Proteomics profiling of ethylene-induced tomato flower pedicel abscission
Proteomics profiling of ethylene-induced tomato flower pedicel abscission
复制标题
乙烯诱导的番茄花蒂脱落的蛋白质组学分析
DOI：
10.1016/j.jprot.2015.03.023
复制
发表时间：
2015-05-21
影响因子：
3.3
通讯作者：
Li, Tian-lai
中科院分区：
文献类型：
--
作者：
Zhang, Xiao-lin;Qi, Ming-fang;Li, Tian-lai
由于脱落对作物的产量和品质有重大影响，因此控制脱落是一个重要的农业问题。基因表达的变化与乙烯介导的脱落有关。然而，针对番茄花梗脱落的大规模蛋白质组学研究很少。采用等压标记法进行相对定量和绝对定量标记，研究了乙烯和1-甲基环丙烯处理后番茄花梗AZ （AZ）蛋白和磷蛋白的变化。在1429个定量蛋白中，166个蛋白对应的383个独特肽丰度变化超过1.5倍。共检测到450个磷酸化肽，其中73个磷酸化蛋白对应的85个磷酸化肽响应乙烯显著调控（>丰度变化1.5倍）。蛋白和磷蛋白组有26个相似蛋白。从138个磷酸化位点中提取了6个磷酸化基序。通过对翻译和修饰水平的分析，我们发现修饰水平并不是由翻译变化引起的。蛋白质和磷蛋白的功能比较表明，蛋白质主要在代谢过程中起作用并具有催化活性，而大部分磷蛋白具有信号传导和转运活性。数据揭示了AZ磷酸蛋白质组学的独特特征，从而表明参与了激酶-底物和磷酸酶-底物相互作用的复杂网络，以响应乙烯。钙依赖性蛋白激酶（CDPKS5523）、CDPKS5527和SRL3（S329）的一些磷酸化位点也被发现对AZ具有保护作用，并有助于乙烯信号转导。器官脱落具有积极和消极的双重作用。脱落有利于成熟果实的掉落和种子的释放和扩散，但一直是制约作物生产力的主要因素。因此，关于这一过程的更多细节可能有助于器官脱落的调节。然而，目前，脱落的具体机制尚不清楚。在番茄中，以花梗为材料进行了转录组分析。然而，目前还没有关于脱落带的大规模蛋白质组学和磷酸化蛋白质组学研究的报道。因此，在本研究中，我们使用等压标签相对和绝对定量（iTRAQ）来测定乙烯诱导的番茄花AZ组织蛋白质组学和磷酸化蛋白质组学的变化。来自两个数据集的蛋白质组学数据揭示了与脱落机制相关的翻译和修饰水平相关的差异表达蛋白。从磷酸化蛋白中获得了乙烯信号转导和防御相关蛋白中的两个关键蛋白CDPK （CDPK5（S523）和CDPK5(S527)）和SRL3（S329）。在这项工作中提出的番茄磷酸化位点至少在两个方面是有用的。首先，作为数据库资源，这些数据将有助于对鉴定的磷酸化蛋白的研究。其次，确定的相关蛋白位点可以为进一步的实验提供足够的知识。因此，我们的研究结果有助于理解植物脱落的机制。(C) 2015 Elsevier B.V.版权所有
The control of abscission is an important agricultural concern because of its substantial effect on crop yield and quality. Changes in gene expression are correlated with the ethylene-mediated execution of abscission. However, only few large-scale proteomic studies focused on tomato pedicel abscission. Isobaric tag for relative and absolute quantification labeling was used to examine the protein and phosphoprotein changes in the tomato pedicel AZ (AZ) treated with ethylene or 1-methylcyclopropene. Among the 1429 quantified proteins, 383 unique peptides corresponding to 166 proteins showed higher than 1.5-fold change in abundance. A total of 450 phosphopeptides were detected, among which 85 phosphopeptides corresponding to 73 phosphoproteins were significantly regulated (>1.5-fold abundance change) in response to ethylene. Protein and phosphoprotein sets showed 26 similar proteins. Six phosphorylation motifs were extracted from the 138 phosphorylation sites. By analyzing translational and modification levels, we found that the modification level was not due to the translational changes. Comparison between the protein and phosphoprotein functions revealed that the proteins acted mainly in the metabolic process and showed catalytic activity, whereas most of the phosphoproteins showed signaling and transporting activities. Data revealed the unique features of the AZ phosphoproteomics, thereby suggesting the involvement of a complex network of kinase-substrate and phosphatase-substrate interactions in response to ethylene. Some phosphorylation sites from calcium-dependent protein kinase (CDPKS5523), CDPKS5527, and SRL3(S329) were also found to perform protective functions for AZ and to be helpful in ethylene signal transduction.Biological significanceOrgan abscission has both positive and negative roles. Abscission is conducive for the fall of ripe fruits and the release and dispersion of seeds, but abscission has been a major limiting factor for crop productivity. Hence, more details about the process may aid in the regulation of organ abscission. However, at present, the detailed mechanism of abscission is still unclear. In tomato, several transcriptome analyses were performed using pedicels as materials. Yet, no large-scale proteomics and phosphoproteomic studies of abscission zone have been reported so far. Hence, in this present study, we determined the ethylene-induced changes in the proteomics and phosphoproteomics of tomato flower AZ tissue using the isobaric tag for relative and absolute quantification (iTRAQ). Proteomics data from both data sets revealed the differentially expressed proteins that are associated with the translational and modification levels relevant to abscission mechanism. Two key proteins (CDPK (CDPK5(S523) and CDPK5(S527)) and SRL3(S329)) among ethylene signal transduction and defense-related proteins were obtained from the phosphoproteins. The set of tomato phosphorylation sites presented in this work is useful in at least two ways. First, as a database resource, the data would facilitate research on the identified phosphoproteins. Second, the identified sites of the related proteins could provide enough knowledge for further experiments. Hence, our results contribute to the understanding of the mechanism of abscission in plants. (C) 2015 Elsevier B.V. All rights reserved.