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对紫外线B辐射后敏感性不同的玉米品系叶片蛋白质组的分析。

Analysis of leaf proteome after UV-B irradiation in maize lines differing in sensitivity.

作者信息

Casati Paula, Zhang Xing, Burlingame Alma L, Walbot Virginia

机构信息

Department of Biological Sciences, Stanford University, Stanford, California 94305-5020, USA.

出版信息

Mol Cell Proteomics. 2005 Nov;4(11):1673-85. doi: 10.1074/mcp.M500173-MCP200. Epub 2005 Jul 25.

Abstract

UV-B radiation causes diverse morphological and physiological responses in plants, but the underlying mechanisms governing these integrated responses are unknown. In this study, we systematically surveyed responses of maize leaves to UV-B radiation using DIGE 2D gels and identified selected proteins by mass spectrometry and immunodetection analysis. To identify changes in protein accumulation in response to UV-B radiation, a line (b, pl W23) deficient in flavonoid sunscreen compounds and hence similar to commercial corn was used. In addition, its proteome in natural UV-B conditions was compared with that of two maize landraces from high altitudes (Cacahuacintle and Confite Puneño) that have improved UV-B tolerance. Protein patterns in adult maize leaves (Zea mays) were documented after growth for 21 days in sunlight depleted of UV-B radiation or growth in sunlight including an 8-h UV-B supplementation during 1 day in the field. We found that there is a very high correlation between previously documented mRNA accumulation assessed by microarray hybridization and quantitative real time reverse transcription-PCR and protein expression after UV-B irradiation in leaves of W23. Multiple isoforms were confirmed for some proteins; at least one protein, pyruvate, phosphate dikinase, is regulated post-translationally by phosphorylation by UV-B exposure. Proteins differentially regulated by UV-B radiation in W23 with higher levels under similar UV-B conditions in high altitude plants were also identified. These could be genetically fixed traits conferring UV-B tolerance and offer clues to specific adaptations to living in high ambient UV-B conditions.

摘要

UV-B辐射会在植物中引发多种形态和生理反应,但其调控这些综合反应的潜在机制尚不清楚。在本研究中,我们使用差异凝胶电泳二维凝胶系统地检测了玉米叶片对UV-B辐射的反应,并通过质谱分析和免疫检测分析鉴定了特定蛋白质。为了确定响应UV-B辐射时蛋白质积累的变化,我们使用了一种缺乏类黄酮防晒化合物且因此与商业玉米相似的品系(b,pl W23)。此外,我们将其在自然UV-B条件下的蛋白质组与两种来自高海拔地区(Cacahuacintle和Confite Puneño)且具有增强的UV-B耐受性的玉米地方品种的蛋白质组进行了比较。在田间,将成年玉米叶片(玉米)在缺乏UV-B辐射的阳光下生长21天,或在包括1天8小时UV-B补充的阳光下生长后,记录其蛋白质模式。我们发现,通过微阵列杂交和定量实时逆转录PCR评估的先前记录的mRNA积累与W23叶片UV-B照射后的蛋白质表达之间存在非常高的相关性。已确认某些蛋白质存在多种同工型;至少一种蛋白质,丙酮酸磷酸双激酶,在UV-B照射下通过磷酸化进行翻译后调控。我们还鉴定了在W23中受UV-B辐射差异调控且在高海拔植物类似UV-B条件下水平较高的蛋白质。这些可能是赋予UV-B耐受性的遗传固定性状,并为适应高环境UV-B条件的特定适应方式提供线索。

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