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Title Characterization of novel LMW-GS genes at Glu-D3 locus on chromosome 1D in Aegilops tauschii
 
Names Zhao Xianlin
Yang Yan
He Zhonghu
Zhensheng Lei
Wujun Ma
Qixin Sun
Xianchun Xia
Date Issued 2008 (iso8601)
Abstract The objectives of this study were to clarify the relationship between LMW-GS Glu-D3 gene of Ae. tauschii registered in GenBank and the six Glu-D3 genes including 12 allelic variants of common wheat characterized in our previous studies, and identify novel Glu-D3 genes and haplotypes from Ae. tauschii using gene specific PCR amplification. By searching the NCBI database, 13 LMW-GS genes/pseudogenes of Ae. tauschii were retrieved and classified into five gene families based on their nucleotide similarity with the six Glu-D3 genes of common wheat. Of them, four Ae. tauschii genes, AY585350, AY585354, AY585355 and AY585356 matched GluD3-4, GluD3-5, GluD3-1 and GluD3-2 of common wheat, respectively, and one pseudogene AY585351 matched to GluD3-6, but none of them matched to GluD3-3. In order to identify the Glu-D3 genes from Ae. tauschii corresponding to GluD3-3 and GluD3-6 of common wheat, gene specific primers were developed to amplify 8–18 Ae. tauschii entries. As a result, two novel Glu-D3 genes, designated as GluDt3-3 and GluDt3-6, were identified. GluDt3-3 showed seven allelic variants or haplotypes at the DNA level in eight Ae. tauschii entries, designated as GluDt3-31, GluDt3-32, GluDt3-33, GluDt3-34, GluDt3-35, GluDt3-36 and GluDt3-37, respectively. Two to eight SNPs were found among the seven haplotypes and 1–4 amino acid substitutions among the deduced peptides. Multiple sequence alignments showed that the DNA similarity was 99.6–99.9% among the seven GluDt3-3 haplotypes, and 99.4–99.7% between these haplotypes and those of common wheat GluD3-3 gene. GluDt3-6 presented seven haplotypes in 18 Ae. tauschii entries, designated as GluDt3-61, GluDt3-62, GluDt3-63, GluDt3-64, GluDt3-65, GluDt3-66 and GluDt3-67, respectively. GluDt3-61 from Ae. tauschii entry Ae38 was the only one haplotype with complete coding sequence, and the other six were all pseudogenes. Compared with GluD3-6 gene of common wheat, GluDt3-61 exhibited a 3-bp insertion, a 42-bp deletion and 11 base substitutions, leading to a glutamine insertion in position 52, 14 amino acid deletion in position 84–97 and 10 amino acid mutations in its deduced peptide; GluDt3-62 and GluDt3-63 showed a 6-bp insertion, a 24-bp deletion and 15–21 base substitutions in coding region, of which a nonsense mutation from C to T at position 622 resulted in pseudogenes; GluDt3-64 had five base substitution, including a nonsense mutation at the position 742. GluDt3-65, GluDt3-66 and GluDt3-67 all had a base deletion at position 247, as well as 7–8 base substitutions, which resulted in frameshift mutations in the three haplotypes. The results indicated that Ae. tauschii also contains six Glu-D3 genes and their allelic variants are even richer than those in common wheat
Genre Article
Access Condition Open Access
Identifier http://hdl.handle.net/10883/2679