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Identification of Rice Genes Associated With Enhanced Cold Tolerance by Comparative Transcriptome Analysis With Two Transgenic Rice Plants Overexpressing DaCBF4 or DaCBF7 Isolated From Antarctic Flowering Plant Deschampsia antarctica

机译:通过比较转录组分析从南极开花植物南极南芥中分离到的两种过表达DaCBF4或DaCBF7的转基因水稻植株通过比较转录组分析鉴定与耐寒性相关的水稻基因

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摘要

Few plant species can survive in Antarctica, the harshest environment for living organisms. Deschampsia antarctica is the only natural grass species to have adapted to and colonized the maritime Antarctic. To investigate the molecular mechanism of the Antarctic adaptation of this plant, we identified and characterized D. antarctica C-repeat binding factor 4 (DaCBF4), which belongs to monocot CBF group IV. The transcript level of DaCBF4 in D. antarctica was markedly increased by cold and dehydration stress. To assess the roles of DaCBF4 in plants, we generated a DaCBF4-overexpressing transgenic rice plant (Ubi:DaCBF4) and analyzed its abiotic stress response phenotype. Ubi:DaCBF4 displayed enhanced tolerance to cold stress without growth retardation under any condition compared to wild-type plants. Because the cold-specific phenotype of Ubi:DaCBF4 was similar to that of Ubi:DaCBF7 (Byun et al., ), we screened for the genes responsible for the improved cold tolerance in rice by selecting differentially regulated genes in both transgenic rice lines. By comparative transcriptome analysis using RNA-seq, we identified 9 and 15 genes under normal and cold-stress conditions, respectively, as putative downstream targets of the two D. antarctica CBFs. Overall, our results suggest that Antarctic hairgrass DaCBF4 mediates the cold-stress response of transgenic rice plants by adjusting the expression levels of a set of stress-responsive genes in transgenic rice plants. Moreover, selected downstream target genes will be useful for genetic engineering to enhance the cold tolerance of cereal plants, including rice.
机译:极少数植物物种可以在南极洲生存,南极洲是生物最恶劣的环境。南极洲Deschampsia是唯一适应并定居于海洋南极洲的天然草种。为了研究该植物对南极适应的分子机制,我们鉴定并鉴定了属于单子叶植物CBF组IV的南极D. C-重复结合因子4(DaCBF4)。低温和脱水胁迫使南极D.DaCBF4的转录水平显着增加。为了评估DaCBF4在植物中的作用,我们生成了过表达DaCBF4的转基因水稻植株(Ubi:DaCBF4),并分析了其非生物胁迫响应表型。与野生型植物相比,Ubi:DaCBF4在任何条件下均显示出增强的对冷胁迫的耐受性,而没有生长迟缓。由于Ubi:DaCBF4的寒冷特异性表型与Ubi:DaCBF7的寒冷特异性表型相似(Byun等人,因此),我们通过在两个转基因水稻品系中选择差异调节的基因,筛选了导致水稻耐寒性提高的基因。通过使用RNA-seq进行比较转录组分析,我们分别在正常和寒冷胁迫条件下鉴定了9个和15个基因,作为两个南极D. CBF的假定下游靶标。总体而言,我们的结果表明,南极毛草DaCBF4通过调节转基因水稻植株中一组胁迫响应基因的表达水平来介导转基因水稻植株的冷胁迫响应。此外,选定的下游靶基因将对遗传工程有用,以增强包括水稻在内的谷物植物的耐寒性。

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