Hypocrealean Fungi Research Articles

Entomophthoralean and hypocrealean fungal pathogens of the sugarcane aphid, Melanaphis sacchari (Hemiptera: Aphididae), on sorghum in Georgia

The sugarcane aphid, Melanaphis sacchari, is a widely distributed insect that attacks grasses in different genera including Miscanthus, Saccharum, and Sorghum. The invasive aphid superclone was first discovered in the U.S. attacking grain sorghum in Texas in 2013. Since then, it has been found in at least 25 states including Georgia. We conducted a survey of naturally occurring fungal pathogens of sugarcane aphids on five farms in Georgia, and identified a hypocrealean fungus, Akanthomyces dipterigenus, and two entomophthoralean fungi, Neoconidiobolus spp. From 2018 to 2020, fungal activity differed across farms but at one farm both major fungal species, A. dipterigenus and N. thromboides, were found each of the 3 years infecting sugarcane aphids, attacking adults, both alatae and apterae, and nymphs.

Fungal Cultivars of Higher Attine Ants Promote Escovopsis Chemotropism.

In varied environments, microorganisms search for partners or nutritional resources using chemical signals. Microbes are drawn (chemotaxis) or grow directionally (chemotropism) towards the chemical source, enabling them to establish and maintain symbiosis. The hypocrealean fungi Escovopsis enhance their growth towards the basidiomycete fungus Leucoagaricus gongylophorus, which is cultivated by leaf-cutting attine ants for food. Although directional growth is well documented in this symbiosis, it is unclear whether non-volatile or volatile organic compounds participate in the interaction between cultivar and Escovopsis, and which specific chemical compounds might attract and induce chemotropism. In this study, we examined the growth responses of Escovopsis isolates to non-volatile and volatile organic compounds produced by fungal cultivars of higher attine ants. We also isolated and identified molecules released by the ant-cultivar and assessed the chemotropism of Escovopsis towards them. Our results indicate that the growth of Escovopsis is stimulated in the presence of both non-volatile and volatile compounds from fungal cultivars. We also identified three isomeric diketopiperazines molecules from crude extracts of the ant cultivar, suggesting that these might play a role in Escovopsis chemotropism. Our findings provide insights into the complex chemical interactions that govern the association between Escovopsis and fungal cultivars.

Fungi in ant-plant interactions: a key to enhancing plant nutrient-acquisition strategies.

This article is a Commentary on Gegenbauer et al. (2023), 238: 2210–2223.

Diversity, molecular dating and ancestral characters state reconstruction of entomopathogenic fungi in Hypocreales

Diversity, molecular dating and ancestral characters state reconstruction of entomopathogenic fungi in Hypocreales

Multigene phylogeny, phylogenetic network, and morphological characterizations reveal four new arthropod-associated Simplicillium species and their evolutional relationship.

Simplicillium species are widely distributed and commonly found on various substrates. A minority of species are associated with arthropods. A spider-associated species Simplicillium araneae, and three insect-associated species, Simplicillium coleopterorum, Simplicillium guizhouense, and Simplicillium larvatum, are proposed as novel species based on a multi-locus phylogenetic analysis and morphological characteristics. These Simplicillium species completely fit the nutritional model of Hypocreales fungi and could be used as a model to study their evolutionary relationship. A phylogenetic network analysis based on ITS sequences suggests that a host jump was common among Simplicillium species, and S. araneae may have originally come from an insect host and then jumped to a spider host. However, the evolutionary relationship of S. coleopterorum, S. guizhouense, and S. larvatum was not clear in the phylogenetic network and more sequencing information should be added to the network. In addition, strain CBS 101267 was identified as Simplicillium subtropicum.

Occurrence of entomopathogenic hypocrealean fungi in mosquitoes and their larval habitats in Central Brazil, and activity against Aedes aegypti

Collecting entomopathogenic fungi associated with mosquitoes and studies on their activity against mosquito developmental stages will improve the understanding of their potential as agents to control important mosquito vectors. Twenty-one strains of entomopathogenic fungi affecting mosquitoes in Central Brazil were studied: 7 of Beauveria bassiana, 7 of Metarhizium humberi, 3 of M. anisopliae, 2 of Cordyceps sp. and one each of Akanthomyces saksenae and Simplicillium lamellicola. These fungi were isolated from field-collected mosquito adults (3 strains) or larvae (a single strain); the other 17 strains were isolated from laboratory-reared Aedes aegypti sentinel larvae set out in partially immersed cages placed in diverse small- to middle-sized aquatic mosquito habitats in or close to areas with secondary tropical forest. The frequent recovery of normally soil-borne Metarhizium spp. and B. bassiana from aquatic habitats is notable. Our laboratory findings indicated that M. anisopliae IP 429 and IP 438 and M. humberi IP 421 and IP 478 were highly active against immature stages and, together with M. anisopliae IP 432, also against adults. These strains appear to be the most promising candidates to develop effective control strategies targeting the different developmental stages of A. aegypti, the most important vector of viral diseases in humans in the tropics.

Virulence of native isolates of entomopathogenic fungi (Hypocreales) against the “sweetpotato whitefly” Bemisia tabaci (Hemiptera: Aleyrodidae), including the effects of temperature and fungicides

Hypocrella, Moelleriella and related species in the Hypocreales (Ascomycota, Sordariomycetes) cause epizootics of whiteflies and scale insects in nature. However, studies on their host specificity, virulence, infection cycles, optimal development under laboratory conditions, and compatibility with other control methods, are unexplored for most species. Under laboratory conditions, the virulence of several isolates of field-collected hypocrealean fungi (Hypocrella, Moelleriella, Regiocrella, and Verticillium) was determined on Bemisia tabaci eggs and 4th instar nymphs. In addition to virulence, the effect of temperature and two commercial fungicides on growth rates and germination of the isolates was evaluated. None of the isolates infected the eggs, while M. libera, M. ochracea, and M. turbinata caused high nymphal mortality. Moelleriella libera was the most virulent isolate. At all temperatures, M. libera, Regiocrella sp. (P17H20), and Verticillium cf. pseudohemipterigenum had the highest germination and growth rates. The optimal growth temperature depended on the isolate, but at 23 °C and 25 °C, the probability of spore germination was higher for most isolates. Finally, the fungicides azoxystrobin and chlorothalonil inhibited growth rates and conidial germination at 24 and 48 h of exposure. This research produces vital knowledge on the virulence and infection cycles of poorly studied native species of entomopathogenic fungi. In addition, the results provide information on the optimal temperature for development in laboratory conditions and susceptibility to fungicides, which could contribute to future biological control strategies.

Activity Against Musca domestica of Hypocrealean Fungi Isolated from Culicids in Central Brazil and Formulated in Vermiculite.

Musca domestica L. is a cosmopolitan nuisance of high sanitary importance. Entomopathogenic fungi are innovative and attractive tools for integrated control of the housefly to overcome insufficient levels of control caused by increasing resistance of this pest against chemical insecticides. High virulence of a fungal strain is a prerequisite to develop a mycoinsecticide, and the present study investigated the potential of hypocrealean fungi from the genera Beauveria, Clonostachys, Cordyceps, Akanthomyces, Metarhizium, and Tolypocladium, isolated from mosquitoes in Central Brazil against M. domestica. The highest mortalities (larvae, pupae, and adults) were caused by Metarhizium humberi IP 478 (98%) and IP 421 (90%), Metarhizium anisopliae IP 432 (85%), Beauveria bassiana IP 433 (82%), and Tolypocladium cylindrosporum IP 425 (68%) after a 23-day exposure of initially pre-pupating third instar larvae to conidia mixed with vermiculite. Lethal concentrations to kill 90% of adults of IP 433 and IP 478 were 5 × 107 and 108 conidia g-1 substrate, respectively. Fifty percent of adults were killed within 4 to 5days of exposure initially as pupae close to emergence to substrate treated with conidia of IP 478 or IP 433 at 1.1 × 108 conidia g-1, respectively. The other fungal strains tested were less virulent. The results demonstrate high potentials for conidial preparations in vermiculite of IP 433 and IP 478 as candidates for the biological control of both pre-pupating larvae, pupae, and emerging adults of houseflies.

Brunneosporopsis yunnanensis gen. et sp. nov. and Allocryptovalsa xishuangbanica sp. nov., New Terrestrial Sordariomycetes from Southwest China.

Three fungal taxa were collected on dead branches of wood during fieldwork in Sichuan and Yunnan Provinces, China. The new generic name Brunneosporopsis gen. nov. and species B. yunnanensis sp. nov. are introduced for a novel taxon characterized by globose to subglobose and dark olivacous-brown conidia. Phylogenetic analyses based on combined LSU, SSU and tef1-α loci strongly support the monophyly of this taxon and place it in the subclass Diaporthomycetidae. It could not be assigned to any currently recognized families in the subclass and was, therefore, placed in the Diaporthomycetidae genera incertae sedis. A second taxon represents a new species in Allocryptovalsa based on an analysis of the sequence datasets of ITS and btub loci of the novel, brown-spored sexual morphic species. This taxon is described here as A. xishuangbanica sp. nov. An interesting hypocrealean fungus producing synnemata, Stilbocrea gracilipes, was collected from dead wood of an unknown host from Sichuan Province and is reported here, with asexual morph from both the host and culture as well as LSU, ITS, tef1-α, rpb2 and rpb1 sequence data.

Multi-locus phylogeny unmasks hidden species within the specialised spider-parasitic fungus, Gibellula (Hypocreales, Cordycipitaceae) in Thailand.

Over 80 species of hypocrealean fungi are reported as pathogens of spiders and harvestmen. Among these fungi, the genus Gibellula is highly regarded as a specialised spider-killer that has never been reported to infect other arthropods. While more than 20 species of Gibellula are known, few attempts to identify the infected spiders have been made despite the fact that the host specificity can help identify the fungal species. Here, we morphologically describe and illustrate eight new species of Gibellula and three new records from Thailand of known species along with the multi-gene phylogeny that clearly showed the segregation among the proposed species. Examination of the Gibellula-infected spider hosts identified Oxyopidae, Uloboridae and, for the first time, the ant-mimicking genus Myrmarachne. Taxonomic novelties: New species: Gibellula brevistipitata Kuephadungphan, Tasanathai & Luangsa-ard, G. longicaudata Tasanathai, Kuephadungphan & Luangsa-ard, G. longispora Kuephadungphan & Luangsa-ard, G. nigelii Kuephadungphan, Tasanathai & Luangsa-ard, G. parvula Kuephadungphan, Tasanathai & Luangsa-ard, G. pilosa Kuephadungphan, Tasanathai & Luangsa-ard, G. solita Kuephadungphan, Tasanathai & Luangsa-ard, G. trimorpha Tasanathai, Khonsanit, Kuephadungphan & Luangsa-ard. Citation: Kuephadungphan W, Petcharad B, Tasanathai K, Thanakitpipattana D, Kobmoo N, Khonsanit A, Samson RA, Luangsa-ard JJ (2022). Multi-locus phylogeny unmasks hidden species within the specialised spider-parasitic fungus, Gibellula (Hypocreales, Cordycipitaceae) in Thailand. Studies in Mycology 101: 245-286. doi: 10.3114/sim.2022.101.04.

Discovery of two hypocrealean fungi infecting spotted lanternflies, Lycorma delicatula: Metarhizium pemphigi and a novel species, Ophiocordyceps delicatula

In the eastern United States, populations of the invasive spotted lanternfly, Lycorma delicatula, can be infected by native fungal entomopathogens, including Batkoa major and Beauveria bassiana. In some areas of southeastern Pennsylvania, localized population collapses have been observed in L. delicatula populations to be caused by these pathogens. Two additional fungal pathogens were discovered infecting L. delicatula at low levels, and these were identified as Metarhizium pemphigi and Ophiocordyceps delicatula, a new species that has not been previously described. Therefore, four species of native entomopathogenic fungi have now been documented infecting this abundant, invasive planthopper that is spreading in the United States.

An Update on Trichoderma Mitogenomes: Complete De Novo Mitochondrial Genome of the Fungal Biocontrol Agent Trichoderma harzianum (Hypocreales, Sordariomycetes), an Ex-Neotype Strain CBS 226.95, and Tracing the Evolutionary Divergences of Mitogenomes in Trichoderma.

Members of the genus Trichoderma (Hypocreales), widely used as biofungicides, biofertilizers, and as model fungi for the industrial production of CAZymes, have actively been studied for the applications of their biological functions. Recently, the study of the nuclear genomes of Trichoderma has expanded in the directions of adaptation and evolution to gain a better understanding of their ecological traits. However, Trichoderma’s mitochondria have received much less attention despite mitochondria being the most necessary element for sustaining cell life. In this study, a mitogenome of the fungus Trichoderma harzianum CBS 226.95 was assembled de novo. A 27,632 bp circular DNA molecule was revealed with specific features, such as the intronless of all core PCGs, one homing endonuclease, and a putative overlapping tRNA, on a closer phylogenetic relationship with T. reesei among hypocrealean fungi. Interestingly, the mitogenome of T. harzianum CBS 226.95 was predicted to have evolved earlier than those of other Trichoderma species and also assumed with a selection pressure in the cox3. Considering the bioavailability, both for the ex-neotype strain of the T. harzianum species complex and the most globally representative commercial fungal biocontrol agent, our results on the T. harzianum CBS 226.95 mitogenome provide crucial information which will be helpful criteria in future studies on Trichoderma.

Comparative genomics reveals a core gene toolbox for lifestyle transitions in Hypocreales fungi.

SummaryFungi have evolved diverse lifestyles and adopted pivotal new roles in both natural ecosystems and human environments. However, the molecular mechanisms underlying their adaptation to new lifestyles are obscure. Here, we hypothesize that genes shared across all species with the same lifestyle, but absent in genera with alternative lifestyles, are crucial to that lifestyle. By analysing dozens of species within four genera in a fungal order, with each genus following a different lifestyle, we find that genus‐specific genes are typically few in number. Notably, not all genus‐specific genes appear to derive from de novo birth, with most instead reflecting recurrent loss across the fungi. Importantly, however, a subset of these genus‐specific genes are shared by fungi with the same lifestyle in quite different evolutionary orders, thus supporting the view that some genus‐specific genes are necessary for specific lifestyles. These lifestyle‐specific genes are enriched for key functional classes and often exhibit specialized expression patterns. Genus‐specific selection also contributes to lifestyle transitions, and is especially associated with intensity of pathogenesis. Our study, therefore, suggests that fungal adaptation to new lifestyles often requires just a small number of core genes, with gene turnover and positive selection playing complementary roles.

Fungal Diversity of Selected Habitat Specific Cynorkis Species (Orchidaceae) in the Central Highlands of Madagascar.

About 90% of Cynorkis species are endemic to the biodiversity hotspot of Madagascar. This terrestrial habitat-specific genus received little study for fungal diversity to support conservation. We evaluated the diversity of culturable fungi of 11 species and soil characteristics from six sites spanning a >40 km radius in and along the region’s inselbergs. Peloton-forming fungi were grown in vitro from root/protocorm slices and positively identified using DNA sequencing. The fungal diversity was then correlated with soil pH, NO3-N, P, and K. All species harbored either putative mycorrhizal associates in the Rhizoctonia complex or Hypocreales fungi. Tulasnella Operational Taxonomic Units (OTUs) were most prevalent in all soil types while Serendipita OTUs were found in species inhabiting granite/rock outcrops in moist soil (seepage areas). Most Cynorkis species were present in soil with low NO3-N and P levels with diversity of mycorrhizal fungi inversely correlated to NO3-N levels. Of the different orchid life stages sampled, only one species (Cynorkis fastigiata) yielded putative mycorrhizal fungi from juvenile stages. As diversity of mycorrhizal fungi of Cynorkis spp. was negatively correlated with NO3-N, and majority of the studied taxa were found in soils with low NO3-N and P contents, reintroduction studies must include analysis of N and P in detail. For the first time, we showed that the assemblage of culturable fungi in the roots of habitat-specific species of Cynorkis (Orchidaceae) are intimately tied to specific soil characteristics.

Ascomycetes from the Qilian Mountains, China - Hypocreales.

To investigate fungi from the Qilian Mountains in Gansu Province, ascomycetous specimens were collected and hypocrealean fungi were examined. Eighteen species belonging to six genera in the families Hypocreaceae and Nectriaceae were identified, including 11 species of Hypomyces and Trichoderma in Hypocreaceae and seven species of Nectria, Stylonectria, Thelonectria, and Thyronectria in Nectriaceae. Among them, Stylonectria qilianshanensis and Trichoderma gansuanum are new to science. DNA sequence analyses of combined ACL1, ITS, RPB2, and TEF1 regions confirmed their taxonomic placements. Morphological distinctions between the new species and their close relatives are discussed. Hypomyces tremellicola is reported for the first time in China.

Mycosis is a Disease State Encountered Rarely in Shore Crabs, Carcinus maenas.

There is a paucity of knowledge regarding the diversity and impact(s) of disease-causing fungi in marine animals, especially shellfish. In efforts to address this knowledge gap for the shore crab Carcinus maenas, a year-long disease screen was carried out across two sites in Swansea Bay (Wales, UK) with a view to characterising putative fungal infections. Crabs were sampled between November 2017 and October 2018, and screened systematically for disease signatures using haemolymph (blood) preparations, targeted PCR and tissue histopathology. Strikingly, mycosis was confirmed in ~0.4% of total crabs tested (n = 1191) and restricted to one location only (Mumbles Pier). Clinical infections were observed in four out of four infected crabs. In these animals, the gills and hepatopancreas were congested with fungal morphotypes. In addition, some evidence indicates haemocyte (immune cell) reactivity toward the fungi. Phylogenetic placement of the partial internal transcribed spacer (ITS1) gene regions amplified from three mycotic crabs revealed the causative agent to be related to hypocrealean fungi, thereby representing a novel species.

Complete Mitochondrial Genome of the Fungal Biocontrol Agent Trichoderma atroviride: Genomic Features, Comparative Analysis and Insight Into the Mitochondrial Evolution in Trichoderma.

The improvement of biopesticides for use in the agriculture industry requires an understanding of the biological- and ecological principles underlying their behavior in natural environments. The nuclear genomes of members of the genus Trichoderma, which are representative fungal biocontrol agents, have been actively studied in relation to the unique characteristics of these species as effective producers of CAZymes/secondary metabolites and biopesticides, but their mitochondrial genomes have received much less attention. In this study, the mitochondrial genome of Trichoderma atroviride (Hypocreales, Sordariomycetes), which targets wood-decaying fungal pathogens and has the ability to degrade chemical fungicides, was assembled de novo. A 32,758 bp circular DNA molecule was revealed with specific features, such as a few more protein CDS and trn genes, two homing endonucleases (LAGLIDADG-/GIY-YIG-type), and even a putative overlapping tRNA gene, on a closer phylogenetic relationship with T. gamsii among hypocrealean fungi. Particularly, introns were observed with several footprints likely to be evolutionarily associated with the intron dynamics of the Trichoderma mitochondrial genomes. This study is the first to report the complete de novo mitochondrial genome of T. atroviride, while comparative analyses of Trichoderma mitochondrial genomes were also conducted from the perspective of mitochondrial evolution for the first time.

Virulence of Commercialized Fungal Entomopathogens Against Asian Longhorned Beetle (Coleoptera: Cerambycidae).

Nine strains of five species of entomopathogenic hypocrealean fungi were tested against adults of the Asian longhorned beetle, Anoplophora glabripennis (Motschulsky). These strains have been developed as commercial biopesticide products in the United States, Brazil, South Korea, or the European Union (EU). Metarhizium anisopliae (Metschnikoff) (Hypocreales: Clavicipitaceae) ESALQ E-9 and Metarhizium brunneum (Petch) F52 (formerly M. anisopliae F52) (Hypocreales: Clavicipitaceae) killed 100% of treated beetles with the shortest survival times. Virulence differed among the five strains of Beauveria bassiana (Balsamo) (Hypocreales: Cordycipitaceae) tested, ranging from 0 to 77.3% mortality within 28 d. Two Isaria fumosorosea (Wize, 1904) (Hypocreales: Cordycipitaceae) (formerly Paecilomyces fumosoroseus) strains and the Lecanicillium muscarium (Petch) Zare & Gams (Hypocreales: Cordycipitaceae) strain used in Mycotal were not pathogenic to A. glabripennis adults. Within the entomopathogenic fungi tested, the Metarhizium strains may be the most appropriate for further evaluation.

Activity of Metarhizium brunneum and Beauveria bassiana against early developmental stages of the false codling moth Thaumatotibia leucotreta

This study evaluated the efficacy of two entomopathogenic Hypocrealean fungi, Metarhizium brunneum (laboratory isolate) and Beauveria bassiana (the commercial product Botanigard), for preventative control of the false codling moth, Thaumatotibia leucotreta. The mortality of eggs and first instar larvae was studied in three different assays. First, fungal virulence was examined under optimal laboratory conditions (25 °C, 85% RH) by placing T. leucotreta eggs on conidia-impregnated filter paper. One-day-old eggs and first instar larvae were susceptible to both fungi. In contrast, 5-day-old eggs (advanced embryo development) were susceptible to M. brunneum, but not to B. bassiana. The activity of both fungi against eggs was assessed under two humidity regimes: 85% RH—optimal for fungal germination, and 60% RH—the average humidity in the laboratory. Pieces of parchment paper serving as oviposition surfaces were treated with each of the fungi and introduced to gravid females at different time points after inoculation (0, 2, 7 and 14 days). Although the tested fungal species differed in their virulence to T. leucotreta eggs, both reduced hatching rate under both humidity regimes to 8.3–58.3%, compared to 71.7–83.3% in the control treatments. To evaluate reduction of T. leucotreta infestation of fruit, ‘Ori’ citrus fruit (easy peeler Citrus sinensis) were treated with each of the fungi. Eggs were placed on the fruit peels and the fruit were maintained under room conditions (25 °C, 60% RH). Between 41.7% and 54.1% of fruit in control groups were infested by the T. leucotreta larvae. Treatments with either of the fungi resulted in about 16% infestation of the fruit with larvae, a marked (3.3-fold) reduction.

Non-Entomopathogenic Roles of Entomopathogenic Fungi in Promoting Plant Health and Growth.

Multiple genera of hypocrealean fungi infect and kill a wide variety of arthropod pests. Several formulations based on these soilborne fungi are commercially available as biopesticides for controlling urban, garden, greenhouse, and agricultural pests. These fungi are an important part of integrated pest management strategies to maintain pest control efficacy, reduce the risk of chemical insecticide resistance, and offer environmentally sustainable pest suppression. While the entomopathogenic or pest management role of these fungi is well documented, several studies in the past decade or two have provided insights into their relationship with plants, soil, and plant pathogens, and their additional roles in promoting plant growth and health. This review highlights these endophytic, mycorrhiza-like, and disease-antagonizing roles of entomopathogenic fungi.