Characterization of the genetic and functional diversity of the NLRome/resistome in melon (Cucumis melo L.) using Nanopore Adaptive sampling (NAS)
Résumé
Nucleotide-binding-site-leucine-rich-repeat (NLR) disease resistance genes encode the most important and one of the most variable family of plant resistance proteins. The characterization of their complete collection in a species (NLRome) would facilitate the creation of varieties with a very broad spectrum of resistance. However, although many efforts have been made, the specific role of each NLR gene remains largely unknown in melon. This lack of information is mainly due to their complex structure and organization, frequently arranged in clusters that include a high level of presence-absence polymorphisms.
These structural singularities of NLR clusters make sequencing methods using short reads often ineffective to decipher them. To solve this problem, Nanopore Adaptive Sampling (NAS) is a cost- and labor-effective approach that provides long reads and selectively sequences pre-defined target regions.
Our project pursues the implementation of NAS for the sequencing and characterization of the NLRome of melon using ≈140 varieties. In a first step, we demonstrated the performance (= increase of enrichment) of the NAS approach against whole genome sequencing (WGS) for the sequencing and assembly of the NLRome in two well-studied melon varieties, obtaining around 30X enrichment. We also assessed the transferability of our design to other varieties, in simplex- and multiplex-sequencing experiments. In the next steps of the project, we plan to continue exploring the possibilities of the NAS approach with a final goal of performing genetic association studies using the NLRome sequence variability of the ≈140 lines and their phenotypic variability against a group of selected pests and pathogens.
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