[1]王沙沙,何宁,汪庆昌,等.小麦γ-醇溶蛋白基因Tagli-γ-11的克隆及其互作蛋白质分析[J].江苏农业学报,2025,(05):833-839.[doi:doi:10.3969/j.issn.1000-4440.2025.05.001]
　WANG Shasha,HE Ning,WANG Qingchang,et al.Cloning of γ-gliadin gene Tagli-γ-11 and interactive proteins analysis in wheat[J].,2025,(05):833-839.[doi:doi:10.3969/j.issn.1000-4440.2025.05.001]
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小麦γ-醇溶蛋白基因Tagli-γ-11的克隆及其互作蛋白质分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2025年05期
页码:
833-839
栏目:
遗传育种·生理生化
出版日期:
2025-05-31

文章信息/Info

Title:
Cloning of γ-gliadin gene Tagli-γ-11 and interactive proteins analysis in wheat
作者:
王沙沙1; 何宁1; 汪庆昌1; 黄超1; 宋晓2; 晁岳恩1
(1.河南省农业科学院小麦研究所/河南省小麦生物学重点实验室,河南郑州450002；2.河南省农业科学院植物营养与资源环境研究所,河南郑州450002)
Author(s):
WANG Shasha1; HE Ning1; WANG Qingchang1; HUANG Chao1; SONG Xiao2; CHAO Yue’en1
(1.Institute of Wheat Research, Henan Academy of Agricultural Sciences/Key Laboratory for Wheat Biology of Henan Province, Zhengzhou 450002, China;2.Institute of Plant Nutrient and Environmental Resources, Henan Academy of Agricultural Science, Zhengzhou 450002, China)
关键词:
小麦; Tagli-γ-11基因; 酵母双杂交; 回转验证
Keywords:
Triticum aestivum L.; Tagli-γ-11 gene; yeast two-hybrid system; rotation validation
分类号:
S512.1;Q756
DOI:
doi:10.3969/j.issn.1000-4440.2025.05.001
文献标志码:
A
摘要:
为进一步解析γ-醇溶蛋白基因在小麦面粉品质中的调控机制,本研究从郑麦158(低蛋白质含量、高面团强度)中克隆了小麦γ-醇溶蛋白基因Tagli-γ-11。生物信息学分析结果表明,该基因具有γ-醇溶蛋白的典型结构特征,含有8个保守的半胱氨酸残基,并在该蛋白质的重复区Ⅱ内第133～141 aa位置发现1个乳糜泻(CD)表位(PQQSFPQQQ)。利用酵母双杂交技术筛选郑麦158的cDNA文库,共筛选了8个可能与Tagli-γ-11互作的蛋白质,它们分别是半胱氨酸蛋白酶、果糖二磷酸醛缩酶、富含半胱氨酸和跨膜结构域蛋白1、(1,3∶1,4)-β-D-葡聚糖酶、生长素响应因子ARF17-like、细胞数目调控因子CNR8-like、泛素结构域蛋白DSK2b和转录因子PIF1-like。3个代表性候选蛋白质[富含半胱氨酸和跨膜结构域蛋白1、果糖二磷酸醛缩酶、(1,3∶1,4)-β-D-葡聚糖酶]的回转验证结果表明,它们与Tagli-γ-11均存在互作关系,推测Tagli-γ-11与这些蛋白质相互作用,主要参与了小麦籽粒中贮藏蛋白(如醇溶蛋白)和淀粉的合成与降解,以及生殖生长过程。
Abstract:
To further analyze the regulation mechanism of γ-gliadin gene in flour quality of wheat, the Tagli-γ-11 gene was successfully cloned from Zhengmai 158 (low protein content and high dough strength). Bioinformatic analysis revealed that this gene possessed the typical structural characteristics of γ-gliadin, containing eight conserved cysteine residues. A celiac disease (CD) epitope (PQQSFPQQQ) was identified within repeat domain Ⅱ of the Tagli-γ-11 protein, located at amino acid positions 133-141. Using the yeast two-hybrid (Y2H) system, the cDNA library of Zhengmai 158 was screened, yielding eight candidate proteins potentially interacting with Tagli-γ-11. These included cysteine protease, fructose-bisphosphate aldolase, cysteine-rich and transmembrane domain-containing protein 1, (1,3∶1,4)-β-D-glucanase, auxin response factor 17-like (ARF17-like), cell number regulator 8-like (CNR8-like), ubiquitin-associated domain-containing protein DSK2b, transcription factor PIF1-like. The rotation validation assays performed on cysteine-rich and transmembrane domain-containing protein 1, fructose-bisphosphate aldolase, and (1,3∶1,4)-β-D-glucanase confirmed physical interaction with Tagli-γ-11. Based on these results, it was hypothesized that Tagli-γ-11 interacted with these proteins and participated in the synthesis and degradation of storage proteins (such as gliadin) and starch within wheat grains, as well as in reproductive growth processes.

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备注/Memo

备注/Memo:
收稿日期：2024-11-08基金项目：河南省科技攻关项目(232102110220)；河南省农业科学院科技攻关项目；河南省农业科学院自主创新项目(2024ZC001)作者简介：王沙沙(1981-),女,河南焦作人,博士,助理研究员,研究方向为小麦分子育种。(E-mail)515605313＠qq.com。何宁为共同第一作者。通讯作者：晁岳恩,(Ｅ-mail)nkychaoyueen@163.com
更新日期/Last Update: 2025-06-24