National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
Sci Rep. 2019 Jan 15;9(1):141. doi: 10.1038/s41598-018-37859-y.
Abiotic stress significantly impacts growth and yield of crop plants. It is imperative for crop improvement to discover and utilize stress-tolerant functional genes. In this study, genes responding to abiotic stresses, such as freezing, salt and osmotic stress, were screened from a cDNA yeast library that was constructed from the drought- and heat-tolerant wheat variety Hanxuan 10. After screening for surviving clones we isolated 7,249, 4,313 and 4,469 raw sequences, corresponding to 4,695, 2,641 and 2,771 genes following each treatment. Venn diagrams revealed 377 overlapping genes. GO analysis suggested that these genes were mainly involved in the metabolic and stress signal pathways. KEGG pathway enrichment analysis indicated that the isolated genes predominantly belonged to pathways concerning energy and metabolism. Overlapping gene TaPR-1-1 within the pathogenesis-related (PR) protein family was selected for detailed characterization. Although previous studies had shown that PR genes function during pathogen attack, our results demonstrated that TaPR-1-1 expression was also induced by freezing, salinity, and osmotic stresses. Overexpression in yeast and Arabidopsis showed that TaPR-1-1 conferred tolerance to these stresses. We concluded that screening cDNA yeast libraries following abiotic stress is an efficient way to identify stress-tolerance genes.
非生物胁迫显著影响作物的生长和产量。发现和利用耐胁迫功能基因对于作物改良至关重要。本研究从耐旱、耐热小麦品种旱选 10 号构建的 cDNA 酵母文库中筛选出响应非生物胁迫（如冷冻、盐和渗透胁迫）的基因。在筛选存活克隆后，我们分别从每种处理中分离出 7249、4313 和 4469 条原始序列，分别对应 4695、2641 和 2771 个基因。Venn 图显示有 377 个重叠基因。GO 分析表明，这些基因主要参与代谢和应激信号途径。KEGG 途径富集分析表明，分离出的基因主要属于与能量和代谢相关的途径。选择与病程相关（PR）蛋白家族重叠的 TaPR-1-1 基因进行详细特征分析。尽管先前的研究表明 PR 基因在病原体攻击时发挥作用，但我们的结果表明 TaPR-1-1 的表达也受到冷冻、盐和渗透胁迫的诱导。在酵母和拟南芥中的过表达表明 TaPR-1-1 赋予了对这些胁迫的耐受性。我们得出结论，筛选 cDNA 酵母文库以应对非生物胁迫是鉴定耐胁迫基因的有效方法。
