Mechanisms of Soybean Roots' Tolerances to Salinity Revealed by Proteomic and Phosphoproteomic Comparisons Between Two Cultivars
Mechanisms of Soybean Roots' Tolerances to Salinity Revealed by Proteomic and Phosphoproteomic Comparisons Between Two Cultivars
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DOI：
10.1074/mcp.m115.051961
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发表时间：
2016-01-01
影响因子：
7
通讯作者：
Du, Liqun
中科院分区：
文献类型：
--
作者：
Pi, Erxu;Qu, Liqun;Du, Liqun
了解植物耐盐性的分子机制为通过基因工程有效改良作物提供了有价值的知识基础。目前的蛋白质组学技术支持可靠和高通量的分析，已被广泛用于探索植物中复杂的分子网络。本研究比较了耐盐大豆品种(文丰07)和盐敏感品种(Union 85140)在盐胁迫下根的磷蛋白质组和蛋白质组的变化。文丰07和UnION 85140在三叶期用150 mM的氯化钠处理后，分别于0h、0.5h、1h、4h、12h、24h和48h采集根样。基于LC-MS/MS的磷酸蛋白质组分析共鉴定出2692个磷酸蛋白和5509个磷酸化位点。对其中含有3744个磷酸化位点的2344个磷酸蛋白进行了定量分析。我们的结果表明，在两个对照品种中，有1163个磷酸化位点存在差异。其中，10个MYB/MYB类转录因子蛋白在不同的时间点具有波动的磷酸化修饰，表明它们在调控黄酮醇积累中的关键作用可能是通过磷酸化修饰来介导的。此外，基于LC-MS/MS的蛋白质组学分析比较了这两个品种的蛋白质表达谱，并通过qRT-PCR独立确认了所有89个差异表达蛋白质的表达模式。有趣的是，查尔酮代谢途径中的酶与耐盐性呈正相关。我们利用大豆复合体和拟南芥突变体证实了查尔酮合成酶、查尔酮异构酶和细胞色素P450单加氧酶基因的功能相关性，发现它们的耐盐性受查尔酮合酶正向调控，而查尔酮异构酶和细胞色素P450单加氧酶负调控。基于我们的发现，提出了一个新的涉及查尔酮代谢的耐盐途径，该途径主要由磷酸化的MYB转录因子介导。(质谱学原始数据可通过标识为PXD002856的ProteomeXchange获得)。
Understanding molecular mechanisms underlying plant salinity tolerance provides valuable knowledgebase for effective crop improvement through genetic engineering. Current proteomic technologies, which support reliable and high-throughput analyses, have been broadly used for exploring sophisticated molecular networks in plants. In the current study, we compared phosphoproteomic and proteomic changes in roots of different soybean seedlings of a salt-tolerant cultivar (Wenfeng07) and a salt-sensitive cultivar (Union85140) induced by salt stress. The root samples of Wenfeng07 and Union85140 at three-trifoliate stage were collected at 0 h, 0.5 h, 1 h, 4 h, 12 h, 24 h, and 48 h after been treated with 150 mM NaCl. LC-MS/MS based phosphoproteomic analysis of these samples identified a total of 2692 phosphoproteins and 5509 phosphorylation sites. Of these, 2344 phosphoproteins containing 3744 phosphorylation sites were quantitatively analyzed. Our results showed that 1163 phosphorylation sites were differentially phosphorylated in the two compared cultivars. Among them, 10 MYB/MYB transcription factor like proteins were identified with fluctuating phosphorylation modifications at different time points, indicating that their crucial roles in regulating flavonol accumulation might be mediated by phosphorylated modifications. In addition, the protein expression profiles of these two cultivars were compared using LC-MS/MS based shotgun proteomic analysis, and expression pattern of all the 89 differentially expressed proteins were independently confirmed by qRT-PCR. Interestingly, the enzymes involved in chalcone metabolic pathway exhibited positive correlations with salt tolerance. We confirmed the functional relevance of chalcone synthase, chalcone isomerase, and cytochrome P450 monooxygenase genes using soybean composites and Arabidopsis thaliana mutants, and found that their salt tolerance were positively regulated by chalcone synthase, but was negatively regulated by chalcone isomerase and cytochrome P450 monooxygenase. A novel salt tolerance pathway involving chalcone metabolism, mostly mediated by phosphorylated MYB transcription factors, was proposed based on our findings. (The mass spectrometry raw data are available via ProteomeXchange with identifier PXD002856).