J Integr Plant Biol. ›› 2010, Vol. 52 ›› Issue (7): 679-691.DOI: 10.1111/j.1744-7909.2010.00954.x

• Molecular Ecology and Evolution • Previous Articles    

Effective Isolation of Retrotransposons and Repetitive DNA Families from the Wheat Genome

Motonori Tomita* *, Munenori Asao and Aya Kuraki   

  1. Molecular Genetics Laboratory, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan

Abstract:

New classes of repetitive DNA elements were effectively identified by isolating small fragments of the elements from the wheat genome. A wheat A genome library was constructed from Triticum monococcum by degenerate cleavage with EcoO109I, the recognition sites of which consisted of 5'-PuGGNCCPy-3' multi-sequences. Three novel repetitive sequences pTm6, pTm69 and pTm58 derived from the A genome were screened and tested for high copy number using a blotting approach. pTm6 showed identity with integrase domains of the barley Ty1-Copia-retrotransposon BARE-1 and pTm58 showed similarity to the barley Ty3-gypsy-like retrotransposon Romani. pTm69, however, constituted a tandem array with useful genomic specificities, but did not share any identity with known repetitive elements. This study also sought to isolate wheat D-genome-specific repetitive elements regardless of the level of methylation, by genomic subtraction. Total genomic DNA of Aegilops tauschii was cleaved into short fragments with a methylation-insensitive 4 bp cutter, MboI, and then common DNA sequences between Ae. tauschii and Triticum turgidum were subtracted by annealing with excess T. turgidum genomic DNA. The D genome repetitive sequence pAt1 was isolated and used to identify an additional novel repetitive sequence family from wheat bacterial artificial chromosomes with a size range of 1 395–1 850 bp. The methods successfully led pathfinding of two unique repetitive families.

Tomita M, Asao M, Kuraki A (2010) Effective isolation of retrotransposons and repetitive DNA families from the wheat genome. J. Integr. Plant Biol. 52(7), 679–691.

Editorial Office, Journal of Integrative Plant Biology, Institute of Botany, CAS
No. 20 Nanxincun, Xiangshan, Beijing 100093, China
Tel: +86 10 6283 6133 Fax: +86 10 8259 2636 E-mail: jipb@ibcas.ac.cn
Copyright © 2022 by the Institute of Botany, the Chinese Academy of Sciences
Online ISSN: 1744-7909 Print ISSN: 1672-9072 CN: 11-5067/Q
备案号:京ICP备16067583号-22