CRISPR mutant rapid identification in B. napus: RNA-Seq functional profiling and breeding technology application

Area of Science:

• Plant Science
• Genetics
• Agricultural Biotechnology

Background:

• Traditional rapeseed breeding methods are often inefficient and lack precision.
• CRISPR genome editing presents a highly precise alternative for targeted trait improvement in crops.
• Understanding gene function is crucial for developing improved crop varieties.

Purpose of the Study:

• To investigate the role of the BnaIDA gene in rapeseed floral organ abscission using CRISPR editing.
• To develop a rapid method for transferring the edited BnaIDA gene into elite rapeseed lines.
• To demonstrate the utility of integrating gene editing with efficient genotyping for crop improvement.

Main Methods:

• CRISPR-Cas9 genome editing was employed to modify the BnaIDA gene in the elite rapeseed cultivar ZS11.
• Phenotypic analyses included measuring petal adhesion time and silique cracking force.
• RNA-sequencing was performed on edited mutants. Backcrossing and locus-specific primers were used for trait introgression and selection.

Main Results:

• Gene editing of BnaIDA resulted in altered phenotypes, including persistent petal adhesion and increased silique integrity.
• Edited BnaIDA alleles were successfully introgressed into the elite line SW1-6 via backcrossing.
• Developed locus-specific primers enabled efficient differentiation of heterozygous and homozygous genotypes for rapid selection.

Conclusions:

• CRISPR editing of BnaIDA offers a precise tool for modifying floral organ abscission in rapeseed.
• The integration of gene editing with marker-assisted backcrossing accelerates the development of improved rapeseed varieties.
• This study provides a framework for future gene editing-assisted breeding strategies in rapeseed and other crops.