State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan, Hubei, PR China.
Institute of Konjac, Enshi Academy of Agricultural Sciences, Enshi, Hubei, PR China.
PLoS One. 2018 Mar 28;13(3):e0194863. doi: 10.1371/journal.pone.0194863. eCollection 2018.
Amorphophallus is a perennial herbaceous plant species mainly distributed in the tropics or subtropics of Asia and Africa. It has been used as a traditional medicine for a long time and now is utilized for the pharmaceutical, chemical and agriculture industries as a valued economic crop. Recently, Amorphophallus has attracted tremendous interest because of its high ceramide content. However, the breeding and genome studies are severely limited by the arduous whole genome sequencing of Amorphophallus. In this study, the transcriptome data of A. muelleri was obtained by utilizing the high-throughput Illumina sequencing platform. Based on this information, the majority of the significant genes involved in the proposed sphingolipid metabolic pathway were identified. Then, the full-length neutral ceramidase cDNA was obtained with the help of its candidate transcripts, which were acquired from the transcriptome data. Furthermore, we demonstrated that this neutral ceramidase was a real ceramidase by eukaryotic expression in the yeast double knockout mutant Δypc1 Δydc1, which lacks the ceramidases-dihydroCDase (YDC1p), phytoCDase (YPC1p). In addition, the biochemical characterization of purified A. muelleri ceramidase (AmCDase) exhibited classical Michaelis-Menten kinetics with an optimal activity ranging from pH 6.5 to 8.0. Based on our knowledge, this study is the first to report the related information of the neutral ceramidase in Amorphophallus. All datasets can provide significant information for related studies, such as gene expression, genetic improvement and application on breeding in Amorphophallus.
魔芋是一种多年生草本植物，主要分布在亚洲和非洲的热带或亚热带地区。它长期以来一直被用作传统药物，现在被用作医药、化工和农业产业的有价值的经济作物。最近，由于魔芋含有高含量的神经酰胺，因此引起了人们的极大兴趣。然而，由于魔芋全基因组测序困难，其繁殖和基因组研究受到严重限制。在这项研究中，利用高通量 Illumina 测序平台获得了魔芋的转录组数据。在此基础上，鉴定了参与拟议神经酰胺代谢途径的大多数重要基因。然后，借助转录组数据获得的候选转录本，获得了全长中性神经酰胺酶 cDNA。此外，我们通过在缺乏神经酰胺酶二氢神经酰胺酶（YDC1p）、植物神经酰胺酶（YPC1p）的酵母双敲除突变体Δypc1 Δydc1 中真核表达，证明了该中性神经酰胺酶是一种真正的神经酰胺酶。此外，纯化的魔芋神经酰胺酶（AmCDase）的生化特性表现出典型的米氏动力学，最适活性范围为 pH6.5 至 8.0。据我们所知，这项研究首次报道了魔芋中性神经酰胺酶的相关信息。所有数据集都可以为相关研究提供重要信息，如基因表达、遗传改良和魔芋育种中的应用。
