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Comparative transcriptomic profiling of Vitis vinifera under high light using a custom-made array and the Affymetrix GeneChip.

机译:使用定制阵列和Affymetrix GeneChip在强光下比较葡萄的转录组谱分析。

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Understanding abiotic stress responses is one of the most important issues in plant research nowadays. Abiotic stress, including excess light, can promote the onset of oxidative stress through the accumulation of reactive oxygen species. Oxidative stress also arises when in vitro propagated plants are exposed to high light upon transfer to ex vitro. To determine whether the underlying pathways activated at the transfer of in vitro grapevine to ex vitro conditions reflect the processes occurring upon light stress, we used Vitis vinifera Affymetrix GeneChip (VvGA) and a custom array of genes responsive to light stress (LSCA) detected by real-time reverse transcriptase PCR (qRT-PCR). When gene-expression profiles were compared, 'protein metabolism and modification', 'signaling', and 'anti-oxidative' genes were more represented in LSCA, while, in VvGA, 'cell wall metabolism' and 'secondary metabolism' were the categories in which gene expression varied more significantly. The above functional categories confirm previous studies involving other types of abiotic stresses, enhancing the common attributes of abiotic stress defense pathways. The LSCA analysis of our experimental system detected strong response of heat shock genes, particularly the protein rescuing mechanism involving the cooperation of two ATP-dependent chaperone systems, Hsp100 and Hsp70, which showed an unusually late response during the recovery period, of extreme relevance to remove non-functional, potentially harmful polypeptides arising from misfolding, denaturation, or aggregation brought about by stress. The success of LSCA also proves the feasibility of a custom-made qRT-PCR approach, particularly for species for which no GeneChip is available and for researchers dealing with a specific and focused problem.Digital Object Identifier http://dx.doi.org/10.1093/mp/ssr027
机译:了解非生物胁迫响应是当今植物研究中最重要的问题之一。非生物胁迫,包括过量的光,可以通过活性氧的积累促进氧化应激的发作。当将体外繁殖的植物转移到体外时,暴露于强光下也会产生氧化胁迫。为了确定在体外葡萄转移到体外条件下激活的潜在途径是否反映了轻胁迫下发生的过程,我们使用了 Vitis vinifera Affymetrix基因芯片(VvGA)和通过实时逆转录酶PCR(qRT-PCR)检测到的对光胁迫(LSCA)响应的基因定制阵列。比较基因表达谱时,“蛋白质代谢和修饰”,“信号传递”和“抗氧化”基因在LSCA中占更多比例,而在VvGA中,“细胞壁代谢”和“次级代谢”属于此类其中基因表达差异更大。以上功能类别确认了先前涉及其他类型的非生物胁迫的研究,从而增强了非生物胁迫防御途径的共同属性。我们的实验系统的LSCA分析检测到了热休克基因的强烈反应,特别是涉及两个ATP依赖性伴侣系统Hsp100和Hsp70协同作用的蛋白质拯救机制,它们在恢复期显示出异常晚的反应,与去除因压力导致的错误折叠,变性或聚集而产生的非功能性,潜在有害的多肽。 LSCA的成功也证明了定制qRT-PCR方法的可行性,特别是对于无法获得GeneChip的物种以及处理特定且关注的问题的研究人员而言。数字对象标识符http://dx.doi.org /10.1093/mp/ssr027

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