Zulkapli Muhammad Mu'izzuddin, Ab Ghani Nur Syatila, Ting Tiew Yik, Aizat Wan Mohd, Goh Hoe-Han
Institute of Systems Biology, Universiti Kebangsaan Malaysia, Bangi, Malaysia.
Front Plant Sci. 2021 Jan 20;11:625507. doi: 10.3389/fpls.2020.625507. eCollection 2020.
is a genus comprising carnivorous tropical pitcher plants that have evolved trapping organs at the tip of their leaves for nutrient acquisition from insect trapping. Recent studies have applied proteomics approaches to identify proteins in the pitcher fluids for better understanding the carnivory mechanism, but protein identification is hindered by limited species-specific transcriptomes for . In this study, the proteomics informed by transcriptomics (PIT) approach was utilized to identify and compare proteins in the pitcher fluids of , , and their hybrid × through PacBio isoform sequencing (Iso-Seq) and liquid chromatography-mass spectrometry (LC-MS) proteomic profiling. We generated full-length transcriptomes from all three species of 80,791 consensus isoforms with an average length of 1,692 bp as a reference for protein identification. The comparative analysis found that transcripts and proteins identified in the hybrid × were more resembling , both of which are insectivorous compared with omnivorous that can derive nutrients from leaf litters. Previously reported hydrolytic proteins were detected, including proteases, glucanases, chitinases, phosphatases, nucleases, peroxidases, lipid transfer protein, thaumatin-like protein, pathogenesis-related protein, and disease resistance proteins. Many new proteins with diverse predicted functions were also identified, such as amylase, invertase, catalase, kinases, ligases, synthases, esterases, transferases, transporters, and transcription factors. Despite the discovery of a few unique enzymes in , we found no strong evidence of adaptive evolution to produce endogenous enzymes for the breakdown of leaf litter. A more complete picture of digestive fluid protein composition in this study provides important insights on the molecular physiology of pitchers and carnivory mechanism of species with distinct dietary habits.
是一个包含肉食性热带猪笼草的属,这些植物在叶片尖端进化出了用于捕捉昆虫以获取营养的捕虫器。最近的研究应用蛋白质组学方法来鉴定猪笼草液中的蛋白质,以便更好地理解食肉机制,但由于该属物种特异性转录组有限,蛋白质鉴定受到阻碍。在本研究中,利用转录组学信息蛋白质组学(PIT)方法,通过PacBio全长转录本测序(Iso-Seq)和液相色谱-质谱(LC-MS)蛋白质组分析,鉴定并比较了、及其杂交种×猪笼草液中的蛋白质。我们从这三个物种中生成了全长转录组,共80,791个共有转录本,平均长度为1,692 bp,作为蛋白质鉴定的参考。比较分析发现,杂交种×中鉴定出的转录本和蛋白质与更相似,与能从落叶中获取营养的杂食性相比二者均为食虫性。检测到了先前报道的水解蛋白,包括蛋白酶、葡聚糖酶、几丁质酶、磷酸酶、核酸酶、过氧化物酶、脂质转移蛋白、类thaumatin蛋白、病程相关蛋白和抗病蛋白。还鉴定出了许多具有不同预测功能的新蛋白质,如淀粉酶、转化酶、过氧化氢酶、激酶、连接酶、合酶、酯酶、转移酶、转运蛋白和转录因子。尽管在中发现了一些独特的酶,但我们没有发现有力证据表明其发生了适应性进化以产生用于分解落叶的内源酶。本研究中对消化液蛋白质组成更完整的描述为猪笼草的分子生理学和具有不同饮食习惯的猪笼草属物种的食肉机制提供了重要见解。