생물적 방제균 Pseudomonas chlororaphis O6 길항 물질 생산 및 유기산 흡수에 관련된 dctA 유전자의 클로닝

한송희(Song-Hee Han),남효송(Hyo-Song Nam),강범룡(Beom-Ryong Kang),김길용(Kil-Yong Kim),구본성(Bon-Sung Koo),조백호(Baik-Ho Cho),김영철(Young-Cheol Kim) 한국토양비료학회 2003 한국토양비료학회지 Vol.36 No.3

생물적 방제균 Bacillus amyloliquefaciens LM11의 유래 생물계면활성물질과 항균활성과의 상관관계

강범용,김용환,남효송,김영철,Kang, Beom Ryong,Kim, Yong Hwan,Nam, Hyo Song,Kim, Young Cheol 한국식물병리학회 2017 식물병연구 Vol.23 No.2

장수풍뎅이 유충의 장내세포에서 분리한 Bacillus amyloliquefaciens LM11은 surfactin, iturin, fengycin 같은 biosurfactants lipopeptide를 생산하여 식물병원성 곰팡이의 성장을 강하게 억제하였다. LM11균주 성장단계에 따라 biosurfactant 생산과 surface tension은 상당히 유의한 차이가 있었다. 항균 물질인 surfactin, iturin, fengycin의 생합성 유전자는 정지기에 도달하면서 집중적으로 발현되었고 그 생산량도 높았다. 또한 LM11균주를 제거한 배양 상등액 함량의 농도에 따라 고추 탄저병원균의 포자발아와 높은 부의 상관관계가 있었다(R=0.761, P<0.001). 식물병원성 곰팡이의 균사 생장억제를 위한 최소 surface tension 수준은 38.5 mN/m였다(R=0.951-0.977, P<0.001). 본 연구 결과는 B. amyloliquefaciens LM11의 biosurfactant가 식물병에 대한 생물학적 방제에 중요한 항진균 대사물질로 작용하며, 배양액의 surface tension 측정은 생물학적 방제제의 최적 사용을 위한 기초 지표로 사용될 수 있음을 보여 주었다. Bacillus amyloliquefaciens LM11 was isolated from the feces of larvae of the rhino beetle and showed strong antifungal activities against various phytopathogenic fungi by producing biosurfactants. In this study, our overall goal was to determine relationship between biosurfactants produced from the LM11 strain and its role in growth inhibition of phytopathogenic fungi. Production and expression levels of B. amyloliquefaciens LM11 biosurfactants were significantly differed depending on growth phases. Transcriptional and biochemical analysis indicated that the biosurfactants of the LM11 strain were greatly enhanced in late log-phase to stationary phase. Inhibitions of phytopathogenic mycelial growth and spore germination were directly correlated (P<0.001, R=0.761) with concentrations of the LM11 cell-free culture filtrates. The minimum inhibitory surface tension of the culture filtrate of the B. amyloliquefaciens LM11 grown in stationary phase to inhibit mycelial growth of the phytopathogenic fungi was 38.5 mN/m (P<0.001, R=0.951-0.977). Our results indicated that the biosurfactants of B. amyloliquefaciens LM11 act as key antifungal metabolites in biocontrol of plant diseases, and measuring surface tension of the cell-free culture fluids can be used as an easy indicator for optimal usage of the biocontrol agents.

The Plant Growth Promotion and Induction of Tolerance to Salinity Stress by Enterobacter intermedium 60-2G in Red Pepper Plants

Ga Ram Baek(백가람),Hyo Song Nam(남효송) 한국토양비료학회 2023 한국토양비료학회 학술발표회 초록집 Vol.2023 No.10

LC-MS/MS를 이용한 멀구슬, 협죽도, 황련 유래 활성성분의 벼 중 잔류양상 연구

박준성 ( Joon Seong Park ),남효송 ( Hyo Song Nam ),김용환 ( Yong Hwan Kim ),김도익 ( Do Ik Kim ),김선암 ( Sun Am Kim ) 한국환경농학회 2015 한국환경농학회지 Vol.34 No.2

BACKGROUND: Plant extracts have been used as environment friendly agricultural materials for organic farming in South Korea. However safety evaluation on the plant extracts was not properly tested. The aim of this study was to evaluate safety of the extracts from Melia azedarach, Nerium indicum and Coptis chinensis on cultivating rice. METHODS AND RESULTS: Pant extarcts 300-fold diluted were treated on rice, and residues of M. azedarach, N. indicum and C. chinensis were determined. The analytes from the rice samples were detected by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The method was validated, and good linearities (r2=0.995-0.998), specificity, and recoveries were obtained. Limits of detection were 0.01 mg/kg for all of the target compounds. Recoveries were 79.3-118.3% at 0.1 mg/kg and 75.2-111.5% at 0.5 mg/kg. The residue levels were below 0.030 mg/kg for azadirachtin, 0.320 mg/kg for oleandrin and 1.460 mg/kg for berberine. CONCLUSION(S): The extracts of M. azedarach, N. indicum and C. chinensis contained azadirachtin, oleandrin and berberine as an active ingredient, respectively. The residue of three active ingredients dramatically decreased after treatment in all fruits, stems and roots of rice.

농업용 저수지에서 Bisphenol A의 잔류실태조사

조일규 ( Il Kyu Cho ),남효송 ( Hyo Song Nam ),전용배 ( Yongbae Jeon ),박준성 ( Jun Sung Park ),나태웅 ( Tae Wong Na ),김백종 ( Back Jong Kim ),( Eunsung Kan ) 한국환경농학회 2016 한국환경농학회지 Vol.35 No.4

BACKGROUND: Significant amount of bisphenol A has been released from the manufacturing process of plastics, epoxy resins, dental material and flame retardants. Bisphenol A has been detected at trace levels in wastewater, surface water, landfill leachate and drinking water. However, the residual survey of bisphenol A has not been performed in agricultural reservoir beside agricultural environment cultivating crops. This study was conducted to monitor the residual bisphenol A in national agricultural reservoirs and understand a level of contamination of bisphenol A in the agricultural environment in Korea. METHODS AND RESULTS: The water and water sediment were collected at agricultural reservoirs in Chungnam, Chungbuk, Kyunggi, Jeonnam, Jeonbuk, Kyungnam and Kyungbuk province. Bisphenol A was analyzed by the LC-MS/MS with triple quad 4500. The recovery of water and water sediment in the agricultural reservoirs showed the level of 95.7~97.2% and 91.5 ~100.3%, respectively. CONCLUSION: Bisphenol A was detected at the level of 0.05 μg/L~0.18 μg/L and 0.1 μg/kg~34 μg/kg in water and water sediment of the reservoirs, respectively. Based on the results, the residue of bisphenol A will be tried in the crops surrounding these agricultural reservoirs where bisphenol A detected above 10 μg/kg of bisphenol A.

토양에서 분리한 Bacillus Siamensis BC-S09의 항진균 활성

서민경 ( Min-kyoung Seo ),양현주 ( Hyun-joo Yang ),남효송 ( Hyo-song Nam ) 한국환경농학회 2021 한국환경농학회 학술대회집 Vol.2021 No.-

A bacterium having antifungal activity was isolated from soil in Gokseong-gun, South Korea and identified as Bacillus siamensis. Antifungal bacteria were isolated from various soil samples. The samples were suspended in phosphate-buffered saline (PBS), spread on Tryptic Soy agar (TSA) medium and allowed to grow at 30℃ for 24 h under aerobic condition. The strain BC-S09 exhibiting the highest antifungal activity was identified as Bacillus siamensis. The strain BC-S09 was identified by 16S rRNA gene sequence analysis after polymerase chain reaction (PCR) amplification using the universal primers 27F and 1492R as described Stackebrandt and Goodfellow. The 16S RNA sequences were compared using “16S-based ID” in the EzBioCloud (https://www.ezbiocloud.net) database. The phylogenetic analysis was performed using the neighbor-joining method in the MEGA program (https://www. megasoftware.net/). Various fungi were kept on potato-dextrose agar (PDA) slants at 25℃. Spore inoculums were prepared by growing the molds on 10-days-old PDA slants and collected after vigorously shaking slants with sterile peptone water (0.2% w/v). Bacterial isolates exhibiting antifungal activity were observed directly on PDA agar plates where the indicator molds test microorganisms were placed with control and experimental group. Bacteria and fungi was incubation of 25℃ for 2 day to 7 day in aerobic incubator. The results showed that Bacillus siamensis BC-S09 suppressed the growth of phytopathogenic fungi including Colletotrichum coccodes (KACC40010), Colletotrichum acutatum (KACC40689), Colletotrichum acutatum (KACC40804 - strong virulence), Altermaria citri (KACC42128), Fusarium oxysporum, Rhizoctonia solani.

탄소 및 질소 공급원과 배양 배지의 비율을 변경하여 Xanthomonas Oryzae pv. Oryzae에 대한 항균 활성을 가진 Bacillus velezensis의 포자 생산 개선

양현주 ( Hyun-joo Yang ),서민경 ( Min-kyoung Seo ),최제연 ( Je Yeon Choi ),남효송 ( Hyo-song Nam ) 한국환경농학회 2021 한국환경농학회 학술대회집 Vol.2021 No.-

Xanthomonas oryzae pv. oryzae (KACC10208) was the causative agent of bacterial blight in rice. Bacillus velezensis has antimicrobial activity to Xanthomonas oryzae pv. oryzae (KACC10208) and antifungal activities Colletotrichum coccodes (KACC40010), Colletotrichum acutatum (KACC40689). The purpose of this study was to develop an economical culture medium for the optimal cell growth and spore forming by submerged fermentation of B. velezensis. B. velezensis was grown in Lysogeny broth(LB) or in Medium Optimal for Antibacterial activity(MOA) at 30℃ and 150 rpm. Antifungal activity were observed directly on PDA agar plates where the indicator molds test microorganisms were placed with control and experimental group. The incubation condition was 25℃ for 48 h to 72 h in aerobic incubator. Experiments were performed at three tines with biological replicates. The data were analyzed using one-way analysis of variance(ANOVA) followed by Duncan’s multiple range test (P<0.05). The antimicrobial activity of the cell-free supernatant of Bacillus velezensis strain grown on each carbon and nitrogen source against the phytopathogenic Xanthomonas oryzae pv. oryzae (KACC10208) was tested on TSA plates using the disc diffusion method. The optimal carbon and nitrogen sources were determined as sucrose and soy bean flour, respectively. Additionally, MgSO4 was significantly involved in the growth and sporulation of Bacillus velezensis in addition to carbon and nitrogen sources. The results showed that bacterial cells of 1.0 x 109cfu/m and sporulation yield of 90%.

벼 세균성 흰잎마름병(Xanthomonas Oryzae pv. Oryzae)의 생물학적 방제를 위한 미생물 사용방법

정종국 ( Jong-kook Jung ),서민경 ( Min-kyoung Seo ),양현주 ( Hyun-joo Yang ),구헌우 ( Heon-woo Ku ),남효송 ( Hyo-song Nam ) 한국환경농학회 2021 한국환경농학회 학술대회집 Vol.2021 No.-

The purpose of this study is to develop eco-friendly microbial agents and application technologies for the safe production of rise by control of rice Bacterial Blight (Xanthomonas oryzae pv. oryzae). Rice seeds (Oryza sativa cv. ShinDongjin) were inoculated with 3 types of microorganisms (Bacillus amyloliquefaciens KB3, Bacillus velezensis LM11, Serratia spp IM2), chemical disinfectant (tebuconazole 12.5 + prochloraz copper chloride 12.5%) and Trichoderma at 25°C for 24 hours. After inoculation, the rice seeds were washed twice with distilled water, placed on PDA medium, and incubated at 25°C for 48 hours. As a result of the culture, microorganisms causing white leaf blight were cultured in the rice seed treated with chemical disinfectant, and each strain was cultured in the rice seed inoculated with Bacillus amyloliquefaciens KB3, Bacillus velezensis LM11, and Trichoderma. The germination rate of the seed inoculated with each sample was confirmed. In the case of rice seeds treated with microorganisms and Trichoderma, the germination rate was about 3.5 times higher than that of chemical disinfectants. As a result of checking the growth level of each sample 2 weeks after sowing, it was confirmed that the Bacillus velezensis LM11 inoculated sample had a higher growth rate than other samples. Each sample was transplanted into a Wagner pot and growth was investigated 2 weeks after transplantation. As a result, the sample inoculated with Bacillus velezensis LM11 showed a higher growth rate than the other samples, and the rest of the samples grew similarly. Each rice seed was immersed in the sample to infect the grown rice with Xanthomonas oryzae pv. oryzae, which causes white leaf blight. As a result of infection, it was confirmed that the infection rate was the lowest in Bacillus velezensis LM11.

포도 과수원에서 살균제 Difenoconazole의 농작업자 노출량 측정

조일규 ( Ll Kyu Cho ),박준성 ( Joon Seong Park ),박소현 ( So Hyun Park ),김수진 ( Su Jin Kim ),김백종 ( Back Jong Kim ),나태웅 ( Tae Wong Na ),남효송 ( Hyo Song Nam ),박경훈 ( Kyung Hun Park ),이지호 ( Jiho Lee ),김정한 ( Jeong-ha 한국환경농학회 2016 한국환경농학회지 Vol.35 No.4

BACKGROUND: 18% of difenoconazole+iminoctadin triacetate microemulsion (3%+15%) formulation were mixed and sprayed as closely as possible to normal practice on the ten of farms located in the Youngju of South Korea. Patches, cotton gloves, socks, masks and XAD-2 resin were used to measure the potential exposure for applicators wearing standardized whole-body outer and inner dosimeter (WBD). This study has been carried out to determine the dermal and inhalation exposure to difenoconazole during preparation of spray suspension and application with a power sprayer on a grape orchard. METHODS AND RESULTS: A personal air monitor equipped with an air pump IOM sampler and cassette and glass fiber filter were used for inhalation exposure. The field studies were carried out in a grape orchard. The temperature and relative humidity were monitored with a thermometer and a hygrometer . Wind speed was measured using a pocket weather meter. All mean field fortification recoveries were between 97.3% and 119.6% in the level of 100 LOQ (limit of quantification) while the LOQ for difenoconazole was 0.025 μg/mL using HPLC-UVD. The arms exposure to difenoconazole for the mixer/loader (0.0794 mg) was higher than other body parts (head, hands, upper body, legs). The exposure to difenoconazole in the legs for applicator (3.78 mg) was highest in the parts of body. The dermal exposure for mixer/loader and applicator were 0.02 and 2.28 mg on a grape orchard, respectively. The inhalation exposure during application was estimated as 0.02 mg. The ratio of inhalation exposure to dermal exposure was equivalent to 0.9% of the dermal exposure. CONCLUSION: The inhalation exposure for applicator indicated 18.8×10^-3 mg, which was level of 0.9% of the dermal exposure (2.28 mg). Operator exposure (0.004 mg/kg bw/day) to difenoconazole during treatment for grape is calculated as 2.5% of the established AOEL (0.16 mg/kg bw/day).