High-Throughput Base Editing-Mediated Artificial Evolution Streamlines Trait Gene Identification in Rice.

Area of Science:

• Agricultural Science
• Molecular Biology
• Genetics

Background:

• Developing novel crop germplasms is crucial for agricultural advancement.
• Identifying functional single nucleotide polymorphisms (SNPs) can accelerate crop improvement.
• Existing gene editing tools may have limitations in generating diverse mutations for evolutionary studies.

Purpose of the Study:

• To describe the Base Editing-Mediated Gene Evolution (BEMGE) method for artificial gene evolution in rice.
• To demonstrate the application of BEMGE in evolving the OsALS1 gene in rice cells.
• To highlight the potential of BEMGE in generating diverse mutations for screening pivotal amino acids and enhancing crop traits.

Main Methods:

• Utilized Cas9n-based cytosine and adenine base editors.
• Employed a sgRNA library tiling the full-length coding region of target genes.
• Applied Agrobacterium-mediated transformation for introducing base editors into rice cells.
• Focused on the artificial evolution of the OsALS1 gene in rice.

Main Results:

• Successfully applied the BEMGE method to induce mutations in the OsALS1 gene in rice cells.
• Demonstrated the capability of BEMGE to generate numerous nucleotide changes within a gene.
• Showcased the potential for screening pivotal amino acids through induced mutations.

Conclusions:

• The BEMGE method offers a powerful approach for generating novel rice germplasms with targeted mutations.
• BEMGE facilitates the discovery of unknown functional SNPs and pivotal amino acids.
• This technique has significant potential to accelerate crop improvement and breeding programs.