has yielded a vast repository of data concerning the genetic underpinnings of pivotal agricultural traits. To harness this wealth of information effectively for the betterment of crops, it is imperative to engender breeding lineages that encapsulate the highest count of desirable genetic variants. The present study aims to develop seedless tomato with improved shelf life using the recently developed CRISPR-Combo orthogonal genome editing system. The CRISPR Combo enables speed breeding of transgene-free crops with increased heritable mutation in hormone-free manner. The technique also allows screening of transgene-free plants with visual differences, further reducing the time and efforts for screening. The proposed project aims to target four genes viz. SlARF7, SlIAA9, SlPAT, and SlEB1 simultaneously through the CRISPR-Combo system. The combined mutations in these genes increase the probability of getting parthenocarpic seeds or seedless fruits on any genetic background. Another gene SlEB1 is involved in an ethylene synthesis and cell wall degradation, suppressing the ethylene synthesis and cell wall degradation, leading to increased shelf life of tomato fruits. The project will also aid adaptation of advances in genome editing systems in agriculturally important crop tomato. |