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Construction of a hexaploid wheat (Triticum aestivum L.) bacterial artificial chromosome library for cloning genes for stripe rust resistance.
Genome. 2005 Dec; 48(6):1028-36.G

Abstract

A hexaploid wheat (Triticum aestivum L.) bacterial artificial chromosome (BAC) library was constructed for cloning Yr5 and other genes conferring resistance to stripe rust (Puccinia striiformis f. sp. tritici). Intact nuclei from a Yr5 near-isogenic line were used to isolate high molecular weight DNA, which was partially cleaved with HindIII and cloned into pECBAC1 and pIndigoBAC-5 vectors. The wheat BAC library consisted of 422,400 clones arrayed in 1100 micro-titer plates (each plate with 384 wells). Random sampling of 300 BAC clones indicated an average insert size of 140 kb, with a size range from 25 to 365 kb. Ninety percent of the clones in the library had an insert size greater than 100 kb and fewer than 5% of the clones did not contain inserts. Based on an estimated genome size of 15,966 Mb for hexaploid wheat, the BAC library was estimated to have a total coverage of 3.58x wheat genome equivalents, giving approximately 96% probability of identifying a clone representing any given wheat DNA sequence. Twelve BAC clones containing an Yr5 locus-specific marker (Yr5STS7/8) were successfully selected by PCR screening of 3-dimensional BAC pools. The results demonstrated that the T. aestivum BAC library is a valuable genomic resource for positional cloning of Yr5. The library also should be useful in cloning other genes for stripe rust resistance and other traits of interest in hexaploid wheat.

Authors+Show Affiliations

U.S. Department of Agriculture, Agricultural Research Service, Wheat Genetics, Quality, Physiology and Disease Research Unit, Pullman, WA 99164-6430, USA.No affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

16391672

Citation

Ling, P, and X M. Chen. "Construction of a Hexaploid Wheat (Triticum Aestivum L.) Bacterial Artificial Chromosome Library for Cloning Genes for Stripe Rust Resistance." Genome, vol. 48, no. 6, 2005, pp. 1028-36.
Ling P, Chen XM. Construction of a hexaploid wheat (Triticum aestivum L.) bacterial artificial chromosome library for cloning genes for stripe rust resistance. Genome. 2005;48(6):1028-36.
Ling, P., & Chen, X. M. (2005). Construction of a hexaploid wheat (Triticum aestivum L.) bacterial artificial chromosome library for cloning genes for stripe rust resistance. Genome, 48(6), 1028-36.
Ling P, Chen XM. Construction of a Hexaploid Wheat (Triticum Aestivum L.) Bacterial Artificial Chromosome Library for Cloning Genes for Stripe Rust Resistance. Genome. 2005;48(6):1028-36. PubMed PMID: 16391672.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Construction of a hexaploid wheat (Triticum aestivum L.) bacterial artificial chromosome library for cloning genes for stripe rust resistance. AU - Ling,P, AU - Chen,X M, PY - 2006/1/5/pubmed PY - 2007/3/9/medline PY - 2006/1/5/entrez SP - 1028 EP - 36 JF - Genome JO - Genome VL - 48 IS - 6 N2 - A hexaploid wheat (Triticum aestivum L.) bacterial artificial chromosome (BAC) library was constructed for cloning Yr5 and other genes conferring resistance to stripe rust (Puccinia striiformis f. sp. tritici). Intact nuclei from a Yr5 near-isogenic line were used to isolate high molecular weight DNA, which was partially cleaved with HindIII and cloned into pECBAC1 and pIndigoBAC-5 vectors. The wheat BAC library consisted of 422,400 clones arrayed in 1100 micro-titer plates (each plate with 384 wells). Random sampling of 300 BAC clones indicated an average insert size of 140 kb, with a size range from 25 to 365 kb. Ninety percent of the clones in the library had an insert size greater than 100 kb and fewer than 5% of the clones did not contain inserts. Based on an estimated genome size of 15,966 Mb for hexaploid wheat, the BAC library was estimated to have a total coverage of 3.58x wheat genome equivalents, giving approximately 96% probability of identifying a clone representing any given wheat DNA sequence. Twelve BAC clones containing an Yr5 locus-specific marker (Yr5STS7/8) were successfully selected by PCR screening of 3-dimensional BAC pools. The results demonstrated that the T. aestivum BAC library is a valuable genomic resource for positional cloning of Yr5. The library also should be useful in cloning other genes for stripe rust resistance and other traits of interest in hexaploid wheat. SN - 0831-2796 UR - https://www.unboundmedicine.com/medline/citation/16391672/Construction_of_a_hexaploid_wheat__Triticum_aestivum_L___bacterial_artificial_chromosome_library_for_cloning_genes_for_stripe_rust_resistance_ L2 - https://cdnsciencepub.com/doi/10.1139/g05-078?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -