Entomopathogenic fungi are natural pathogens of insects and contribute to the regulation of host insect populations in the environment. Several these fungi produce a wide range of secreted enzymes, secreted protein toxins and secondary metabolites to overcome host defenses and ultimately kill the host, and to defend host resources against competing pathogens and saprophytes. This study was performed to evaluate the antimicrobial activity of 207 entomopathogenic fungi form Korea soils against plant pathogenic bacteria Ralstonia solanacearum and plant pathogenic fungi Botrytis cinerea using dual culture technique on SDYA. As results, twelve isolates (5.7%) and six isolates (2.8%) showing the greatest inhibition against R. solanacearum and B. cinerea, respectively. The culture supernatant of these selected isolates completely suppressed the growth of the pathogen, indicating that suppression was due to the presence of antimicrobial compound in the culture filtrate. The stability test of the culture filtrate showed that the antimicrobial component was heat stable and not protein. These entomopathogenic fungal metabolites may be a good feature to be used in the development of a new biocontrol method of R. solanacearum and B. cinerea.

To collect entomopathogenic fungi we sampled soils from two different cultural types of farm which are conducting conventional culture and organic culture, and non-crop land. Forty-seven soil samples were collected from 9 provinces. Among them, 7 samples were collected from non-crop land, 6 samples were from organic farm and 34 samples were from conventional culture farm. Insect-bait method using Galleria mellonella and agar plating method were conducted to collect entomopathogenic fungi. Collection of entomopathogenic fungi using insect-bait method was highest in non-crop land as 7.1 mycotic cadavers per soil sample followed by organic farm with 3.6 and conventional farm with 3.3. Result from agar plating method showed the highest colony forming unit (cfu) was in organic farm followed by non-crop land, but cfu in conventional cultivating farm was a half compared with the non-crop land.

Entomopathogenic fungi have high potential in controlling harmful pests in agriculture and forest, but their slow progress in insect killing and low thermotolerance are major impediments to successful industrialization. Two possible efforts were done to overcome these problems. First the use of supernatant of a Beauveria bassiana isolate reduced the population of cotton aphid with a dosage-dependant manner, which allowed a quality control factor to be determined for the evaluation of the supernatant as the first step of a development. Chitinase was one of the major pathogenesis-related enzymes in the supernatant. A mineral material-mediated chitinase precipitation method was established to enhance the thermotolerance of chitinase. The use of supernatant can be a quicker way to control aphids. Secondly, to increase thermotolerance the effects of nutritional manipulation of culture media and oil-coating of conidia of B. bassiana and Metarhizium anisopliae isolates were investigated, followed by pairing of similar isolates. Recently, thermo-susceptible B.bassiana mutants were generated by a fungal transformation platform to discover thermotolerance-related genes, which are now underdetermination. Genetic information on fungal thermotolerance is going to be clear in near the future.

The green peach aphid, Myzus persicae Sulzer, is one of the most important pests affecting protected and open-grown crops, because they cause direct damage by feeding on crops and indirect damage as virus vectors. It has recently become a serious problem because of the continuous use of insecticide resulting in resistance among green peach aphid population. Thus, the development of entomopathogenic fungi as aphid biocontrol agents has received increasing interest as part of integrated control strategies. In this study, we report the screening result of pathogenic fungi for the control of green peach aphid. Initial screenings were performed using 347 isolates of putative pathogenic fungi from Korea soils. As results, 20 isolates of entomopathogenic fungi were isolated from cadavers of green peach aphid supporting fungal conidiation. These isolates were identified as three strains of Lecanicillium attenuatum, nine strains of Beauveria bassiana, one strain of Metarhizium anisopliae, one strain of Metarhizium flavoviride, five strains of Paecilomyces lilacinus, one strain of Aspergillus sp. by microscopic examination, genetic sequencing of the ITS region and Universally Primed PCR (UP-PCR). Based on the screening results, twenty isolates were tested for their pathogenicity against adult green peach aphid. All fungal isolates were pathogenic to green peach aphid but mortality varied with isolates. These entomopathogenic fungi may be useful to develop eco-friendly insecticide to control green peach aphid.

Recently, the Q biotype of tobacco whitefly has been recognized as the most hazardous strain of worldwide because of increased resistance to some insecticide groups requiring alternative strategies for its control. We studied the susceptibility of this biotype of B. tabaci to 21 isolates of Beauveria bassiana, 3 isolates of Isaria spp., 1 isolate of P. cateni，3 isolates of Lecanicillium lecanii, 1 isolate of L. attenuatum and 1 isolate of Aschersonia aleyrodes. These isolates were evaluated on pruned egg plant seedlings, at a concentration of 108conidia/ml (deposited at 6000±586 conidia mm-2). The mortality based on mycosis varied from 18 to 97% after 6 days. An Isaria sp. isolate, two B. bassiana isolates and a L. lecanii isolate were found the most effective. Furthermore, five isolates were chosen for concentration-mortality response assays and compared to B. bassiana GHA as a standard. The numbers of nymphs infected by fungi were correlated with the spore concentration. A L. lecanii isolate and Isaria sp. had the short LT50 at 6000±586 conidia mm-2. Mortality reduced and LT50s were longer as the concentration of conidia was reduced. The LD50 values were differed in isolates from 87 to 269 conidia mm-2. These results indicated that the Q biotype of sweetpotato whitefly was susceptible to the five isolates of entomopathogenic fungi and these isolates have potential to be developed as a microbial pesticide for whitefly control.

The entomopathogenic fungi were an important natural pathogenic of insects that has been developed as potential biological control agents for many important agricultural, forest and medical pests. Several these fungi produce a wide range of secondary metabolites with high therapeutic value as antibiotics, cytotoxic substances, insecticides, compounds that promote or inhibit growth, attractor and repellent. Therefore, this study was performed to select the antibacterial activity of liquid culture filtrates of 347 entomopathogenic fungi form Korea soils against two pathogenic bacteria including Ralstonia solanacearum and Escherichia coli using novel method which represents a quick and easily applicable tool obtaining large number of samples. As results, eight-five strains (24%) and seventy-six strains (22%) of these fungal metabolites produced anti-R. solanacearum and anti-E. coli compounds, respectively. The preferential antibacterial activity against R. solanacearum and E. coli gives evidence that these entomopathogenic fungal metabolites might be useful as an agent for bacteria control and the technique was simple to operate and allowed a large number of samples to be handled concurrently.

Reported herein is the use of mycotized millet grains with entomopathogenic fungi applied to soil to control western flower thrips (WFT), Franklinella occidentalis. The majority of mature larvae move from plants to soil for pupation. Mycotized millet grains with Beauveria bassiana ERL1170 and Metarhizium anisopliae ERL1171 were mixed in the upper layer of potting soil containing a blossomed marigold as a banker plant. B. bassiana GHA, a commercial isolate, served as a control. One week post applicaion 3 mated WFT females were added per plant. At 8 wks post infestation WFT mortality in pots with ERL1170 and ERL1171 treatments was 90% and 81% respectively compared to GHA with 15% mortality. Plants in the two ERL treatments had lower levels of damage. The insecticidal activities were positively associated with the fungal soil colonization. No significant release of fungal innoculum from the potting medium was observed. These results suggest that soil applications of entomopathogenic fungi can reduce WFT populations to significantly low levels by targeting pupae. Application timing and the life cycle of WFT are important considerations.

Cultural characteristics Lecanicillium lecani Btab01 and its insecticidal activity against tobacco whitefly (Bemisia tabaci) were investigated. On potato dextrose agar, tryptic soy agar and SDA+Y media, mycelial growth of L. lecani Btab01 was best at 20~25℃ and suppressed above 28℃. Both solid culture and liquid culture of L. lecani Btab01 showed high insecticidal activity, 93.9 and 98.3% respectively, against nymph of tobacco whitefly, but there is no significant difference. When culture of L. lecani Btab01 was treated at the concentration of 10⁵, 10⁶, 10⁷ and 10⁸ cfu/ml, their insecticidal activity were 5.8%, 33.8%, 77.3% and 98.5% respectively, and LT<SUB>50</SUB> values were 16.1 days, 7.3 days, 5.1 days and 3.5 days respectively. When nymphs were treated by the cultures of L. lecani Btab01 and maintained under saturated condition for zero hour, 24 hours and 168 hours, their control activities were 0%, 20.3% and 100% respectively. Spore germination of L. lecani Btab01 was increased about two times by adding edible oil. When L. lecani Btab01 was treated to control nymph with 0.1% edible oil, it showed high control activity (98.6%) compared to single treatment of L. lecani Btab01 (79.9%).

In agricultural fields, the entomopathogenic fungal species have been investigated for their potential as the biological control agents due to their role of natural enemies for insects. To address the requirements of a potential South Korea based biocontrol effort using entomopathogenic fungi, we investigated the occurrence of various entomopathogenic fungi in 1080 soil samples representing from various area and locations in South Korea. Entomopathogenic fungi were isolated from soils using semiselective medium SDA-D50 contained saboraund dextrose agar, 50 ug/ml dodine, 100 ug/ml chloramphenicol and 50 ug/ml streptomycin. The isolated putative fungi were identified by the determination of internal transcribed spacer (ITS) region sequences of the nuclear ribosomal analysis. As a result, entomopathogenic fungi were found to occur in 30.8% of the soil samples studied. The most abundant species were Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae (Metschn.) Sorok. Isolates of B. brongniartii, Cordyceps sp., Lecanicillium sp., Isaria sp. and Tolypocladium cylindrosporum were also found. The occurrence of entomopathogenic fungi was analyzed by the area and soil types. These positive entomopathogenic fungi may have potential against variety pests in agriculture and forest

In agricultural fields, the entomopathogenic fungal species have been investigated for their potential as the biological control agents due to their role of natural enemies for insects. To address the requirements of a potential South Korea based biocontrol effort using entomopathogenic fungi, we investigated the occurrence of various entomopathogenic fungi in 1080 soil samples representing from various area and locations in South Korea. Entomopathogenic fungi were isolated from soils using semiselective medium SDA-D50 contained saboraund dextrose agar, 50 ug/ml dodine, 100 ug/ml chloramphenicol and 50 ug/ml streptomycin. The isolated putative fungi were identified by the determination of internal transcribed spacer (ITS) region sequences of the nuclear ribosomal analysis. As a result, entomopathogenic fungi were found to occur in 30.8% of the soil samples studied. The most abundant species were Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae (Metschn.) Sorok. Isolates of B. brongniartii, Cordyceps sp., Lecanicillium sp., Isaria sp. and Tolypocladium cylindrosporum were also found. The occurrence of entomopathogenic fungi was analyzed by the area and soil types. These positive entomopathogenic fungi may have potential against variety pests in agriculture and forest

Sweet potato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is a serious pest of many economically important crops. The insect has developed resistance to chemical insecticides. Therefore, the development of microbial agent such as entomopathogenic fungi, Lecanicillium lecanii is necessary. Spores of L. lecanii Btab01 was collected after cultivation on solid PDA and liquid RB (rice bran amended with 2% molasses) media. The bioassay was carried out with B. taabci nymphs for 7 days at 25℃ and 60% relative humidity. Further, mortality was corrected with appropriate controls. The results revealed that spores obtained from RB medium caused high mortality (98.31%) compared to PDA medium (93.94%). Spore concentrations 105, 106, 107, and 108 colony forming units (c. f. u) ml-1 caused 5.81, 33.80, 77.27, and 98.54% mortality, respectively. The mortality (100%) was observed for 4 days when L. lecanii spores was mixed with 0.1 - 0.3% soybean oil. Hence, it is concluded from this study, L. lecanii Btab01 cultivated on RB medium can be recommended to control the nymphs of B. tabaci. Spore suspension can be expected to high efficacy when soybean oil was blended.

In agricultural fields, the entomopathogenic fungal species have been investigated for their potential as the biological control agents due to their role of natural enemies for insects. Until recent times, most of the studies for these fungi have been based on isolation from insect cadaver or soil. However, these methods, especially isolation from soil, might cause a problem involving differential isolation of the each entomopathogenic fungi. The purpose of this study is to determine the optimal isolation medium for entomopathogenic fungi using dodine, chitin, and skim milk. The growth rates of entomopathogenic fungi and non-entomopathogenic fungi were compared on dodine agar medium. The medium for this experiment was modified Veen semiselective medium which consisted of based on SDA (Sabouraund Dextrose Agar), 100 mg/ml for chloramphenicol, 50 mg/ml for streptomycin and adjusted dodine to 40, 50, 70 and 100 mg/ml. As a result, optimal concentration of dodine for isolation of entomopathogenic fungi was 50 mg/ml and 168 positive entomopathogenic fungi were isolated in 470 soil samples and 11 cadavers of insect. In addition, the isolates had significantly greater chitinase and protease activity than non-entomopathogenic fungi. The isolation method described represents a valuable tool for rapid and simple isolation of entomopathogenic fungi. These positive entomopathogenic fungi may have potential against variety pests in agriculture.

This study was conducted to investigate the total phenol contents, antioxidative activities and antibacterial activities of twenty species of entomopathogenic fungi. The total phenol content was highest in Aspergillus flavus (553.0±52.15 ㎍/g) and A. parasiticus (529.9±60.10 ㎍/g). On the other hand those in other strains were within the range of 26.6～121.9 ㎍/g. The antioxdative activity was shown in the most of strains and the highest DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging activity was observed in A. flavus (90.9±2.90%) and A. parasiticus (77.9±4.13%). This result indicated that the antioxidative activities were very correlated with the total phenol contents. The antibacterial activitiy was found in the every tested pathogenic bacteria. Especially, the antibacterial activity was strongest against Listeria monocytogenes and Escherchia coli.

Among the cultured products of Beauveria bassiana SFB-205 (KCCM 10892P), the supernatant showed the highest insecticidal activity against 2nd instars of Aphis gossypii (Aphididae) nymphs under glasshouse condition. The enzymes in the supernatant were confirmed as active materials, and the chitinase was finally selected as a QC factor for commercial production. However, the chitinase activity in the supernatant decreased by 11-folds due to the thermal stress at 50℃ for 2 h. To obtain thermal stability, the chitinase in the supernatant was adsorbed to a precipitable material and the pellet was freeze-dried (PCT/KR2007/005886). The adsorbent-A showed about 92.7% of harvesting efficiency which was higher than the other candidates. The chitinase activity of the freeze-dried powder was kept up about 82.0% of initial activity for the same thermal stress. Subsequently, an optimal formulation recipe was established to maximize long-term storage stability and efficacy. SFB-205 oil-based formulation was stable up to 18 months at room temperature. It showed 96.1% efficacy against 2nd instars of A. gossypii nymphs at 1 day after the treatment in the glasshouse. This novel approach can be a promising method to develop competitive biopesticies in the entomopathogenic fungi, even though it needs to be intensively studied.

농약이 한국산 곤충병원성곰팡이 Beauveria bassiana와 Metarhizium anisopliae에 미치는 영향을 알아본 결과, SMAY배지상에서의 농약농도에 따른 곰팡이 포자형성수는 metalaxyl과 tolclofos-methyl 처리에서는 대조구와 큰 차이가 없었으나. fenitrothion과 mepronil 처리에서는 권장량의 배 처리에서도 포자형성이 미약하였다. 농약과의 동시 처리가 곤충병원성 곰팡이에 미치는 영향을 알아보기 위하여 농약희석액에 병원성 곰팡이를 1-5시간 동안 침지시켜 병원성을 알아본 결과 전 약종에서 100%의 병원성을 나타내었다. 배추밭에서 농약전처리가 M. anisopliae의 병원성에 끼치는 영향을 알아본 결과는 Bt나 teflubenzuron, metalaxyl, carbofuran 처리구는 대조구와 병원성에서 차이를 보이지 않았으나 alachlol 처리구에서는 병원성이 80% 이하로 떨어졌다. 잔디밭에서 농약의 후처리가 병원성곰팡이의 병원성과 지속성에 미치는 영향을 알아본 바 iprodione+thiram과 tolclofos-methyl 처리구에서는 4달째까지 병원성이 지속되었고 fenitrothion처리구에서는 3개월 후부터 B. bassiana와 M. anisopliae의 병원성이 상실되었으며 mepronil처리구에서는 M. anisopliae가 병원성을 상실하였다.

우리나라의 곤충병원성곰팡이 분포를 조사하기 위하여 5개도와 3개 광역시에서 200개소의 토양 sample을 채집하여 꿀벌부채명나방 유충을 이용하여 분리하였다. 8종의 곤충 사체로 부터는 직접 분리하였다. 그 결과 40개소(20.0%)의 토양과 8종의 곤충 사체에서 곰팡이가 분리되었는데, 토양에서 분리된 균종별로는 Bequveria가 21개소(10.5%), Metarhizium이 19개소(9.5%)이었다. 그외 7종의 곤충에서 Beauvria가 분리되었다. 서식처별로는 산림 110개소중 25개소(22.7%), 경작지 20개소중 2개소(11.0%), 휴한지 11개소중 6개소(54.5%), 하천 제방 19개소중 7개소(36.8%)에서 곤충병원성곰팡이가 검출되었다. 한편, 곤충 사체에서는 긴수염 대벌레에서 Metarhizium이 분리된 것을 제외하고는 모두 Beauveria가 분리되었다.