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. 2017 May 23;7(1):2308.
doi: 10.1038/s41598-017-02393-w.

Characterization of Juvenile Hormone Related Genes Regulating Cantharidin Biosynthesis in Epicauta chinensis

Ming Jiang  1 , Shumin Lü  1 , Yalin Zhang  2
Affiliations

Characterization of Juvenile Hormone Related Genes Regulating Cantharidin Biosynthesis in Epicauta chinensis

Ming Jiang et al. Sci Rep. .

Abstract

Cantharidin is a defensive toxin biosynthesized by blister beetles. It is well known for medical applications and toxicity. However, the biosynthesis process of cantharidin is still not well understood. In the present study, three genes (methyl farnesoate epoxidase (EcMFE), juvenile hormone acid O-methyltransferase (EcJHAMT) and juvenile hormone epoxide hydrolase (EcJHEH)) were identified from Epicauta chinensis. The temporal and spatial expression patterns of these three genes revealed that the expression levels of EcMFE and EcJHEH were high in the first instar larval stage of E. chinensis with EcJHEH transcripts highest in the fifth larval instar. The expression level of EcJHAMT was significantly higher in the 2nd and 3rd larval instars. The transcripts of EcMFE, EcJHEH and EcJHAMT showed a similar tendency with the cantharidin production in male blister beetles after mating. We verified the functions of these three genes in cantharidin biosynthesis using the RNA interference method. Interference of EcMFE and EcJHEH significantly inhibited the biosynthesis of cantharidin in male E. chinensis after mating, but interference of EcJHAMT has no apparent influence on cantharidin biosynthesis. We propose that EcMFE and EcJHEH may be involved in the biosynthesis of cantharidin, but JH III might not be the direct precursor of cantharidin.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Relative expression profiles of EcMFE, EcJEHE and EcJHAMT mRNA and cantharidin content at developmental stages in Epicauta chinensis. (A,C and E) are expression levels on the 0 to 7th day after mating in E. chinensis. (B,D and F) are expression levels in different instar larvae and eggs, (G,H and L) are cantharidin content at different developmental stages. Asterisks above represent statistically significant differences by Student’s t-test (P < 0.05), and different letters on error bars indicate significant differences in each group by one-way ANOVA analysis (P < 0.05).
Figure 2
Figure 2
Relative expression profiles of EcMFE, EcJEHE and EcJHAMT mRNA and cantharidin content in different tissues from adult E. chinensis. (A) Relative expression profiles in male tissues. (B) Relative expression profiles in female tissues. (C) cantharidin content in male tissues. (D) cantharidin content in female tissues. Different letters on error bars indicate significant differences in each group by one-way ANOVA analysis (P < 0.05).
Figure 3
Figure 3
Relative expression profiles of genes in male E. chinensis at different times after dsRNA injection. (A) EcMFE. (B) EcJHEH. (C) EcJHAMT. Asterisks above represent statistically significant differences by Student’s t-test (*P < 0.05; **P < 0.01).
Figure 4
Figure 4
The cantharidin content in male E. chinensis at different times after dsRNA injection. (A) EcMFE. (B) EcJHEH. (C) EcJHAMT. Asterisks above represent statistically significant differences by Student’s t-test (*P < 0.05; **P ≤ 0.01).
Figure 5
Figure 5
Diagram of the process of EcMFE, EcJHAMT and EcJHEH regulating cantharidin biosynthesis. Reactions that could occur normally are shown in black. Reactions that could not occur after dsRNA knockdown are shown in gray.
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