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Transcriptome and metabolome profiling of Narcissus pseudonarcissus ‘King Alfred’ reveal components of Amaryllidaceae alkaloid metabolism

机译:水仙假水仙国王阿尔弗雷德的转录组和代谢组谱分析揭示了金莲花科生物碱代谢的成分

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摘要

Amaryllidaceae alkaloids (AAs) represent a diverse class of plant specialized metabolites and many display potent pharmacological activities. The AA metabolic pathway is poorly understood and resources are minimal. To enable AA pathway elucidation and novel biosynthetic enzymes discovery, we generated comprehensive metabolomic and corresponding transcriptomic datasets from different tissues of Narcissus pseudonarcissus ‘King Alfred’. In this study, we performed untargeted UPLC-QTOF-MS metabolite analysis from different tissues, which generated exhaustive list of compounds, including several AAs, most predominant and diverse in bulbs. RNA sequencing of N. pseudonarcissus ‘King Alfred’ bulbs yielded 195,347 transcripts, after assembly. Top expressed genes belong to process like metabolism, survival, and defense including alkaloid biosynthetic genes. The transcriptome contained complete sequences for all proposed genes encoding AA-biosynthetic enzymes such as tyrosine decarboxylase (TYDC1 and TYDC2), phenylalanine ammonia-lyase (PAL1 and PAL2) and phenolic acids hydroxylases (C4H and C3H) to name a few. Furthermore, transcriptome data were validated using RT-qPCR analysis and expression study in different tissues of N. pseudonarcissus ‘King Alfred’ was performed. Here, we present the first comprehensive metabolome and transcriptome study from N. pseudonarcissus ‘King Alfred’ providing invaluable resources for metabolic engineering and biotechnological applications.
机译:石蒜科生物碱(AAs)代表了多种植物特有的代谢产物,并且显示出强大的药理活性。 AA代谢途径了解甚少,资源很少。为了阐明AA途径并发现新的生物合成酶,我们从水仙假水仙“阿尔弗雷德王”的不同组织中生成了综合的代谢组学和相应的转录组数据集。在这项研究中,我们对来自不同组织的UPLC-QTOF-MS代谢物进行了非目标分析,生成了详尽的化合物清单,包括几种AA,其中最主要的是灯泡中的多种。组装后,假水仙“ King Alfred”鳞茎的RNA测序产生195,347个转录本。表达最高的基因属于代谢,存活和防御等过程,包括生物碱生物合成基因。转录组包含所有提议的编码AA生物合成酶的基因的完整序列,这些基因包括酪氨酸脱羧酶(TYDC1和TYDC2),苯丙氨酸氨裂合酶(PAL1和PAL2)和酚酸羟化酶(C4H和C3H)等。此外,转录组数据已通过RT-qPCR分析进行了验证,并在假水仙猪笼草的不同组织“ King Alfred”中进行了表达研究。在这里,我们展示了来自假水仙念珠菌“阿尔弗雷德国王”的首个综合代谢组和转录组研究,为代谢工程和生物技术应用提供了宝贵的资源。

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