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Fine particulate matter is classified as a first-class carcinogen and is receiving global attention. In particular, particulate matter (PM2.5, and PM10) are more fatal in students at a growing age while their respiratory system develops than in ordinary adults. During school hours, the main entrance is often left open for the convenience of students and staff. In this situation, the inflow of PM2.5 through the main entrance increases significantly. Thus, air curtains are widely used at the main entrance to block the inflow of PM2.5. However, few experimental studies have examined the effect of blocking PM2.5 through such air curtains. This study analyzed the effectiveness of reducing PM2.5 inflow by installing an air curtain at the main entrance of a school. The efficiency in blocking PM2.5 was determined by analyzing the average I/O ratio of PM2.5 at the respiratory height through experimental measurements. The average I/O ratio of PM2.5 in the school entrance was 0.84 before the installation of the air curtain, which was reduced to 0.46 after installing the air curtain. The blockage efficiency of the air curtain was 42%. This study quantitatively confirms that air curtains effectively reduce the inflow of PM2.5 in schools.

The task of assigning internationally accepted commodity codes (aka HS codes) to traded goods is a critical function of customs offices. Like court decisions made by judges, this task follows the doctrine of precedent and can be nontrivial even for experienced officers. Together with the Korea Customs Service (KCS), we propose a first-ever explainable decision supporting model that suggests the most likely subheadings (i.e., the first six digits) of the HS code. The model also provides reasoning for its suggestion in the form of a document that is interpretable by customs officers. We evaluated the model using 5,000 cases that recently received a classification request. The results showed that the top-3 suggestions made by our model had an accuracy of 93.9% when classifying 925 challenging subheadings. A user study with 32 customs experts further confirmed that our algorithmic suggestions accompanied by explainable reasonings, can substantially reduce the time and effort taken by customs officers for classification reviews. [ABSTRACT FROM AUTHOR]

The species of Beauveria bassiana is widely used for the management of agricultural insect pests. In this study, we integrated egfp‐double‐stranded RNA (dsRNA) to a previously generated egfp‐expressing B. bassiana transformant (Bb‐egfp#3) using a protoplast integration method. The Bb‐egfp#3 protoplast was mixed with the dsRNA under PEG/CaCl2 conditions and liquid‐cultured in Sabouraud dextrose broth for 5 days. A control culture followed the same procedure without dsRNA. Bb‐egfp#3/egfp‐dsRNA cultures showed very low fungal growth (OD630 = 0.2) compared to the control culture, Bb‐egfp#3 only (OD630 = 1.1). Screening of possible transformants on Sabouraud dextrose agar revealed a transformant T3, without egfp signal. T3 was confirmed as B. bassiana through sequencing of conserved genes and insect bioassays. Interestingly, the genomic egfp fragment of T3 was disrupted, and the egfp signal was not detected over four subcultures, which was also confirmed by RNA‐seq of Bb‐egfp#3 and T3. This study provides an interesting observation that protoplast integration with dsRNA could possibly generate significantly reduced gene expression in B. bassiana and it is stable across several generations. [ABSTRACT FROM AUTHOR]

• Metarhizium anisopliae JEF-197 and JEF-279 were selected for the Japanese pine sawyer beetle control. • M. anisopliae JEF-197 and JEF-279 showed high insecticidal activity in both laboratory and semi-field conditions. • Fungal granules may also be used for the Japanese pine sawyer beetle control. Chemical control is widely used to control the Japanese pine sawyer beetle, Monochamus alternatus , but strong chemical regulations require an environmentally sound management strategy. In this work, we investigated the use of entomopathogenic fungi and their application as a means of practical pest management. Thirty-two diverse species of fungal isolates were assayed against adult pine sawyer beetles using a contact method under laboratory conditions, and four isolates showed over 70% virulence consequently. These isolates, two each of Beauveria bassiana and Metarhizium anisopliae were sprayed on the adult beetles at 1 × 107 conidia/ml in plastic containers, respectively. The M. anisopliae -treated adult beetles showed 67% mortality. M. anisopliae isolates JEF-197 and JEF-279 demonstrated dosage-dependent insecticidal activity. Following the laboratory experiments, semi-field trials were conducted in young pine trees under high (RH 94%) and low (RH 35%) humidity conditions. In the high humidity conditions, most of the adult beetles stayed on the top of the branches. When the two M. anisopliae isolates were sprayed on the beetles, they showed ca. 50–70% insecticidal activity 11 days after application. In contrast, in low humidity conditions, the adult beetles tried to move off the branches and onto the soil. When the beetles reached the JEF-197 and JEF-279-treated soil, we measured >90% insecticidal activity. This work suggests that M. anisopliae was the most virulent entomopathogenic fungus against adult Japanese pine sawyer beetles, and this forest insect could be ecologically controlled by the spray and soil application of the M. anisopliae isolates. [ABSTRACT FROM AUTHOR]

Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae) ERL836 has been commercialized under the name ChongchaeSak to control an agricultural insect pest, the western flower thrips Frankliniella occidentalis Pergande (Thysanoptera: Thripidae), in the Republic of Korea. As soon as it was launched in 2017, it became a popular product and has received a positive response. However, study of the storage stability of the fungus ERL836 has yet to be investigated. To determine the optimum conditions for long-term storage, we assessed conidial viability and insecticidal activity of B. bassiana ERL836 according to storage temperature and culture substrate. Viability of B. bassiana ERL836 conidia from mycotized grains (millet and rice) stored at low (4 °C) and moderate (25 and 30 °C) temperatures was maintained at >85% for 24 and 18 months, respectively, along with insecticidal activity. In contrast, the samples stored at 37 °C showed low germination rate (about 80% germination rate for only 5 months). This result suggests that low and moderate temperatures (4 to 30 °C) conserve B. bassiana ERL836 viability and virulence. Unlabelled Image • Viability of ERL836 conidia stored at low (4 °C) temperatures was maintained at >85% for 24 months. • Long-term storage of ERL836 conidia also maintained insecticidal activity. • This study can serve as a guide to commercializing the entomopathogenic fungus ERL836. [ABSTRACT FROM AUTHOR]

We established a fungal production platform by focusing on substrates of solid culture for conidial productivity and thermotolerance, and focusing on surfactants to effectively deliver fungal conidia to the Riptortus pedestris cuticles. First, to produce thermotolerant fungal conidia, 2 of each Beauveria bassiana and Metarhizium anisopliae isolates were cultured on 13 cereal substrates for 10 days. Overall, five substrates (millet, non-glutinous Italian foxtail millet, barley, glutinous Italian foxtail millet, and brown rice) produced greatest number of conidia with thermotolerant conidia. When the selected substrates were mixed with minerals, zeolite, perlite or vermiculite to reduce the amount of cereal grains, vermiculite combination showed relatively high conidial production compared to the other mineral combinations. Next, to efficiently deliver the fungal conidia to the cuticles of R. pedestris, six surfactants, CO-2.5, CO-12, LE-7, PE-61, TED-3, and siloxane were each combined with the fungal conidia. The 0.01% combination showed significantly increased insect mortality, which varied depending on isolate. Virulence tests against R. pedestris were performed with conidial suspensions of isolates to assess their virulence. As a result, isolates showed the highest virulence when a virulence test was conducted at 25°C, rather than 20°C, 30°C and 35°C. This work suggests that the combination of cereal grain substrates and vermiculite could be considered for economic conidial production with high thermotolerance, and the CO-12 surfactant is the most suitable for effective delivery to target insects, followed by the information on optimal temperature for virulence against R. pedestris. [ABSTRACT FROM AUTHOR]

BACKGROUND Entomopathogenic Beauveria bassiana has been used as a biocontrol agent for insect pests, but its effect at the molecular level on the hosts has not been studied in detail. Herein, we performed transcriptome analysis of bean bug, Riptortus pedestris (Hemiptera: Alydidae) in response to infection with a highly virulent strain of B. bassiana JEF‐007 (Bb JEF‐007). RESULTS: Based on RNA‐seq data from R. pedestris infected with Bb JEF‐007 compared with non‐infected bean bugs, infection was assumed to strongly activate (i) the energy production pathway by expressing dehydrogenases, (ii) metabolic pathways by expressing secreted proteins, GTPase, MBF2 transcription factor family, pigment‐dispersing factor, antioxidants, and cuticle proteins, and (iii) the immune response pathway by expressing serine‐threonine kinase in Toll pathway of bean bug. CONCLUSION: We have established the platform for functional studies of the genes required for an immune response against entomopathogenic fungi like B. bassiana in the bean bug, R. pedestris. Moreover, this study also paves the way for genetic modification of B. bassiana to combat with the defense mechanism of R. pedestris. © 2018 Society of Chemical Industry The fungal infection was assumed to strongly activate some of the insect pathways, such as: (i) energy production pathway, (ii) metabolic pathways, and (iii) the immune response pathway. [ABSTRACT FROM AUTHOR]

Abstract Entomopathogenic fungi have great potential to control agricultural and horticultural insect pests, however optimizing conidial production systems to demonstrate high productivity and stability still needs additional efforts for successful field application and industrialization. Although many virulent entomopathogenic fungal isolates have been viewed as potential candidates in a laboratory environment, very few of the isolates are being used in practice for application in agricultural fields as commercial products. I. javanicus is an entomopathogenic fungus that is parasitic to various diverse coleopteran and lepidopteran insects and thought good candidate as biopesticdes. In this work, the basic characteristics of two entomopathogenic fungi, I. javanica FG340 and Pf04, were investigated in morphological examinations, genetic identification, and virulence against Thrips palmi , and then the feasibility of various grains substrates for conidial production was assessed, particularly focusing on conidial productivity and thermotolerance. Isaria javanica FG340 and Pf04 conidia were solid-cultured on 12 grains for 14 days in a Petri dish. Of the tested Italian millet, perilla seed, millet and barley-based cultures showed high conidial production. The four-grain media yielded >1 × 109 conidia/g of I. javanica FG340 and Pf04. Pf04 strain had enhanced thermotolerance up to 45 °C when cultured on Italian millet. In application, it was easy to make a conidial suspension using the cultured grains, and several surfactants were tested to release the conidia. This work suggests several possible inexpensive grain substrates by which to promote conidial production combined with enhanced stability against exposure to high temperature. Graphical abstract Unlabelled Image Highlights • Entomopathogenic fungi FG340 and Pf04 were identified as Isaria javanica. • Italian millet, perilla seed, millet and barley-based cultures showed high conidial production. • Highly thermotolerant conidia were produced on Italian millet as substrates in the Pf04. [ABSTRACT FROM AUTHOR]

The Toll signaling pathway is responsible for defense against both Gram‐positive bacteria and fungi. Gram‐negative binding protein 3 (GNBP3) has a strong affinity for the fungal cell wall component, β‐1,3‐glucan, which can activate the prophenoloxidase (proPO) cascade and induce the Toll signaling pathway. Myeloid differentiation factor 88 (MyD88) is an intracellular adaptor protein involved in the Toll signaling pathway. In this study, we monitored the response of 5 key genes (TmGNBP3, TmMyD88, and Tenecin 1, 2, and 3) in the Toll pathway of the mealworm Tenebrio molitor immune system against the fungus Beauveria bassiana JEF‐007 using RT‐PCR. TmGNBP3, Tenecin 1, and Tenecin 2 were significantly upregulated after fungal infection. To better understand the roles of the Toll signaling pathway in the mealworm immune system, TmGNBP3 and TmMyD88 were knocked down by RNAi silencing. Target gene expression levels decreased at 2 d postknockdown and were dramatically reduced at 6 d post‐dsRNA injection. Therefore, mealworms were compromised by B. bassiana JEF‐007 at 6 d post‐dsRNA injection. Silencing of TmMyD88 and TmGNBP3 resulted in reduced resistance of the host to fungal infection. Particularly, reducing TmGNBP3 levels obviously downregulated Tenecin 1 and Tenecin 2 expression levels, whereas silencing TmMyD88 expression resulted in decreased Tenecin 2 expression. These results indicate that TmGNBP3 is essential to induce downstream antifungal peptide Tenecin 1 expression against B. bassiana JEF‐007. [ABSTRACT FROM AUTHOR]

Entomopathogenic fungi are soil-dwelling microorganisms that infect host insects, and some have been used as biological control agents. Construction of an entomopathogenic fungal library provides a strong platform for the development of highly effective biopesticides, but the traditional method using antibiotic media has low isolation efficiency. Herein, to increase the efficiency of isolation, a Tenebrio molitor pathogenicity-based fungal collection method was used for the construction of a library. The isolation efficiency using the T . molitor larvae-based pathogenicity assay was 55.4%, which was significantly higher than the method of antibiotic medium with dodine (6.0%). We named it the Jeonbuk (Chonbuk) National University Entomopathogenic Fungi (JEF) library. It consisted of 279 isolates belonging to 12 genera and 29 species. Main features of the species-representative 15 isolates were characterized in terms of morphology, virulence, conidiogenesis, thermotolerance and production of biologically active materials such as virulence-related enzymes. Some of the species-representative isolates showed higher virulence against T . molitor , Riptortus pedestris and Plutella xylostella in laboratory conditions. The production of enzymes such as chitinase, Pr1 protease and lipase all related to pathogenesis in LB medium was higher than in the SDB and PDB media. In the thermotolerance assay, a Purpureocillium lilacinum isolate showed higher thermotolerance than other isolates. This work reports an efficient library method using T . molitor for building a library for development of fungal biopesticides for pest management. [ABSTRACT FROM AUTHOR]

Most entomopathogenic fungal biopesticides have been produced in wettable powder and liquid concentrate formulations. Separation of fungal conidia from granular substrates to manufacture these formulations is costly and labor-intensive, which has hampered growth of the fungal biopesticide industry. Herein to overcome this challenge, we developed a novel screen bag formulation using mycotized granules produced from solid-substrate cultures of Beauveria bassiana JEF-007 and Metarhizium anisopliae JEF-003. The isolates were solid-cultured on Italian millet grains and packed in screen bags (pore size: ~ 15 μm). The bags containing the mycotized granules were immersed in water and agitated to disperse the conidia, after which the number of conidia released into the water was determined. These results were compared to the number of conidia released after vortexing the granules in water as a control. Various surfactants were added to the water to assess their effect on conidial release. Silwet L-77 yielded the highest conidial release. The insecticidal activity against Riptortus pedestris nymphs of suspensions produced from the screen bag formulation and vortexed conidial suspensions was similar, demonstrating that our novel formulation could be a cost-effective replacement for standard WP and ES formulations. The screen bag formulation has several advantages over other standard fungal-based formulations, such as a shorter and lower-cost production process. This formulation technology could also be applied to other conidial-based biopesticides, such as fungal fungicides, herbicides, and fertilizers. [ABSTRACT FROM AUTHOR]

In the present study, an enhanced green fluorescent protein gene ( egfp ) was expressed in Beauveria bassiana ERL836 based on the Agrobacterium tumefaciens -mediated transformation (AtMT) method. The ERL836 transformants were generated with pCambia-egfp binary vectors. Ten transformants were randomly selected and analyzed for the T-DNA insertion and gene expression. The results revealed that 60% ( egfp ) of the fungal putative transformants were inserted by the T-DNA fragment. The expressions of egfp in putative transformants were rapidly detected by reverse transcription PCR (RT-PCR) method. Among these transformants, 33.33% ( egfp , 2 transformants) expressed the target genes. Two selected egfp transformants showed strong green fluorescence with different expression levels. The results of this study could provide a successful procedure for foreign protein production in B. bassiana by using the AtMT method and accomplished with a rapid selection of fungal transformants using RT-PCR. [ABSTRACT FROM AUTHOR]

Mosquitoes have been becoming serious vectors worldwide thus effective and safe control strategies should be established. Entomopathogenic fungi can be alternative controlling agents by substituting chemical insecticides. Herein we assayed 12 soil-borne entomopathogenic fungi against Asian tiger mosquito ( Aedes albopictus ) larvae in laboratory conditions and tried to establish an effective application method using millet granular formulation (GR). Twelve fungal isolates which belong to 6 genera ( Beauveria , Cordyceps , Metarhizium , Paecilomyces , Purpureocillium and Verticillium ) were assayed; M. anisopliae JEF-003 showed the fastest mosquitocidal activity, approximately 73% mortality rate at 2 days post-inoculation (dpi.) and > 90% mortality rate at 5 dpi. Conidia of M. anisopliae JEF-003 also showed a dosage dependent activity at 1 × 10 5 , 1 × 10 6 and 1 × 10 7 conidia ml − 1 . Hyphal growth of M. anisopliae JEF-003 in the Ae. albopictus larvae was observed by infection of an M. anisopliae JEF-003 EGFP-transformant, which was generated by restriction enzyme-mediated integration (REMI) method. GR of M. anisopliae JEF-003 showed high virulence to Ae. albopictus larvae (> 90% mortality) after 5 days of application. These results suggest that M. anisopliae JEF-003 has a potential to control A. albopictus larvae and GR can be practically used for management of the serious vector in water environment. [ABSTRACT FROM AUTHOR]

Beauveria bassiana is an entomopathogenic fungi used in environmentally mindful pest management. Its main active ingredient, conidia, is commercially available as a fungal biopesticide. Many studies of conidia production have focused on how to optimize culture conditions for maximum productivity and stability against unfavorable abiotic factors. However, understanding of how conidiogenesis-related genes provide improved conidial production remains unclear. In this study, we focus on identifying conidiogenesis-related genes in B. bassiana ERL1170 using a random mutagenesis technique. Transformation of ERL1170 using restriction enzyme-mediated integration generated one morphologically different transformant, ERL1170-pABeG #163. The transformant was confirmed to represent B. bassiana , and the binary vector was successfully integrated into the genome of ERL1170. Compared to the wild type, transformant #163 showed very slow hyphal growth and within 6 days only produced <1 × 10 6  conidia/0.28 cm 2 agar block (wild type: 6.2 × 10 7  conidia/agar block). Transformant #163 also exhibited different morphology than the wild type, including thicker hyphae with some club-shaped parts. In contrast, the typical morphology of wild type B. bassiana exhibits thread-like hyphae and conidiophore structures and circular conidia. To determine the location of the randomly inserted DNA, we conducted thermal asymmetric interlaced (TAIL) PCR and Escherichia coli cloning to clearly sequence the disrupted region. We identified one colony (colony No. 7) with an insertion site identified as DNA photolyase. This was confirmed through a gene knock-out study. It is possible the gene that encodes for DNA photolyase was disrupted during the insertion process and might be involved in fungal conidiogenesis. This work serves as a platform for exploring the function of a variety of B. bassiana genes involved in pest management and their downstream processing. [ABSTRACT FROM AUTHOR]

Infectious laryngotracheitis (ILT), fowlpox (FP), and reticuloendotheliosis are important poultry diseases caused by gallid herpesvirus 1 (ILTV), fowlpox virus (FWPV), and reticuloendotheliosis virus (REV), respectively. Coinfections with ILTV and FWPV occur naturally in chickens, and FP in its more virulent wet form is characterized by diphtheritic lesions and easily confused with ILT. Moreover, the insertion of only partial REV-LTR or a nearly full-length REV into the FWPV genome, located between the ORF 201 and ORF 203, has increased recently in wild-type field FWPV isolates. Therefore, it is critical to detect ILTV, FWPV, REV-integrated FWPV, and REV early and accurately. In this study, we successfully developed a multiplex PCR assay for the simultaneous detection of ILTV, FWPV, REV-integrated FWPV, and REV, and the detection limits was 1 × 54 copies/tube. When used to test clinical samples, the results of the multiplex PCR were in 100% agreement with singleplex PCRs and sequencing. This new multiplex PCR is a simple, rapid, sensitive, specific, and cost-effective method for detection of 4 viruses in clinical specimens. [ABSTRACT FROM AUTHOR]

We investigated the colonization potential of five Metarhizium anisopliae isolates on pine tree surfaces under laboratory conditions, determined the influence of the pine bark extract on fungal growth and evaluated the insecticidal activity following colonization on the Japanese pine sawyer. Finally, the effect of colonization on adults pine sawyer was evaluated using the top three performing isolates (JEF-197, JEF-271 and JEF-279) under laboratory and field conditions. As a result, isolate JEF-197 showed the highest conidial production on the pine surfaces, and five isolates, including JEF-197, showed higher hyphal growth on autoclaved pine bark extract agar, compared to a water agar. Pine bark treated with the isolates showed 40–70 % mortality of adults pine sawyer. Under mimicked overwintering conditions, in the JEF-197 treatment group, 40 % of the inserted larvae became adults and all were dead after 59 d. In a field test, colonized isolate JEF-197 also showed 37 % insecticidal activity against emerged adults from the pine logs as overwintering sites. This work suggests that M. anisopliae isolate JEF-197 possibly colonized the pine surface and application of a conidial suspension on the pine logs as overwintering sites could be an effective strategy to control the pine sawyer. Image 1 • Metarhizium anisopliae isolates could colonize pine bark surface. • Pine bark extract promoted the growth and colonization of M. anisopliae. • Colonized JEF-197 induced 37 % mortality of emerged adults in pine logs. • The isolates can be applied on pine logs to control the pine sawyer. [ABSTRACT FROM AUTHOR]

A viral genome was assembled de novo from next-generation sequencing (NGS) data from bean bugs, Riptortus pedestris , infected with an entomopathogenic fungus, Beauveria bassiana ( Bb ), and was further confirmed via the RACE method. This is a novel insect positive-sense single-stranded RNA virus, which we named Riptortus pedestris virus-1 (RiPV-1) (GenBank accession no. KU958718). The genome of RiPV-1 consists of 10,554 nucleotides (nt), excluding the poly(A) tail, which contains a single large open reading frame (ORF) of 10,371 nt encoding a polyprotein (3456 aa) and flanked by 71 and 112 nt at the 5′ and 3′ untranslated regions (UTR), respectively. RiPV-1 genome organization from the 5′ end contains a consensus organization of picorna-like RNA virus helicase, cysteine protease, and RNA-dependent RNA polymerase (RdRp), in addition to two putative structural proteins located at the 3′ region and a poly(A) tail at the 3′ end. The viral particles were approximately 30 nm in diameter with some dispersal distinctive surface projections. Based on the phylogenetic analysis of the RdRp sequences, RiPV-1 was clustered in the unassigned insect RNA viruses with two other viruses, APV and KFV. These three viruses were suggested to constitute a new group of insect RNA viruses. RiPV-1 could be found in all stages of lab-reared bean bugs and was detected abundantly in the thorax, abdomen, midgut and fat body, but not in the reproductive organs and muscle. Interestingly, RiPV-1 replication was increased dramatically in bean bugs 2–6 days after fungal infection. In conclusion, a novel insect RNA virus was found by NGS data assembly. This virus can provide further insight into the interaction between virus, fungus and the host. [ABSTRACT FROM AUTHOR]

Beauveria bassiana ( Bb ) is used as an environment-friendly biopesticide. However, the molecular mechanisms of Bb -host interactions are not well understood. Herein, RNA isolated from B. bassiana ( Bb JEF-007) and Riptortus pedestris (Hemiptera: Alydidae) infected with this strain were firstly subjected to high-throughput next generation sequencing (NGS) to analyze and compare transcriptomes. Due to lack of fungal and host genome information, fungal transcriptome was processed to partially exclude non-infection specific genes and host-flora. Differentially Expressed Gene (DEG) analysis showed that 2381 genes were up-regulated and 2303 genes were down-regulated upon infection. Most DEGs were classified into the categories of single-organism, cellular and metabolism processes by Gene Ontology analysis. Most DEGs were involved in metabolic pathways based on Kyoto Encyclopedia of Genes and Genomes pathway mapping. Carbon metabolism-related enzymes in the glyoxylate cycle were significantly up-regulated, suggesting a possible role for them in Bb growth in the host. Additionally, transcript levels of several fungal genes were dramatically increased after infection, such as cytotoxic lectin-like protein, bacterial-like toxin, proteins related to cell wall formation, hyphal growth, nutrient uptake, and halogenated compound synthesis. This work provides insight into how entomopathogenic B. bassiana grows in agriculturally harmful bean bug at 6 d post infection. [ABSTRACT FROM AUTHOR]

• Beauveria , Metarhizium , Isaria , and Akanthomyces were screened against H. longicornis. • M. anisopliae isolates were highly virulent against nymphs in a dose-dependent manner. • Spray application of M. anisopliae to nymphs on soil caused 90% mortality in 30 days. The longhorned tick (bush tick), Haemaphysalis longicornis (Ixodida: Ixodidae), is a serious pest; it transmits the severe fever with thrombocytopenia syndrome (SFTS) virus to humans and has a wide distribution. The use of chemical control is not favored for environmental and health reasons, so more environmentally sound management methods need to be developed. Herein, we describe the use of an entomopathogenic fungal library to develop a fungus-mediated tick management system. Field-collected nymphs were assayed for their susceptibility to entomopathogenic fungi belonging to genera Beauveria , Metarhizium , Cordyceps , and Akanthomyces. Three M. anisopliae s.l. isolates, JEF-214, -279, and -290 showed high virulence in a dose-dependent manner. One Cordyceps isolate was pathogenic but virulence was much lower than the M. anisopliae isolates. Beauveria isolates were not pathogenic to the tick. Because the longhorned tick dwells on the soil surface except for blood-feeding periods, the soil surface was sprayed with conidial suspensions of the isolates after the release of longhorned ticks. The treatments resulted in 60–90% mortality after 30 days. M. anisopliae s.l. isolates were highly virulent against longhorned tick, and the application of fungus-based biopesticides on the soil surface could be an effective control strategy to reduce the tick population for long-term tick management. [ABSTRACT FROM AUTHOR]

The Japanese pine sawyer (JPS) beetle, Monochamus alternatus Hope (Coleoptera: Cerambycidae), damages pine trees and transmits the pine wilt nematode, Bursaphelenchus xylophilus Nickle. Chemical agents have been used to control JPS beetle, but due to various issues, efforts are being made to replace these chemical agents with entomopathogenic fungi. We investigated the expression of immune-related genes in JPS beetle in response to infection with JEF-197, a Metarhizium anisopliae isolate, using RNA-seq. RNA samples were obtained from JEF-197, JPS adults treated with JEF-197, and non-treated JPS adults on the 8th day after fungal treatment, and RNA-seq was performed using Illumina sequencing. JPS beetle transcriptome was assembled de novo and differentially expressed gene (DEG) analysis was performed. There were 719 and 1953 up- and downregulated unigenes upon JEF-197 infection, respectively. Upregulated contigs included genes involved in RNA transport, ribosome biogenesis in eukaryotes, spliceosome-related genes, and genes involved in immune-related signaling pathways such as the Toll and Imd pathways. Forty-two fungal DEGs related to energy and protein metabolism were upregulated, and genes involved in the stress response were also upregulated in the infected JPS beetles. Together, our results indicate that infection of JPS beetles by JEF-197 induces the expression of immune-related genes.

Root-feeding Scarabaeidae, particularly white grubs are considered among the most harmful coleopteran insect pests in turfgrass. In this work, sixteen entomopathogenic fungal species were assayed against flower chafer beetle, Protaetia brevitarsis (Coleoptera: Scarabaeidae) and Metarhizium anisopliae JEF-314 showed high virulence. The control ability of the isolate JEF-314 has been in detail tested for a model insect flower chafer beetle. Further analyses showed insect stage-dependent virulence where the fungal virulence was the highest against smaller instar larvae. Additionally, we confirmed that millet-based solid cultured granule was effective against the soil-dwelling larval stage. The isolate also showed a similar ability for a representative pest ( Popillia spp.) in laboratory conditions. Our results clearly suggest a high potential of M. anisopliae JEF-314 to control the flower chafer beetle, possibly resulting in controlling of root-feeding white grubs in turfgrass. Based on the insect life cycle and susceptibility to the fungus, late spring and summer time would be the optimum time to apply JEF-314 granules for an effective control. Further characterization of the efficacy of the fungus under field conditions against the Scarabaeidae beetles might provide an efficient tool to control this beetle in an environment-friendly way., (© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of the Korean Society of Mycology.)