Research Projects

Post-harvest losses in tomato production significantly reduce farmer profitability and contribute to food insecurity (FAO, 2019). To address this challenge, this study proposes a sustainable, non-transgenic bioengineering strategy using CRISPR activation (CRISPRa) technology to enhance endogenous stress- and defence-related gene expression. Key target genes include LEA (late embryogenesis abundant proteins), LEC (lectins), WAK (wall-associated kinases), and DEFL (defensin-like proteins), which are implicated in dehydration tolerance, pathogen resistance, and cell wall reinforcement (Tiwari et al., 2022; Ali et al., 2018). Objective 1: Development of CRISPRa-Engineered Tomato Lines: A dCas9-based transcriptional activation system will be utilised to enhance the expression of native stress-responsive genes by targeting their natural promoter regions. Unlike traditional genetic modification, this method avoids inserting foreign DNA, thereby maintaining a non-GMO status —a key factor for regulatory approval and consumer acceptance (Puchta, 2017). Objective 2: Physiological and Molecular Characterisation Under Simulated Supply Chain Conditions: Engineered lines will undergo controlled stress assays mimicking post-harvest supply chain conditions (storage, transport, and packaging). Evaluations will include assessments of fruit firmness (via penetrometer testing), water retention capacity (gravimetric analysis), pathogen resistance (challenge assays with Botrytis cinerea and Colletotrichum spp.), and shelf-life extension (marketability assessment over time).