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1.
Microb Biotechnol ; 16(11): 2011-2014, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37462982

RESUMO

Extensive use of chemical insecticides to control insect pests in agriculture has improved yields and production of high-quality food products. However, chemical insecticides have been shown to be harmful also to beneficial insects and many other organisms like vertebrates. Thus, there is a need to replace those chemical insecticides by other control methods in order to protect the environment. Insect pest pathogens, like bacteria, viruses or fungi, are interesting alternatives for production of microbial-based insecticides to replace the use of chemical products in agriculture. Organic farming, which does not use chemical pesticides for pest control, relies on integrated pest management techniques and in the use of microbial-based insecticides for pest control. Microbial-based insecticides require precise formulation and extensive monitoring of insect pests, since they are highly specific for certain insect pests and in general are more effective for larval young instars. Here, we analyse the possibility of using microbial-based insecticides to replace chemical pesticides in agricultural production.


Assuntos
Inseticidas , Praguicidas , Animais , Controle Biológico de Vetores/métodos , Agricultura/métodos , Insetos/microbiologia
2.
Microbiol Spectr ; 10(5): e0289122, 2022 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-35972281

RESUMO

Succinate dehydrogenase (SDH), also known as respiratory chain complex II, plays a crucial role in energy production in which SdhC functions as an anchored subunit in the inner membrane of mitochondria. In this study, domain annotation analyses revealed that two SdhC domain-containing proteins were present in the filamentous insect-pathogenic fungus Beauveria bassiana, and they were named BbSdhC1 and BbSdhC2, respectively. Only BbSdhC1 localized to mitochondria; hence, this protein is considered the ortholog of SdhC in B. bassiana. Ablation of BbSdhC1 led to significantly reduced vegetative growth on various nutrients. The ΔBbsdhc1 mutant displayed the significantly reduced ATP synthesis and abnormal differentiation under aerial and submerged conditions. Notably, the BbSdhC1 loss resulted in enhanced intracellular levels of reactive oxygen species (ROS) and impaired growth of mycelia under oxidative stress. Finally, insect bioassays (via cuticle and intrahemocoel injection infection) revealed that disruption of BbSdhC1 significantly attenuated fungal virulence against the insect hosts. These findings indicate that BbSdhC1 contributes to vegetative growth, resistance to oxidative stress, differentiation, and virulence of B. bassiana due to its roles in energy generation and maintaining the homeostasis of the intracellular ROS levels. IMPORTANCE The electron transport chain (ETC) is critical for energy supply by mediating the electron flow along the mitochondrial membrane. Succinate dehydrogenase (SDH) is also known as complex II in the ETC, in which SdhC is a subunit anchored in mitochondrial membrane. However, the physiological roles of SdhC remain enigmatic in filamentous fungi. In filamentous insect-pathogenic fungus B. bassiana, SdhC is required for maintaining mitochondrial functionality, which is critical for fungal stress response, development, and pathogenicity. These findings improve our understanding of physiological mechanisms of ETC components involved in pathogenicity of the entomopathogenic fungi.


Assuntos
Beauveria , Animais , Beauveria/genética , Beauveria/metabolismo , Virulência , Espécies Reativas de Oxigênio/metabolismo , Esporos Fúngicos , Succinato Desidrogenase/genética , Succinato Desidrogenase/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Insetos/microbiologia , Crescimento e Desenvolvimento , Trifosfato de Adenosina/metabolismo
3.
Sci Rep ; 11(1): 19624, 2021 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-34608174

RESUMO

The putative ferricrocin synthetase gene ferS in the fungal entomopathogen Beauveria bassiana BCC 2660 was identified and characterized. The 14,445-bp ferS encodes a multimodular nonribosomal siderophore synthetase tightly clustered with Fusarium graminearum ferricrocin synthetase. Functional analysis of this gene was performed by disruption with the bar cassette. ΔferS mutants were verified by Southern and PCR analyses. HPLC and TLC analyses of crude extracts indicated that biosynthesis of ferricrocin was abolished in ΔferS. Insect bioassays surprisingly indicated that ΔferS killed the Spodoptera exigua larvae faster (LT50 59 h) than wild type (66 h). Growth and developmental assays of the mutant and wild type demonstrated that ΔferS had a significant increase in germination under iron depletion and radial growth and a decrease in conidiation. Mitotracker staining showed that the mitochondrial activity was enriched in ΔferS under both iron excess and iron depletion. Comparative transcriptomes between wild type and ΔferS indicated that the mutant was increased in the expression of eight cytochrome P450 genes and those in iron homeostasis, ferroptosis, oxidative stress response, ergosterol biosynthesis, and TCA cycle, compared to wild type. Our data suggested that ΔferS sensed the iron excess and the oxidative stress and, in turn, was up-regulated in the antioxidant-related genes and those in ergosterol biosynthesis and TCA cycle. These increased biological pathways help ΔferS grow and germinate faster than the wild type and caused higher insect mortality than the wild type in the early phase of infection.


Assuntos
Beauveria/crescimento & desenvolvimento , Beauveria/metabolismo , Ferricromo/análogos & derivados , Interações Hospedeiro-Patógeno , Insetos/microbiologia , Ferro/metabolismo , Animais , Beauveria/classificação , Beauveria/patogenicidade , Biologia Computacional , Ferricromo/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Perfilação da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Homeostase , Mutação , Estresse Oxidativo , Filogenia , Virulência/genética
4.
Microbiol Spectr ; 9(1): e0056421, 2021 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-34378960

RESUMO

Beauveria bassiana is an insect pathogenic fungus that serves as a model system for exploring the mechanisms of fungal development and host-pathogen interactions. Clinical and experimental studies have indicated that SND1 is closely correlated with the progression and invasiveness of common cancers as a potential oncogene, but this gene has rarely been studied in fungi. Here, we characterized the contributions of an SND1 ortholog (Tdp1) by constructing a BbTdp1 deletion strain and a complemented strain of B. bassiana. Compared with the wild-type (WT) strain, the ΔBbTdp1 mutant lost conidiation capacity (∼87.7%) and blastospore (∼96.3%) yields, increased sensitivity to chemical stress (4.4 to 54.3%) and heat shock (∼44.2%), and decreased virulence following topical application (∼24.7%) and hemocoel injection (∼40.0%). Flow cytometry readings showed smaller sizes of both conidia and blastospores for ΔBbTdp1 mutants. Transcriptomic data revealed 4,094 differentially expressed genes (|log2 ratio| > 2 and a q value of <0.05) between ΔBbTdp1 mutants and the WT strain, which accounted for 41.6% of the total genes, indicating that extreme fluctuation in the global gene expression pattern had occurred. Moreover, deletion of BbTdp1 led to an abnormal cell cycle with a longer S phase and shorter G2/M and G0/G1 phases of blastospores, and enzyme-linked immunosorbent assay confirmed that the level of phosphorylated cyclin-dependent kinase 1 (Cdk1) in the ΔBbTdp1 strain was ∼31.5% lower than in the WT strain. In summary, our study is the first to report that BbTdp1 plays a vital role in regulating conidia and blastospore yields, fungal morphological changes, and pathogenicity in entomopathogenic fungi. IMPORTANCE In this study, we used Beauveria bassiana as a biological model to report the role of BbTdp1 in entomopathogenic fungi. Our findings indicated that BbTdp1 contributed significantly to cell development, the cell cycle, and virulence in B. bassiana. In addition, deletion of BbTdp1 led to drastic fluctuations in the transcriptional profile. BbTdp1 can be developed as a novel target for B. bassiana development and pathogenicity, which also provides a framework for the study of Tdp1 in other fungi.


Assuntos
Beauveria/crescimento & desenvolvimento , Beauveria/metabolismo , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Insetos/microbiologia , Animais , Beauveria/genética , Beauveria/patogenicidade , Ciclo Celular , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , Transcriptoma , Domínio Tudor , Virulência
5.
Fitoterapia ; 153: 104973, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34157376

RESUMO

As part of our program to discover new bioactive agents from endophytic fungi, three new indole alkaloids (1-2, 4) along with twelve known compounds were isolated from an inset derived endophytic strain Aspergillus lentulus. Their structures were determined by comprehensive spectroscopic analyses of 1D/2D NMR and HR-ESI-MS data. The absolute configurations were confirmed by ECD calculation using Time-dependent Density functional theory (TD-DFT) at the B3LYP/6-31 + g (d, p) level and Rh2(OCOCF3)4-induced ECD experiments. Compounds 2, 4, 5, 13 and 15 exhibited moderate cytotoxic effects on A549 cell line with IC50 in the range of 17.92-48.29 µM. Compounds 1, 2 and 13-15 displayed the anti-bacterial activity against Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola with MIC values ranging from 25 to 100 µg/mL.


Assuntos
Antibacterianos/farmacologia , Antineoplásicos/farmacologia , Aspergillus/química , Alcaloides Indólicos/farmacologia , Insetos/microbiologia , Células A549 , Animais , Antibacterianos/isolamento & purificação , Antineoplásicos/isolamento & purificação , Humanos , Alcaloides Indólicos/isolamento & purificação , Estrutura Molecular , Xanthomonas/efeitos dos fármacos
6.
Artigo em Inglês | MEDLINE | ID: mdl-34186180

RESUMO

Larvae of some insect species (Coleoptera and Lepidoptera) can consume and biodegrade synthetic polymers, including polyethylene, polystyrene, polyvinyl chloride, and polypropylene. Multiple chemical (polymer mass loss and shift of the molecular weight, alterations in chemical functionality, formation of biodegraded intermediates, CO2 production), physical (surface hydrophobicity, thermal analysis), and biological approaches (antibiotic treatment, gut dysbiosis, isolation of plastic microbial degraders) have provided evidence for polymer biodegradation in the larva digestive tract. However, the extent and rate of biodegradation largely depend on the physicochemical structure of the polymer as well as the presence of additives. Additionally, toxicology associated with plastic biodegradation has not been investigated. This knowledge gap is critical to understand the gut symbiont-host interaction in the biodegradation process, its viability in the long term, the effects of plastic additives and their metabolites, and the phenotypic traits linked to a plastic-rich diet might be transferred in successive generations. Likewise, plastic-eating larvae represent a unique case study for elucidating the mechanisms of toxic action by micro- and nanoplastics because of the high concentration of plastics these organisms may be intentionally exposed to. This perspective review graphically summarizes the current knowledge on plastic biodegradation by insect larvae and describes the physiological processes (digestive and immune systems) that may be disrupted by micro- and nanoplastics. It also provides an outlook to advance current knowledge on the toxicity assessment of plastic-rich diets and the environmental risks of plastic-containing by-products (e.g., insect manure used as fertilizer).


Assuntos
Bactérias/metabolismo , Plásticos Biodegradáveis/metabolismo , Biodegradação Ambiental , Insetos/metabolismo , Animais , Plásticos Biodegradáveis/toxicidade , Insetos/microbiologia
7.
J Biosci ; 452020.
Artigo em Inglês | MEDLINE | ID: mdl-33361627

RESUMO

Tobacco is one of the major industrial crops cultivated worldwide. Chemical control is the main method employed to reduce damage by insect pests. The use of entomopathogenic fungi represents an alternative to replace insecticides. The search for effective strains in the field constitutes a first step when developing a formulation. The objective of this work was to study genetic differences among isolates of entomopathogenic fungi obtained from tobacco grown soils using ISSR markers. The pathogenicity of the strains towards Helicoverpa gelotopoeon and Diabrotica speciosa was also assessed in order to search for a relationship between virulence and genetic diversity. Nineteen isolates were identified according to morphological features and molecular techniques as Beauveria bassiana (11) and Purpureocillium lilacinum (8). The diversity tree generated by ISSR analysis showed a high diversity among the strains. The pathogenicity towards H. gelotopoeon and D. speciosa was assessed and the logistic models generated showed that B. bassiana isolates LPSc1215 and LPSc1364 were the most pathogenic against both insect pests tested. In the diversity tree, these strains were grouped in a same cluster with a similarity level of approximately 85%, indicating a possible relationship between virulence and the band pattern generated.


Assuntos
Fungos/genética , Nicotiana/crescimento & desenvolvimento , Controle Biológico de Vetores , Microbiologia do Solo , Animais , Beauveria/genética , Beauveria/patogenicidade , Fungos/crescimento & desenvolvimento , Fungos/patogenicidade , Variação Genética/genética , Hypocreales/genética , Hypocreales/patogenicidade , Insetos/microbiologia , Insetos/parasitologia , Mariposas/microbiologia , Mariposas/parasitologia , Filogenia , Nicotiana/microbiologia
8.
Mol Biochem Parasitol ; 238: 111298, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32621939

RESUMO

Endosymbiotic bacteria that obligately associate with entomopathogenic nematodes as a complex are a unique model system to study competition. These nematodes seek an insect host and provide entry for their endosymbionts. Through their natural products, the endosymbionts nurture their nematodes by eliminating secondary infection, providing nutrients through bioconversion of the insect cadaver, and facilitating reproduction. On one hand, they cooperatively colonize the insect host and neutralize other opportunistic biotic threats. On the other hand, inside the insect cadaver as a fighting pit, they fiercely compete for the fittest partnership that will grant them the reproductive dominance. Here, we review the protective and nurturing nature of endosymbiotic bacteria for their nematodes and how their selective preference shapes the superior nematode-endosymbiont pairs as we know today.


Assuntos
Bactérias/metabolismo , Fatores Biológicos/biossíntese , Insetos/parasitologia , Nematoides/microbiologia , Infecções por Nematoides/parasitologia , Simbiose/fisiologia , Animais , Bactérias/crescimento & desenvolvimento , Catecol Oxidase/metabolismo , Precursores Enzimáticos/metabolismo , Proteínas de Helminto/metabolismo , Hemolinfa/microbiologia , Hemolinfa/parasitologia , Insetos/microbiologia , Nematoides/enzimologia , Nematoides/patogenicidade , Infecções por Nematoides/microbiologia , Fosfolipases A2/metabolismo
10.
PLoS Pathog ; 16(4): e1008518, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32324832

RESUMO

Competition is one of the fundamental driving forces of natural selection. Beauveria bassiana is a soil and plant phylloplane/root fungus capable of parasitizing insect hosts. Soil and plant environments are often enriched with other fungi against which B. bassiana competes for survival. Here, we report an antifungal peptide (BbAFP1), specifically expressed and localized to the conidial cell wall and is released into the surrounding microenvironment inhibiting growth of competing fungi. B. bassiana strains expressing BbAFP1, including overexpression strains, inhibited growth of Alternaria brassicae in co-cultured experiments, whereas targeted gene deletion of BbAFP1 significantly decreased (25%) this inhibitory effect. Recombinant BbAFP1 showed chitin and glucan binding abilities, and growth inhibition of a wide range of phytopathogenic fungi by disrupting membrane integrity and eliciting reactive oxygen species (ROS) production. A phenylalanine residue (F50) contributes to chitin binding and antifungal activity, but was not required for the latter. Expression of BbAFP1 in tomato resulted in transgenic plants with enhanced resistance to plant fungal pathogens. These results highlight the importance of fungal competition in shaping primitive competition strategies, with antimicrobial compounds that can be embedded in the spore cell wall to be released into the environment during the critical initial phases of germination for successful growth in its environmental niche. Furthermore, these peptides can be exploited to increase plant resistance to fungal pathogens.


Assuntos
Antifúngicos/metabolismo , Beauveria/metabolismo , Esporos Fúngicos/metabolismo , Animais , Antifúngicos/farmacologia , Beauveria/genética , Parede Celular/metabolismo , Quitina/metabolismo , Proteínas Fúngicas/metabolismo , Glucanos/metabolismo , Insetos/microbiologia , Peptídeos , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Espécies Reativas de Oxigênio , Estresse Fisiológico/efeitos dos fármacos , Virulência
11.
Arch Microbiol ; 202(6): 1275-1284, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32185410

RESUMO

Insect and mite pests are damaging stressors that are threatening the cultivation of tea plants, which result in enormous crop loss. Over the years, the effectiveness of synthetic pesticides has allowed for its prominent application as a control strategy. However, the adverse effects of synthetic pesticides in terms of pesticide residue, environmental contamination and insect pest resistance have necessitated the need for alternative strategies. Meanwhile, microbial pesticides have been applied to tackle the damaging activities of the insect and mite pests of tea plants, and their performances were scientifically adjudged appreciable and environmental friendly. Herein, entomopathogenic microbes that were effective against tea geometrid (Ectropis obliqua Prout), tea green leafhopper (Empoasca onukii Matsuda), paraguay tea ampul (Gyropsylla spegazziniana), tea mosquito bug (Helopeltis theivora Waterhouse) and red spider mite (Oligonychus coffea Nietner) have been reviewed. The current findings revealed that microbial pesticides were effective and showed promising performances against these pests. Overall, this review has provided the basic and integrative information on the integrated pest management (IPM) tool(s) that can be utilized towards successful control of the aforementioned insect and mite pests.


Assuntos
Camellia sinensis/parasitologia , Insetos , Ácaros , Controle Biológico de Vetores/normas , Animais , Insetos/microbiologia , Insetos/virologia , Praguicidas/normas
12.
Subcell Biochem ; 94: 81-121, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32189297

RESUMO

The composition of insect hemolymph can change depending on many factors, e.g. access to nutrients, stress conditions, and current needs of the insect. In this chapter, insect immune-related polypeptides, which can be permanently or occasionally present in the hemolymph, are described. Their division into peptides or low-molecular weight proteins is not always determined by the length or secondary structure of a given molecule but also depends on the mode of action in insect immunity and, therefore, it is rather arbitrary. Antimicrobial peptides (AMPs) with their role in immunity, modes of action, and classification are presented in the chapter, followed by a short description of some examples: cecropins, moricins, defensins, proline- and glycine-rich peptides. Further, we will describe selected immune-related proteins that may participate in immune recognition, may possess direct antimicrobial properties, or can be involved in the modulation of insect immunity by both abiotic and biotic factors. We briefly cover Fibrinogen-Related Proteins (FREPs), Down Syndrome Cell Adhesion Molecules (Dscam), Hemolin, Lipophorins, Lysozyme, Insect Metalloproteinase Inhibitor (IMPI), and Heat Shock Proteins. The reader will obtain a partial picture presenting molecules participating in one of the most efficient immune strategies found in the animal world, which allow insects to inhabit all ecological land niches in the world.


Assuntos
Antibacterianos/imunologia , Antibacterianos/metabolismo , Proteínas de Insetos/imunologia , Proteínas de Insetos/metabolismo , Insetos/imunologia , Peptídeos/imunologia , Peptídeos/metabolismo , Animais , Hemolinfa/imunologia , Hemolinfa/metabolismo , Insetos/microbiologia
13.
Virulence ; 11(1): 222-237, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32079481

RESUMO

A diverse family of metalloproteases (MPs) is distributed in eukaryotes. However, the functions of MPs are still understudied. We report that seven MPs belonging to the M35 family are encoded in the genome of the insect pathogenic fungus Metarhizium robertsii. By gene deletions and insect bioassays, we found that one of the M35-family MPs, i.e. MrM35-4, is required for fungal virulence against insect hosts. MrM35-4 is a secretable enzyme and shows a proteolytic activity implicated in facilitating fungal penetration of insect cuticles. After gene rescue and overexpression, insect bioassays indicated that MrM35-4 contributes to inhibiting insect cuticular and hemocyte melanization activities. Enzymatic cleavage assays revealed that the recombinant prophenoloxidases PPO1 and PPO2 of Drosophila melanogaster could be clipped by MrM35-4 in a manner differing from a serine protease that can activate PPO activities. In addition, it was found that MrM35-4 is involved in suppressing antifungal gene expression in insects. Consistent with the evident apoptogenic effect of MrM35-4 on host cells, we found that the PPO mutant flies differentially succumbed to the infections of the wild-type and mutant strains of M. robertsii. Thus, MrM35-4 plays a multifaceted role beyond targeting PPOs during fungus-insect interactions, which represents a previously unsuspected strategy employed by Metarhizium to outmaneuver insect immune defenses.


Assuntos
Catecol Oxidase/metabolismo , Precursores Enzimáticos/metabolismo , Insetos/microbiologia , Metaloproteases/genética , Metarhizium/genética , Metarhizium/metabolismo , Animais , Apoptose , Drosophila melanogaster/microbiologia , Feminino , Proteínas Fúngicas/genética , Deleção de Genes , Regulação Fúngica da Expressão Gênica , Interações Hospedeiro-Patógeno , Larva/microbiologia , Metaloproteases/metabolismo , Metarhizium/enzimologia , Virulência/genética
14.
Anticancer Agents Med Chem ; 20(4): 417-428, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31830896

RESUMO

BACKGROUND: In our previous study, we have isolated a new compound, named Fumosorinone (FU) from insect pathogenic fungi, and was found to inhibit proliferation, migration, and invasion of breast cancer MDA-MB-231 cells. OBJECTIVE: The aim of this study was to identify the underlying molecular mechanisms for FU effects on MDAMB- 231 cells. METHODS: After MDA-MB-231 cells were treated with FU for 48h, RNA sequencing was used to identify the effect of FU on the transcriptome of MDA-MB-231 cells. The validation of the relative expression of the selective genes was done using quantitative real-time PCR (qRT-PCR). RESULTS: The transcriptome results showed that 2733 genes were differentially expressed between the untreated and the FU-treated cells, including 1614 up-regulated and 1119 down-regulated genes. The multiple genes are associated with cancer cell growth, migration, and invasion. Functional analysis identified multitude of pathways related to cancer, such as cell cycle, ECM-receptor interaction, p53 signaling pathway. We selected 4 upregulated and 9 downregulated genes, which are associated with breast cancer to verify their expression using qRT-PCR. The validation showed that HSD3B1, ALOX5, AQP5, COL1A2, CCNB1, CCND1, VCAM-1, PTPN1 and PTPN11 were significantly downregulated while DUSP1, DUSP5, GADD45A, EGR1 were upregulated in FU-treated MDA-MB-231cells. CONCLUSION: These aberrantly expressed genes and pathways may play pivotal roles in the anti-cancer activity of FU, and maybe potential targets of FU treatments for TNBC. Further investigations are required to evaluate the FU mechanisms of anti-cancer action in vivo.


Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Ácidos Hidroxâmicos/farmacologia , Piridonas/farmacologia , Transcriptoma/efeitos dos fármacos , Animais , Antineoplásicos/química , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Feminino , Fungos/química , Perfilação da Expressão Gênica , Humanos , Ácidos Hidroxâmicos/química , Insetos/microbiologia , Piridonas/química
15.
Acta sci., Biol. sci ; 42: e52919, fev. 2020. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1460943

RESUMO

Few studies try to explain the effects in tropical lotic ecosystems of an increase in water temperature on the shredding activity of invertebrate shredders, particularly in association with the quality of the leaf litter and the degree of litter conditioning. Therefore, the aims of this study were as follows: i) to better understand how this key invertebrate shredder group affects the decomposition of different species of leaf litter under gradual increases in temperature and microbial conditioning; and ii) to verify the possible consequences on leaf mass loss (LML). Three species of leaf litter were used in two experiments. Inexperiment I, the litters of three species (Protium spruceanum, Richeria grandisand Ingalaurina) at three conditioning levels (1, 7, 14 days) were tested under five different temperatures (20, 22, 24, 26 and28°C). In experiment II, the leaf litters of three species were used, without conditioning, under four temperatures (20, 22, 26 and27°C). The shredding performed by Phylloicussp. was largely dependent on the lignin and cellulose concentrations in each leaf species, independent of conditioning. The presence or absence of conditioning may cause the shredders to use different energy compensation strategies in response to the temperature increases.


Assuntos
Ecossistema , Insetos/microbiologia , Trituração de Resíduos Sólidos , Viabilidade Microbiana
16.
New Phytol ; 223(4): 2002-2010, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31002383

RESUMO

Herbaceous plants harbour species-rich communities of asymptomatic endophytic fungi. Although some of these endophytes are entomopathogenic, many are not, and remarkably little is known about how the presence of these fungi in plant tissues affects phytophagous insects. Here we show through a meta-analysis that both entomopathogenic and nonentomopathogenic endophytes have a negative effect on insect herbivores. Growth and performance of polyphagous and sucking insects are reduced by nonentomopathogenic endophytes, but monophages are unaffected, likely because the latter are better adapted to secondary metabolites produced or induced by the fungi. Furthermore, studies using excised leaves report weaker effects than those with intact plants, likely caused by chemical changes being masked by leaf excision. Most surprisingly, endophyte infection of seeds produces the greatest effect on insect herbivores in subsequent mature plants, even though the usual mode of fungal transmission is infection of leaves by airborne spores. We conclude that these ubiquitous hidden fungi may be important bodyguards of plants. However, in order to fully understand their roles in plant protection, we must be aware that minor differences in experimental design can lead to contradictory results.


Assuntos
Endófitos/fisiologia , Insetos/microbiologia , Plantas/microbiologia , Animais , Comportamento Alimentar , Folhas de Planta/fisiologia , Pupa/fisiologia
17.
PLoS One ; 14(2): e0212505, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30817757

RESUMO

The digestive system of selected phytophagous insects has been examined as a potential prospecting resource for identification of novel cellulolytic enzymes with potential industrial applications. In contrast to other model species, however, limited detailed information is available that characterizes cellulolytic activity and systems in basal hexapod groups. As part of a screening effort to identify insects with highly active cellulolytic systems, we have for the first time, identified species of Zygentoma that displayed the highest relative cellulase activity levels when compared to all other tested insect groups under the experimental conditions, including model species for cellulolytic systems such as termite and cockroach species in Rhinotermitidae (formerly Isoptera) and Cryptocercidae (formerly Blattodea). The goal of the present study was to provide a morphohistological characterization of cellulose digestion and to identify highly active cellulase enzymes present in digestive fluids of Zygentoma species. Morphohistological characterization supported no relevant differences in the digestive system of firebrat (Thermobia domestica) and the gray silverfish (Ctenolepisma longicaudata). Quantitative and qualitative cellulase assays identified the foregut as the region with the highest levels of cellulase activity in both T. domestica and C. longicaudata. However, T. domestica was found to have higher endoglucanase, xylanase and pectinase activities compared to C. longicaudata. Using nano liquid chromatography coupled to tandem mass spectrometry (nanoLC/MS/MS) and a custom gut transcriptome we identified cellulolytic enzymes from digestive fluids of T. domestica. Among the identified enzymes we report putative endoglucanases matching to insect or arthropod enzymes and glucan endo-1,6-ß-glucosidases matching bacterial enzymes. These findings support combined activities of endogenous and symbiont-derived plant cell wall degrading enzymes in lignocellulose digestion in Zygentoma and advance our understanding of cellulose digestion in a primitive insect group.


Assuntos
Celulase/metabolismo , Proteínas de Insetos/metabolismo , Insetos/enzimologia , Animais , Celulase/genética , Baratas/enzimologia , Baratas/genética , Baratas/microbiologia , Sistema Digestório/anatomia & histologia , Sistema Digestório/enzimologia , Sistema Digestório/microbiologia , Endo-1,4-beta-Xilanases/metabolismo , Proteínas de Insetos/genética , Insetos/genética , Insetos/microbiologia , Isópteros/enzimologia , Isópteros/genética , Isópteros/microbiologia , Lepisma/enzimologia , Lepisma/genética , Lepisma/microbiologia , Modelos Biológicos , Poligalacturonase/metabolismo , Especificidade da Espécie , Transcriptoma
18.
J Nat Prod ; 81(8): 1752-1759, 2018 08 24.
Artigo em Inglês | MEDLINE | ID: mdl-30024750

RESUMO

Four new highly oxygenated p-terphenyls, hawaiienols A-D (1-4), have been isolated from cultures of Paraconiothyrium hawaiiense, a fungus associated with the Septobasidium-infected insect Diaspidiotus sp.; their structures were elucidated primarily by NMR experiments. The absolute configurations of 1 and 2-4 were assigned by single-crystal X-ray diffraction analysis using Cu Kα radiation and via electronic circular dichroism calculations, respectively. Compound 1 incorporated the first naturally occurring 4,7-dioxatricyclo[3.2.1.03,6]octane unit in its p-terphenyl skeleton and showed cytotoxicity toward six human tumor cell lines.


Assuntos
Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/farmacologia , Ascomicetos/metabolismo , Insetos/microbiologia , Compostos de Terfenil/química , Compostos de Terfenil/farmacologia , Animais , Ascomicetos/química , Linhagem Celular Tumoral , Dicroísmo Circular , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Espectrometria de Massas por Ionização por Electrospray , Difração de Raios X
19.
Appl Environ Microbiol ; 84(15)2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-29802184

RESUMO

Fungal ß-1,3-glucanosyltransferases are cell wall-remodeling enzymes implicated in stress response, cell wall integrity, and virulence, with most fungal genomes containing multiple members. The insect-pathogenic fungus Beauveria bassiana displays robust growth over a wide pH range (pH 4 to 10). A random insertion mutant library screening for increased sensitivity to alkaline (pH 10) growth conditions resulted in the identification and mapping of a mutant to a ß-1,3-glucanosyltransferase gene (Bbgas3). Bbgas3 expression was pH dependent and regulated by the PacC transcription factor, which activates genes in response to neutral/alkaline growth conditions. Targeted gene knockout of Bbgas3 resulted in reduced growth under alkaline conditions, with only minor effects of increased sensitivity to cell wall stress (Congo red and calcofluor white) and no significant effects on fungal sensitivity to oxidative or osmotic stress. The cell walls of ΔBbgas3 aerial conidia were thinner than those of the wild-type and complemented strains in response to alkaline conditions, and ß-1,3-glucan antibody and lectin staining revealed alterations in cell surface carbohydrate epitopes. The ΔBbgas3 mutant displayed alterations in cell wall chitin and carbohydrate content in response to alkaline pH. Insect bioassays revealed impaired virulence for the ΔBbgas3 mutant depending upon the pH of the media on which the conidia were grown and harvested. Unexpectedly, a decreased median lethal time to kill (LT50, i.e., increased virulence) was seen for the mutant using intrahemocoel injection assays using conidia grown at acidic pH (5.6). These data show that BbGas3 acts as a pH-responsive cell wall-remodeling enzyme involved in resistance to extreme pH (>9).IMPORTANCE Little is known about adaptations required for growth at high (>9) pH. Here, we show that a specific fungal membrane-remodeling ß-1,3-glucanosyltransferase gene (Bbgas3) regulated by the pH-responsive PacC transcription factor forms a critical aspect of the ability of the insect-pathogenic fungus Beauveria bassiana to grow at extreme pH. The loss of Bbgas3 resulted in a unique decreased ability to grow at high pH, with little to no effects seen with respect to other stress conditions, i.e., cell wall integrity and osmotic and oxidative stress. However, pH-dependent alternations in cell wall properties and virulence were noted for the ΔBbgas3 mutant. These data provide a mechanistic insight into the importance of the specific cell wall structure required to stabilize the cell at high pH and link it to the PacC/Pal/Rim pH-sensing and regulatory system.


Assuntos
Álcalis/metabolismo , Beauveria/enzimologia , Proteínas Fúngicas/metabolismo , Transferases/metabolismo , Adaptação Fisiológica , Animais , Beauveria/genética , Beauveria/patogenicidade , Beauveria/fisiologia , Parede Celular/enzimologia , Parede Celular/genética , Parede Celular/metabolismo , Meios de Cultura/química , Meios de Cultura/metabolismo , Proteínas Fúngicas/genética , Insetos/microbiologia , Esporos Fúngicos/enzimologia , Esporos Fúngicos/genética , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/fisiologia , Estresse Fisiológico , Transferases/genética , Virulência
20.
Nat Prod Rep ; 35(5): 434-454, 2018 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-29644346

RESUMO

Covering: up to 2018 Insects live in a world full of toxic compounds such as plant toxins and manmade pesticides. To overcome the effects of these toxins, herbivorous insects have evolved diverse, elaborate mechanisms of resistance, such as toxin avoidance, target-site alteration, and detoxification. These resistance mechanisms are thought to be encoded by the insects' own genomes, and in many cases, this holds true. However, recent omics analyses, in conjunction with classic culture-dependent analyses, have revealed that a number of insects possess specific gut microorganisms, some of which significantly contribute to resistance against phytotoxins and pesticides by degrading such chemical compounds. Here, we review recent advances in our understanding on the symbiont-mediated degradation of natural and artificial toxins, with a special emphasis on their underlying genetic basis, focus on the importance of environmental microbiota as a resource of toxin-degrading microorganisms, and discuss the ecological and evolutionary significance of these symbiotic associations.


Assuntos
Insetos/efeitos dos fármacos , Insetos/microbiologia , Praguicidas/farmacocinética , Simbiose/fisiologia , Toxinas Biológicas/farmacocinética , Animais , Evolução Biológica , Enzimas/genética , Enzimas/metabolismo , Inativação Metabólica/genética , Isotiocianatos/farmacocinética , Oxalatos/farmacocinética , Fenóis/farmacocinética , Simbiose/efeitos dos fármacos , Terpenos/farmacocinética
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