Gidekel Manuel, Destefano-Beltrán Luis, García Patricia, Mujica Lorena, Leal Pamela, Cuba Marely, Fuentes Lida, Bravo León A, Corcuera Luis J, Alberdi Miren, Concha Ilona, Gutiérrez Ana
Laboratorio de Fisiología y Biología Molecular Vegetal, Instituto de Agroindustria, Facultad de Ciencias Agropecuarias y Forestales, Universidad de La Frontera, Temuco, Chile.
Extremophiles. 2003 Dec;7(6):459-69. doi: 10.1007/s00792-003-0345-4. Epub 2003 Sep 2.
Deschampsia antarctica Desv. is the only monocot that thrives in the harsh conditions of the Antarctic Peninsula and represents an invaluable resource for the identification of genes associated with freezing tolerance. In order to identify genes regulated by low temperature, we have initiated a detailed analysis of its gene expression. Preliminary 2-D gels of in vivo-labeled leaf proteins showed qualitative and quantitative differences between cold-acclimated and non-acclimated plants, suggesting differential gene expression. Similarly, cold-acclimation-related transcripts were screened by a differential display method. Of the 38 cDNAs initially identified, three cDNA clones were characterized for their protein encoding, expression pattern, response to several stresses, and for their tissue-specific expression. Northern blot analysis of DaGrx, DaRub1, and DaPyk1 encoding a glutaredoxin, a related-to-ubiquitin protein, and a pyruvate kinase-like protein, respectively, showed a distinct regulation pattern during the cold-acclimation process, and in some cases, their cold response seemed to be tissue specific. All three transcripts seem to be responsive to water stress as their levels were up-regulated with polyethyleneglycol treatment. DaRUB1 and DaPyk1 expression was up-regulated in leaf and crown, but down-regulated in roots from cold-acclimated plants. The significance of these results during the cold-acclimation process will be discussed.
南极发草(Deschampsia antarctica Desv.)是唯一能在南极半岛恶劣条件下茁壮成长的单子叶植物,是鉴定与耐冻性相关基因的宝贵资源。为了鉴定受低温调控的基因,我们已开始对其基因表达进行详细分析。体内标记叶蛋白的初步二维凝胶显示,冷驯化植物和未冷驯化植物之间存在定性和定量差异,表明基因表达存在差异。同样,通过差异显示法筛选了与冷驯化相关的转录本。在最初鉴定的38个cDNA中,对三个cDNA克隆进行了蛋白质编码、表达模式、对几种胁迫的反应以及组织特异性表达方面的表征。对分别编码谷氧还蛋白、泛素相关蛋白和丙酮酸激酶样蛋白的DaGrx、DaRub1和DaPyk1进行Northern印迹分析,结果显示在冷驯化过程中存在明显的调控模式,在某些情况下,它们的冷反应似乎具有组织特异性。所有这三种转录本似乎都对水分胁迫有反应,因为用聚乙二醇处理后它们的水平上调。冷驯化植物的叶片和冠部中DaRUB1和DaPyk1的表达上调,但根部中的表达下调。将讨论这些结果在冷驯化过程中的意义。