an entomopathogenic fungus, infects and leads to high mortality of
larvae, which die soon after the infection of
, usually before the colonization of
owing to competitive inhibition and fruiting body ...formation. Therefore, monitoring
in the
cultivation environment is critical for minimizing the
infection-induced losses. In this study, we initially designed a PCR primer pair (Tar-1F/Tar-1R) through open reading frame prediction and homology comparison of the
genome sequence. This primer pair can detect both
and
. To further distinguish, primers (ITS5-172/ITS4-95) were then designed to selectively amplify the large ribosomal subunit sequences in the
genome. All these primers were applied in combination for detection of
in soil samples. The sensitivity reached a detection limit of 1 × 10
spores/g soil. In addition, these primers can detect the presence of
in dead
larval samples. This newly established rapid detection method provides important information for
control during
cultivation.
Entomopathogenic fungi comprise an ecologically important group of specialized pathogens infecting other fungi, invertebrates, and plants. These fungi are species-rich with high diversity and broad ...distribution worldwide. The majority of entomopathogenic fungi belong to clavicipitoids, which consist of the hypocrealean families, Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae, and Polycephalomycetaceae. The latter is a newly established entomopathogenic family that recently separated from the family Ophiocordycipitaceae to accommodate the genera,
, and
. In recent years, Polycephalomycetaceae has been enriched with parasitic and hyperparasitic fungi. With 16 species spread across China, Ecuador, Japan, and Thailand,
is the most speciose genus in the family. In this study, we expand the number of taxa in the genus by introducing four new
species from China, namely,
, and
. We provide detailed descriptions and illustrations and infer genus-level phylogenies based on a combined 6-loci gene sequence dataset comprising the internal transcribed spacer gene region (ITS), small subunit ribosomal RNA gene region (SSU), large subunit rRNA gene region (LSU), translation elongation factor 1-alpha gene region (TEF-1α), RNA polymerase II largest subunit gene region (RPB1), and RNA polymerase II second largest subunit (RPB2). This study contributes to knowledge with regard to the diversity of
specifically and entomopathogenic
more broadly.
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The two-spotted spider mite (TSSM) Tetranychus urticae, is regarded as one of the most dangerous pests responsible for great losses in most of agricultural crops. It is a persistent ...pest in Saudi Arabia, especially in greenhouses where T. urticae is primarily controlled by chemical pesticides. The main problem for the two-spotted spider mites is its high resistance to pesticides and high fertility rate. In the long term, chemical pesticides cause health problems and economic losses, so it was necessary to search for a safe alternative method for human health and the environment. One of these alternative methods was the selection of plant varieties resistant to the TSSM, in addition to biological control that includes mites or predatory insects and entomopathogenic fungi. The growth, reproduction, and life-table parameters of T. urticae were examined in a laboratory setting with a 16L:8D photoperiod at 28 ± 1 °C and 65 ± 5% RH, in the presence of three major members of Family: Solanaceae tomato, eggplant, and pepper. Pepper was shown to be less conducive to T. urticae growth and reproduction compared to eggplant and tomato. Tetranychus urticae proceeded through all five stages of its life cycle (egg, larva, protonymph, deutonymph, and adult) on tested solanaceous plants, and these plants significantly influenced its growth, reproduction, and Life-table parameters. Additionally, entomopathogenic fungi have been used against insects that have proven highly effective in controlling and reducing the density of two-spotted spider mites. Eight fungi were isolated from 80 insect and mite samples collected from several Saudi Arabia regions. Analysis of the 18S rRNA sequences revealed that the fungal strains identified as Beauveria bassiana, Fusarium sp. F. equiseti, F. oxysporium, Scopulariopsis brevicaulis1, S. brevicaulis2, Aspergillus sclerotiorum, and Penicillium citrinum. The ability of isolated fungi to secrete enzymes degrading the two-spotted spider mite cuticle, namely lipase, protease, and chitinase, were studied.
A newly discovered entomopathogenic fungus Cordyceps javanica (Friedrichs & Bally) Samson & Hywel-Jones (Hypocreales: Cordycipitaceae) strain Wf GA17 was compared with the commercial Cordyceps ...fumosorosea Wize (Hypocreales: Cordycipitaceae) Apopka 97 strain for liquid-culture production, formulation, insecticidal efficacy, and storage stability under laboratory conditions. We compared culture media with carbon:nitrogen (C:N) ratios of 10:1, 30:1, and 50:1 for these two isolates. A third strain, C. fumosorosea strain ARSEF 3581, had previously been optimized for liquid-culture production of blastospores at 10:1 C:N served as an added control. These seven cultures were processed by spray drying with skim milk powder, stored at 25°C to evaluate storage stability, and assayed for insecticidal activity against Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) neonates. Final blastospore concentrations were not significantly different among cultures, ranging from 4.47 to 9.88 × 108 spores/ml. Fungal biomass decreased and final glucose concentrations increased with increasing C:N ratios, indicating better fungal growth with higher nitrogen concentrations. Product yields from the spray dryer (grams per liter culture) increased with increased C:N ratios while spore concentrations decreased, ranging from 2.27 to 7.17 × 109 spores/g. There were no significant differences for insecticidal efficacy among the seven treatments. Spores produced in 10:1 C:N ratio media retained viability longer than spores produced in other media. Cost of ingredients decreased with increasing C:N ratios, such that the 30:1 media may yield the most economical product. The raw material cost needed for application was 1.4× greater for Wf GA17 compared with Apopka 97, a difference that could be erased by optimization of culture conditions.
This research aimed to assess the effectiveness of
Metarhizium robertsii
, diatomaceous earth (Protect-It), and lambda-cyhalothrin, for the long-term protection of stored wheat against three ...destructive grain insect pests,
Rhyzopertha dominica
,
Tribolium castaneum
, and
Trogoderma granarium
. Different treatments were applied, both alone and in paired combinations in laboratory and persistence trials. Single treatments exhibited significantly lower mortality rates in comparison to the paired treatments for all tested insect species. Among the single treatments, lambda-cyhalothrin (Lamb) resulted in significantly higher mortality rates in laboratory trials, followed by diatomaceous earth (DE) and
M. robertsii
(Mr), with insignificant differences between Mr and DE. Evidently, DE exhibited the highest persistence after 120 days of storage for all insect species and initial exposures, although variations in mortality rates among treatments were mostly insignificant. Overall, the most effective treatment in terms of mortality in laboratory, and persistence trials, and progeny production was DE + Lamb, followed by Mr + Lamb, and Mr + DE for all tested insect species. In general, the most susceptible insect species was
R. dominica
, followed by
T. castaneum
and
T. granarium
. This research highlights the effectiveness of
M. robertsii
, DE, and lambda-cyhalothrin in providing prolonged protection of stored wheat against all the examined grain insect species.
The effect of inoculation of strawberry roots by two entomopathogenic fungal isolates,
Metarhizium robertsii
(ESALQ 1622) and
Beauveria bassiana
(ESALQ 3375), on naturally occurring arthropod pests ...and plant diseases was investigated in four commercial strawberry fields during two growing seasons in Brazil. Three locations represented open-field production while strawberries were grown in low tunnels at the fourth location. Population responses of predatory mites to the fungal treatments were also assessed. Plants inoculated by the fungal isolates resulted in significantly fewer
Tetranychus urticae
adults compared to control plants at all four locations. The mean cumulative numbers ± SE of
T. urticae
per leaflet were:
M. robertsii
(225.6 ± 59.32),
B. bassiana
(206.5 ± 51.48) and control (534.1 ± 115.55) at the three open-field locations, while at the location with tunnels numbers were:
M. robertsii
(79.7 ± 10.02),
B. bassiana
(107.7 ± 26.85) and control (207.4 ± 49.90). Plants treated with
B. bassiana
had 50% fewer leaves damaged by Coleoptera, while there were no effects on numbers of whiteflies and thrips. Further, lower proportions of leaflets with symptoms of the foliar plant pathogenic fungi
Mycosphaerella fragariae
and
Pestalotia longisetula
were observed in the
M. robertsii
(4.6% and 1.3%)- and
B. bassiana
(6.1% and 1.3%)-treated plots compared to control plots (9.8% and 3.7%). No effect was seen on numbers of naturally occurring predatory mites. Our results suggest that both isolates tested may be used as root inoculants of strawberries to protect against foliar pests, particularly spider mites, and also against foliar plant pathogenic fungi without harming naturally occurring and beneficial predatory mites.
Chitin is one of the major components of the fungal cell wall and contributes to the mechanical strength and shape of the fungal cell. Zn(II)2Cys6 transcription factors are unique to the fungal ...kingdom and have a variety of functions in some fungi. However, the mechanisms by which Zn(II)2Cys6 proteins affect entomopathogenic fungi are largely unknown. Here, we characterized the Zn(II)2Cys6 transcription factor BbTpc1 in the insect pathogenic fungus Beauveria bassiana. Disruption of BbTpc1 resulted in a distinct changes in vegetative growth and septation patterns, and a significant decrease in conidia and blastospore yield. The ΔBbTpc1 mutant displayed impaired resistance to chemical stresses and heat shock and attenuated virulence in topical and intrahemocoel injection assays. Importantly, the ΔBbTpc1 mutant had an abnormal cell wall with altered wall thickness and chitin synthesis, which were accompanied by transcriptional repression of the chitin synthetase family genes. In addition, comparative transcriptomics revealed that deletion of BbTpc1 altered fungal asexual reproduction via different genetic pathways. These data revealed that BbTpc1 regulates fungal development, chitin synthesis and biological control potential in B. bassiana.
•Plant-derived lipids support the fungus Metarhizium robertsii to develop symbiotic relationship with plants.•Fatty acid binding protein genes acquired via horizontal gene transfer are involved in ...utilization of plant-derived lipids.•Utilization of plant-derived sugars and lipids are coupled during colonization of rhizoplane and rhizosphere by M. robertsii.
Plant-derived sugars and lipids are key nutritional sources for plant associated fungi. However, the relationship between utilization of host-derived sugars and lipids during development of the symbiotic association remains unknown. Here we show that the fungus Metarhizium robertsii also needs plant-derived lipids to develop symbiotic relationship with plants. The fatty acid binding proteins FABP1 and FABP2 are important for utilization of plant-derived lipids as the deletion of Fabp1 and Fabp2 significantly reduced the ability of M. robertsii to colonize rhizoplane and rhizosphere of maize and Arabidopsis thaliana. Deleting Fabp1 and Fabp2 increased sugar utilization by upregulating six sugar transporters, and this explains why deleting the monosaccharide transporter gene Mst1, which plays an important role in utilization of plant-derived sugars, had no impact on the ability of the double-gene deletion mutant ΔFabp1::ΔFabp2 to colonize plant roots. FABP1 and FABP2 were also found in other plant-associated Metarhizium species, and they were highly expressed in the medium using the tomato root exudate as the sole carbon and nitrogen source, suggesting that they could be also important for these species to develop symbiotic relationship with plants. In conclusion, we discovered that utilization of plant-derived sugars and lipids are coupled during colonization of rhizoplane and rhizosphere by M. robertsii.
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•Entomophthorales overwinter as fungus-killed Rhopalosiphum padi on Prunus padus.•Zoophthora cf. aphidis formed resting spores; Entomophthora planchoniana modified hyphal bodies.•A ...negative correlation between eggs and fungus-killed cadavers per branch was found.•Numbers of eggs and cadavers varied greatly between years and location.•Prunus padus may act as a reservoir for fungi infecting aphids in cereals.
In Scandinavia, the bird cherry-oat aphid Rhopalosiphum padi overwinter as eggs on the bird cherry tree Prunus padus. Branches of P. padus were collected at the late February / early March from 17 locations in Norway over a three-year period. We found 3599 overwintering aphid eggs, 59.5% of which were dead. Further, a total of 879 overwintering fungus-killed cadavers were observed. These cadavers were found close to bud axils, where overwintering eggs were also usually attached. Cadavers were infected with either Zoophthora cf. aphidis or Entomophthora planchoniana. All the fungal-killed cadavers were filled with overwintering structures of Z. cf. aphidis (as resting spores) or E. planchoniana (as modified hyphal bodies). We found a significant negative correlation between eggs and cadavers per branch. However, both numbers of eggs and cadavers varied greatly between years and among tree locations. This is the first report of E. planchoniana overwintering in R. padi cadavers as modified hyphal bodies. We discuss whether P. padus may act as an inoculum reservoir for fungi infecting aphids in cereals in spring.
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•Multigene phylogeny was employed to identify Brazilian anamorphic Cordyceps species.•Morphological features are generally inadequate for species delimitations in this group.•C. ...amoenerosea, C. fumosorosea, C. javanica and C. tenuipes were confirmed.•C. brevistroma and C. spegazzinii are reported in Brazil for the first time.•Four other taxa could not be confidently identified to species level.
A total of 53 anamorphic strains of Brazilian Cordyceps species currently maintained in a government-owned culture collection, were reassessed for diversity and species identity using multi-loci-based phylogenetic methods. The strains used in this study were originally obtained from soil samples or were isolated from insects of the orders Hemiptera, Lepidoptera, Coleoptera and Diptera, mostly from agricultural sites. A Bayesian phylogenetic tree was constructed based on a concatenation of five loci (ITS, LSU, RPB1, RPB2 and TEF). In a few cases of ambiguity, morphological traits were also considered for species delimitations. Considerable variability within the set of strains was detected and six Cordyceps species were identified: C. amoenerosea, C. fumosorosea, C. javanica, C. tenuipes and, for the first time, C. brevistroma and C. spegazzinii are reported in Brazil. Four other taxonomically equivocal groups, closely related to other known taxa (C. amoenerosea, C. cateniannulata, C. polyarthra and C. spegazzinii), were also recognized, although further studies will be required to confirm their identifications or their descriptions as new species. Cordyceps javanica was the most common species in our dataset, originally isolated from soil and several different insect orders, and includes 17 strains from the whitefly, Bemisia tabaci. Interestingly, strains previously identified as C. fumosorosea based on morphology and growth characteristics, were shown to be C. javanica, including the active ingredients of some commercial mycoinsecticides. Cordyceps farinosa, usually mentioned in the literature as occurring in Brazil, was not found in our study. Since most strains were from insect crop pests, further studies with hosts from non-agricultural settings or from environmental samples would be advisable for a deeper understanding of the occurrence of anamorphic Cordyceps in Brazil.
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