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从不同葡萄品种(Vitis vinifera L.)中分离和表达盐胁迫诱导基因。

Isolation and expression analysis of salt induced genes from contrasting grapevine (Vitis vinifera L.) cultivars.

机构信息

Centre de Biotechnologie de Borj cédria, Laboratoire de Physiologie Moléculaire des Plantes, B.P.901, 2050 Hammam-Lif, Tunisia; RLP-Agroscience GmbH/Alplanta-Institute for Plant Research, Breitenweg 71, 67435 Neustadt and der Weinstraße, Germany.

出版信息

Plant Sci. 2010 Nov;179(5):489-98. doi: 10.1016/j.plantsci.2010.07.017. Epub 2010 Aug 3.

Abstract

Salt stress adversely affects the growth of grapevine plants. In order to understand the molecular basis of salt stress response in grapevine plants, suppression subtractive hybridization (SSH) and microarray based screening approaches were combined. Two leaf-specific subtractive cDNA libraries were constructed from grapevine plants subjected to a moderate, incremental salt stress treatment. SSH were performed 6h and 24h after NaCl peaked at 100mM using cDNAs prepared from leaves of a salt tolerant cultivar (Razegui) as testers and cDNAs from unstressed leaves as drivers. Then, a pre-screened subset of cDNA clones from these SSH libraries were used to construct a Vitis vinifera cDNA array, in order to verify the expression changes of the genes upon salt treatment. Expression profiles were compared between the salt tolerant and a susceptible cultivar (Syrah) under both control conditions and after salt stress treatment. Seven cDNA clones were identified which were up-regulated by salt stress in two independent growth experiments and confirmed by RNA blot analysis. The transcript expression patterns of the selected genes differed between the contrasting grapevine cultivars tested with respect to stress-regulation. The possible relationship of individual cDNAs with salinity tolerance mechanisms is discussed.

摘要

盐胁迫会对葡萄植株的生长造成不利影响。为了深入理解葡萄植株盐胁迫响应的分子基础,我们将抑制差减杂交(SSH)与基于微阵列的筛选方法相结合。利用耐盐品种(Razegui)叶片的 cDNA 作为测试物,以未胁迫叶片的 cDNA 作为驱动物,从受到中度递增盐胁迫处理的葡萄植株中构建了两个叶片特异性的差减 cDNA 文库。在 NaCl 达到 100mM 后 6h 和 24h 进行 SSH。然后,我们使用这些 SSH 文库中的预筛选 cDNA 克隆子集构建了一个葡萄 cDNA 微阵列,以验证基因在盐处理后的表达变化。在对照条件和盐胁迫处理后,我们比较了耐盐品种(Razegui)和易感品种(Syrah)之间的表达谱。在两个独立的生长实验中,有 7 个 cDNA 克隆因盐胁迫而上调,并通过 RNA 印迹分析得到了验证。在受测试的不同葡萄品种中,所选基因的转录表达模式在应激调控方面存在差异。我们讨论了个别 cDNA 与耐盐性机制的可能关系。

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