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Nam, Jin-Sik,Lee, Dong-Hun,Lee, Kang Hyun,Park, Hee-Moon,Bae, Kyung Sook 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-
Degenerated PCR primers were used to amplify chitin synthase genes from genomic DNA of Metarhizium anisopliae var. anisopliae. Through cloning and sequencing of approximately 600-bp fragments amplified by PCR, we found three genes encoding different types of chitin synthases, designated MaCHS1, MaCHS2, and MaCHS3. Southern blot analysis performed on genomic DNA showed that each of the chitin synthases MaCHS1, MaCHS2, and MaCHS3 if encoded by a single copy gene. Alignment of their deduced amino acid sequences with those of other euascomycetes separated the sequences into three distinct classes. MaCHS1 was identified as a gene for classⅠ chitin synthase, MaCHS2 for classⅡ, and MaCHS3 for classⅢ. The UPGMA dendrogram and phylogenetic tree of the deduced amino acid sequences revealed the taxonomic and evolutionary position of Metarhizium anisopliae var. anisopliae. ⓒ 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V.
살충성곰팡이 Metarhizium anisopliae의 ura5 유전자의 분리동정
박인철,이동규,강선철,황철원,Park, In-Cheol,Lee, Dong-Kyu,Kang, Sun-Cheol,Hwang, Cher-Won 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.43 No.1
환경친화형 생물농약개발을 위한 방안의 일환으로, 벼별구 등 농해충병원사상균 Metarhizium anisopliae의 분자생물학적 육종을 위해 영양요구성 돌연변이를 상보하는 선택유전자, ura5 (Orotate phosphoribosyl transferase)를 cloning하였다. Cloning방법으로는 기존에 알려진 사상균의 ura5 유전자들간에 확인된 상보성 염기배열을 합성하여, 이것을 primer로 사용하여 PCR기법에 의해 부분적으로 cloning하였다 또한, PCR기법에 의해cloning된 uras유전자단편의 염기배열을 결정한 결과, Trichoderma resei의 ura5유전자와는 아미노산수준에서 약 85%의 상동성을 나타내었으며, 이 단편을 이용하여 Metarhizium anisopliae의 genomic library로 부터 ura5유전자가 포함된 약 4.4 kb의 DNA단편을 cloning 하였다. 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.
M.T. Islam,D. Omar,M. Shabanimofrad 한국응용곤충학회 2014 Journal of Asia-Pacific Entomology Vol.17 No.3
The aimof this studywas to investigate the molecular identification and virulence of the entomopathogenic fungusMetarhizium anisopliae. Initially, the alignments of nucleotide sequences of the internal transcribed spacer(ITS) 1-5.8S-ITS4 regions of six isolates–TFFH1, TFFH3, PR1, MAP, GT3, and GT2–were investigated using GenBankITS sequences for the same region and species of fungus. Thereafter, the in vitro (germination, vegetative growth,and sporulation) and the in vivo (%mortality of Bemisia tabaci) virulence of the funguswere investigated. A BLASTsearch of the ITS1 region revealed a match to a sequence registered under the accession number forM. anisopliae. The submitted sequence datawere registered and providedwith the accession numbers JX041507 to JX0415012,which were released in August 2012 (GenBank). In the virulence experiments, the highest germination percentage,vegetative growth, and sporulation of M. anisopliae were observed in the isolate PR1, with mean values of97.2%, 4.6 cm, and 7.2 × 106 conidia/ml, respectively; while the highest mortality percentage of B. tabaci was observedin the isolate GT3,with an mean value of 84.3%. This study concludes that all the six tested isolates–TFFH1,TFFH3, PR1, MAP, GT3, and GT2–are M. anisopliae and the isolates PR1 and GT3 are more virulent to B. tabaci thanthat by the other four isolates.
김종철,Se Jin Lee,김시현,이미롱,Sehyeon Baek,박소은,김준헌,Tae Young Shin,Jae Su Kim 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.1
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 × 10 7 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.
Kwak, Kyu-Won,Nam, Sung-Hee,Park, Kwan-Ho,Lee, Heuisam,Han, Myung-Sae Korean Society of Sericultural Science 2018 International Journal of Industrial Entomology Vol.37 No.1
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.
Outbreak of insect diseases in domestic insect farms
Eunsun Kim,Jihyun Im,Han Mun Hee,Myung-Ha Song,Seokhyun Lee,Kwan-Ho Park,Wontae Kim,Ji-Young Choi 한국응용곤충학회 2017 한국응용곤충학회 학술대회논문집 Vol.2017 No.04
The frequency and time of the outbreak of insect diseases were investigated among the insect-rearing farms by regional groups. The purpose of this study was to predict the insect diseases and reduce the spread of diseases. From February to October 2016, 87 of diseased insects were collected in seven regions. It turned out that the pathogens for fungal diseases were identified as Metarhizium anisopliae. But in case of bacterial diseases, the pathogenicity of the bacteria isolated from the diseased insects were not confirmed. Bacterial diseases occurred mainly in June and August in Gyeonggi-do and Gangwon-do, while in Chungcheong-do, bacterial and viral disease raged. In Jeolla-do, both fungal and bacterial diseases occurred in June. Generally, bacterial diseases occurred mainly from June to October, while the frugal diseases occurred from January to March. It was concluded that the entomopathogenic fungi, Metarhizium anisopliae, tended to cause diseases during late-winter and early spring and bacteria, uncertain, was prone to cause diseases during summer. Therefore, it necessary to adequately control depending on the type of the entomopathogen.
Kook-Il Han,Young Ho Nam,Jum Oc Jung,Eunsun Kim,Mi-Hwa Lee 한국응용곤충학회 2023 한국응용곤충학회 학술대회논문집 Vol.2023 No.10
Bacillus velezensis TJS119 was isolated from the freshwater, and antagonistic activity against of pathogenic fungi. Strain TJS119 showed a broad spectrum of antagonistic activities many fungal pathogens, including the green muscardine fungus Metarhizium anisopliae. The whole-genome sequence of B. velzensis TJS119 was analyzed using the illumina platform. The genome comprises a 3,809,913 bp chromosome with a G + C content of 46.43%, 3,834 total genes, 10 rRNA and 73 tRNA genes. The genome contained a total of 8 candidate gene clusters (difficidin, fengycin, bacillaene, macrolactin, bacillibactin, bacilysin, surfactin and butirosin) to synthesize secondary metabolite biosynthesis. Overall, our data will aid future studies of the biocontrol mechanisms of B. velezensis TJS119 and promote its application in insect disease control.