Abstract
Mutagenesis is the only process by which unpredicted biological gene function can be identified. Despite that several macroalgal developmental mutants have been generated, their causal mutation was never identified, because experimental conditions were not gathered at that time. Today, progresses in macroalgal genomics and judicious choices of suitable genetic models make mutated gene identification possible. This article presents a comparative study of two methods aiming at identifying a genetic locus in the brown alga Ectocarpus siliculosus: positional cloning and Next-Generation Sequencing (NGS)-based mapping. Once necessary preliminary experimental tools were gathered, we tested both analyses on an Ectocarpus morphogenetic mutant. We show how a narrower localization results from the combination of the two methods. Advantages and drawbacks of these two approaches as well as potential transfer to other macroalgae are discussed.
Highlights
This article presents a comparative study of two methods aiming at identifying a genetic locus in the brown alga Ectocarpus siliculosus: positional cloning and Next-Generation Sequencing (NGS)-based mapping
Several morphological mutants of macroalgae such as the green Ulva, the red Gracilaria and the brown Ectocarpus have been generated by UV or chemical mutagenesis and summarily studied at the genetic level
The work reported here describes how to identify a single mutation within a 214 Mbp macroalgal genome
Summary
This article presents a comparative study of two methods aiming at identifying a genetic locus in the brown alga Ectocarpus siliculosus: positional cloning and Next-Generation Sequencing (NGS)-based mapping. A genetic map was built by performing segregation analyses of SSR markers present on the super-contigs (sctgs) and shown to be polymorphic between the two Ectocarpus strains used for the cross.
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