| 何 宁，王沙沙，黄 超，汪庆昌，宋 晓，晁岳恩.小麦燕麦类似蛋白TaALP-7D自激活活性检测及其互作蛋白分析[J].麦类作物学报,2025,(12):1599 |
| 小麦燕麦类似蛋白TaALP-7D自激活活性检测及其互作蛋白分析 |
| Detection of Self-Activation Activity and Analysis of Interacting Proteins of Avenin-Like Protein TaALP-7D in Bread Wheat |
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| DOI： |
| 中文关键词: 小麦 燕麦类似蛋白 TaALP-7D基因 表达分析 酵母双杂交 回转验证 |
| 英文关键词:Triticum aestivum L. Avenin-like proteins TaALP-7D gene Expression analysis Yeast two-hybrid system Rotation validation |
| 基金项目:河南省重大科技专项(241100110300)；河南省中央引进地方科技发展资金项目(Z20241471126)；河南省农科院自主创新项目(2025ZC03) |
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| 中文摘要: |
| 为解析燕麦类似蛋白(avenin-like proteins，ALPs)在小麦品质中的调控机制，从小麦品种郑麦158中克隆了小麦燕麦类似蛋白基因TaALP-7D，并对其进行表达分析及其互作蛋白质筛选，确定与其互作的关键候选蛋白。结果表明，在小麦不同组织中，TaALP-7D基因在籽粒不同发育时期的相对表达量均最高，推测TaALP-7D基因可能参与小麦种子发育过程中面团强度相关基因的调控。利用酵母双杂交技术共筛选了6个可能与TaALP-7D互作的蛋白，分别是半胱氨酸水解酶1(XM_044496235.1)、F-box蛋白SKP2A-like (XM_044518121.1)、非特异性脂转移蛋白LTP1-like (XM_044488525.1)和1,3-β-葡聚糖苷酶(XM_044596055.1)以及与植物激素迫响应有关的乙烯响应转录因子TaERF1-like (XM_044549609.1) 和茉莉酸诱导的氧化酶JOX2-like (XM_044519612.1)。经回转验证，5个代表性候选蛋白(半胱氨酸水解酶 1、F-box蛋白SKP2A-like、非特异性脂转移蛋白LTP1-like、1,3-β-葡聚糖苷酶和乙烯响应转录因子TaERF1-like)与TaALP-7D均存在互作关系，推测TaALP-7D可能通过与这些蛋白互作，参与了小麦籽粒中贮藏蛋白(如醇溶蛋白)的降解、淀粉的合成以及其他与激素响应有关的过程。 |
| 英文摘要: |
| In order to further analyze the regulation mechanism of avenin-like proteins in flour quality of wheat, the TaALP-7D gene was successfully cloned from Zhengmai 158(strong dough strength). Relative expression patterns of different tissues showed that the TaALP-7D gene was highly expressed at different developmental stages of the kernels. It was suggested that TaALP-7D gene may be involved in the regulation of genes related to dough strength during wheat seed development. Six possible candidate interactive proteins of TaALP-7D were screened from Zhengmai 158 cDNA library by the yeast two-hybrid system: cysteine hydrolase 1(XM_044496235.1), F-box protein SKP2A-like(XM_044518121.1), non specific lipid transfer protein LTP1-like(XM_044488525.1), and 1,3-β-glucosidase(XM_044596055.1),and also the ethylene response transcription factor TaERF1-like(XM_044549609.1) and jasmonic acid-induced oxidase JOX2-like(XM_044519612.1) related to plant hormone response. The rotation validation assays of the five representative candidate proteins(cysteine hydrolase 1, F-box protein SKP2A like, non-specific lipid transfer protein LTP1-like, 1,3-β-glucosidase, and ethylene response transcription factor TaERF1-like) indicated that all of them interacted with TaALP-7D. Therefore, it is speculated that TaALP-7D probably interacts with these candidate proteins and be mainly involved in the degradation of storage proteins(such as gliadins), synthesis of starch in wheat kernels, as well as other processes of plant hormone response |
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