Cloning and Expression of a LMW-i Glutenin Gene

Cloutier, Sylvie; Rampitsch, Christof; Penner, G.B.; Lukow, O. M.

doi:10.1006/jcrs.2000.0359

Cited by 97 publications

(73 citation statements)

References 31 publications

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“…Additionally, a single locus could encode different types of LMW-GS genes; for example, both LMW-s and LMW-m subunit genes appeared to be present at the Glu-B3 locus in the common wheat cultivar Norin 61 (Ikeda et al 2006). The Glu-A3 locus could encode m-type (Lee et al 1999) and i-type LMW-GS (Cloutier et al 2001;Zhang et al 2004;An et al 2006;Li et al 2008). Consequently, it could be deduced that each Glu-3 locus might encode more than one type of LMW-GS.…”

Section: Pcr Amplification and Cloning Of Lmw-gs Genesmentioning

confidence: 99%

A Novel Chimeric Low-Molecular-Weight Glutenin Subunit Gene From the Wild Relatives of Wheat Aegilops kotschyi and Ae. juvenalis: Evolution at the Glu-3 Loci

Gao

et al. 2008

Genetics

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Four LMW-m and one novel chimeric (between LMW-i and LMW-m types) low-molecular-weight glutenin subunit (LMW-GS) genes from Aegilops neglecta (UUMM), Ae. kotschyi (UUSS), and Ae. juvenalis (DDMMUU) were isolated and characterized. Sequence structures showed that the 4 LMW-m-type genes, assigned to the M genome of Ae. neglecta, displayed a high homology with those from hexaploid common wheat. The novel chimeric gene, designed as AjkLMW-i, was isolated from both Ae. kotschyi and Ae. juvenalis and shown to be located on the U genome. Phylogentic analysis demonstrated that it had higher identity to the LMW-m-type than the LMW-i-type genes. A total of 20 single nucleotide polymorphisms (SNPs) were detected among the 4 LMW-m genes, with 13 of these being nonsynonymous SNPs that resulted in amino acid substitutions in the deduced mature proteins. Phylogenetic analysis demonstrated that it had higher identity to the LMW-m-type than the LMW-i-type genes. The divergence time estimation showed that the M and D genomes were closely related and diverged at 5.42 million years ago (MYA) while the differentiation between the U and A genomes was 6.82 MYA. We propose that, in addition to homologous recombination, an illegitimate recombination event on the U genome may have occurred 6.38 MYA and resulted in the generation of the chimeric gene AjkLMW-i, which may be an important genetic mechanism for the origin and evolution of LMW-GS Glu-3 alleles as well as other prolamin genes.

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Section: Pcr Amplification and Cloning Of Lmw-gs Genesmentioning

confidence: 99%

A Novel Chimeric Low-Molecular-Weight Glutenin Subunit Gene From the Wild Relatives of Wheat Aegilops kotschyi and Ae. juvenalis: Evolution at the Glu-3 Loci

Gao

et al. 2008

Genetics

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“…0 open reading fragment and promoter sequence Genomic DNA was extracted from W958 using the CTAB method as described by Cloutier (Cloutier et al 2001). The primers for cloning the complete coding region sequences of the genes for x-type subunits (both 1Dx5 and 1Dx2) at the Glu-D1 locus (Sugiyama et al 1985;Anderson and Greene 1989) were as follows: P1, 5…”

Section: Cloning and Sequencing Of 1dx5mentioning

confidence: 99%

Molecular cloning of a novel chimeric HMW glutenin subunit gene 1Dx5′ from a common wheat line W958

Feng

et al. 2010

Mol Breeding

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A novel chimeric high-molecular-weight (HMW) glutenin subunit gene from a new common wheat line W958 (2n = 6x = 42) was isolated and characterized. SDS-PAGE analysis revealed that this glutenin subunit has similar electrophoretic mobility to 1Dx5, so it was designated 1Dx5 0 . Genomic DNA from W958 was amplified and a 2,505-bp fragment was obtained. The 1Dx5 0 subunit showed a chimeric primary structure of 1Dx5 and 1Dx2, with the 1Dx5 sequence in the 5 0 and middle repetitive regions and the 1Dx2 sequence in the repetitive domain and 3 0 region. MALDI-TOF-MS analysis demonstrated that 1Dx5 0 had a molecular weight of 86815.1 Da, close to that of an x-type glutenin subunit. Secondary structure analysis showed that this subunit had six helixes and one strand, including four helixes in the repetitive domain which could enhance the dough properties. Additionally, the promoter of 1Dx5 0 was obtained and showed the same sequence as 1Dx5 or 1Dx2 except for a few base conversions. The promoter analysis indicated that the cis-acting regulatory elements of 1Dx5 0 were the same as those of 1Dx5 and/or 1Dx2. Previously, we have demonstrated that this novel glutenin subunit is associated with good bread-making quality and comprises a very large proportion of the F2 segregation population.Electronic supplementary material The online version of this article

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“…It provides the possibility to detect the genetic variations of a particular group of LMW-GS genes at a specific locus. Among the classified nine groups of LMW-GS genes, the A-genome specific group 1, the so-called LMW-i gene (or i-type LMW-GS gene) [26], possessed the longest repetitive domain and seems to be more variable than the other gene groups [21]. In this study, a pair of specific PCR primers was employed to reveal length variation of LMW-i genes in the diploid wheats, the A-genome progenitor of hexaploid wheat.…”

Section: Discussionmentioning

confidence: 99%

Length variation of i-type low-molecular-weight glutenin subunit genes in diploid wheats

et al. 2008

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-Allelic variation of the low-molecular-weight glutenin subunit (LMW-GS) is associated with the significant differences of dough quality in bread and durum wheat, and has been widely evaluated at protein level in wheat and its relatives. In this study, a PCR primer set, targeting the high variable repetitive domains, was employed to assay the length variation of i-type LMW-GS genes in the A-genomes of diploid wheats, the diploid progenitors of tetraploid and hexaploid wheat. A total of 71 accessions of diploid wheats, belonging to two wild and one cultivated species, were investigated. The higher variations of repetitive length in i-type LMW-GS genes were found in diploid wheats with Nei's genetic variation index ( H ) of 0.834. The two wild species, T. boeoticum and T. urartu , were found to possess the similar degree of variability, with the Nei's genetic variation index of 0.806 and 0.783, respectively. Less variation was detected in T. monococcum ( H = 0.680), a cultivated species domesticated from T. boeoticum. The sufficient variation found in this study could be used as valuable source for the enrichment of genetic variations and the alteration of flour-processing properties of the cultivated wheat. To our knowledge, it was the first time that an analysis of length variation targeting a particular group of genes of LMW-GS complex multigene families was conducted.

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Cloning and Expression of a LMW-i Glutenin Gene

Cited by 97 publications

References 31 publications

A Novel Chimeric Low-Molecular-Weight Glutenin Subunit Gene From the Wild Relatives of Wheat Aegilops kotschyi and Ae. juvenalis: Evolution at the Glu-3 Loci

A Novel Chimeric Low-Molecular-Weight Glutenin Subunit Gene From the Wild Relatives of Wheat Aegilops kotschyi and Ae. juvenalis: Evolution at the Glu-3 Loci

Molecular cloning of a novel chimeric HMW glutenin subunit gene 1Dx5′ from a common wheat line W958

Length variation of i-type low-molecular-weight glutenin subunit genes in diploid wheats

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