Isocitrate dehydrogenase-mediated metabolic disorders disrupt active immunization against fungal pathogens in eusocial termites

Abstract

Active immunization is a crucial colony-level pathogen defense to improve the survival of eusocial termites, which limits the lethal effect of biopesticides for termite control. Here, we report a possible strategy for RNAi-mediated metabolic disorders, which causes reduction in insect pathogen defense and might facilitate the foundation of nontoxic and sustainable pest control methods. Isocitrate dehydrogenase (IDH) was significantly upregulated by active immunization in termites, but its regulatory mechanism has yet to be well understood. We found that dsIDH-injected termites exhibited significantly reduced IDH at mRNA and protein levels and altered levels of isocitrate and NADH, indicating the impaired NAD+-IDH reaction. IDH-silenced termites displayed metabolic disorders, which was implied by significant changes in several metabolites from the carbohydrates and amino acids. When grooming toward fungus-exposed termites, IDH-silenced nestmates showed a significant upregulation of four apoptosis-related genes and caspase 3 activity, and hence the significantly increased rate of apoptosis. These physiological changes led to more apoptotic cell death in different body parts of the nestmates. In particular, more apoptotic cells increased disease susceptibility of the nestmates as revealed by growth of the significantly increased number of colony-forming units from dissected gut contents. Furthermore, the IDH-silenced nestmates exhibited lower antifungal activity and higher mortality, suggesting that IDH downregulation disrupted active immunization against fungal pathogens in termites. These findings illustrated the metabolic regulation of active immunization and provide a sound foundation for enhancing the impact of biopesticides by reducing the pathogen defense of hosts.

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Acknowledgements

We thank Pengdong Sun, Yongyong Gao and Ganghua Li for their assistance with the field collections. We thank Drs. Sylvia Cremer, Barbara Milutinovic and Megan Kutzer for valuable suggestions on an earlier draft of this manuscript. This work was supported by the National Natural Science Foundation of China (31572322) and the Fundamental Research Funds for the Central Universities (2662016PY062).

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Correspondence to Qiu-Ying Huang.

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Video S1 Five nestmates are grooming toward one fungus-exposed termite in a cell Petri dish (WMV 10207 kb)

Text S1 Experimental protocol (DOCX 33 kb)

Fig. S1 Immune responses in the dsIDH-injected naïve termites. (A) IDH silencing had no influence on GNBP1, GNBP2 and termicin expressions in naïve termites. (B) The expression of four apoptosis-related genes was significantly increased in the dsIDH-injected termites as compared to those of the dsGFP-injected termites. The data are shown as the mean ± SEM. *, P < 0.05 (TIFF 980 kb)

Fig. S2 The dead dsIDH-injected nestmates of the fungus-exposed termites due to infection with the pathogenic fungus M. anisopliae (black scale bar, 1 mm) (TIFF 5713 kb)

Table S1 Distribution of the 19 R. chinensis colonies for each experiment (XLSX 12 kb)

Table S2 Primers used for real-time RT-PCR in this study (XLSX 12 kb)

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Liu, L., Wang, CC., Zhao, XY. et al. Isocitrate dehydrogenase-mediated metabolic disorders disrupt active immunization against fungal pathogens in eusocial termites. J Pest Sci 93, 291–301 (2020). https://doi.org/10.1007/s10340-019-01164-y

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Keywords

  • Isocitrate dehydrogenase
  • Reticulitermes chinensis
  • Metarhizium anisopliae
  • Social immunity
  • Glucose metabolism
  • Apoptotic lesions