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. 2018 Aug 10;19(8):2364.
doi: 10.3390/ijms19082364.

Transcriptome Analysis of JA Signal Transduction, Transcription Factors, and Monoterpene Biosynthesis Pathway in Response to Methyl Jasmonate Elicitation in Mentha canadensis L

Xiwu Qi  1   2 , Hailing Fang  3 , Xu Yu  4 , Dongbei Xu  5 , Li Li  6 , Chengyuan Liang  7 , Hongfei Lu  8 , Weilin Li  9 , Yin Chen  10 , Zequn Chen  11
Affiliations

Transcriptome Analysis of JA Signal Transduction, Transcription Factors, and Monoterpene Biosynthesis Pathway in Response to Methyl Jasmonate Elicitation in Mentha canadensis L

Xiwu Qi et al. Int J Mol Sci. .

Abstract

Mentha canadensis L. has important economic value for its abundance in essential oils. Menthol is the main component of M. canadensis essential oils, which is certainly the best-known monoterpene for its simple structure and wide applications. However, the regulation of menthol biosynthesis remains elusive in M. canadensis. In this study, transcriptome sequencing of M. canadensis with MeJA treatment was applied to illustrate the transcriptional regulation of plant secondary metabolites, especially menthol biosynthesis. Six sequencing libraries were constructed including three replicates for both control check (CK) and methyl jasmonate (MeJA) treatment and at least 8 Gb clean bases was produced for each library. After assembly, a total of 81,843 unigenes were obtained with an average length of 724 bp. Functional annotation indicated that 64.55% of unigenes could be annotated in at least one database. Additionally, 4430 differentially expressed genes (DEGs) with 2383 up-regulated and 2047 down-regulated transcripts were identified under MeJA treatment. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment indicated that "Monoterpenoid biosynthesis" was one of the most significantly enriched pathways in metabolism. Subsequently, DEGs involved in JA signal transduction, transcription factors, and monoterpene biosynthesis were analyzed. 9 orthologous genes involved in menthol biosynthesis were also identified. This is the first report of a transcriptome study of M. canadensis and will facilitate the studies of monoterpene biosynthesis in the genus Mentha.

Keywords: JA signaling; Mentha canadensis L.; menthol biosynthesis; transcription factors; transcriptome sequencing.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Statistics of DEGs induced by MeJA in M. canadensis transcriptomes. (A) Volcano plots of the unigenes in the comparisons of MeJA-treated and CK samples; (B) DEG numbers in the comparisons of MeJA-treated and CK samples. RPKM: Reads per Kb per Million Reads, FDR: False Discovery Rate, DEG: Differentially Expressed Gene.
Figure 2
Figure 2
KEGG pathway enrichment of DEGs induced by MeJA.
Figure 3
Figure 3
Heat maps of the DEGs in JA signal transduction pathway. JAZ, Jasmonate ZIM domain protein; MYC2, bHLHzip transcription factor MYC2; TPL, TOPLESS; NINJA, Novel interactor of JAZ; SKP1, S-phase kinase-associated protein 1.
Figure 4
Figure 4
Expression validations of 12 selected JA signal transduction related genes in control and MeJA-treated samples using qRT-PCR. The RPKM values obtained from RNA-Seq data were indicated on the top of each graph.
Figure 5
Figure 5
Statistics of differentially expressed TFs under MeJA treatment.
Figure 6
Figure 6
Heat maps of the DEGs in the monoterpenoid biosynthesis pathway. TPS, Terpene synthase; IPR, (−)-Isopiperitenone reductase; MR, (−)-Menthol dehydrogenase; NMR, (+)-Neomenthol dehydrogenase.
Figure 7
Figure 7
Expression validations of 9 menthol biosynthetic genes in control and MeJA-treated samples using qRT-PCR. The RPKM values obtained from RNA-Seq data are indicated on the top of each graph. GPPS, Geranyl diphosphate synthase; LS, (−)-Limonene synthase; L3OH, (−)-Limonene-3-hydroxylase; iPD, (−)-trans-Isopiperitenol dehydrogenase; iPR, (−)-Isopiperitenone reductase; iPI, (+)-cis-Isopulegone isomerase; PR, (+)-Pulegone reductase; MFS, Menthofuran synthase; MR, (−)-Menthol dehydrogenase.
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