• Applied Biological Chemistry
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• v.40 no.1
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• pp.30-33
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• 1997

About 250 bp ura5 gene (Orotate phosphoribosyl transferase) fragment was cloned from genomic DNA of entomopathogenic fungus Metarhizium anisopliae by using PCR method. Entire nucleotide sequences of cloned DNA fragment were determined and analysed as compared with other fungus ura5 genes. The amino acid sequence deduced from the nucleotide sequence showed 85.5% homology to ura5 protein of Trichoderma reesei. Using this 250 bp PCR fragment we have isolated full ura5 gene of M. anisopliae by genomic Southern hybridization and the isolated 4.4 kb DNA fragments were mapped by restrictional enzyme.

• The Korean Journal of Pesticide Science
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• v.2 no.2
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• pp.90-96
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• 1998

The purpose of this study is to investigate the liquid culture conditions for mass production of Metarhizium anisopliae var. majus ARS 978 which is a potential microbiological pesticide. The temperature and pH range for optimal cultivation were $28^{\circ}C$ and pH 7.0, respectively. For M. anisopliae ARS 978, 1.0 %(v/v) molasses, 1.0 %(w/v) distiller's dry solubles, and 0.3 %(w/v) $CaCO_{3}$ were found to be the proper nutrients, considering cell mass, enzyme activities and spore concentration.

• Journal of Microbiology and Biotechnology
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• v.9 no.3
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• pp.334-339
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• 1999

More innovative molecular markers were investigated for rapid and consistent differentiation of Metarhizium anisopliae var. majus from M. anisopliae var. anisopliae. A total of 28 isolates were obtained from various countries and hosts: 13 isolates of M. anisopliae var. anisopliae, 12 isolates of M. anisopliae var. majus, and 3 isolates of M. anisopliae collected in Korea. This study involved restriction enzyme digestions of a PCR product amplified from nuclear internally transcribed spacer (ITS) and a portion of the 28S rDNA regions. Among 11 different restriction enzymes used in this study, MboⅠ digestion particularly produced a restriction pattern that had characteristics of M. anisopliae var. majus. This restriction pattern was consistent in all isolates of M. anisopliae var. majus regardless of their geographic origins and insect hosts. Mapping experiments revealed that MboⅠ sites of M. anisopliae var. majus are identical to those of M. anisopliae var. anisopliae with an exception for the presence of an additional site in the PCR product. Results from this study provide an additional method for identification and differentiation of isolates of these two varieties of M. anisopliae for use in the field and laboratory experiments.

• Asian Journal of Turfgrass Science
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• v.11 no.3
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• pp.185-191
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• 1997

Biological control of turfgrass insect pest Blitopertha orientalis, forest insect pests, Agelastica coerulea, Meganola melancholia, and Glyphodes perspectalis,vegetable insect pests, Plutella xylostella and Agrotis segetum were conducted with entonopathogenic fungus, Metarhizium anisopliae J-22 isolated from black pine forest soil in Cheju province. Mortality of B. orientalis larvae was 53.3% at the rate of 3.4 $\times$ 1O 7 conidia /ml. A. coerulea and M melancholia larvae showed 100% mortalities at 9.6 $\times$ 106 conidia /ml and 2.7 $\times$ 10 7 conidia /ml as well. However, G. perspectalis larvae were not dead even at 4$\times$ 1O 7 conidia /ml. On the other hand, M anisopliae J-22 was effective against P. xylostella larvae showing 100% mortality at 4 $\times$ 10 7 conidia /ml. KEy words:Entomopathogenic fungi, Biological control, Metarhizium anisopliae, Pathogenicity,Blitopertha orientalis, Agelastica coerulea, Meganola melancholia, Glyphodes perspectalis,Plutella xylostella, Agrotis segetum.

• Applied Biological Chemistry
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• v.41 no.3
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• pp.219-223
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• 1998

We have established a transformation system for entomopathogenic fungus, Metarhizium anisopliae, in order to develop mycoinsecticide by recombinant DNA techniques. Protoplasts of M. anisopliae would be transformed to a benomyl-resistant by introducing pBRG-4 plasmid DNA, which contains a ${\beta}-tubulin$ gene of Aspergillus flavus conferring resistance to benomyl and a pyr4 gene of Neurospora crassa, in the presence of 5% polyethylene glycol and 10 mM calcium chloride. Transformants occuring at a frequency of 10 colonies per $50\;{\mu}g$ pBRG-4 DNA grew on the $5\;{\mu}g/ml$ concentrations of benamyl, while the wild type was inhibited by $2.5\;{\mu}g/ml$. From the Southern analysis using genomic DNAs isolated from M. anisopliae transformants, the positive signals suggested that the ${\beta}-tubulin$ gene had integrated in the M. anisopliae genome by homologous recombination.

• Korean journal of applied entomology
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• v.36 no.2
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• pp.179-184
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• 1997

This study was conducted to observe the effects of pesticides on the pathogenicity of entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae isolated from Korea. Spomlation of entomopathogenic fungi in SMAY medium that mixed different concentrations of pesticides was similar to control in metalaxyl and tolclofos- methyl treatment but in half recommanded treatment of fenitrothion and mepronil very poor spomlation was showed. Pathogenicity of entomopathogenic fungi dipped in all tested pesticide solution for 1 to 5hr was 100% pathogenicity of M. aniropliae in the pesticide treated chinese cabage fields were similar to control, Bt, teflubenzuron, metalaxyl, and carbofuran treatment but alachlol was below 80%. Pathogenicity and persistence of entomopathogenic fungi in turfgrass when pesticides were posttreatmented were continued to 4 months in iprodione+thiram and tolclofos-methyl treated plots but those of B. bassiana and M. anisopliae was losted in fenitrothion pathogenicity and persistence of in mepronil were nil M. anisopliae from 3 months after treatment.

• The Korean Journal of Mycology
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• v.24 no.1 s.76
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• pp.49-55
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• 1996

This study was conducted to find out antifungal activities of entomopathogenic fungus, Metarhizium anisopliae, against phytopathogenic fungi, Fusarium oxysporum, Botrytis cinerea and Alternaria solani. M. anisopliae was confirmed its antagonistic effect through mycelial inhibition zone of phytopathogenic fungi by culture filtrate of the antagonist. The filtrate (30%: v/v) inhibited the conidial germination of B. cinerea and F. oxysporum to 21.5% (control: 88.2%) and 53.0% (control: 78.6%), respectively and delayed the start of spore germination about 8hours. Microscopic observations proved that the addition of 10% culture filtrate of M. anisopliae restricted the growth of phytopathogenic fungus, F. oxysporum, to the formation of chlamydospore. From these results, we concluded that an addition effect of the filtrate from M. anisopliae on culturing F. oxysporum was fungistatic.

• Korean Journal of Ecology and Environment
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• v.53 no.1
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• pp.73-79
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• 2020

Metarhizium anisopliae is one of the entomopathogenic fungi infecting Protaetia brevitarsis seulensis. Environmental conditions strongly affect the virulence of entomopathogenic fungus. To test the susceptibility of P. brevitarsis seulensis larvae to M. anisopliae, we determined level mortality rates under different rearing conditions: temperatures (20℃, 25℃, 30℃), relative humidities (RH; RH 40%, 50%, 60%, 70%), and water content in sawdust (40%, 50%, 60%). The mortality rate of larvae treated with 5×10⁶ conidia mL^-1 M. anisopliae was not significantly affected by rearing temperature, but was affected by low RH (40%) and low water content in sawdust (40%), both of which resulted in high mortality rates. Mortality rated among M. anisopliae-treated 1^st, 2^nd, and 3^rd instar larvae were highest in 1^st instar larvae.

• The Korean Journal of Pesticide Science
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• v.18 no.4
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• pp.417-421
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• 2014

The beet armyworm, Spodoptera exigua is pest which is difficult to control. For eco-friendly beet armyworm managements, we isolated entomopathogenic fungi from soil samples by insect-bait method using Tenebrio molitor and conducted bioassay to larvae of beet armyworm. The result of bioassay, a selected strain Metarhizium anisopliae FT83 caused 100% mortality against first ~ third instar larva of S. exigua at $1{\times}10^8conidia/ml$ and medial lethal time ($LT_{50}$) were 0.5 days, 2.6 days and 2.5 days respectively. Mortality against fourth and fifth larvae were $83.3{\pm}6.2%$ and $86.0{\pm}5.7%$ and medial lethal time ($LT_{50}$) were 4.2 days and 3.6 days respectively. Mortality against pupae and eggs of S. exigua were 100%. M. anisopliae FT83 showed high virulence at all developmental stages of S. exigua.

• Mycobiology
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• v.40 no.2
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• pp.111-116
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• 2012

Ten strains of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana were evaluated to find the most effective strain for optimization studies. The first criterion tested for strain selection was the mortality (> 50%) of Spodoptera litura larvae after inoculation of the fungus for 4 days. Results on several bioassays revealed that B. bassiana BNBCRC showed the most virulence on mortality S. litura larvae (80% mortality). B. bassiana BNBCRC also showed the highest germination rate (72.22%). However, its conidia yield ($7.2{\times}10^8$ conidia/mL) was lower than those of B. bassiana B 14841 ($8.3{\times}10^8$ conidia/mL) and M. anisopliae M6 ($8.2{\times}10^8$ conidia/mL). The highest accumulative radial growth was obtained from the strain B14841 (37.10 mm/day) while the strain BNBCRC showed moderate radial growth (24.40 mm/day). M. anisopliae M6 possessed the highest protease activity (145.00 mU/mL) while M. anisopliae M8 possessed the highest chitinase activity (20.00 mU/mL) during 96~144 hr cultivation. Amongst these criteria, selection based on virulence and germination rate lead to the selection of B. bassiana BNBCRC. B. bassiana B14841 would be selected if based on growth rate while M. anisopliae M6 and M8 possessed the highest enzyme activities.