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| 基因敲除揭示TaMAPK1的三个部分同源基因以叠加效应正调控小麦苗期耐旱性 |
| Gene knockout reveals that the three homeologous genes of TaMAPK1 positively regulate drought tolerance in wheat seedlings through an additive effect |
| 投稿时间：2026-01-14 修订日期：2026-02-03 |
| DOI： |
| 中文关键词: 小麦 TaMAPK1 耐旱性 基因编辑 同源基因 叠加效应 |
| 英文关键词:Wheat TaMAPK1 Drought tolerance Gene editing Homoeologs Additive effect |
| 基金项目:陕西省重点研发项目（2025NC-YBXM-002）；国家自然基金项目（32372103和32072003）；陕西省自然科学基础研究计划项目(No.2024JC-YBMS-171) |
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| 中文摘要: |
| 为解析小麦丝裂原活化蛋白激酶基因TaMAPK1-6A、-6B和-6D在苗期耐旱性中的功能及其同源基因之间的叠加效应,本研究以春小麦品种Fielder和通过CRISPR/Cas9介导的小麦基因组编辑技术创制的在Fielder遗传背景下的TaMAPK1的单基因突变体mapk1-aa、mapk1-bb和mapk1-dd单突变体为试材,通过杂交、自交及其后代分离群体的基因型检测,创制了双基因敲除突变体和三基因敲除纯合突变体(分别简称双突变体和三突变体)；并对这些单突、双突和三突变体以及对照(野生型小麦Fielder)苗期耐旱性进行了系统评价。结果表明,TaMAPK1单基因功能缺失突变导致小麦幼苗耐旱性显著降低,表现为叶片保水能力下降、渗透调节物质积累减少、膜脂过氧化加剧。进一步研究发现双突变体与三突变体的耐旱性进一步降低,生理生化指标恶化程度随突变基因拷贝数增加而加剧。综上,TaMAPK1基因正调控小麦苗期耐旱性,其同源基因具有累加调控作用,研究结果为小麦抗旱分子育种提供了重要候选基因,并为多倍体作物同源基因功能解析提供了理论参考。 |
| 英文摘要: |
| To elucidate the functions of the wheat mitogen-activated protein kinase genes TaMAPK1-6A, -6B, and -6D in seedling drought tolerance and the additive effects among these homeologous genes, this study utilized the spring wheat cultivar Fielder and its CRISPR/Cas9 mediated single-gene mutants-mapk1-aa, mapk1-bb, and mapk1-dd-generated in the Fielder genetic background. Through hybridization, self-pollination, and genotyping of segregating progeny, double-gene knockout mutants and triple-gene knockout homozygous mutants (hereafter referred to as double and triple mutants) were successfully developed. The drought tolerance at the seedling stage was systematically evaluated for these single, double, and triple mutants, along with the wild-type Fielder control. The results demonstrated that loss-of-function mutations in single TaMAPK1 genes significantly reduced drought tolerance in wheat seedlings, manifested as decreased leaf water retention, reduced accumulation of osmotic adjustment substances, and enhanced membrane lipid peroxidation. Further investigation revealed that drought tolerance was further impaired in double and triple mutants, with the deterioration of physiological and biochemical indicators intensifying as the number of mutated gene copies increased. In summary, the TaMAPK1 gene positively regulates drought tolerance at the wheat seedling stage, and its homeologous genes exhibit cumulative regulatory effects. This study provides an important candidate gene for drought-resistant molecular breeding in wheat and offers theoretical insights for functional analysis of homeologous genes in polyploid crops. |
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