Efficient DNA fingerprinting based on the targeted sequencing of active retrotransposon insertion sites using a bench-top high-throughput sequencing platform

Monden Yuki, Yamamoto Ayaka, Shindo Akiko, Makoto Tahara

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

In many crop species, DNA finger printing is required for the precise identification of cultivars to protect the rights of breeders. Many families of retrotransposons have multiple copies throughout the eukaryotic genome and their integrated copies are inherited genetically. Thus, their insertion polymorphisms among cultivars are useful for DNA finger printing. In this study, we conducted a DNA fingerprinting based on the insertion polymorphisms of active retro transposon families (Rtsp-1 and LIb) in sweet potato. Using 38 cultivars, we identified 2,024 insertion sites in the two families with an Illumina MiSeq sequencing platform. Of these insertion sites, 91.4% appeared to be polymorphic among the cultivars and 376 cultivar-specific insertion sites were identified, which were converted directly into cultivar-specific sequence-characterized amplified region (SCAR) markers. Aphylogenetic tree was constructed using these insertion sites, which corresponded well with known pedigree information, thereby indicating their suitability for genetic diversity studies. Thus, the genome-wide comparative analysis of active retrotransposon insertion sites using the bench-top MiSeq sequencing platform is highly effective for DNA fingerprinting without any requirement for whole genome sequence information. This approach may facilitate the development of practical polymerase chain reaction-based cultivar diagnostic system and could also be applied to the determination of genetic relationships.

Original languageEnglish
Pages (from-to)491-498
Number of pages8
JournalDNA Research
Volume21
Issue number5
DOIs
Publication statusPublished - 2014

Keywords

  • DNA fingerprinting
  • High-throughput sequencing
  • Molecular marker
  • Retrotransposon
  • Sweet potato

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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