The Application of RenSeq for Pan-NLRome Analysis

Area of Science:

• Plant genetics
• Molecular biology
• Genomics

Background:

• Resistance gene enrichment sequencing (RenSeq), developed in 2013, has been pivotal in cloning plant nucleotide-binding leucine-rich repeat (NLR) genes.
• It has significantly contributed to resistance breeding in major crops like potato, wheat, and rice.
• Various RenSeq-based strategies (e.g., cDNA-RenSeq, MutRenSeq) have expanded its applications across diverse plant species.

Purpose of the Study:

• To provide a detailed protocol for applying RenSeq in gene cloning and Pan-NLRome analysis.
• To make RenSeq methodology more accessible to researchers across different crop systems.
• To deepen the understanding of plant resistance genes within the context of pan-genome research.

Main Methods:

• The protocol outlines essential steps including bait design, library preparation, and target enrichment.
• It also details downstream bioinformatic analysis procedures.
• The methodology integrates various RenSeq applications for comprehensive analysis.

Main Results:

• The presented protocol facilitates efficient gene cloning and Pan-NLRome analysis using RenSeq.
• It demonstrates the broad applicability of RenSeq across various crop plants.
• The enhanced accessibility of RenSeq promotes wider adoption in plant resistance research.

Conclusions:

• This RenSeq protocol empowers researchers to clone resistance genes and conduct Pan-NLRome analyses more effectively.
• It supports advancements in crop resistance breeding and genetic improvement.
• The methodology contributes to a more comprehensive understanding of plant immune systems in the pan-genome era.