Evaluation of Blast Disease Resistance in Rice Plants (Oryza sativa) Resulting from BSR-D1 Gene Editing Through In-Planta Transformation With CRISPR/Cas9 Technology
Abstract
Rice blast disease, caused by the fungus Magnaporthe oryzae, poses a significant threat to rice productivity
worldwide. To enhance rice resistance to this disease, CRISPR/Cas9 gene-editing technology has been used to
modify the Bsr-d1 gene, which is known to regulate plant resistance to fungal infections negatively. This study
aims to evaluate the blast disease resistance of the T2 generation of the Bsr-d1 gene-edited rice variety MR 297.
The transformed plants were inoculated with a P. oryzae isolate (Isolate MR297 3.1), and disease symptoms were
assessed nine days post-inoculation using the IRRI scale (0–9). The results demonstrated that several gene-edited
plant lines exhibited significantly milder disease symptoms compared to susceptible control plants. These findings
indicate that editing the Bsr-d1 gene effectively enhances rice resistance to blast disease without compromising
key agronomic traits. By eliminating the need for tissue culture, this protocol offers a simpler and more efficient
approach with the potential for widespread adoption in disease-resistant rice breeding programs. The primary
advantage of this study is the application of CRISPR/Cas9 technology to specifically target the Bsr-d1 gene, a
negative regulator of blast resistance. This method enhances rice plant resistance without introducing foreign
genes, increasing its likelihood of regulatory approval and acceptance in field applications. Resistance testing
using local isolates of Magnaporthe oryzae enhances the relevance of the findings to local agroecosystem
conditions. Furthermore, the study demonstrates that increased resistance does not compromise key agronomic
traits, supporting the development of superior rice varieties that are both high-yielding and disease-resistant. It is
recommended that users conduct multi-location and multi-season trials to evaluate the stability of gene-edited
plant resistance under varying environmental conditions and pathogen pressures. Additionally, a more in-depth
analysis of the impact of the Bsr-d1 gene on the expression of other genes related to plant growth and yield is
necessary.
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