CRISPR-Mediated Molecular Breeding in Tropical Agriculture: Insights and Results

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2024

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This manuscript discusses crop breeding as a developing science, particularly in addressing specific challenges in the context of climate change. Crop breeding has evolved from domestication and selection of major food crops to precision gene editing for improvement. Molecular breeding in maize, rice, wheat, and other globally important food crops is well documented, but improvement in tropical agriculture lags in comparison due to unique breeding challenges and limited genomic resources. Regional crops play a crucial role in diversified food markets, but tropical and subtropical countries are especially vulnerable to environmental changes and require novel tools for improving crops. CRISPR-Cas, a customizable gene editing tool, represents a powerful development in molecular breeding for reintroducing genetic diversity lost due to domestication and selection. In addition to genetic bottlenecks, tropical crops have many breeding challenges that limit improvement strategies like CRISPR-Cas. DNA-free strategies for introducing CRISPR-Cas systems exist but have yet to be demonstrated in tropical agriculture. The goal of this project is to optimize biolistic delivery of CRISPR-Cas as a ribonucleic protein complex to determine editing efficiencies of two systems: Cas9 and Cas12a. Since editing occurs in the absence of selection, a second goal is to develop a high-throughput screening protocol. Chapter II focuses on key considerations for developing CRISPR-Cas gene editing projects and investigates papaya and sugarcane as tropical crop models. Chapters III and IV discuss papaya and sugarcane experiments, respectively. Overall, this project underscores the challenges for adapting modern tools in tropical agriculture and highlights an area in need of scientific exploration.

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Genetics, Agriculture, CRISPR-Cas, Crop Improvement, Gene Editing, Tropical Agriculture

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96 pages

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