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New insights into the organization, recombination, expression and functional mechanism of low molecular weight glutenin subunit genes in bread wheat.

Dong L, Zhang X, Liu D, Fan H, Sun J, Zhang Z, Qin H, Li B, Hao S, Li Z, Wang D, Zhang A, Ling HQ - PLoS ONE (2010)

Bottom Line: Fourteen unique LMW-GS genes were identified for Xiaoyan 54 (with superior bread-making quality).This work provides substantial new insights into the genomic organization and expression of LMW-GS genes, and molecular genetic evidence suggesting that these genes contribute quantitatively to bread-making quality in hexaploid wheat.Our analysis also indicates that selection for high numbers of active LMW-GS genes can be used for improvement of bread-making quality in wheat breeding.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Plant Cell and Chromosome Engineering, National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

ABSTRACT
The bread-making quality of wheat is strongly influenced by multiple low molecular weight glutenin subunit (LMW-GS) proteins expressed in the seeds. However, the organization, recombination and expression of LMW-GS genes and their functional mechanism in bread-making are not well understood. Here we report a systematic molecular analysis of LMW-GS genes located at the orthologous Glu-3 loci (Glu-A3, B3 and D3) of bread wheat using complementary approaches (genome wide characterization of gene members, expression profiling, proteomic analysis). Fourteen unique LMW-GS genes were identified for Xiaoyan 54 (with superior bread-making quality). Molecular mapping and recombination analyses revealed that the three Glu-3 loci of Xiaoyan 54 harbored dissimilar numbers of LMW-GS genes and covered different genetic distances. The number of expressed LMW-GS in the seeds was higher in Xiaoyan 54 than in Jing 411 (with relatively poor bread-making quality). This correlated with the finding of higher numbers of active LMW-GS genes at the A3 and D3 loci in Xiaoyan 54. Association analysis using recombinant inbred lines suggested that positive interactions, conferred by genetic combinations of the Glu-3 locus alleles with more numerous active LMW-GS genes, were generally important for the recombinant progenies to attain high Zeleny sedimentation value (ZSV), an important indicator of bread-making quality. A higher number of active LMW-GS genes tended to lead to a more elevated ZSV, although this tendency was influenced by genetic background. This work provides substantial new insights into the genomic organization and expression of LMW-GS genes, and molecular genetic evidence suggesting that these genes contribute quantitatively to bread-making quality in hexaploid wheat. Our analysis also indicates that selection for high numbers of active LMW-GS genes can be used for improvement of bread-making quality in wheat breeding.

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Transcriptional profiles of LMW-GS genes in developing grains of Jing 411 and Xiaoyan 54.Total RNA samples, extracted from developing grains at 7, 14 and 21 days post anthesis (DPA), were used for evaluating transcript levels of the LMW-GS gene members. The 11 LMW-GS genes were all highly transcribed in the developing grains of Xiaoyan 54 at 14 and 21 DPA. By contrast, in the developing grains of Jing 411, only 6 members (A3-4, B3-1, B3-2, D3-1, D3-2, D3-3) were highly transcribed at 14 and 21 DPA; two members (A3-1, D3-4) were weakly transcribed, and the transcripts of 3 members (A3-2, D3-6, D3-7) were undetectable by RT-PCR at the same time points. The amplification of wheat tubulin gene transcripts served as an internal control for normalizing the cDNA contents before PCR and checking the kinetics of thermamplification during PCR.
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pone-0013548-g003: Transcriptional profiles of LMW-GS genes in developing grains of Jing 411 and Xiaoyan 54.Total RNA samples, extracted from developing grains at 7, 14 and 21 days post anthesis (DPA), were used for evaluating transcript levels of the LMW-GS gene members. The 11 LMW-GS genes were all highly transcribed in the developing grains of Xiaoyan 54 at 14 and 21 DPA. By contrast, in the developing grains of Jing 411, only 6 members (A3-4, B3-1, B3-2, D3-1, D3-2, D3-3) were highly transcribed at 14 and 21 DPA; two members (A3-1, D3-4) were weakly transcribed, and the transcripts of 3 members (A3-2, D3-6, D3-7) were undetectable by RT-PCR at the same time points. The amplification of wheat tubulin gene transcripts served as an internal control for normalizing the cDNA contents before PCR and checking the kinetics of thermamplification during PCR.

Mentions: Further to the experiments described above, it was relevant and important to study the transcription profiles of LMW-GS genes in the developing seeds, and to compare the transcriptional patterns of these genes in wheat varieties differing in bread making quality. To achieve these goals, semiquantitative RT-PCR experiments were performed using gene-specific oligonucleotide primers (Table S1) designed for the 11 active LMW-GS genes of Xiaoyan 54. Figure 3 shows that the 11 members were all highly transcribed in the developing grains of Xiaoyan 54. By contrast, in the developing grains of Jing 411, the transcripts of A3-2, D3-6 and D3-7 were undetectable, and the relative transcript levels of A3-1 and D3-4 were both substantially lower than those in Xiaoyan 54, especially at 21 days post anthesis (DPA) (Figure 3). The data displayed in Figure 3 are representative of four independent RT-PCR experiments.


New insights into the organization, recombination, expression and functional mechanism of low molecular weight glutenin subunit genes in bread wheat.

Dong L, Zhang X, Liu D, Fan H, Sun J, Zhang Z, Qin H, Li B, Hao S, Li Z, Wang D, Zhang A, Ling HQ - PLoS ONE (2010)

Transcriptional profiles of LMW-GS genes in developing grains of Jing 411 and Xiaoyan 54.Total RNA samples, extracted from developing grains at 7, 14 and 21 days post anthesis (DPA), were used for evaluating transcript levels of the LMW-GS gene members. The 11 LMW-GS genes were all highly transcribed in the developing grains of Xiaoyan 54 at 14 and 21 DPA. By contrast, in the developing grains of Jing 411, only 6 members (A3-4, B3-1, B3-2, D3-1, D3-2, D3-3) were highly transcribed at 14 and 21 DPA; two members (A3-1, D3-4) were weakly transcribed, and the transcripts of 3 members (A3-2, D3-6, D3-7) were undetectable by RT-PCR at the same time points. The amplification of wheat tubulin gene transcripts served as an internal control for normalizing the cDNA contents before PCR and checking the kinetics of thermamplification during PCR.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2958824&req=5

pone-0013548-g003: Transcriptional profiles of LMW-GS genes in developing grains of Jing 411 and Xiaoyan 54.Total RNA samples, extracted from developing grains at 7, 14 and 21 days post anthesis (DPA), were used for evaluating transcript levels of the LMW-GS gene members. The 11 LMW-GS genes were all highly transcribed in the developing grains of Xiaoyan 54 at 14 and 21 DPA. By contrast, in the developing grains of Jing 411, only 6 members (A3-4, B3-1, B3-2, D3-1, D3-2, D3-3) were highly transcribed at 14 and 21 DPA; two members (A3-1, D3-4) were weakly transcribed, and the transcripts of 3 members (A3-2, D3-6, D3-7) were undetectable by RT-PCR at the same time points. The amplification of wheat tubulin gene transcripts served as an internal control for normalizing the cDNA contents before PCR and checking the kinetics of thermamplification during PCR.
Mentions: Further to the experiments described above, it was relevant and important to study the transcription profiles of LMW-GS genes in the developing seeds, and to compare the transcriptional patterns of these genes in wheat varieties differing in bread making quality. To achieve these goals, semiquantitative RT-PCR experiments were performed using gene-specific oligonucleotide primers (Table S1) designed for the 11 active LMW-GS genes of Xiaoyan 54. Figure 3 shows that the 11 members were all highly transcribed in the developing grains of Xiaoyan 54. By contrast, in the developing grains of Jing 411, the transcripts of A3-2, D3-6 and D3-7 were undetectable, and the relative transcript levels of A3-1 and D3-4 were both substantially lower than those in Xiaoyan 54, especially at 21 days post anthesis (DPA) (Figure 3). The data displayed in Figure 3 are representative of four independent RT-PCR experiments.

Bottom Line: Fourteen unique LMW-GS genes were identified for Xiaoyan 54 (with superior bread-making quality).This work provides substantial new insights into the genomic organization and expression of LMW-GS genes, and molecular genetic evidence suggesting that these genes contribute quantitatively to bread-making quality in hexaploid wheat.Our analysis also indicates that selection for high numbers of active LMW-GS genes can be used for improvement of bread-making quality in wheat breeding.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Plant Cell and Chromosome Engineering, National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

ABSTRACT
The bread-making quality of wheat is strongly influenced by multiple low molecular weight glutenin subunit (LMW-GS) proteins expressed in the seeds. However, the organization, recombination and expression of LMW-GS genes and their functional mechanism in bread-making are not well understood. Here we report a systematic molecular analysis of LMW-GS genes located at the orthologous Glu-3 loci (Glu-A3, B3 and D3) of bread wheat using complementary approaches (genome wide characterization of gene members, expression profiling, proteomic analysis). Fourteen unique LMW-GS genes were identified for Xiaoyan 54 (with superior bread-making quality). Molecular mapping and recombination analyses revealed that the three Glu-3 loci of Xiaoyan 54 harbored dissimilar numbers of LMW-GS genes and covered different genetic distances. The number of expressed LMW-GS in the seeds was higher in Xiaoyan 54 than in Jing 411 (with relatively poor bread-making quality). This correlated with the finding of higher numbers of active LMW-GS genes at the A3 and D3 loci in Xiaoyan 54. Association analysis using recombinant inbred lines suggested that positive interactions, conferred by genetic combinations of the Glu-3 locus alleles with more numerous active LMW-GS genes, were generally important for the recombinant progenies to attain high Zeleny sedimentation value (ZSV), an important indicator of bread-making quality. A higher number of active LMW-GS genes tended to lead to a more elevated ZSV, although this tendency was influenced by genetic background. This work provides substantial new insights into the genomic organization and expression of LMW-GS genes, and molecular genetic evidence suggesting that these genes contribute quantitatively to bread-making quality in hexaploid wheat. Our analysis also indicates that selection for high numbers of active LMW-GS genes can be used for improvement of bread-making quality in wheat breeding.

Show MeSH
Related in: MedlinePlus