Combining a New Exome Capture Panel With an Effective varBScore Algorithm Accelerates BSA-Based Gene Cloning in...

Area of Science:

• Plant genetics
• Molecular biology
• Agricultural science

Background:

• Identifying functional genes is crucial for enhancing wheat agronomic traits.
• Existing methods for gene mapping and identification can be hindered by background noise.

Purpose of the Study:

• To develop an efficient method for discovering causal mutations and cloning genes in wheat.
• To improve gene mapping and identification by reducing background noise.

Main Methods:

• Designed a new wheat exome capture probe panel based on IWGSC RefSeq v1.0.
• Developed a robust algorithm, varBScore, to reduce background noise in gene mapping.
• Established bulked segregant exome capture sequencing (BSE-Seq) combining the probe panel, sequencing, and varBScore.

Main Results:

• Validated varBScore's effectiveness on SNP calling for mapping the yellow rust resistance gene *Yr7*.
• Successfully identified the wheat yellow leaf mutant gene *ygl1* using BSE-Seq.
• Confirmed *ygl1* as the causal gene through CRISPR/Cas9 gene editing, demonstrating its role in leaf yellowing.

Conclusions:

• The developed BSE-Seq method is highly efficient for discovering causal mutations and cloning genes in wheat.
• This approach significantly aids in identifying genes underlying important agronomic traits for wheat improvement.