Selection of Flax Genotypes for Pan-Genomic Studies by Sequencing Tagmentation-Based Transcriptome Libraries

Area of Science:

• Plant Genomics
• Agricultural Science
• Biotechnology

Background:

• Flax (Linum usitatissimum L.) is a versatile crop with applications across food, pharmaceutical, textile, and industrial sectors.
• The development of a flax pan-genome is essential for advancing flax research and breeding efforts.
• Selecting diverse flax genotypes is a critical prerequisite for comprehensive pan-genome construction.

Purpose of the Study:

• To adapt and validate a cost-effective method for preparing transcriptome libraries for high-throughput sequencing.
• To assess the genetic relationships among flax accessions and identify representative genotypes for pan-genome analysis.
• To ensure the selection of diverse flax genotypes for robust pan-genome construction.

Main Methods:

• Adapted a tagmentation-based (using Tn5 transposase) Illumina sequencing method for cDNA library preparation.
• Generated RNA-seq data from 192 flax plants across 44 accessions of varied morphology and origin.
• Utilized sequencing data to evaluate genetic relationships and identify potential misidentifications.

Main Results:

• The tagmentation-based approach proved efficient and cost-effective for preparing numerous cDNA libraries.
• Genetic relationship analysis revealed misidentification in five flax accessions, leading to their exclusion.
• Representative genotypes covering significant genetic diversity were selected from the remaining samples for pan-genome construction.

Conclusions:

• Tagmentation-based transcriptome library preparation is a highly efficient method for assessing genetic diversity in flax.
• This approach facilitates the selection of optimal genotypes for constructing a comprehensive flax pan-genome.
• The study provides a validated methodology for future flax genomics and breeding initiatives.