High-throughput polymorphism detection and genotyping in Brassica napus using next-generation RAD sequencing

BMC Genomics

Research Article

Jochen Hecht¹  Bruno Huettel²  Richard Reinhardt²  Benjamin Stich³  Anja Bus³ 

[1] Berlin-Brandenburg Center for Regenerative Therapies, Charité-Universitaetsmedizin Berlin, Augustenburger Platz 1, 13353, Berlin, Germany;Max Planck Genome Centre Cologne, Carl-von-Linné-Weg 10, 50829, Cologne, Germany;Max Planck Institute for Plant Breeding Research, Carl-von-Linné-Weg 10, 50829, Cologne, Germany;

关键词: Brassica napus;    Restriction-site associated DNA;    Next-generation sequencing;    Single nucleotide polymorphism;    Genotyping by sequencing;    Genetic diversity;   

DOI  :  10.1186/1471-2164-13-281

 received in 2012-03-16, accepted in 2012-06-24,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundThe complex genome of rapeseed (Brassica napus) is not well understood despite the economic importance of the species. Good knowledge of sequence variation is needed for genetics approaches and breeding purposes. We used a diversity set of B. napus representing eight different germplasm types to sequence genome-wide distributed restriction-site associated DNA (RAD) fragments for polymorphism detection and genotyping.ResultsMore than 113,000 RAD clusters with more than 20,000 single nucleotide polymorphisms (SNPs) and 125 insertions/deletions were detected and characterized. About one third of the RAD clusters and polymorphisms mapped to the Brassica rapa reference sequence. An even distribution of RAD clusters and polymorphisms was observed across the B. rapa chromosomes, which suggests that there might be an equal distribution over the Brassica oleracea chromosomes, too. The representation of Gene Ontology (GO) terms for unigenes with RAD clusters and polymorphisms revealed no signature of selection with respect to the distribution of polymorphisms within genes belonging to a specific GO category.ConclusionsConsidering the decreasing costs for next-generation sequencing, the results of our study suggest that RAD sequencing is not only a simple and cost-effective method for high-density polymorphism detection but also an alternative to SNP genotyping from transcriptome sequencing or SNP arrays, even for species with complex genomes such as B. napus.

【 授权许可】

Unknown   
© Bus et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.