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Molecular characterization and genomic organization of low molecular weight glutenin subunit genes at the Glu-3 loci in hexaploid wheat (Triticum aestivum L.).
Theor Appl Genet. 2008 May; 116(7):953-66.TA

Abstract

In this study, we report on the molecular characterization and genomic organization of the low molecular weight glutenin subunit (LMW-GS) gene family in hexaploid wheat (Triticum aestivum L.). Eighty-two positive BAC clones were identified to contain LMW-GS genes from the hexaploid wheat 'Glenlea' BAC library via filter hybridization and PCR validation. Twelve unique LMW glutenin genes and seven pseudogenes were isolated from these positive BAC clones by primer-template mismatch PCR and subsequent primer walking using hemi-nested touchdown PCR. These genes were sequenced and each consisted of a single-open reading frame (ORF) and untranslated 5' and 3' flanking regions. All 12 LMW glutenin subunits contained eight cysteine residues. The LMW-m-type subunits are the most abundant in hexaploid wheat. Of the 12 LMW-GS, 1, 2 and 9 are i-type, s-type and m-type, respectively. The phylogenetic analysis suggested that the LMW-i type gene showed greater differences to LMW-s and LMW-m-type genes, which, in turn, were more closely related to one another. On the basis of their N-terminal sequences, they were classified into nine groups. Fingerprinting of the 82 BAC clones indicated 30 BAC clones assembled into eight contigs, while the remaining clones were singletons. BAC end sequencing of the 82 clones revealed that long terminal repeat (LTR) retrotransposons were abundant in the Glu-3 regions. The average physical distance between two adjacent LMW-GS genes was estimated to be 81 kb. Most of LMW-GS genes are located in the D: -genome, suggesting that the Glu-D3 locus is much larger than the Glu-B3 locus and Glu-A3 locus. Alignments of sequences indicated that the same type (starting with the same N-terminal sequence) LMW-GS genes were highly conserved in the homologous genomes between hexaploid wheat and its donors such as durum wheat and T. tauschii.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Huang XQ
Cereal Research Centre, Agriculture and Agri-Food Canada, Winnipeg, MB, Canada.
Cloutier S
No affiliation info available

MeSH

Amino Acid SequenceChromosomes, Artificial, BacterialGene LibraryGenes, PlantGlutensMolecular Sequence DataMolecular WeightPhylogenyPloidiesPolymerase Chain ReactionRetroelementsSequence Homology, Amino AcidTriticum

Pub Type(s)

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

Language

eng

PubMed ID

18305921

Citation

Huang, Xiu-Qiang, and Sylvie Cloutier. "Molecular Characterization and Genomic Organization of Low Molecular Weight Glutenin Subunit Genes at the Glu-3 Loci in Hexaploid Wheat (Triticum Aestivum L.)." TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik, vol. 116, no. 7, 2008, pp. 953-66.
Huang XQ, Cloutier S. Molecular characterization and genomic organization of low molecular weight glutenin subunit genes at the Glu-3 loci in hexaploid wheat (Triticum aestivum L.). Theor Appl Genet. 2008;116(7):953-66.
Huang, X. Q., & Cloutier, S. (2008). Molecular characterization and genomic organization of low molecular weight glutenin subunit genes at the Glu-3 loci in hexaploid wheat (Triticum aestivum L.). TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik, 116(7), 953-66. https://doi.org/10.1007/s00122-008-0727-1
Huang XQ, Cloutier S. Molecular Characterization and Genomic Organization of Low Molecular Weight Glutenin Subunit Genes at the Glu-3 Loci in Hexaploid Wheat (Triticum Aestivum L.). Theor Appl Genet. 2008;116(7):953-66. PubMed PMID: 18305921.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular characterization and genomic organization of low molecular weight glutenin subunit genes at the Glu-3 loci in hexaploid wheat (Triticum aestivum L.). AU - Huang,Xiu-Qiang, AU - Cloutier,Sylvie, Y1 - 2008/02/28/ PY - 2007/10/05/received PY - 2008/02/01/accepted PY - 2008/2/29/pubmed PY - 2008/8/1/medline PY - 2008/2/29/entrez SP - 953 EP - 66 JF - TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik JO - Theor Appl Genet VL - 116 IS - 7 N2 - In this study, we report on the molecular characterization and genomic organization of the low molecular weight glutenin subunit (LMW-GS) gene family in hexaploid wheat (Triticum aestivum L.). Eighty-two positive BAC clones were identified to contain LMW-GS genes from the hexaploid wheat 'Glenlea' BAC library via filter hybridization and PCR validation. Twelve unique LMW glutenin genes and seven pseudogenes were isolated from these positive BAC clones by primer-template mismatch PCR and subsequent primer walking using hemi-nested touchdown PCR. These genes were sequenced and each consisted of a single-open reading frame (ORF) and untranslated 5' and 3' flanking regions. All 12 LMW glutenin subunits contained eight cysteine residues. The LMW-m-type subunits are the most abundant in hexaploid wheat. Of the 12 LMW-GS, 1, 2 and 9 are i-type, s-type and m-type, respectively. The phylogenetic analysis suggested that the LMW-i type gene showed greater differences to LMW-s and LMW-m-type genes, which, in turn, were more closely related to one another. On the basis of their N-terminal sequences, they were classified into nine groups. Fingerprinting of the 82 BAC clones indicated 30 BAC clones assembled into eight contigs, while the remaining clones were singletons. BAC end sequencing of the 82 clones revealed that long terminal repeat (LTR) retrotransposons were abundant in the Glu-3 regions. The average physical distance between two adjacent LMW-GS genes was estimated to be 81 kb. Most of LMW-GS genes are located in the D: -genome, suggesting that the Glu-D3 locus is much larger than the Glu-B3 locus and Glu-A3 locus. Alignments of sequences indicated that the same type (starting with the same N-terminal sequence) LMW-GS genes were highly conserved in the homologous genomes between hexaploid wheat and its donors such as durum wheat and T. tauschii. SN - 0040-5752 UR - https://www.unboundmedicine.com/medline/citation/18305921/Molecular_characterization_and_genomic_organization_of_low_molecular_weight_glutenin_subunit_genes_at_the_Glu_3_loci_in_hexaploid_wheat__Triticum_aestivum_L___ DB - PRIME DP - Unbound Medicine ER -