Sen-Miao Tong

The high costs of mycoinsecticides restrain their extensive application in green agriculture. Two six-week field trials were performed in spring to test synergistic effects of fungal–chemical interactions against faba bean aphid (Aphis fabae) populations in Zhejiang, China. The treatments (three 50-m² plots each) in each trial included Beauveria bassiana ZJU435 (Bb) and imidacloprid (ImD) applied biweekly at recommended rates (1.5 × 10¹³ conidia and 45 a.i. g ha⁻¹) and co-applied at reduced rates of 2/3Bb, 1/2Bb, and 1/3Bb plus 1/5ImD, which reduced the cost of the mycoinsecticide by 32–65%. During the first three weeks at 1.3–12.5 °C (daily means), the aphid population steadily increased to a peak in the blank control, and it was much more effectively suppressed by ImD than by Bb and Bb + ImD treatments. As the weather subsequently warmed to 8.7–21.0 °C, the aphid population was increasingly suppressed by the Bb and Bb + ImD treatments, which showed suppressive effects superior or close to those of ImD on days 35 and 42. Percent efficacy values (n = 36) of ImD, 2/3Bb + 1/5ImD, 1/2Bb + 1/5ImD, Bb, and 1/3Bb + 1/5ImD in two six-week trials averaged respectively 86%, 66%, 62.2%, 59.4%, and 58.4%, which significantly differed from one another (p < 0.01). Conclusively, low-rate co-applications of fungal/chemical insecticides offer a ‘low-cost–high-efficacy’ strategy to promote extensive mycoinsecticide application for sustainable aphid control.

Thechemical control of rice planthoppers (RPH)is prohibited in annual rice–shrimp rotation paddy fields. Here, the fungal insecticides Beauveria bassiana ZJU435 and Metarizhium anisoplae CQ421 were tested for control of RPH populations dominated by Nilaparvata lugens in three field trials. During four-week field trials initiated from the harsh weather of high temperatures and strong sunlight, the rice crop at the stages from tillering to flowering was effectively protected by fungal sprays applied at 14-day intervals. The sprays of either fungal insecticide after 5:00 p.m. (solar UV avoidance) suppressed the RPH population better than those before 10 a.m. The ZJU435 and CQ421 sprays for UV avoidance versus UV exposure resulted in mean control efficacies of 60% and 56% versus 41% and 45% on day 7, 77% and 78% versus 63% and 67% on day 14, 84% and 82% versus 80% and 79% on day 21, and 84% and 81% versus 79% and 75 on day 28, respectively. These results indicate that fungal insecticides can control RPH in the rice–shrimp rotation fields and offer a novel insight into the significance of solar-UV-avoiding fungal application for improved pest control during sunny summers.

FluG is a core regulator upstream of central developmental pathway (CDP) in Aspergillus nidulans but multiple FluG-like regulators (FLRs) remain functionally uncharacterized in ascomycetes. Our previous study revealed no role for FluG in the CDP activation and an existence of sole FLR (FlrA) in an insect-pathogenic fungus.

Resistance to solar ultraviolet (UV) irradiation is crucial for field‐persistent control efficacies of fungal formulations against arthropod pests, because their active ingredients are formulated conidia very sensitive to solar UV wavelengths. This review seeks to summarize advances in studies aiming to quantify, understand and improve conidial UV resistance. One focus of studies has been on the many sets of genes that have been revealed in the postgenomic era to contribute to or mediate UV resistance in the insect pathogens serving as main sources of fungal insecticides. Such genetic studies have unveiled the broad basis of UV‐resistant molecules including cytosolic solutes, cell wall components, various antioxidant enzymes, and numerous effectors and signaling proteins, that function in developmental, biosynthetic and stress‐responsive pathways. Another focus has been on the molecular basis and regulatory mechanisms underlying photorepair of UV‐induced DNA lesions and photoreactivation of UV‐impaired conidia. Studies have shed light upon a photoprotective mechanism depending on not only one or two photorepair‐required photolyases, but also two white collar proteins and other partners that play similar or more important roles in photorepair via interactions with photolyases. Research hotspots are suggested to explore a regulatory network of fungal photoprotection and to improve the development and application strategies of UV‐resistant fungal insecticides. © 2021 Society of Chemical Industry.

Summary: Biological control potential of insect‐pathogenic fungi against pests is an overall output of various cellular processes regulated by signalling and epigenetic networks. In Beauveria bassiana , mono/di/trimethylation of histone H3 Lys 4 (H3K4me1/me2/m3) was abolished by inactivation of the histone lysine methyltransferase SET1/KMT2, leading to marked virulence loss, reductions in conidial hydrophobicity and adherence to insect cuticle, impeded proliferation in vivo , severe defects in growth and conidiation, and increased sensitivities to cell wall perturbation, H ₂ O ₂ and heat shock. Such compromised phenotypes correlated well with transcriptional abolishment or repression of carbon catabolite‐repressing transcription factor Cre1, classes I and II hydrophobins Hyd1 and Hyd2 required for cell hydrophobicity, key developmental regulators, and stress‐responsive enzymes/proteins. Particularly, expression of cre1 , which upregulates hyd4 upon activation by KMT2‐mediated H3K4me3 in Metarhizium robertsii , was nearly abolished in the Δ set1 mutant, leading to abolished expression of hyd1 and hyd2 as homologues of hyd4 . These data suggest that the SET1‐Cre1‐Hyd1/2 pathway function in B . bassiana like the KMT2‐Cre1‐Hyd4 pathway elucidated to mediate pathogenicity in M . robertsii . Our findings unveil not only a regulatory role for the SET1‐cored pathway in fungal virulence but also its novel role in mediating asexual cycle in vitro and stress responses in B . bassiana .

Csn5 is a subunit ofthe COP9/signalosome complex in model fungi. Here, we report heavier accumulation of orthologous Csn5 in the nucleus than in the cytoplasm and its indispensability to insect pathogenicity and virulence-related cellular events of Beauveria bassiana. Deletion of csn5 led to a 68% increase in intracellular ubiquitin accumulation and the dysregulation of 18 genes encoding ubiquitin-activating (E1), -conjugating (E2), and -ligating (E3) enzymes and ubiquitin-specific proteases, suggesting the role of Csn5 in balanced ubiquitination/deubiquitination. Consequently, the deletion mutant displayed abolished insect pathogenicity, marked reductions in conidial hydrophobicity and adherence to the insect cuticle, the abolished secretion of cuticle penetration-required enzymes, blocked haemocoel colonisation, and reduced conidiation capacity despite unaffected biomass accumulation. These phenotypes correlated well with sharply repressed or abolished expressions of key hydrophobin genes required for hydrophobin biosynthesis/assembly and of developmental activator genes essential for aerial conidiation and submerged blastospore production. In the mutant, increased sensitivities to heat shock and oxidative stress also correlated with reduced expression levels of several heat-responsive genes and decreased activities of antioxidant enzymes. Altogether, Csn5-reliant ubiquitination/deubiquitination balance coordinates the expression of those crucial genes and the quality control of functionally important enzymes, which are collectively essential for fungal pathogenicity, virulence-related cellular events, and asexual development.

Mono-, di- and tri-methylation of histone H3 Lys 9, Lys 4, and Lys 36 (H3K_me1/me2/me3) required for mediation of DNA-based cellular events in eukaryotes usually rely upon the activities of histone lysine methyltransferases (KMTs) classified to the KMT1, KMT2, and KMT3 families, respectively. Here, an H3K9-specific DIM5/KMT1 orthologue, which lacks a C-terminal post-SET domain and localizes mainly in nucleus, is reported to have both conserved and noncanonical roles in methylating the H3 core lysines in Beauveria bassiana, an insect-pathogenic fungus serving as a main source of wide-spectrum fungal insecticides. Disruption of dim5 led to abolishment of H3K9me3 and marked attenuation of H3K4me1/me2, H3K9me1/me2 and H3K36me2. Consequently, the Δdim5 mutant lost the whole insect pathogenicity through normal cuticle infection, and was compromised severely in virulence through cuticle-bypassing infection (hemocoel injection) and also in a series of cellular events critical for the fungal virulence and lifecycle in vivo and in vitro, including reduced hyphal growth, blocked conidiation, impeded proliferation in vivo, altered carbohydrate epitopes, disturbed cell cycle, reduced biosynthesis and secretion of cuticle-degrading enzymes, and increased sensitivities to various stresses. Among 1,201 dysregulated genes (up/down ratio: 712:489) associated with those phenotypic changes, 92 (up/down ratio: 59:33) encode transcription factors and proteins or enzymes involved in posttranslational modifications, implying that the DIM5-methylated H3 core lysines could act as preferential marks of those transcription-active genes crucial for global gene regulation. These findings uncover a novel scenario of DIM5 and its indispensability for insect-pathogenic lifestyle and genome stability of B. bassiana.

Summary: The upstream developmental activation (UDA) pathway comprises three fluG ‐cored cascades ( fluG‐flbA , fluG‐flbE/B/D and fluG‐flbC ) that activate the key gene brlA of central developmental pathway (CDP) to initiate conidiation in aspergilli. However, the core role of fluG remains poorly understood in other fungi. Here, we report distinctive role of fluG in the insect‐pathogenic lifecycle of Beauveria bassiana . Disruption of fluG resulted in limited conidiation defect, which was mitigated with incubation time and associated with time‐course up‐regulation/down‐regulation of all flb and CDP genes and another fluG ‐like gene (BBA_06309). In Δ fluG , increased sensitivities to various stresses correlated with repression of corresponding stress‐responsive genes. Its virulence through normal cuticle infection was attenuated greatly due to blocked secretion of cuticle‐degrading enzymes and delayed formation of hyphal bodies (blastospores) to accelerate proliferation in vivo and host death. In submerged Δ fluG cultures mimicking insect haemolymph, largely increased blastospore production concurred with drastic up‐regulation of the CDP genes brlA and abaA , which was associated with earlier up‐regulation of most flb genes in the cultures. Our results unveil an essentiality of fluG for fungal adaptation to insect‐pathogenic lifecycle and suggest the other fluG ‐like gene to act as an alternative player in the UDA pathway of B . bassiana .

Opposite nuclear rhythms of two distinct FRQ proteins (Frq1 and Frq2) coexisting in an asexual fungal insect pathogen have been shown to orchestrate the fungal nonrhythmic conidiation in vitro in a circadian day independent of photoperiod change. This paper reports essential roles of both Frq1 and Frq2, but a greater role for Frq2, in sustaining the fungal virulence and infection cycle since either frq1 or frq2 deletion led to marked delay of lethal action against a model insect and drastic reduction of conidial yield on insect cadavers.