Mao Rongjia, Chen Jing, Chen Yuejun, Guo Zhigang
Department of Chemical Engineering, Institute of Biochemical Engineering, Tsinghua University, Beijing, China.
Physiol Mol Biol Plants. 2018 Sep;24(5):715-727. doi: 10.1007/s12298-018-0527-2. Epub 2018 May 4.
Jasmonate, an effective elicitor, can induce the biosynthesis of paclitaxel, a well-known anticancer drug, in cell culture. The jasmonate signaling pathway has been well studied in Arabidopsis, and many early jasmonate-responsive genes have been found to be involved in signaling pathway. In , only a few late jasmonate-responsive genes related to paclitaxel biosynthesis were identified. So, identification of early responsive genes and knowledge of the jasmonate signaling pathway are essential for understanding the effects of jasmonate on paclitaxel biosynthesis and for improving paclitaxel production in cells. In this study, total RNA of × cells cultured in liquid medium was extracted after 0, 0.5, 3, and 24 h of methyl jasmonate treatment. Three biological independent repetitions were performed. The 12 extracted RNA samples were integrated and sequenced on an Illumina HiSeq 2500 platform using the paired-end method. A total of 45,583 transcript clusters were obtained by de novo assembly of the sequenced reads. Based on the transcriptome data, the digital gene expressions of each RNA sample were investigated. We found that after 0.5, 3, and 24 h of methyl jasmonate treatment; 134, 1008, and 987 unigenes were differentially expressed. For the secondary metabolism pathways, phenylalanine pathway unigenes were responsive to jasmonate after 3 h of treatment, while genes related to paclitaxel biosynthesis were induced after 0.5 h of treatment. The digital gene expression levels of candidate genes related to paclitaxel biosynthesis were confirmed by qRT-PCR. Transcriptome sequencing and digital gene expression profiling identified early jasmonate-responsive genes in cultured × cells. The comprehensive time series jasmonate-responsive gene expression data have provided transcriptome-wide information about the mechanism of paclitaxel biosynthesis regulation by jasmonate signaling.
茉莉酸是一种有效的诱导子，可在细胞培养中诱导著名抗癌药物紫杉醇的生物合成。茉莉酸信号通路在拟南芥中已得到充分研究，许多早期茉莉酸响应基因被发现参与信号通路。在[具体植物名称]中，仅鉴定出少数与紫杉醇生物合成相关的晚期茉莉酸响应基因。因此，鉴定早期响应基因并了解茉莉酸信号通路对于理解茉莉酸对紫杉醇生物合成的影响以及提高[具体植物名称]细胞中紫杉醇的产量至关重要。在本研究中，在茉莉酸甲酯处理0、0.5、3和24小时后，提取在液体培养基中培养的[具体植物名称]×细胞的总RNA。进行了三次生物学独立重复。将提取的12个RNA样品整合，并使用双末端方法在Illumina HiSeq 2500平台上进行测序。通过对测序读数进行从头组装，共获得45,583个转录本簇。基于转录组数据，研究了每个RNA样品的数字基因表达。我们发现，在茉莉酸甲酯处理0.5、3和24小时后，分别有134、1008和987个单基因差异表达。对于次生代谢途径，苯丙氨酸途径单基因在处理3小时后对茉莉酸有响应，而与紫杉醇生物合成相关的基因在处理0.5小时后被诱导。通过qRT-PCR证实了与紫杉醇生物合成相关的候选基因的数字基因表达水平。转录组测序和数字基因表达谱分析鉴定了培养的[具体植物名称]×细胞中的早期茉莉酸响应基因。全面的时间序列茉莉酸响应基因表达数据提供了关于茉莉酸信号调控紫杉醇生物合成机制的全转录组信息。
