Insect fungal symbionts: A promising source of detoxifying enzymes

Summary Many species of insects cultivate, inoculate, or contain symbiotic fungi. Insects feed on plant materials that contain plant-produced defensive toxins, or are exposed to insecticides or other pesticides when they become economically important pests. Therefore, it is likely that the symbiotic fungi are also exposed to these toxins and may actually contribute to detoxification of these compounds. Fungi associated with bark beetles, ambrosia beetles, termites, leaf-cutting ants, long-horned beetles, wood wasps, and drug store beetles can variously metabolize/detoxify tannins, lignins, terpenes, esters, chlorinated hydrocarbons, and other toxins. The fungi (Attamyces) cultivated by the ants and the yeast (Symbiotaphrina) contained in the cigarette beetle gut appear to have broad-spectrum detoxifying abilities. The present limiting factor for using many of these fungi for large scale detoxification of, for example, contaminated soils or agricultural commodities is their slow growth rate, but conventional strain selection techniques or biotechnological approaches should overcome this problem.Presented at the Symposium on Fungal Detoxification at the 48th Annual Meeting of the Society for Industrial Microbiology, Philadelphia, PA, August 4–9, 1991.     

The contribution of symbiotic yeast to toxin resistance of the cigarette beetle (Lasioderma serricorne)

Representative plant allelochemicals were tested for toxicity to larvae of the cigarette beetle Lasioderma serricorne (F.) that were either untreated, treated with fungicides, or treated to render them free of symbiotic yeast (aposymbiotic) through surface sterilization of eggs. Insects rendered symbiont-free had higher mortality and/or developmental rates than controls when fed diets containing flavone, resorcinol or tannic acid. Fungicides reduced symbiont populations and/or caused morphological abnormalities. Two hydrolytic enzymes, which made up a significant protion of mycetome hydrolytic activity, were absent when symbionts were not present, as indicated by gel electrophoresis. This information indicates symbionts do contribute to the survival of their host by detoxifying toxins.

---

Résumé La toxicité relative de substances allélochimiques végétales typiques a été examinée sur des larves de L. serricorne F. traitées aux fongicides et rendues aposymbiotiques par stérilisation superficielle des ufs. La toxicité de la nicotine est relativement inaffectée chez les larves aposymbiotiques; cependant les toxicités des resorcinol, flavone et acide tannique sont significativement augmentées chez les insectes ayant reçu un traitement pour éliminer leurs symbiontes. L'électrophorèse sur gel a montré que quelques enzymes responsables de l'hydrolyse du 1-naphthyl acétate, réaction enzymatique de la détoxification enzymatique, sont absentes chez les insectes aposymbiotiques. L'acide sorbique a réduit significativement l'effectif de symbiontes, tandis que le bénomyl a réduit leur gamme provoquant la multiplication anormale d'autres espèces. Ces résultats montrent que les symbiontes contribuent aux possibilités de détoxification de l'insecte. L'élimination des symbiontes peut être une technique efficace pour lutter contre les insectes qui en contiennent, puisqu'elle les priverait de leur contribution à l'alimentation et à la détoxification. Cette stratégie est vraisemblablement compatible avec des programmes de lutte intégrée, puisque les prédateurs et parasites ne contiennent pas de symbiontes.

     

Biological consequences of a reduction in the non-target DNA scanning capacity of a DNA repair enzyme

Numerous DNA-interactive proteins have been shown to locate specific sequences within large domains of non-target DNA in vitro and in vivo by a one-dimensional diffusion mechanism; however, the biological significance of this process has not been evaluated. We have examined the biological consequences of sliding for the pyrimidine dimer-specific DNA repair enzyme T4 endonuclease V, an enzyme which scans non-target DNA both in vitro and in vivo. An endonuclease V mutant was constructed whose only altered biochemical characteristic, measured in vitro, was a loss in its ability to slide on non-target DNA. In contrast to the native enzyme, when the mutated endonuclease V was expressed in DNA repair-deficient Escherichia coli, no enhanced ultraviolet survival was conferred. These results suggest that the mechanisms which DNA-interactive proteins employ to enhance the probability of locating their target sequences are of significant biological importance.     

Inhibitory effects of gossypol-related compounds on growth of Aspergillus flavus

Aims: The objective of this study was to test a series of gossypol‐related compounds for growth inhibition against Aspergillus flavus. Methods and Results: A series of chiral and achiral gossypol derivatives, some natural products of the cotton plant and others prepared by synthesis from gossypol, were incorporated into agar plates to follow the rate of A. flavus isolate AF13 colony growth. All tested compounds exhibited some growth inhibition against this organism. The synthetic compounds, gossypolone and apogossypolone, exhibited greater activity than either racemic or chiral gossypol. Methylated derivatives (i.e. 6‐methoxy and 6,6′‐dimethoxy derivatives) generally exhibited less activity than the nonmethylated parent compounds. The (?)‐optical form of gossypol was found to be slightly more active than the (+)‐optical form, and this trend was observed regardless of the presence of methoxy groups at the 6‐position. Growth inhibition of gossypolone and apogossypolone was concentration dependent. For gossypolone, the 50% effective dose was 90 μg ml^?1 of medium (165 μmol l^?1). For apogossypolone, the most active compound in the study, the 50% effective dose was 19 μg ml^?1 (38·7 μmol l^?1). The presence of gossypol‐related terpenoids appeared to stimulate production of A. flavus sclerotia, although replicate variability was so large that it was not possible to determine a significant correlation between the mass of sclerotia formed and compound growth inhibition. Conclusions: The quinone derivatives of gossypol, gossypolone and apogossypolone demonstrated significant fungal growth inhibitory activity against A. flavus. Significance and Impact of the Study: These gossypol derivatives may provide a new class of fungicide for use against the mycotoxigenic fungus A. flavus.     

Effects of dietary fat on cholesterol movement between tissues in CBA/J and C57BR/cdJ mice.

Differences in dietary fats cause differences in cholesterol metabolism in mice. CBA/J mice are resistant to diet-induced hypercholesterolemia and atherosclerosis; they adjust hepatic hydroxymethyl-glutaryl-CoA reductase activity (HMGR) to maintain homeostasis; C57BR/cdJ mice are susceptible, but young animals are thought to maintain homeostasis by changing fecal excretion of sterols. Compartmental modelling of movement of [4-14C]cholesterol was used to analyze movement of cholesterol between serum and liver, heart, and carcass in mice fed 40 en% fat, polyunsaturated to saturated fatty acid ratio (P/S) = 0.24 (US74) or 30 en% fat, P/S = 1 (MOD). Dietary effects were quite pronounced, while strain effects were more subdued. The C57/cdJ animals appear to regulate the overall cholesterol balance by reducing synthesis, as do the CBA/J animals, even though synthesis is not reduced to the same degree as in the CBA/J animals. Both diet and strain influence the whole-animal turnover rate, with slower turnover occurring for C57BR/cdJ animals and animals fed the US74 diet.