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面包小麦中的α-醇溶蛋白:氮处理对两个RNA干扰低醇溶蛋白品系中主要乳糜泻免疫原性复合物表达的影响

The α-Gliadins in Bread Wheat: Effect of Nitrogen Treatment on the Expression of the Major Celiac Disease Immunogenic Complex in Two RNAi Low-Gliadin Lines.

作者信息

Sánchez-León Susana, Giménez María José, Barro Francisco

机构信息

Department of Plant Breeding, Institute for Sustainable Agriculture - Spanish National Research Council, Córdoba, Spain.

出版信息

Front Plant Sci. 2021 Apr 29;12:663653. doi: 10.3389/fpls.2021.663653. eCollection 2021.

Abstract

Celiac Disease (CD) is an autoimmune disorder that affects approximately 1% of the worldwide population. The α-gliadins of wheat contain the 33-mer peptide, the most active peptide in CD both in adults and pediatric patients. In this study, we have characterized the variants and expression profile of an α-gliadins amplicon, harboring the 33-mer peptide, in two low-gliadin RNAi wheat lines, under two different Nitrogen (N) treatments. We estimated that the amplicon expands 45 different α-gliadin variants with high variability due to length, randomly distributed SNPs, and the presence of encoded CD epitopes. Expression of this amplicon is reduced in both RNAi lines in comparison to the wild type. High N treatment significantly increases transcripts of the amplicon in the wild type, but not in the transgenic lines. Classification of α-gliadin variants, considering the number of epitopes, revealed that amplicon variants containing the full complement of 33-mer peptide were affected by N treatment, increasing their expression when N was increased. Line D793 provided higher and more stable silencing through different N fertilization regimes, expressing fewer CD epitopes than D783. Results of this study are important for better understanding of RNAi α-gliadin silencing in response to N treatments, and for undertaking new strategies by RNAi or CRISPR/Cas toward obtaining new varieties suitable for people suffering gluten intolerances.

摘要

乳糜泻(CD)是一种自身免疫性疾病,影响着全球约1%的人口。小麦的α-麦醇溶蛋白含有33聚体肽,这是在成人和儿童患者中CD最具活性的肽。在本研究中,我们对两个低麦醇溶蛋白RNA干扰小麦品系中含有33聚体肽的α-麦醇溶蛋白扩增子的变体和表达谱进行了表征,这两个品系处于两种不同的氮(N)处理条件下。我们估计该扩增子扩展出45种不同的α-麦醇溶蛋白变体,由于长度、随机分布的单核苷酸多态性(SNP)以及编码的CD表位的存在,这些变体具有高度变异性。与野生型相比,该扩增子在两个RNA干扰品系中的表达均降低。高氮处理显著增加了野生型中该扩增子的转录本,但在转基因品系中没有增加。考虑到表位数量对α-麦醇溶蛋白变体进行分类,结果显示含有完整33聚体肽互补序列的扩增子变体受到氮处理的影响,当氮含量增加时其表达增加。品系D793通过不同的氮肥施用方式提供了更高且更稳定的沉默效果,其表达的CD表位比D783少。本研究结果对于更好地理解RNA干扰α-麦醇溶蛋白在氮处理响应中的沉默作用,以及通过RNA干扰或CRISPR/Cas采取新策略以获得适合麸质不耐受人群的新品种具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2933/8116895/03c24acb8149/fpls-12-663653-g001.jpg

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