• Title/Summary/Keyword: B. bassiana

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Genetic Relationships of Internal Transcribed Spacer (ITS) Regions on Entomopathogenic Fungi by RFLP (Entomopathogenic Fungi의 ITS 영역에 대한 RFLP 분석)

  • Choi, In-Young;You, Young-Jin;Choi, Joung-Sik;Lee, Wang-Hyu
    • The Korean Journal of Mycology
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    • v.28 no.2
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    • pp.112-117
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    • 2000
  • A similarity coefficient were analyzed by RFLP of fourteen species of entomopathogenic fungi, isolated from inhabiting pupa and adult insect at forest. Each rDNA ITS I and ITS II with primers of ITS 1 and ITS 4 was amplified by PCR. The amplified products were conserved to 500 bp were not demarcated between genus and species. Four Paeciliomyces tenuipes, two Beauveria bassiana and six Cordyceps militaris were treated by seven restriction enzymes and confirmed in species except JB3 by electrophoresis band. However, the band of C. scarabaeicola showed the identity with B. bassiana. The result of this experiment indicated that the teleomorph of C. scarabaeicola was the same as that of B. bassiana. CfoI and HpaII of seven restricted enzymes were easily discriminating in the genus between Paecilomyees and Cordyceps. Especially, CfoI was more effective to classify the genera of Paecilomyees, Cordyceps and Beauveria than other restriction enzymes. The band patterns of RFLP of P. tenuipes, C. militaris, C. scarabaeicola and B. bassiana were also analyzed by UPGMA program of NTSYS-pc and showed 100% significance. Thus, the similarity coefficient tended to be lower between genera by RFLP analysis, but was higher between species.

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Identification of Beauveria spp. Isolated from Mulberry Longicorn Beetle (Apriona germari Hope) using Polymerase Chain Reaction (뽕나무 하늘소(Apriona germari Hope)로부터 Beauveria속 사상균의 분리 및 PCR에 의한 동정)

  • 서종복;진병래
    • Journal of Sericultural and Entomological Science
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    • v.37 no.2
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    • pp.167-171
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    • 1995
  • To develope a microbial pesticide for the control of mulberry longicorn beetle, Apriona germari, Beauveria spp. were isolated from the infected Apriona germari larvae. The morphology of Beauveria spp. was observed by phase contrast and scanning electron microscope. In addition, the Beauveria spp. isolated from Apriona germari were identified by the random amplification of polymorphic DNA using polymerase chain reaction. The results showed that the Beauveria spp., SFB-1A and SFB-3A, isolated from Apriona germari were identified with B. bassiana and B. brongniartii, respectively, suggesting that the random amplification of polymorphic DNA is effective for the identification of Beauveria spp.

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Development of Liquid Culture Media for the Mass Production of Beauveria bassiana Blastospore (흰굳음병균(Beauveria basiana) 단균사의 대량배양생산을 위한 액체배지 개발)

  • 서종복;진병래
    • Journal of Sericultural and Entomological Science
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    • v.37 no.2
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    • pp.172-175
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    • 1995
  • To develope a liquid culture media for the mass culture of Beauveria bassiana blastospore, wheat bran and soybean cake were selected as a carbon and nitrogen source, respectively. The composition and final concentration of wheat bran and soybean cake in liquid culture media were determined by the growth of B. bassiana blastospore. The results showed that the growth of B. bassiana blastospore was effective in the rate of 3 wheat bran: 2 soybean cake and in 5% of final concentration of the liquid culture media, suggesting that the liquid culture media using wheat bran and soybean cake are available.

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Cordyceps bassiana and Production of Stromata in vitro Showing Beauveria Anamorph in Korea

  • Sung, Jae-Mo;Lee, Je-O;Humber Richard A.;Sung, Gi-Ho;Shrestha Bhushan
    • Mycobiology
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    • v.34 no.1
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    • pp.1-6
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    • 2006
  • A Cordyceps species was found with a Beauveria anamorph state on larval insect cadavers on Obong mountains in Gangwon Provinces, Republic of Korea. Cultures from discharged ascospores formed an anamorph identifiable as Beauveria bassiana. This teleomorph-anamorph connection was also confirmed by the in vitro production of fertile ascomata from conidial cultures with morphology like that of field-collected specimen. This is the first report of in vitro production of a teleomorph for any Beauveria species. The Cordyceps species has been conspecified as Cordyceps bassiana, a species described from China with B. bassiana anamorph.

Analysis of Genetic Relationship of Entomogenous Fungi in Korea by Morphological Characteristics and RAPD (한국에서 채집한 Entomogenous fungi의 형태와 RAPD에 의한 유연관계 분석)

  • Choi, In-Young;Choi, Joung-Sik;Yu, Young-Jin;Lee, Wang-Hyu
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.34-40
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    • 2001
  • RAPD test and the observation of morphological, cultural characteristics of fourteen selected entomogenous fungi were conducted to investigate the analysis of their internal relationships. Paecilomyces tenuipes showed snow flower form attached to numerous white conidiophores, produced globular and semi-egg types on the club types of phialides. Cordyceps militaris formed globosely conidiophores, dark yellow fruiting body on pupa. The phialide as on Acremonium-type in global conidiophores. Beauveria bassiana covered with conidia was not formed fruiting body and adhered conidia on conidiophore of zigzag type. The PDA and SDAY medium were confirmed as an optimum growth of them. P. tenuipes showed to velvet and plane types in several media whereas C. militaris was belong to centrally raised and floccose in the morphological type. In contrast, B. bassiana covered with conidia on velvet shape. The size of amplified products were analyzed by RAPD using URP primer and were from 100 bp to 2.0 kb with $10{\sim}14$ geuomic DNA. Total similarities of two groups were by dendrogram of UPGMA analysis. The homology of P. tenuipes groups was 94.8 to 100%. It also showed 70.1 to 96.6% in C. militaris group and B. bassiana was higher similarity than any other. The internal change of C. militaris, produced telemorph fruiting body, was higher seperated in species than P. tenuipes and B. bassiana in the RAPD.

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Evolutionary Explanation for Beauveria bassiana Being a Potent Biological Control Agent Against Agricultural Pests

  • Han, Jae-Gu
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.27-28
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    • 2014
  • Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

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Effect of Saccharomyces cerevisiae consumption on the pathogenicity of Beauveria bassiana in Protaetia brevitarsis

  • Kwak, Kyu-Won;Han, Myung-Sae;Nam, Sung-Hee;Park, Kwan-Ho;Kim, Eun-Sun;Lee, Seokhyun;Song, Myung-Ha;Kim, Wontae;Choi, Ji-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • v.33 no.1
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    • pp.1-5
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    • 2016
  • Beauveria bassiana is a common fungal pathogen of Protaetia brevitarsis larvae, and although it is less common than Metarhizium anisopliae , the pathogen still poses a great risk to humans and animals that consume infected insects, owing to B. bassiana's production of toxins like beauvericin and mycotoxin. Interestingly, the beneficial microorganism Saccharomyces cerevisiae possesses antifungal properties. In the present study, we found that S. cerevisiae inhibited the growth of B. bassiana by 97% and that S. cerevisiae failed to harm P. brevitarsis when administered via intracoelomic injection (1×107 cfu/mL). In addition, we also found that S. cerevisiae consumption increased the survival time of percutaneously infected P. brevitarsis larvae by 5 d and reduced the mortality of infected larvae by 12%. Therefore, S. cerevisiae is expected to be useful in the prevention and control of B. bassiana in the production of P. brevitarsis larvae.

Inhibitory Activity of Cordyceps bassiana Extract on LPS-induced Inflammation in RAW 264.7 Cells by Suppressing NF-κB Activation

  • Yoon, Deok Hyo;Han, Changwoo;Fang, Yuanying;Gundeti, Shankariah;Han Lee, In-Sook;Song, Won O;Hwang, Ki-Chul;Kim, Tae Woong;Sung, Gi-Ho;Park, Haeil
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.162-168
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    • 2017
  • Cordyceps bassiana has long been used as an oriental medicine and reported to possess diverse biological activities. The fruiting bodies of Cordyceps bassiana was extracted with ethanol and then further fractionated with n-hexane, ethyl acetate, n-butanol and water. The butanol fraction from Cordyceps bassiana (CBBF) exhibited the most effective in anti-inflammatory activity in RAW 264.7 macrophages and the roles of CBBF on the anti-inflammation cascade in LPS-stimulated RAW 264.7 cells were studied. To investigate the mechanism by which CBBF inhibits NO, iNOS and COX-2, the activation of $I{\kappa}B$ and MAPKs in LPS-activated macrophage were examined. Our present results demonstrated that CBBF inhibits NO production and iNOS expression in LPS-stimulated RAW 264.7 macrophage cells, and these effects were mediated through the inhibition of $I{\kappa}B-{\alpha}$, JNK and p38 phosphorylation. Also, CBBF suppressed activation of MAPKs including p38 and SAPK/JNK. Furthermore, CBBF significantly suppressed LPS-induced intracellular ROS generation. Its inhibition on iNOS expression, together with its antioxidant activity, may support its anti-inflammatory activity. Thus Cordyceps bassiana can be used as a useful medicinal food or drug for further studies.

A Technique for the Prevention of Greenhouse Whitefly (Trialeurodes vaporariorum) Using the Entomopathogenic Fungus Beauveria bassiana M130

  • Kim, Chang-Su;Lee, Jung-Bok;Kim, Beam-Soo;Nam, Young-Ho;Shin, Kee-Sun;Kim, Jin-Won;Kim, Jang-Eok;Kwon, Gi-Seok
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.1-7
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    • 2014
  • The possibility of using hyphomycete fungi as suitable biocontrol agents against greenhouse whitefly has led to the isolation of various insect pathogenic fungi. Among them is Beauveria bassiana, one of the most studied entomopathogenic fungi. The objective of this study was to use B. bassiana M130 as an insecticidal agent against the greenhouse whitefly. M130 isolated from infected insects is known to be a biocontrol agent against greenhouse whitefly. Phylogenetic classification of M130 was determined according to its morphological features and 18S rRNA sequence analysis. M130 was identified as B. bassiana M130 and showed chitinase (342.28 units/ml) and protease (461.70 units/ml) activities, which were involved in the invasion of the host through the outer cuticle layer, thus killing them. The insecticidal activity was 55.2% in petri-dish test, 84.6% in pot test, and 45.3% in field test. The results of this study indicate that B. bassiana has potential as a biological agent for the control of greenhouse whitefly to replace chemical pesticides.

Identification of an Entomopathogenic Fungus, Beauveria bassiana SFB-205 toxic to the Green Peach Aphid, Myzus persicae

  • Kim, Jae-Su;Roh, Jong-Yul;Choi, Jae-Young;Shin, Sang-Chul;Jeon, Mun-Jang;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.17 no.2
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    • pp.211-215
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    • 2008
  • To select entomopathogenic fungi controlling aphids effectively, several isolates were screened against second instars of Myzus persicae nymphs in the glasshouse using conidia suspension at $1.0{\times}10^5$ conidia/ml. Among these isolates, SFB-205 conidia showed the highest insecticidal activity about 32.7% efficacy to M. persicae at 4 days after application in the glasshouse. The attachment of SFB-205 conidia on the surface of M. persicae nymphs, and germination and penetration were observed using scanning electron microscopy. SFB-205 was identified as Beauveria bassiana species through the comparison of 5.8 s rRNA genes. There were 24 polymorphisms between SFB-205 and the previously reported isolate, B. bassiana ATCC74040 using six kinds of primer combinations in amplified fragment length polymorphism (AFLP) analysis. The B. bassiana SFB-205 might be used as a practical biological control agent for the green peach aphid, M. persicae in the field.