Introduction: As a result of climate change, increased drought incidence significantly affects the crop yield of rice, Oryza sativa. Given that rice serves as a staple food, adaptation strategies to combat climate change-induced drought are critical. Water retention is regulated by stomata size, stomata density, and the opening and closing of the stomata central pore. Previous studies have identified relevant developmental genes in the Arabidopsis thaliana model system, encoding for epidermal patterning factor (EPFs) and EPF-like (EPFL) signaling peptides, and their orthologs across various plant species. In barley (Hordeum vulgare), genetic manipulation of EPF1 has been shown to reduce stomatal density, resulting in improved drought tolerance. In rice, overexpression of OsEPF1 yields a similar phenotype. The purpose of our study is to develop a proposal for a method to increase drought tolerance of Oryza sativa in an effort to battle climate change.
Methods: It has been shown that CRISPR-mediated editing successfully generated knockouts (KOs) of EPFL9—a positive regulator of stomatal development—in Oryza sativa. As such, we propose to downregulate EPFL9 via CRISPR-Cpfb1 gene editing in Oryza sativa. Our proposal includes the growth of genetically altered and control Oryza sativa under specific conditions, including drought conditions, in order to simulate a natural environment. Following the growth of the plants, we propose conducting tests to determine yield and growth in order to assess drought tolerance.
Discussion: We expect to observe reduced stomatal densities and better drought tolerance in the mutant Oryza sativa samples. This should be observed in increased yield and growth from genetically altered samples. Potential implications of our proposal could include improvements in proto-plants developed in the agricultural sector, as well as providing a foundation for future studies to be conducted on drought tolerance.
Conclusion: Our proposal uniquely addresses the impact of climate change on rice by potentially providing an opportunity to scale-up, generating a drought-tolerant rice plant for comparison with previous prototypes, and secondarily, the elucidation of stomatal development. Our proposal may open further opportunities to address and alter plant resistance to climate change.
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