Agroforestry and Plant Biochemistry, Proteomics and Systems Biology, Department of Biochemistry and Molecular Biology, University of Cordoba, UCO-CeiA3, 14014 Cordoba, Spain.
Agricultural Microbiology, Faculty of Agricultural Science, National University of Cordoba, CONICET, 5001 Cordoba, Argentina.
Int J Mol Sci. 2021 Mar 21;22(6):3191. doi: 10.3390/ijms22063191.
Drought is one of the main causes of mortality in holm oak () seedlings used in reforestation programs. Although this species shows high adaptability to the extreme climate conditions prevailing in Southern Spain, its intrinsic genetic variability may play a role in the differential response of some populations and individuals. The aim of this work was to identify proteins and derived proteotypic peptides potentially useful as putative markers for drought tolerance in holm oak by using a targeted post-acquisition proteomics approach. For this purpose, we used a set of proteins identified by shotgun (LC-MSMS) analysis in a drought experiment on seedlings from four different provenances (viz. the Andalusian provinces Granada, Huelva, Cadiz and Seville). A double strategy involving the quantification of proteins and target peptides by shotgun analysis and post-acquisition data analysis based on proteotypic peptides was used. To this end, an initial list of proteotypic peptides from proteins highly represented under drought conditions was compiled that was used in combination with the raw files from the shotgun experiment to quantify the relative abundance of the fragment's ion peaks with the software Skyline. The most abundant peptides under drought conditions in at least two populations were selected as putative markers of drought tolerance. A total of 30 proteins and 46 derived peptides belonging to the redox, stress-related, synthesis,-folding and degradation, and primary and secondary metabolism functional groups were thus identified. Two proteins (viz., subtilisin and chaperone GrpE protein) were found at increased levels in three populations, which make them especially interesting for validation drought tolerance markers in subsequent experiments.
干旱是造林计划中使用的欧洲栓皮栎()幼苗死亡率的主要原因之一。尽管该物种对西班牙南部盛行的极端气候条件表现出高度的适应性，但它内在的遗传变异性可能在一些种群和个体的差异响应中发挥作用。本工作旨在通过采用靶向采集后蛋白质组学方法，鉴定可能作为欧洲栓皮栎耐旱性的潜在标记物的蛋白质和衍生的肽段。为此，我们使用了在来自四个不同起源地（即格拉纳达、韦尔瓦、卡迪斯和塞维利亚的安达卢西亚省）的幼苗的干旱实验中通过鸟枪法（LC-MSMS）分析鉴定的一组蛋白质。采用涉及通过鸟枪法分析和基于衍生肽的采集后数据分析来定量蛋白质和目标肽的双重策略。为此，我们编写了一个来自在干旱条件下高度表达的蛋白质的衍生肽的初始列表，然后将其与鸟枪实验的原始文件结合使用，以便使用 Skyline 软件定量片段的离子峰的相对丰度。在至少两个种群中，选择在干旱条件下最丰富的肽段作为耐旱性的潜在标记物。总共鉴定出 30 种蛋白质和 46 种衍生肽，它们属于氧化还原、应激相关、合成-折叠和降解以及初级和次级代谢功能组。发现两种蛋白质（即枯草杆菌蛋白酶和伴侣蛋白 GrpE 蛋白）在三个种群中水平升高，这使得它们特别有趣，可作为后续实验中验证耐旱性标记物的候选物。
