• Mycobiology
• /
• v.45 no.2
• /
• pp.84-89
• /
• 2017

Fungi of the Metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. To establish a co-transformation system for the transformation of multiple M. robertsii genes using Agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. Subsequently, in the two vectors containing the nourseothricin and phosphinothricin resistance cassettes were inserted eGFP and mCherry expression cassettes, respectively. These new vectors were then introduced independently into A. tumefaciens and used to transform M. robertsii either in independent events or in one single co-transformation event using an equimolar mixture of A. tumefaciens cultures. The number of transformants obtained by co-transformation was similar to that obtained by the individual transformation events. This method provides an additional strategy for the simultaneous insertion of multiple genes into M. robertsii.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.41 no.2
• /
• pp.28-35
• /
• 2020

Entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide, and have high potential in industrialization. However, their thermo-susceptibility limits long-term storage under high temperature conditions and insecticidal activity after application in target pests. The virulences of M. anisopliae JEF-003 and JEF-004, and B. bassiana JEF-006 and JEF-007 against bean bug (Riptortus pedestris), and the thermotolerance of conidia produced on three kinds of grains, sorghum, millet and Italian millet as substrates for solid cultures were investigated. Of the three grains, Italian millet was the suitable grains in the production of thermotolerant conidia, and a significant relationship between conidial thermotolerance and the media was demonstrated. This work suggests that biological characteristics of entomopathogenic fungi are altered by culture media, and these results will be a chance to understand the factors in the media which make such changes.

• Journal of Microbiology and Biotechnology
• /
• v.8 no.3
• /
• pp.284-286
• /
• 1998

A ura5 gene encoding Orotate Phosphoribosyl Transferase (OPRTase) of Metarhizium anisopliae was cloned by PCR methods and sequenced. The sequenced ura 5 gene encodes a polypeptide of 234 amino acid residues. This deduced amino acid sequence showed high similarity to other fungi OPRTase and there was no intron sequence between ATG starting codon and TGA ending codon.

• Korean Journal of Microbiology
• /
• v.36 no.2
• /
• pp.97-102
• /
• 2000

In order to use Metarhizizmn~ anisopliae as a biological pesticide, effect of envrionmental factors on nlycelial growth, spore formation, and extracellular enzyme activity in culture broth of M. anisopliae DGUM 35001 was determined. Optimal temperature was 26^{\circ}C.$ and optimal pH ranged from 5 to 9. Among the complex media tested, MCM and SDPY media were the most favorable for mycelial growth. When Czapek-Dox agar was used as a mnimal medium, glucose and sucrose among the saccharides were very excellent source of carbohydrate. Among the biopolyners tested. chitin was the most favorable source for mycelial growth and produced high aerial inycelia. Urea and ammonium phosphate as an inorganic nitrogen source and bacto-peptone and soytone as an organic nitrogen source enhanced the mycelial growth When serine as a source of amino acid was supplemented, excellent mycelial growth was shown. Large amount of spores could be obtained from the aerial mycelia of starch medium. When the culture broth was filtrated and then the concentrate with ammonium sulfate was used as a crnde enzyme solution, high enzyme activities of amylase and protease were shown. However, lipase and chitinase activities were comparatively low.

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

• International Journal of Industrial Entomology and Biomaterials
• /
• v.37 no.1
• /
• pp.9-14
• /
• 2018

Bacterial species, Bacillus amyloliquefaciens and Lactobacillus species (L. sp.5-1), are known to inhibit the growth of pathogenic bacteria and fungi. Metarhizium anisopliae is a pathogenic fungal species which causes fatal damage to P. brevitarsis populations. Therefore, we investigated the inhibitory effect of B. amyloliquefaciens and L. sp. 5-1 on M. anisopliae induced fatality rates in P. brevitarsis. Samples of M. anisopliae-infected sawdust were treated with strain B. amyloliquefaciens KACC10116, strain L. sp. 5-1 KACC19351, and a combination of the two. P. brevitarsis were fed treated sawdust samples, and their subsequent fatality rate was monitored. The fatality rate fell below 1.5% after 10 days and decreased by approximately 40% after 15 days. On average, the fatality rate decreased by 20%, compared to the control. The difference in the decrease in fatality rate between B. amyloliquefaciens treatment and L. sp. 5-1 treatment was not significant. Results indicate that both strains exhibit high anti-fungal activity, which may be useful in environmental purification efforts. These strains may be used for effective prevention of fungal infection in P. brevitarsis.

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

• The Korean Journal of Mycology
• /
• v.49 no.3
• /
• pp.315-324
• /
• 2021

In this study, the fungal strains KNU-Gunwi 2B and KNU-SOT5 were isolated from root-soil in a hillside and the cherry tree bark (Prunus serrulata), respectively from Gyeongbuk province in Korea. The strain KNU-SOT5 produced dark brown chlamydospores that were smooth to lightly rough-walled, globose to ellipsoidal, and the conidia were aseptate, guttulate, mostly fusiform with a diameter of 5.3-17.6×4.2-7.0 ㎛. Strain KNU-Gunwi 2B produced phialides that were smooth-walled, cylindrical with semi-papillate apices and the conidia were pale-green, broadly ellipsoid, and sometimes cylindrical with a diameter of 4.4-8.0×2.3-4.0 ㎛. The strain KNU-SOT5 and KNU-Gunwi 2B were resolved based on cultural and morphological characteristics, along with the phylogenetic analysis using the small subunit (SSU), large subunit (LSU), and internal transcribed spacers (ITS) regions. The fungal strains KNU-SOT5 and KNU-Gunwi 2B were identified as Dothidea insculpta and Metarhizium rileyi, which have not been reported in Korea.

• The Korean Journal of Pesticide Science
• /
• v.17 no.4
• /
• pp.454-459
• /
• 2013

Beet armyworm, Spodoptera exigua is difficult to control using chemical insecticides because of the fast development of insecticide resistance. For eco-friendly beet armyworm managements, various control agents are required. Entomopathogenic fungus is one of the promise control agents as an alternative to chemical control agent. We isolated entomopathogenic fungi from soil samples of Yangpyeong, Gyeonggi-do by insect-bait method using Tenebio molitor and conducted bioassay to larva of beet armyworm. The result of bioassay, a selected strain FT83 showed 100% mortality against third instar larva of S. exigua. On the basis of morphological characteristics and analysis of 18srRNA sequence for ITS, the strain FT83 was identified as a Metarhizium anisopliae. The mortality of beet armyworm showed $81.6{\pm}9.3%$ at $1{\times}10^6$ conidia/ml, 100% at $1{\times}10^7$ conidia/ml and 100% at $1{\times}10^8$ conidia/ml respectively. Therefore, we recommend to proper control efficacy against S. exigua in which more than $1{\times}10^7$ conidia/ml suspension of M. anisopliae FT83.

• The Korean Journal of Mycology
• /
• v.43 no.2
• /
• pp.77-87
• /
• 2015

Insects are commonly infected by fungal diseases and are mostly susceptible to them. Increasing levels of insecticide resistance has recently become an issue for control programs; thus, research has focused on mosquito control using entomopathogenic fungi, including fungal pathogens such as Beauveria bassiana, Metarhizium anisopliae, and Lagenidium giganteum. Review discusses entomopathogenic fungi related to control programs for mosquito transmitted vector-borne diseases such as dengue, filariasis, malaria, and yellow fever, and how to use entomopathogenic fungi for mosquito control.