Protein replenishment in pitcher fluids of Nepenthes × ventrata revealed by quantitative proteomics (SWATH-MS) informed by transcriptomics
Protein replenishment in pitcher fluids of Nepenthes × ventrata revealed by quantitative proteomics (SWATH-MS) informed by transcriptomics
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DOI：
10.1007/s10265-019-01130-w
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发表时间：
2019-08
影响因子：
2.8
通讯作者：
W. W. Wan Zakaria-W.;W. Aizat;Hoe-Han Goh;Normah Mohd Noor
中科院分区：
文献类型：
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作者：
W. W. Wan Zakaria-W.;W. Aizat;Hoe-Han Goh;Normah Mohd Noor
肉食植物捕获并消化昆虫以获取营养，使它们能够在缺乏氮营养的土壤中生存。猪笼草属植物产生独特的捕虫笼，其中含有分泌腺，可将酶分泌到消化液中。我们对猪笼草的捕虫笼组织进行了RNA-seq分析，并对猪笼草捕虫笼液体进行了液相色谱-串联质谱（LC-MS/MS）分析，研究了该食肉器官在捕虫笼开放初期的蛋白质表达。该转录组为捕虫笼液蛋白鉴定提供了序列数据库。成功鉴定出总共32种不同功能的蛋白质，其中19种蛋白质可以基于无标记定量蛋白质组学（SWATH-MS）分析进行定量，而16种蛋白质以前未报道过。我们的研究结果表明，即使没有任何猎物或几丁质诱导，捕虫笼液中的某些蛋白质在捕虫笼打开后也会持续分泌或补充。我们还发现了一种新的天冬氨酸蛋白酶 Nep6，它分泌到捕虫笼液中。这是首次使用物种特异性参考转录组对猪笼草液体中的蛋白质表达进行 SWATH-MS 分析。总而言之，我们使用转录组学 (PIT) 方法提供的无凝胶鸟枪蛋白质组学研究显示了 N.x ventrata 消化器官中内源蛋白质分泌的动态，并提供了在捕获猎物之前捕虫笼早期打开过程中蛋白质调节的见解。
Carnivorous plants capture and digest insects for nutrients, allowing them to survive in soil deprived of nitrogenous nutrients. Plants from the genusNepenthesproduce unique pitchers containing secretory glands, which secrete enzymes into the digestive fluid. We performed RNA-seq analysis on the pitcher tissues and liquid chromatography-tandem mass spectrometry (LC–MS/MS) analysis on the pitcher fluids ofNepenthes×ventratato study protein expression in this carnivory organ during early days of pitcher opening. This transcriptome provides a sequence database for pitcher fluid protein identification. A total of 32 proteins of diverse functions were successfully identified in which 19 proteins can be quantified based on label-free quantitative proteomics (SWATH-MS) analysis while 16 proteins were not reported previously. Our findings show that certain proteins in the pitcher fluid were continuously secreted or replenished after pitcher opening, even without any prey or chitin induction. We also discovered a new aspartic proteinase, Nep6, secreted into pitcher fluid. This is the first SWATH-MS analysis of protein expression inNepenthespitcher fluid using a species-specific reference transcriptome. Taken together, our study using a gel-free shotgun proteomics informed by transcriptomics (PIT) approach showed the dynamics of endogenous protein secretion in the digestive organ ofN.×ventrataand provides insights on protein regulation during early pitcher opening prior to prey capture.