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Deletion of high-molecular-weight glutenin subunits in wheat significantly reduced dough strength and bread-baking quality

BACKGROUND: High-molecular-weight glutenin subunits (HMW-GS) play important roles in the elasticity of dough made from wheat. The HMW-GS null line is useful for studying the contribution of HMW-GS to the end-use quality of wheat. METHODS: In a previous work, we cloned the Glu-1E(b)x gene from Thinop...

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Autores principales: Zhang, Yingjun, Hu, Mengyun, Liu, Qian, Sun, Lijing, Chen, Xiyong, Lv, Liangjie, Liu, Yuping, Jia, Xu, Li, Hui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6276161/
https://www.ncbi.nlm.nih.gov/pubmed/30509162
http://dx.doi.org/10.1186/s12870-018-1530-z
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author Zhang, Yingjun
Hu, Mengyun
Liu, Qian
Sun, Lijing
Chen, Xiyong
Lv, Liangjie
Liu, Yuping
Jia, Xu
Li, Hui
author_facet Zhang, Yingjun
Hu, Mengyun
Liu, Qian
Sun, Lijing
Chen, Xiyong
Lv, Liangjie
Liu, Yuping
Jia, Xu
Li, Hui
author_sort Zhang, Yingjun
collection PubMed
description BACKGROUND: High-molecular-weight glutenin subunits (HMW-GS) play important roles in the elasticity of dough made from wheat. The HMW-GS null line is useful for studying the contribution of HMW-GS to the end-use quality of wheat. METHODS: In a previous work, we cloned the Glu-1E(b)x gene from Thinopyrum bessarabicum and introduced it into the wheat cultivar, Bobwhite. In addition to lines expressing the Glu-1E(b)x gene, we also obtained a transgenic line (LH-11) with all the HMW-GS genes silenced. The HMW-GS deletion was stably inherited as a dominant and conformed to Mendel’s laws. Expression levels of HMW-GS were determined by RT-PCR and epigenetic changes in methylation patterns and small RNAs were analyzed. Glutenins and gliadins were separated and quantitated by reversed-phase ultra-performance liquid chromatography. Measurement of glutenin macropolymer, and analysis of agronomic traits and end-use quality were also performed. RESULTS: DNA methylation and the presence of small double-stranded RNA may be the causes of post-transcriptional gene silencing in LH-11. The accumulation rate and final content of glutenin macropolymer (GMP) in LH-11 were significantly lower than in wild-type (WT) Bobwhite. The total protein content was not significantly affected as the total gliadin content increased in LH-11 compared to WT. Deletion of HMW-GS also changed the content of different gliadin fractions. The ratio of ω-gliadin increased, whereas α/β- and γ-gliadins declined in LH-11. The wet gluten content, sedimentation value, development time and stability time of LH-11 were remarkably lower than that of Bobwhite. Bread cannot be made using the flour of LH-11. CONCLUSIONS: Post-transcriptional gene silencing through epigenetic changes and RNA inhibition appear to be the causes for the gene expression deficiency in the transgenic line LH-11. The silencing of HMW-GW in LH-11 significantly reduced the dough properties, GMP content, wet gluten content, sedimentation value, development time and stability time of flour made from this wheat cultivar. The HMW-GS null line may provide a potential material for biscuit-making because of its low dough strength. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12870-018-1530-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-62761612018-12-06 Deletion of high-molecular-weight glutenin subunits in wheat significantly reduced dough strength and bread-baking quality Zhang, Yingjun Hu, Mengyun Liu, Qian Sun, Lijing Chen, Xiyong Lv, Liangjie Liu, Yuping Jia, Xu Li, Hui BMC Plant Biol Research Article BACKGROUND: High-molecular-weight glutenin subunits (HMW-GS) play important roles in the elasticity of dough made from wheat. The HMW-GS null line is useful for studying the contribution of HMW-GS to the end-use quality of wheat. METHODS: In a previous work, we cloned the Glu-1E(b)x gene from Thinopyrum bessarabicum and introduced it into the wheat cultivar, Bobwhite. In addition to lines expressing the Glu-1E(b)x gene, we also obtained a transgenic line (LH-11) with all the HMW-GS genes silenced. The HMW-GS deletion was stably inherited as a dominant and conformed to Mendel’s laws. Expression levels of HMW-GS were determined by RT-PCR and epigenetic changes in methylation patterns and small RNAs were analyzed. Glutenins and gliadins were separated and quantitated by reversed-phase ultra-performance liquid chromatography. Measurement of glutenin macropolymer, and analysis of agronomic traits and end-use quality were also performed. RESULTS: DNA methylation and the presence of small double-stranded RNA may be the causes of post-transcriptional gene silencing in LH-11. The accumulation rate and final content of glutenin macropolymer (GMP) in LH-11 were significantly lower than in wild-type (WT) Bobwhite. The total protein content was not significantly affected as the total gliadin content increased in LH-11 compared to WT. Deletion of HMW-GS also changed the content of different gliadin fractions. The ratio of ω-gliadin increased, whereas α/β- and γ-gliadins declined in LH-11. The wet gluten content, sedimentation value, development time and stability time of LH-11 were remarkably lower than that of Bobwhite. Bread cannot be made using the flour of LH-11. CONCLUSIONS: Post-transcriptional gene silencing through epigenetic changes and RNA inhibition appear to be the causes for the gene expression deficiency in the transgenic line LH-11. The silencing of HMW-GW in LH-11 significantly reduced the dough properties, GMP content, wet gluten content, sedimentation value, development time and stability time of flour made from this wheat cultivar. The HMW-GS null line may provide a potential material for biscuit-making because of its low dough strength. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12870-018-1530-z) contains supplementary material, which is available to authorized users. BioMed Central 2018-12-03 /pmc/articles/PMC6276161/ /pubmed/30509162 http://dx.doi.org/10.1186/s12870-018-1530-z Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Zhang, Yingjun
Hu, Mengyun
Liu, Qian
Sun, Lijing
Chen, Xiyong
Lv, Liangjie
Liu, Yuping
Jia, Xu
Li, Hui
Deletion of high-molecular-weight glutenin subunits in wheat significantly reduced dough strength and bread-baking quality
title Deletion of high-molecular-weight glutenin subunits in wheat significantly reduced dough strength and bread-baking quality
title_full Deletion of high-molecular-weight glutenin subunits in wheat significantly reduced dough strength and bread-baking quality
title_fullStr Deletion of high-molecular-weight glutenin subunits in wheat significantly reduced dough strength and bread-baking quality
title_full_unstemmed Deletion of high-molecular-weight glutenin subunits in wheat significantly reduced dough strength and bread-baking quality
title_short Deletion of high-molecular-weight glutenin subunits in wheat significantly reduced dough strength and bread-baking quality
title_sort deletion of high-molecular-weight glutenin subunits in wheat significantly reduced dough strength and bread-baking quality
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6276161/
https://www.ncbi.nlm.nih.gov/pubmed/30509162
http://dx.doi.org/10.1186/s12870-018-1530-z
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