Proteomic analysis of ubiquitinated proteins in 'Xiangshui' lemon [Citrus limon (L.)] pistils after self- and cross-pollination
Proteomic analysis of ubiquitinated proteins in 'Xiangshui' lemon [Citrus limon (L.)] pistils after self- and cross-pollination
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DOI：
10.1016/j.jprot.2022.104631
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发表时间：
2022-05-28
影响因子：
3.3
通讯作者：
He，Xinhua
中科院分区：
文献类型：
--
作者：
Lin，Wei;Li，Yuze;He，Xinhua
‘响水’柠檬配子体自交不亲和，自花授粉后，花柱中的花粉管生长受到抑制。本研究旨在研究授粉对‘响水’柠檬雌蕊泛素组和蛋白质组的影响。本文采用无标记定量、抗双甘氨肽赖氨酸抗体琼脂糖微球富集、高分辨液相色谱串联质谱（LC-MS/MS）和鸟枪蛋白质组学方法，研究了“响水”柠檬授粉后雌蕊的定量蛋白质组、泛素化组以及两个数据集之间的相互影响。总共定量了4682种蛋白质，1956种泛素化蛋白质和6922个泛素化位点。主成分分析结果表明，对照、自花授粉和异花授粉的泛素和蛋白质组存在一定差异。串扰结果表明，蛋白质组和泛素组呈负相关。自花授粉后，与细胞程序性死亡、细胞衰老和鞘脂代谢途径相关的泛素化蛋白种类的丰度发生了变化。此外，我们分析了自花授粉后雌蕊中的蛋白酶体途径，并进一步讨论了泛素化事件影响花柱中S-RNase的机制。授粉显著改变了香水柠檬雌蕊中泛素化蛋白的丰度。因此，本研究对“响水”柠檬自交不亲和雌蕊中的蛋白质组和泛素化组进行了系统的分析，为进一步研究“响水”柠檬自交不亲和雌蕊中泛素化的功能和作用机制提供了理论依据。虽然在果树中已经发现了大量的SI相关基因，但关于影响和参与果树对SI反应的PTM的研究很少。本研究重点研究授粉对“响水”柠檬雌蕊蛋白质组和泛素化组的影响，探讨转录组与蛋白质组、泛素化组与蛋白质组的相关性，分析自花授粉和异花授粉后SI相关蛋白和途径的表达和泛素化水平的变化，以及异花授粉后26 S蛋白酶体途径的泛素化水平的变化。本研究为“响水”柠檬中SI的泛素化途径提供了新的认识。
‘Xiangshui’ lemon exhibits gametophyte self-incompatibility: after self-pollination, the growth of a pollen tube in the style is inhibited. The aim of this work was to study the effect of pollination on the ubiquitylome and proteome in ‘Xiangshui’ lemon pistil. Here, by using label-free quantification, enrichment by anti-diglycine lysine antibody conjugated agarose beads, high-resolution liquid chromatography tandem mass spectrometry (LC-MS/MS) and the method of shotgun proteomics, we investigated the quantitative proteome, ubiquitylome, and crosstalk between the two datasets in the ‘Xiangshui’ lemon pistil after pollination. In total, 4682 protein species, 1956 ubiquitinated protein species and 6922 ubiquitination sites were quantified. A principal component analysis revealed that there were some differences in the ubiquitylome and proteome of CK, self-pollination and cross-pollination. The results of the crosstalk showed that the proteome and ubiquitylome were negatively correlated. After self-pollination, the abundance of the ubiquitinated protein species related to programmed cell death, cell aging and sphingolipid metabolic pathways was changed. Furthermore, we analyzed the proteasome pathway in the pistil after self-pollination and further discussed the mechanism by which ubiquitination events affect S-RNase in the style. Overall, pollination greatly changed the ubiquitinated protein species abundance in the pistils of ‘Xiangshui’ lemon. Accordingly, our work provides a systematic analysis of the proteome and ubiquitylome in ‘Xiangshui’ lemon pistils under self-incompatibility and sheds light for future studies on the function and mechanism of ubiquitination during self-incompatibility in ‘Xiangshui’ lemon.SignificanceAs a mechanism to prevent self-pollination, self-incompatibility (SI) exists widely in plant species. Although a large number of SI-related genes have been found in fruit trees, there are few studies on the PTMs that affect and are involved in fruit tree responses to SI. This study highlights the fact that the effects of pollination on proteome and ubiquitylome in the ‘Xiangshui’ lemon pistil, we discussed the correlation between transcriptome and proteome, ubiquitylome and proteome, and we analyzed the expression and the changes of ubiquitination levels of SI related proteins and pathway after self- and cross pollination, and the changes of ubiquitination level of 26S proteasome pathway after cross-pollination. This study provides new insights into the ubiquitination pathway of SI in ‘Xiangshui’ lemon.