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‘King Alfred’ 水仙转录组和代谢组分析揭示石蒜科生物碱代谢成分。

Transcriptome and metabolome profiling of Narcissus pseudonarcissus 'King Alfred' reveal components of Amaryllidaceae alkaloid metabolism.

机构信息

Department of Chemistry, Biochemistry and Physics, Université du Québec à Trois-Rivières, 3351 boul. des Forges, Trois-Rivières, QC, G9A 5H7, Canada.

Plant Biology Research Group, Université du Québec à Trois-Rivières, 3351 boul. des Forges, Trois-Rivières, QC, G9A 5H7, Canada.

出版信息

Sci Rep. 2017 Dec 11;7(1):17356. doi: 10.1038/s41598-017-17724-0.

Abstract

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.

摘要

石蒜科生物碱(AAs)代表了一类多样化的植物次生代谢物,其中许多具有强大的药理学活性。AA 代谢途径知之甚少,资源也很少。为了阐明 AA 途径并发现新的生物合成酶,我们从水仙‘King Alfred’的不同组织中生成了全面的代谢组学和相应的转录组学数据集。在这项研究中,我们对不同组织进行了非靶向 UPLC-QTOF-MS 代谢物分析,生成了包括几种 AA 在内的化合物的详尽清单,其中大多数在鳞茎中含量最高且种类最多。水仙‘King Alfred’鳞茎的 RNA 测序组装后产生了 195,347 个转录本。表达量最高的基因属于代谢、生存和防御等过程,包括生物碱生物合成基因。转录组包含了编码 AA 生物合成酶的所有预期基因的完整序列,例如酪氨酸脱羧酶(TYDC1 和 TYDC2)、苯丙氨酸解氨酶(PAL1 和 PAL2)和酚酸羟化酶(C4H 和 C3H)等。此外,还使用 RT-qPCR 分析对转录组数据进行了验证,并对水仙‘King Alfred’的不同组织进行了表达研究。在这里,我们首次对水仙‘King Alfred’进行了全面的代谢组学和转录组学研究,为代谢工程和生物技术应用提供了宝贵的资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be9f/5725579/4b476bd47bbe/41598_2017_17724_Fig1_HTML.jpg

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