Entomopathogenic Fungus Cordyceps Research Articles

Pyricularia oryzae enhances Streptomyces griseus growth via non-volatile alkaline metabolites.

Chemical compounds that affect microbial interactions have attracted wide interest. In this study, Streptomyces griseus showed enhanced growth when cocultured with the rice blast fungus Pyricularia oryzae on potato dextrose agar (PDA) medium. An improvement in S. griseus growth was observed before contact with P. oryzae, and no growth-promoting effect was observed when the growth medium between the two microorganisms was separated. These results suggested that the chemicals produced by P. oryzae diffused through the medium and were not volatile. A PDA plate supplemented with phenol red showed that the pH of the area surrounding P. oryzae increased. The area with increased pH promoted S. griseus growth, suggesting that the alkaline compounds produced by P. oryzae were involved in this growth stimulation. In contrast, coculture with the soilborne plant pathogen Fusarium oxysporum and entomopathogenic fungus Cordyceps tenuipes did not promote S. griseus growth. Furthermore, DL-α-Difluoromethylornithine, a polyamine biosynthesis inhibitor, prevented the increase in pH and growth promotion of S. griseus by P. oryzae. These results indicated that P. oryzae increased pH by producing a polyamine.

Identification of a novel member of the genus Laulavirus (family Phenuiviridae) from the entomopathogenic ascomycete fungus Cordyceps javanica.

The virus family Phenuiviridae (order Hareavirales, comprising segmented negative-sense single stranded RNA viruses) has highly diverse members that are known to infect animals, plants, protozoans, and fungi. In this study, we identified a novel phenuivirus infecting a strain of the entomopathogenic fungus Cordyceps javanica isolated from a small brown plant hopper (Laodelphax striatellus), and this virus was tentatively named "Cordyceps javanica negative-strand RNA virus 1" (CjNRSV1). The CjNRSV1 genome consists of three negative-sense single stranded RNA segments (RNA1-3) with lengths of 7252, 2401, and 1117 nt, respectively. The 3'- and 5'-terminal regions of the RNA1, 2, and 3 segments have identical sequences, and the termini of the RNA segments are complementary to each other, reflecting a common characteristic of viruses in the order Hareavirales. RNA1 encodes a large protein (∼274 kDa) containing a conserved domain for the bunyavirus RNA-dependent RNA polymerase (RdRP) superfamily, with 57-80% identity to the RdRP encoded by phenuiviruses in the genus Laulavirus. RNA2 encodes a protein (∼79 kDa) showing sequence similarity (47-63% identity) to the movement protein (MP, a plant viral cell-to-cell movement protein)-like protein (MP-L) encoded by RNA2 of laulaviruses. RNA3 encodes a protein (∼28 kDa) with a conserved domain of the phenuivirid nucleocapsid protein superfamily. Phylogenetic analysis using the RdRPs of various phenuiviruses and other unclassified phenuiviruses showed CjNRSV1 to be grouped with established members of the genus Laulavirus. Our results suggest that CjNRSV1 is a novel fungus-infecting member of the genus Laulavirus in the family Phenuiviridae.

Improved control of Trialeurodes vaporariorum using mixture combinations of entomopathogenic fungi and the chemical insecticide spiromesifen

Greenhouse whitefly (Trialeurodes vaporariorum) is a major global pest, causing direct damage to plants and transmitting viral plant diseases. Management of T. vaporariorum is problematic because of widespread pesticide resistance, and many greenhouse growers rely on biological control agents to regulate T. vaporariorum populations. However, these are often slow and vary in efficacy, leading to subsequent application of chemical insecticides when pest populations exceed threshold levels. Combining chemical and biological pesticides has great potential but can result in different outcomes, from positive to negative interactions. In this study, we evaluated co-applications of the entomopathogenic fungi (EPF) Beauveria bassiana and Cordyceps farinosa and the chemical insecticide spiromesifen in laboratory bioassays. Complex interactions between the EPFs and insecticide were described using an ecotoxicological mixtures model, the MixTox analysis. Depending on the EPF and chemical concentrations applied, mixtures resulted in additivity, synergism, or antagonism in terms of total whitefly mortality. Combinations of B. bassiana and spiromesifen, compared to single treatments, increased the rate of kill by 5 days. Results indicate the potential for combined applications of EPF and spiromesifen as an effective integrated pest management strategy and demonstrate the applicability of the MixTox model to describe complex mixture interactions.

Development of SCAR Markers for Genetic Authentication of Metarhizium acridum.

In this study, molecular typing using Randomly Amplified Polymorphic DNA (RAPD-PCR) was conducted on 16 original isolates of Metarhizium acridum obtained from locusts (Schistocerca piceifrons ssp. piceifrons.) in Mexico (MX). The analysis included reference strains of the genus Metarhizium sourced from various geographical regions. The isolates were identified by phenotypic (macro and micromorphology) and genotypic methods (RAPD-PCR and Amplified Fragment Length Polymorphisms (AFLP), through a multidimensional analysis of principal coordinates (PCoA) and a minimum spanning network (MST). Subsequently, Sequences-Characterized Amplified Region (SCAR) markers were developed for the molecular detection of M. acridum, these markers were chosen from polymorphic patterns obtained with 14 primers via RAPD-PCR. Phenotypic and genotypic characterization identified the MX isolates as M. acridum. Of all the polymorphic patterns obtained, only OPA04 and OPA05 were chosen, which presented species-specific bands for M. acridum, and further utilized to create SCAR markers through cloning and sequencing of the specific bands. The specificity of these two markers was confirmed via Southern hybridization. The SCAR markers (Ma-160OPA-05 and Ma-151OPA-04) exhibit remarkable sensitivity, detecting down to less than 0.1 ng, as well as high specificity, as evidenced by their inability to cross-amplify or generate amplification with DNAs from other strains of Metarhizium (as Metarhizium anisopliae) or different genera of entomopathogenic fungi (Cordyceps fumosorosea and Akanthomyces lecanii). These SCAR markers yield readily detectable results, showcasing high reproducibility. They serve as a valuable tool, especially in field applications.

DC-electrical penetration graph waveforms for Dalbulus maidis (Hemiptera: Cicadellidae) and the effects of entomopathogenic fungi on its probing behavior

The corn leafhopper Dalbulus maidis is an important transmitter of viruses and bacteria to maize plants. Data on the stylet probing and ingestion behavior of D. maidis, gathered using the DC-electrical penetration graph (DC-EPG) technique, are limited. The increasing prevalence of this pest and the diseases that it transmits in maize crops heighten the importance of studying how control tools affect the probing behavior of these insects, to reduce or prevent the chances of pathogen transmission and the damage from feeding. Our study recorded stylet activities of D. maidis, using a DC-EPG system and compared the appearances of waveforms with those from published AC and AC-DC EPG studies. All types of systems produced similar waveform appearances; therefore, we used the waveform nomenclature previously published. We also determined the effects of the entomopathogenic fungus Cordyceps javanica on the probing behavior of D. maidis at different time points after the fungus was applied by spraying. Forty-eight hours after the insects were sprayed, the effects were pronounced, with significant disruption of the stylet activities in phloem and non-phloem phases. Our study indicated that this commercial microbiological product, with the active ingredient C. javanica, can alter the probing behavior of D. maidis and may be helpful in management of the vector.

Antitumor activity of cordycepin in murine malignant tumor cell line: An in vitro and in silico study

Cordycepin, also known as 3′-deoxyadenosine, is a substance similar to adenosine found in the entomopathogenic fungus Cordyceps militaris. This fungus is recognized for its remarkable therapeutic properties. This study seeks to investigate the antitumor activity of cordycepin in Dalton's lymphoma (DL) malignant cancer cell line using cytotoxicity and apoptosis as key parameters. Molecular docking (MD) and molecular dynamic simulation (MDS) were used to further validate the wet lab findings. Cytotoxicity study revealed that cordycepin induced significant cytotoxicity (P ≤ 0.05) in the DL cell line after 24 h of treatment. The efficacy of cordycepin as an apoptotic agent was substantiated by the highly promising results obtained through the apoptotic assay, utilizing the acridine orange/ethidium bromide (AO/EB) dual staining method. Cordycepin interacted with the active sites of antiapoptotic target proteins (BCL-XL, SURVIVIN, BCL-2, BCL-B, HSP90, and IAP1), as demonstrated by MD study that corroborated the findings of cytotoxicity and apoptotic assays. The comparative examination of docking outcomes indicated that cordycepin exhibits higher affinity for HSP90 in comparison to other targets. Molecular dynamics simulation for 120 ns further validated the findings of MD as cordycepin remained bound with HSP90 throughout the entire simulation time (120 ns) with favourable binding energy.

Auto-dissemination of Cordyceps fumosorosea amongst adult females of the two-spotted spider mite.

Tetranychus urticae is an important pest worldwide. The auto-dissemination of spores of entomopathogenic fungi from an infected individual to conspecifics may be important for controlling pests that can build high populations. The current study was carried out to determine the auto-dissemination of the entomopathogenic fungus Cordyceps fumosorosea strain PFs-1 (Priority®) between T. urticae females. The study consisted of four experiments. First, the efficacy of entomopathogenic fungus bioassays was assessed in Petri dishes (experiment 1) and on potted bean plants (experiment 2). In the auto-dissemination trials (experiments 3 and 4, in Petri dishes and on potted plants, respectively), contaminated adult females (1-5) were released among uncontaminated females (10 individuals). All experiments were carried out separately, and observations were made on days 3, 5, and 7. In exp. 1, the control was different from Priority on all observation days. In exp. 2, the average number of surviving individuals in the control was significantly higher than in the Priority treatment. In the auto-dissemination experiments, as the number of contaminated individuals increased, the mortality rate of uncontaminated individuals also increased, in exp. 3 (Petri dishes) on all observation days, and in exp. 4 (potted plants) only on days 5 and 7. The median lethal time (LT50)decreased as the number of individuals contaminated with Priority increased in both Petri dish and pot trials. Consequently, the effectiveness of biological control may increase with the occurrence of indirect contamination from infected to uncontaminated individuals.

Bifusicoumarins A-D: Cytotoxic 3S-dihydroisocoumarins from the entomopathogenic fungus Cordyceps bifusispora (NBRC 108997)

Cordyceps is a genus of ascomycete fungi with some of them being edible and/or having a long tradition in Chinese medicine. The chemical characterization of a solvent extract of the entomopathogenic fungus Cordyceps bifusispora afforded four undescribed coumarins, bifusicoumarin A-D (1–4), along with previously reported metabolites (5–8). Structural elucidation was performed via NMR, UV and HRMS analyses, X-ray single crystal diffraction and experimental ECD. A high throughput resazurin reduction assay, that measures cell viability, indicated that 5 has a IC50 between 1 and 15 μM for several assayed tumor lines. Moreover, a protein-interaction network indicated that C. bifusispora is a promising source of additional antitumor metabolites based on SwissTargetPrediction software predictions.

Antimicrobial metabolite of Cordyceps tenuipes targeting MurE ligase and histidine kinase via in silico study

Cordyceps spp. are widely healthy foods around the world with several traditional uses and bio-functionalities. The chemical characterization of ethyl acetate-soluble extract of the entomopathogenic fungus Cordyceps tenuipes NBRC 111,630 afforded two new metabolites with 1,6-dioxaspiro[4.4]nonane motif, tenuipesone A (1) and tenuipesone B (2), along with four well-known metabolites (3-6). The elucidation of the chemical structures was carried out via extensive spectroscopic experiments including FTIR, HRMS, 1D-NMR, and 2D-NMR. Theprobable biosynthetic pathway of 1 and 2 was hypothesized. From the six isolates, beauvericin (6) exhibited antimicrobial activity against Bacillus subtilis and Staphylococcus aureus with respective MIC of 6.25 and 12.5μM. Docking results exhibited that beauvericin (6) has significant binding affinities against MurE and HK proteins with ΔG = - 8.021 and - 8.585kcal/mol, respectively. KEY POINTS: • Six compounds, including two new, were isolated from the entomopathogenic fungus Cordyceps tenuipes. • Plausible biosynthetic pathway of compounds 1, 2, 4, and 5 was hypothesized. • Beauvericin (6) exhibited significant antimicrobial activity against Bacillus subtilis and Staphylococcus aureus alongside binding affinities against MurE and HK proteins in MOE study.

Toxicity of crude toxin protein produced by Cordyceps fumosorosea IF-1106 against Myzus persicae (Sulze)

The entomopathogenic fungus Cordyceps fumosorosea IF-1106 is a potential biocontrol agent with high pathogenicity to the aphid Myzus persicae. We extracted the crude toxin protein from a liquid culture broth of an isolated C. fumosorosea strain using the ammonium sulfate precipitation method, and its toxicity to Myzus persicae was measured by injection, oral exposure, and topical exposure. The crude toxin protein of C. fumosorosea had insecticidal activity against M. persicae. Body cavity injection and oral exposure had significantly higher insecticidal activity against adults than contact sprays. The highest cumulative corrected mortality of adults after injection was 81.85 ± 13.45 %, and the highest cumulative corrected mortality of adults after ingestion was 85.45 ± 11.88 %. The proportion of plasmatocytes in adult blood lymphocytes reached the highest at 3 days after injection and feeding, and the proportion of granulocytes was the highest at 2 days after injection and feeding. These data confirmed the toxicity of the crude toxin protein of C. fumosorosea toxin to M. persicae and helped clarify the pathogenic mechanism of the strain. Population management of M. persicae may be possible by using a natural toxic compound produced by C. fumosorosea that is selective to this pest species.

Bt protein hasten entomopathogenic fungi-induced death of nontarget pest whitefly by suppressing protective symbionts

The risk assessment of Bacillus thuringiensis (Bt) crops on nontarget pests has received much attention. Despite the knowledge of various beneficial bacterial symbionts in pests, whether Bt proteins affect these symbionts and subsequently alter the pest's ecology remains largely unknown. The whitefly Bemisia tabaci is one of the most serious nontarget pests in Bt cotton. Here, we explored the Bt Cry1Ac protein-induced changes in whitefly symbiont abundance and the subsequent effects on whitefly response against a naturally prevalent entomopathogenic fungus Cordyceps javanica. The obligate symbiont ‘Candidatus Portiera aleyrodidarum’ (hereafter P. aleyrodidarum) as well as facultative symbionts ‘Candidatus Hamiltonella defensa’ (hereafter H. defensa), ‘Candidatus Cardinium hertigii’ (hereafter C. hertigii) and ‘Candidatus Rickettsia bellii’ (hereafter R. bellii) dominate the microbial community of whiteflies. The Bt exposure had no effects on H. defensa infected (H) and H. defensa-C. hertigii doubly infected (HC) whiteflies, but decreased the total copy number of symbionts as well as the R. bellii proportion in H. defensa-C. hertigii- R. bellii triply infected whiteflies (HCR). C. javanica caused whitefly adults 100 % mortality within 8 days. Without Bt protein exposure, HCR whiteflies survived significantly longer than H and HC whiteflies sprayed by C. javanica, suggesting that R. bellii confers protection. However, in Bt-exposed groups, C. javanica generated synchronous death of H, HC and HCR whiteflies. Specifically, in H and HC whiteflies, Bt protein-exposure showed no significant difference in progress of death caused by C. javanica. But in HCR whiteflies, Bt exposure hastened death induced by C. javanica, suppressing the R. bellii-conferred protection. This is the first report revealing that Bt protein altered symbiont community conferred adverse effects on nontarget pests, providing a new perspective for Bt risk assessment and biocontrol strategies of nontarget pests.

Entomopathogenic fungus disrupts the phloem-probing behavior of Diaphorina citri and may be an important biological control tool in citrus

Citrus is among the most important fruit crops worldwide; however, numerous pests and diseases affect the orchards, increasing production costs. The psyllid Diaphorina citri, is a vector of the phloem-limited bacteria ‘Candidatus Liberibacter spp.’, the causal agent of Huanglongbing (HLB) disease. The lack of a cure for HLB requires management of the vector, mainly by intensive use of chemical insecticides, leading to the selection of resistant populations. Our study determined the effects of the entomopathogenic fungus Cordyceps fumosorosea on the probing behavior of D. citri at different time points after the fungus was applied by spraying. The electrical penetration graph technique was used to monitor the stylet activities of D. citri after application of the microbiological product. The effects were more pronounced between 30 and 96 h after the insects were sprayed, with significant disruption of the stylet activities related to the phloem and directly associated with the transmission of HLB. Our study indicated that the microbiological product Challenger®, with the active ingredient C. fumosorosea fungus, can significantly change the probing behavior of D. citri, may be helpful in more-sustainable management of the vector, and can be used to reduce the spread of HLB.

Molecular characterization of a novel alternavirus infecting the entomopathogenic fungus Cordyceps chanhua.

In this study, a novel double-stranded (ds) RNA mycovirus, named Cordyceps chanhua alternavirus 1 (CcAV1), was detected in the entomogenous fungus Cordyceps chanhua in China and characterized. The complete genome of CcAV1 is composed of three dsRNA segments: dsRNA 1 (3,512 bp), dsRNA 2 (2,655 bp), and dsRNA 3 (2,415 bp). Each of the three dsRNAs possesses a single open reading frame (ORF). dsRNA 1 encodes a putative RNA-dependent RNA polymerase (RdRp), and dsRNA 2 and dsRNA 3 encode hypothetical protein 1 (HP 1) and hypothetical protein 2 (HP 2), respectively. The predicted amino acid sequences of the putative RdRp, HP 1, and HP 2 had the highest identity of 66.99%, 49.30%, and 56.91%, respectively, to those of Aspergillus foetidus dsRNA mycovirus. A maximum-likelihood phylogenetic tree based on RdRp amino acid sequences showed that CcAV1 clustered with members of the proposed family "Alternaviridae". Hence, we propose that Cordyceps chanhua alternavirus 1 is a novel member of the proposed family "Alternaviridae".

Comparing Production and Efficacy of Cordyceps javanica With Cordyceps fumosorosea.

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 25oC 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.

Field persistence and pathogenicity of Cordyceps fumosorosea for management of Diaphorina citri

ABSTRACT The commercial fungal biopesticide product PFR-97 20% WDG, based on the entomopathogenic fungus Cordyceps fumosorosea (Cfr), is registered in the U.S.A. for foliar sprays on citrus. The persistence of Cfr suspensions (107 blastospores/mL) applied to grapefruit plants was evaluated under summer field conditions in southern Florida. Concurrently leaf disk bioassays were conducted to evaluate the pathogenicity of fungal residues on field-aged leaves against the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama. The results demonstrated that Cfr persisted for at least 35 days under our field conditions and caused up to 40% ACP mortality when used in bioassays. There was a general trend of higher colony forming unit counts and a correspondingly higher ACP mortality with the residue age of field-treated leaves. We hypothesise that under certain environmental conditions the Cfr propagule residues will germinate, produce hyphae, phialides and develop conidia on the leaf phylloplane over time. During this study, an adult citrus leafminer Phyllocnistis citrella (Stainton), another important pest of citrus was observed mycosed with Cfr hyphae and subsequently attached to the citrus leaf. Our study shows that Cfr can potentially provide supplemental longer-term management for ACP. Abiotic and biotic factors that may affect the field activity of Cfr and other mycoinsecticides over time are discussed.

Field Efficacy of Cordyceps javanica, White Oil and Spinetoram for the Management of the Asian Citrus Psyllid, Diaphorina citri.

Simple SummaryCitrus greening is devastating the citrus industry in Florida, and conventional synthetic pesticide applications are rapidly becoming unsustainable for the control of the vector, the Asian citrus psyllid (AsCP), Diaphorina citri. Laboratory experiments indicate that the entomopathogenic fungus Cordyceps javanica, alone and in combination with horticultural oils, may offer a more sustainable strategy to manage AsCP. The field study in this paper indicated that C. javanica alone, C. javanica mixed with white oil, and spinetoram (the chemical standard) mixed with white oil significantly suppressed AsCP populations by 60–90% and 61–83% 7 and 14 days after treatment, respectively, in 2018, compared to white oil only and the untreated control treatments. Only spinetoram + oil suppressed AsCP 100% up to 7 days after treatment in 2019. AsCP’s natural enemies, mostly lady beetles, were observed on fungal-treated citrus trees and the untreated control. Overall, C. javanica was as effective in suppressing AsCP adults as spinetoram for up to 14 days in field conditions and was compatible with the psyllids’ natural enemies. The use of C. javanica in citrus-integrated pest management is suggested.Citrus greening disease is devastating the citrus industry in Florida, and the conventional synthetic pesticide applications used to control the vector, the Asian citrus psyllid (AsCP), Diaphorina citri, are rapidly becoming unsustainable. Various laboratory experiments indicate that the entomopathogenic fungus Cordyceps javanica, alone and in combination with horticultural oils, may offer a more sustainable strategy for the management of AsCP. Field studies conducted in 2018 and 2019 in mature citrus indicated that C. javanica alone, C. javanica mixed with white oil, and the chemical standard spinetoram mixed with white oil significantly suppressed AsCP adult populations by 61–83% up to 14 days after treatment in 2018, although colony-forming units of C. javanica were still present on the leaves 21 days after treatment (DAT). Only spinetoram + oil significantly suppressed AsCP, by 100%, up to 7 DAT in 2019. Natural enemies of AsCP, including lady beetles, lacewing larvae and the parasitoid Tamarixia radiata, were observed in the fungal treatments and the untreated control. The AsCP suppression by C. javanica and its compatibility with beneficial organisms suggest the potential use of this entomopathogenic fungus in citrus-integrated pest management.

Efficacy of entomophatogenic fungi Beauveria bassiana and Cordyceps sp against Crocidolomia pavonana (FABR.) Lepidoptera: Pyralidae) as possible pest management strategy on pepper multi-cropping plantation

Black and white pepper have been considered as the most important perennial crops in some regions of Indonesia yet it took about 3-5 years before the plant start producing. One of the possible approaches to maintain the income of the farmer in the perennial plantation is multi-cropping with high-value annual crops, such as vegetables. However, many pests of vegetables already showed resistance to the common synthetic pesticide applied on the black pepper plant. Thus, it is necessary to apply another strategy for pest control, by applying entomopathogenic fungi. In this study, bioassays and field efficacy of two local isolates of entomopathogenic fungi (Beauveria bassiana and Cordyceps sp.) against one of the most insecticide-resistant pests, larvae of Crocidolomia pavonana (Fabr.) was conducted. In this study, exposure to 1 x 104 conidia mL−1 to 1 x 107 conidia mL−1 resulted in mean larval mortalities from 23.33 to 100% in two days. The probit analysis showed that the LC50 of B. bassiana was estimated at 4.26 x 104 conidia mL−1 which significantly more toxic than Cordyceps sp. (1.57 x 105 conidia mL−1) although the lethal time was similar. From the field tests, cabbage was sprayed with suspension contained 107 conidia mL−1. Means percent of mortality for all treatments was more than 75% and significantly higher (P < 0.05) than the control (tween 80). Conidia of Cordyceps sp. in palm oil cooking oil offered the most promising result against the C. pavonana.

Improving Hypoxia Adaption Causes Distinct Effects on Growth and Bioactive Compounds Synthesis in an Entomopathogenic Fungus Cordyceps militaris

Cordyceps militaris is an entomopathogenic fungus producing a variety of bioactive compounds. To meet the huge demand for medicinal and edible products, industrialized fermentation of mycelia and cultivation of stromata have been widely developed in China. The content of bioactive metabolites of C. militaris, such as cordycepin, is higher when cultivated on silkworm pupae than on rice or in broth. However, compared with other cultivation methods, C. militaris grows more slowly and accumulates less biomass. The hypoxic environment in pupa hemocoel is one of environmental factor which is not existed in other cultivation methods. It is suggested that hypoxia plays an important role on the growth and the synthesis of bioactive compounds in C. militaris. Here, we demonstrated that the distinct effects on the growth and synthesis of bioactive compounds employing different strategies of improving hypoxia adaption. The introduction of Vitreoscilla hemoglobin enhanced growth, biomass accumulation, and crude polysaccharides content of C. militaris. However, cordycepin production was decreased to 9–15% of the control group. Meanwhile, the yield of adenosine was increased significantly. Nonetheless, when the predicted bHLH transcription factor of sterol regulatory element binding proteins (SREBPs) was overexpressed in C. militaris to improve the hypoxia adaption of fungal cells, cordycepin content was significantly increased more than two-fold. These findings reveal the role of SREBPs on growth and bioactive compounds synthesis. And it also provides a scientific basis for rationally engineering strains and optimization strategies of air supply in cultivation and fermentation.

Host-pathogen interaction between Asian citrus psyllid and entomopathogenic fungus (Cordyceps fumosorosea) is regulated by modulations in gene expression, enzymatic activity and HLB-bacterial population of the host

The host-pathogen interaction has been explored by several investigations, but the impact of fungal pathogens against insect resistance is still ambiguous. Therefore, we assessed the enzymatic activity and defense-related gene expression of Asian citrus psyllid (ACP) nymphal and adult populations on Huanglongbing-diseased citrus plants under the attack of Cordyceps fumosorosea. Overall, five enzymes viz. superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), glutathione S-transferase (GST), carboxylesterase (CarE), and four genes, namely SOD, 16S, CYP4C68, CYP4BD1, were selected for respective observations from ACP populations. Enzymatic activity of four enzymes (SOD, POD, GST, CarE) was significantly decreased after 5-days post-treatment (dpt) and 3-dpt fungal exposure in fungal treated ACP adult and nymphal populations, respectively, whereas the activity of CAT was boosted substantially post-treatment time schedule. Besides, we recorded drastic fluctuations in the expression of CYP4 genes among fungal treated ACP populations. After 24 hours post-treatment (hpt), expression of both CYP4 genes was boosted in fungal treated populations than controlled populations (adult and nymph). After 3-dpt, however, the expression of CYP4 genes was declined in the given populations. Likewise, fungal attack deteriorated the resistance of adult and nymphal of ACP population, as SOD expression was down-regulated in fungal-treated adult and nymphs after 5-dpt and 3-dpt exposure, respectively. Moreover, bacterial expression via the 16S gene was significantly increased in fungal-treated adult and nymphal ACP populations with increasing post-treatment time. Overall, our data illustrate that the fungal application disrupted the insect defense system. The expression of these genes and enzymes suppress the immune function of adult and nymphal ACP populations. As it is reported first time that the applications of C. fumosorosea against ACP reduce insect resistance by interfering with the CYP4 and SOD system. Therefore, we propose new strategies to discover the role of certain toxic compounds from fungus, which can reduce insect resistance, focusing on resistance-related genes and enzymes.

Phototoxicity of Ultraviolet-A against the Whitefly Bemisia tabaci and Its Compatibility with an Entomopathogenic Fungus and Whitefly Parasitoid.

Ultraviolet (UV) radiation significantly affects insect life and, as a result, has been widely used to control different invertebrate pests. The current results demonstrate that when Bemisia tabaci first instar nymphs are exposed to UV-A light for 12, 24, 48, and 72 h, their developmental and biological parameters are negatively affected by UV-A exposure; the effect increased with an increase in exposure time. We hypothesized that UV-A light is compatible with other biological control agents. Results showed that when the entomopathogenic fungus Cordyceps fumosorosea was applied to third instar nymphs of B. tabaci previously exposed to UV-A light, the LC50 was 3.4% lower after 72 h of exposure to UV-A light compared to the control. However, when the fungus was exposed to UV-A light, its virulence decreased with an increase in UV-A exposure time. The parasitism rate of Encarsia formosa against 24 h UV-A-exposed third instar nymphs of B. tabaci increased while the adult emergence from parasitized nymphs was not affected after UV-A light exposure. Parasitism rate was significantly reduced however following E. formosa exposure to UV-A light; but again, adult emergence was not affected from parasitized nymphs. The percentage mortality of E. formosa increased with increasing exposure time to UV-A light. The enzyme activity of SOD, CAT, GST, and AChE and the energy reserve contents were negatively affected due to UV-A exposure. Collectively, this study has demonstrated that UV-A light significantly suppresses the immune system of B. tabaci and that UV-A light is compatible with other biological control agents if it is applied separately from the biological agent.