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对两种在体外渗透胁迫处理后呈现不同反应的马铃薯基因型(Solanum tuberosum L.)进行蛋白质组学分析。

Proteomic analysis of two divergently responding potato genotypes (Solanum tuberosum L.) following osmotic stress treatment in vitro.

作者信息

Bündig Christin, Jozefowicz Anna Maria, Mock Hans-Peter, Winkelmann Traud

机构信息

Leibniz Universität Hannover, Institute of Horticultural Production Systems, Herrenhaeuser Strasse 2, 30419 Hannover, Germany.

Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), OT Gatersleben, Corrensstraße 3, 06466 Stadt Seeland, Germany.

出版信息

J Proteomics. 2016 Jun 30;143:227-241. doi: 10.1016/j.jprot.2016.04.048. Epub 2016 May 3.

Abstract

UNLABELLED

Starch potatoes (Solanum tuberosum L.) are of interest for production of starch, ethanol, and biopolymers. Due to the predicted increase in drought periods, the breeding of starch potatoes for drought tolerance is essential. This study aims to elucidate the physiological mechanisms that give rise to drought tolerance. Two genotypes contrasting in drought tolerance were compared. We applied osmotic stress which is a known component of drought stress under in vitro conditions. Shoot tips were harvested after 11days of culture on control medium and medium supplied with 0.2M sorbitol. Their proteomes were analyzed using two-dimensional isoelectric focussing sodium dodecyl sulphate polyacrylamide gel electrophoresis (2D-IEF/SDS-PAGE). Of a total of 679 distinct protein spots, 118 and 20 spots with differential abundance were found in the sensitive and the tolerant genotype, respectively, after the application of stress. Using mass spectrometry, the proteins in 100 differentially abundant spots were identified; a majority of these proteins were from the chloroplast. For the sensitive genotype, an increase in the abundance of proteinase inhibitors and their precursors, changes in stress responsive proteins and an altered RNA/DNA-binding response were observed. The differentially abundant spots of the tolerant genotype comprised one chaperone and one hydrogen peroxide detoxifying protein.

BIOLOGICAL SIGNIFICANCE

Our findings reveal that the two genotypes have different responses to osmotic stress in terms of protein degradation and reactive oxygen species (ROS) scavenging and production. Our data suggest that the tolerant genotype might adjust to the applied stress more quickly. A comparative temporal analysis might provide further insights into these rapid changes and assist in the development of biomarkers.

摘要

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淀粉土豆(Solanum tuberosum L.)在淀粉、乙醇和生物聚合物生产方面具有重要意义。由于预计干旱期会增加,培育耐旱淀粉土豆至关重要。本研究旨在阐明产生耐旱性的生理机制。比较了两种耐旱性不同的基因型。我们在体外条件下施加了渗透胁迫,这是干旱胁迫的一个已知组成部分。在对照培养基和添加0.2M山梨醇的培养基上培养11天后收获茎尖。使用二维等电聚焦十二烷基硫酸钠聚丙烯酰胺凝胶电泳(2D-IEF/SDS-PAGE)分析它们的蛋白质组。在施加胁迫后,在敏感基因型和耐受基因型中分别发现了118个和20个丰度有差异的不同蛋白质斑点,总共679个不同蛋白质斑点。使用质谱法鉴定了100个丰度有差异的斑点中的蛋白质;这些蛋白质大多数来自叶绿体。对于敏感基因型,观察到蛋白酶抑制剂及其前体丰度增加、应激反应蛋白发生变化以及RNA/DNA结合反应改变。耐受基因型中丰度有差异的斑点包括一种伴侣蛋白和一种过氧化氢解毒蛋白。

生物学意义

我们的研究结果表明,这两种基因型在蛋白质降解以及活性氧(ROS)清除和产生方面对渗透胁迫有不同反应。我们的数据表明,耐受基因型可能更快地适应所施加的胁迫。比较性的时间分析可能会进一步深入了解这些快速变化,并有助于生物标志物的开发。

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