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Identification of early jasmonate-responsive genes in Taxus × media cells by analyzing time series digital gene
Rongjia Mao1, Jing Chen1, Yuejun Chen1
1Department of Chemical Engineering, Institute of Biochemical Engineering, Tsinghua University, Beijing, China.
Abstract:
Jasmonate, an effective elicitor, can induce the biosynthesis of paclitaxel, a well-known anticancer drug, in Taxus 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 Taxus, 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 Taxus cells. In this study, total RNA of Taxus × media 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 Taxus × media 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.
Insights
Jasmonate treatment rapidly induces early genes involved in paclitaxel biosynthesis in Taxus cells. This study identifies key early responsive genes, advancing our understanding of jasmonate signaling for improved anticancer drug production.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Pharmacology
Background:
- Jasmonate is a known inducer of paclitaxel biosynthesis in Taxus cell cultures.
- Understanding jasmonate signaling is crucial for enhancing paclitaxel production, an important anticancer drug.
- Limited knowledge exists on early jasmonate-responsive genes in Taxus, hindering pathway elucidation.
Purpose of the Study:
- To identify early jasmonate-responsive genes in Taxus × media cells.
- To investigate the temporal dynamics of gene expression following methyl jasmonate treatment.
- To provide transcriptome-wide data on jasmonate signaling in paclitaxel biosynthesis regulation.
Main Methods:
- Extraction of total RNA from Taxus × media cells at 0, 0.5, 3, and 24 hours post-methyl jasmonate treatment.
- Illumina HiSeq 2500 sequencing of 12 RNA samples (3 biological replicates).
- De novo assembly, transcriptome analysis, and digital gene expression profiling, with qRT-PCR validation for key genes.
Main Results:
- Identification of 134, 1008, and 987 differentially expressed unigenes at 0.5, 3, and 24 hours, respectively.
- Phenylalanine pathway genes responded after 3 hours, while paclitaxel biosynthesis genes were induced as early as 0.5 hours.
- Digital gene expression profiling revealed early jasmonate-responsive genes critical for paclitaxel production.
Conclusions:
- Transcriptome sequencing successfully identified early jasmonate-responsive genes in cultured Taxus × media cells.
- The study provides comprehensive time-series gene expression data, illuminating jasmonate's regulatory role in paclitaxel biosynthesis.
- Findings are essential for improving paclitaxel production strategies in Taxus cell cultures.
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