Biosurfactants Induce Antimicrobial Peptide Production through the Activation of TmSpatzles in Tenebrio molitor
Abstract
:1. Introduction
2. Results
2.1. AMP Activity in the Hemolymph of Biosurfactant-Injected T. molitor Larvae
2.2. Biosurfactants Induced AMP Expression in T. molitor Hemocytes
2.3. Biosurfactants Increased T. molitor Larvae Survivability against E. coli
2.4. Biosurfactants Activated the Expression of Spätzle Genes in T. molitor
2.5. Effect of TmSpz Gene Silencing on the Expression of AMPs in T. molitor
2.6. T. molitor Larvae Survivability Study
3. Discussion
4. Materials and Methods
4.1. Insect Culture
4.2. Source of Biosurfactants
4.3. Preparation of Microorganisms
4.4. AMPs Activity Study
4.5. RNA Extraction, cDNA Synthesis, and Expression of AMPs
4.6. Biosurfactant-Triggered Immune Survivability of T. molitor
4.7. Screening of Biosurfactant-Activated T. molitor Spatzle Genes
4.8. Effect of Tm Spätzle Gene Silencing on the Expression of AMPs
4.9. Effect of TmSpz Genes Silencing on T. molitor Survivability
4.10. Data Analyses
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Biosurfactants. | Type | Biological Source | Type | Purity |
---|---|---|---|---|
Bacitracin | cyclic lipopeptide | Bacillus licheniformis | Gram-positive | 90% |
Fengycin | lipopeptide complex | Bacillus subtilis | Gram-positive | >90% |
Iturin A | cyclic lipopeptide | Bacillus subtilis | Gram-positive | >95% |
Rhamnolipid | Glycolipids | Pseudomonas aeruginosa | Gram-negative | >95% |
Name | Primer Sequences |
---|---|
TmTencin-1_Fw | 5′-CAGCTGAAGAAATCGAACAAGG-3′ |
TmTencin-1_Rv | 5′-CAGACCCTCTTTCCGTTACAGT-3′ |
TmTencin-2_Fw | 5′-CAGCAAAACGGAGGATGGTC-3′ |
TmTencin-2_Rv | 5′-CGTTGAAATCGTGATCTTGTCC-3′ |
TmTencin-3_Fw | 5′-GATTTGCTTGATTCTGGTGGTC-3′ |
TmTencin-3_Rv | 5′-CTGATGGCCTCCTAAATGTCC-3′ |
TmTencin-4_Fw | 5′-GGACATTGAAGATCCAGGAAAG-3′ |
TmTencin-4_Rv | 5′-CGGTGTTCCTTATGTAGAGCTG-3′ |
TmDefensin-1_Fw | 5′-AAATCGAACAAGGCCAACAC-3′ |
TmDefencin-1_Rv | 5′-GCAAATGCAGACCCTCTTTC-3′ |
TmDefencin-2_Fw | 5′-GGGATGCCTCATGAAGATGTAG-3′ |
TmDefencin-2_Rv | 5′-CCAATGCAAACACATTCGTC-3′ |
TmColoptericin-1_Fw | 5′-GGACAGAATGGTGGATGGTC-3′ |
TmColoptericin-1_Rv | 5′-CTCCAACATTCCAGGTAGGC-3 |
TmColoptericin-2_Fw | 5′-GGACGGTTCTGATCTTCTTGAT-3′ |
TmColoptericin-2_Rv | 5′-CAGCTGTTTGTTTGTTCTCGTC-3′ |
TmAttacin-1a_Fw | 5′-GAAACGAAATGGAAGGTGGA-3′ |
TmAttacin-1a_Rv | 5′-TGCTTCGGCAGACAATACAG-3′ |
TmAttacin-1b_Fw | 5′-GAGCTGTGAATGCAGGACAA-3′ |
TmAttacin-1b_Rv | 5′-CCCTCTGATGAAACCTCCAA-3′ |
TmAttacin-2_Fw | 5′-AACTGGGATATTCGCACGTC-3′ |
TmAttacin-2_Rv | 5′-CCCTCCGAAATGTCTGTTGT-3 |
TmCecropin-2_Fw | 5′-TACTAGCAGCGCCAAAACCT-3′ |
TmCecropin-2_Rv | 5′-CTGGAACATTAGGCGGAGAA-3′ |
TmThaumatin-like protein-1_Fw | 5′-CTCAAAGGACACGCAGGACT-3′ |
TmThaumatin-like protein-1_Rv | 5′-ACTTTGAGCTTCTCGGGACA-3′ |
TmThaumatin-like protein-2_Fw | 5′-CCGTCTGGCTAGGAGTTCTG-3′ |
TmThaumatin-like protein-2_Rv | 5′-ACTCCTCCAGCTCCGTTACA-3′ |
TmL27a_qPCR_Fw | 5′-TCATCCTGAAGGCAAAGCTCCAGT-3′ |
TmL27a_qPCR_Rv | 5′-AGGTTGGTTAGGCAGGCACCTTTA-3′ |
TmSpz4-qPCR-Fw | 5′-GGCGATGCTCTTCCAGGAC-3′ |
TmSpz4-qPCR-Rv | 5′-CGCGTTCACTCCTTTCATTTGG-3′ |
TmSpz4-T7-Fw | 5′-TAATACGACTCACTATAGGGT CCAGATGTACTGTCGCGATG-3′ |
TmSpz4-T7-Rv | 5′-TAATACGACTCACTATAGGGT TTCCTTCTGTACCAGTCGGG-3′ |
TmSpz6-qPCR-Fw | 5′-GCACAACTCCAAGACGACCT-3′ |
TmSpz6-qPCR-Rv | 5′-TCTCTTCACCCGATCGTTGC-3′ |
TmSpz6-T7-Fw | 5′-TAATACGACTCACTATAGGGT ACCGCGCAAGAGAGTAAAAA-3′ |
TmSpz6-T7-Rv | 5′-TAATACGACTCACTATAGGGT ACGTATCTCCACACCCCTTG-3′ |
TmSpz3-qPCR-Fw | 5′-TCTCAACAACGGGACCTTCG-3′ |
TmSpz3-qPCR-Rv | 5′-GGGACGCCCCGTATGTATTC-3′ |
TmSPZ3-T7-270bp-Fw | 5′-TAATACGACTCACTATAGGGT CGAGAACAAGGCACTGATGA-3′ |
TmSPZ3-T7-270bp-Rv | 5′-TAATACGACTCACTATAGGGT GCGGTGCCATTTGTACTTCT-3′ |
TmSPZ-like-qPCR-Fw | 5′-CAGTTGAGGGTGCCTGTTCA-3′ |
TmSPZ-like-qPCR-Rv | 5′-TTGTTGGCATCGTCCCTTGA-3′ |
TmSpz-like-T7-Fw | 5′-TAATACGACTCACTATAGGGT ATGTTCCCAAAATCAACGGA-3′ |
TmSpz-like-T7-Rv | 5′-TAATACGACTCACTATAGGGT AATCACACGCAGATCCTTCC-3′ |
dsEGFP_Fw | 5′-TAATACGACTCACTATAGGGT CGTAAACGGCCACAAGTTC-3′ |
dsEGFP_Rv | 5′-TAATACGACTCACTATAGGGT TGCTCAGGTAGTGTTGTCG-3′ |
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Edosa, T.T.; Jo, Y.H.; Keshavarz, M.; Kim, I.S.; Han, Y.S. Biosurfactants Induce Antimicrobial Peptide Production through the Activation of TmSpatzles in Tenebrio molitor. Int. J. Mol. Sci. 2020, 21, 6090. https://doi.org/10.3390/ijms21176090
Edosa TT, Jo YH, Keshavarz M, Kim IS, Han YS. Biosurfactants Induce Antimicrobial Peptide Production through the Activation of TmSpatzles in Tenebrio molitor. International Journal of Molecular Sciences. 2020; 21(17):6090. https://doi.org/10.3390/ijms21176090
Chicago/Turabian StyleEdosa, Tariku Tesfaye, Yong Hun Jo, Maryam Keshavarz, In Seon Kim, and Yeon Soo Han. 2020. "Biosurfactants Induce Antimicrobial Peptide Production through the Activation of TmSpatzles in Tenebrio molitor" International Journal of Molecular Sciences 21, no. 17: 6090. https://doi.org/10.3390/ijms21176090