• The Korean Journal of Mycology
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• v.32 no.1
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• pp.31-38
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• 2004

An endophytic bacterial strain EB215 that was isolated from cucumber (Cucumis sativus) roots displayed a potent in vivo antifungal activity against Colletotrichum species. The strain was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, and 16S rDNA gene sequence. Optimal medium and incubation period for the production of antifungal substances by B. cepacia EB215 were nutrient broth (NB) and 3 days, respectively. An antifungal substance was isolated from the NB cultures of B. cepacia EB215 strain by centrifugation, n-hexane partitioning, silica gel column chromatography, preparative TLC, and in vitro bioassay. Its chemical structure was determined to be pyrrolnitrin by mass and NMR spectral analyses. Pyrrolnitrin showed potent disease control efficacy of more than 90% against pepper anthracnose (Colletotrichum coccodes), cucumber anthracnose (Colletotrichum orbiculare), rice blast (Magnaporthe grisea) and rice sheath blight (Corticium sasaki) even at a low concentration of $11.1\;{\mu}g/ml$. In addition, it effectively controlled the development of tomato gray mold (Botrytis cinerea) and wheat leaf rust (Puccinia recondita) at concentrations over $33.3\;{\mu}g/ml$. However, it had no antifungal activity against Phytophthora infestans on tomato plants. Further studies on the development of microbial fungicide using B. cepacia EB215 are in progress.

• Microbiology and Biotechnology Letters
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• v.33 no.1
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• pp.16-23
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• 2005

In order to develop a new microbial fungicide using endophytic bacteria for the control of anthracnoses occurring on various crops, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth medium, their antifungal activities were tested for in vivo antifungal activity against cucumber anthracnose caused by Colletotrichum orbiculare. As the results, liquid cultures of 28 strains showed potent antifungal activities more than $90\%$ against cucumber anthracnose. At 3-fold dilutions of liquid cultures, 18 strains inhibited the development of cucumber anthracnose of more than $70\%$. They were further tested for in vivo antifungal activity against red pepper anthracnose caused by C. coccodes and in vitro antifungal activity against C. acutatum, a fungal agent causing red pepper anthracnose. Among 18 strains, a bacterial strain EB215 isolated from cucumber roots displayed the most potent antifungal activity against Colletotrichum species. It was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, Biolog test and 16S rDNA gene sequence. It also controlled effectively the development of rice blast (Magnaporthe grisea), rice sheath blight (Corticium sasaki), tomato gray mold (Botrytis cinerea), and tomato late blight (Phytophthora infestans). Studies on the characterization of antifungal substances produced by B. cepacia EB215 are in progress.

• Journal of Life Science
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• v.20 no.9
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• pp.1373-1377
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• 2010

Coastal sand-dune plants can survive very effectively with the help of various microbes, especially ecto- and endomycorrihizae. Penicillium citrinum KACC43900 was screened according to growth promoting activity on sand-dune plants Calystegia soldanella and Ischaemum anthephoroides. In this study, coastal sand-dune plants were treated with a fungal culture filtrate of endophytic fungus P. citrinum KACC43900 to confirm the promotion of plant growth. C. soldanella and I. anthephoroides were used as representative coastal sand-dune plants, and their shoot length, plant length, and dry weight were analyzed. The shoot lengths of control C. soldanella and I. anthephoroides were 8.60 cm and 8.12 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 16.30 cm and 10.56 cm, respectively. The plant lengths of control C. soldanella and I. anthephoroides were 14.90 cm and 14.80 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 24.06 cm and 17.06 cm, respectively. The dry weight of C. soldanella and I. anthephoroides treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 0.163 g and 0.032 g, respectively. It was shown that the growth of shoots in C. soldanella and I. anthephoroides was promoted, 89.54% and 29.60%, by culture filtrate of P. citrinum KACC43900.

• BMB Reports
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• v.40 no.5
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• pp.773-782
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• 2007

Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 ${\mu}g/mL$ and 5 ${\mu}g/mL$, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil.

• Korean Journal of Environment and Ecology
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• v.28 no.3
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• pp.293-301
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• 2014

15 artificial ponds in Seoul have been examined to find out the characteristics of Odonata community according to age and size of pond. We found 6 Families 36 Species of Odonata community, and Libellulidae was the most frequent and 19 Species were observed among Libellulidae. Considering relative importance with frequency of appearance and density, Ischnura asiatica and Orthetrum albistylum was the most frequent. Also, 16 Species including Lestes temporaris, Sympetrum parvulum, and Anax nigrofasciatus were relatively rare. The result of dominant species tells that Ischnura asiatica in 1 ~ 3 year old ponds, Pantala flavescens-Orthetrum albistylum-Crocothemis servilia mariannae in order of mention in 4 ~ 6 year old ponds, Platycnemis phyllopoda-Ischnura asiatica in order of mention in 10 or more year old ponds are the dominant species. Species diversity index regarding age of pond has been researched, and species diversity index in 10 year old pond was higher than that of in 1 ~ 3 year old pond. Having done the analysis of variance for types of spawn, there are many species Endophytic egg-layers and pasting in mud or sand in 10 year old artificial ponds. The wider ponds get, the higher species diversity index gets. And, we suggest that desirable size for making artificial ponds to increase the species diversity index of Odonata is $100{\sim}300m^2$.

• Korean Journal of Soil Science and Fertilizer
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• v.50 no.3
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• pp.135-149
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• 2017

In order to obtain promising rice growth-promoting microbial strains that can be used as substitutes for chemical fertilizers, 172 bacterial strains were isolated from rice roots grown in Korean and Russian soils. Out of them, the strains KR076, KR083, KR181 and RRj228 showed plant growth-promoting activities on maize seedlings. Bacillus megaterium KR076 and Bacillus sp. KR083 showed both nitrogen-fixing and plant growth-promoting activities, while Rhizobium sp. KR181 and Pseudomonas sp. RRj228 appeared to support only plant growth-promotion, but not $N_2$ fixation. Especially, RRj228 showed high growth promoting activity at low concentrations. Inoculation studies with KR083 and RRj228 revealed a high affinity to the Japonica rice variety such as Junambyeo than the Korean Tongil type variety such as Arumbyeo. Both KR083 and RRj228 strains showed rhizoplane and/or endophytic colonization in Japonica and Tongil types rice when soaked with the bacterial suspension of $1.1{\times}10^5cfu\;ml^{-1}$ for six and twelve hours. However, the total bacterial cell numbers were higher in the roots of Japonica variety than in the Tongil type. In inoculation trials with Daesanbyeo rice variety, the seedlings inoculated with KR181 and RRj228 at the rate of $2.0{\times}10^6cfu\;ml^{-1}$ showed yield increment of 35% and 33% (p < 0.01), respectively, so that they contributed to the replacement of chemical fertilizer at half doses of N, $P_2O_5$, and $K_2O$ in pots. In Junambyeo rice seedlings, the strain RRj228, when inoculated with a cell suspension of $1.8{\times}10^6cfu\;ml^{-1}$, promoted 3.4% higher yield at 70% dose than at a full dose level of N $110kg\;ha^{-1}$ in field. These results suggest that the rhizobacteria KR181 and RRj228 are prospective strains for enhancing rice performance.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1428-1440
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• 2017

Phomopsis sp. XP-8 (an endophytic fungus) was previously found to produce pinoresinol diglucoside (PDG), a major antihypertensive compound of Tu-Chung (the bark of Eucommia ulmoides Oliv.), which is widely used in Chinese traditional medicines. In the present study, two bioconversion systems were developed for the production of PDG in Tris-HCl buffer containing glucose and Phomopsis sp. XP-8 cells (both resting and freeze-dried). When other factors remained unchanged, the bioconversion time, glucose concentration, cell ages, cell dosage, pH, temperature, and stirring speed influenced PDG production in a similar and decreasing manner after an initial increase with increasing levels for each factor. Considering the simultaneous change of various factors, the optimal conditions for PDG production were established as 70 g/l cells (8-day-old), 14 g/l glucose, $28^{\circ}C$, pH 7.5, and 180 rpm for systems employing resting cells, and 3.87 g/l cells, 14.67 g/l glucose, $28^{\circ}C$, pH 7.5, and 180 rpm for systems employing freeze-dried cells. The systems employing freeze-dried cells showed lower peak PDG production ($110.28{\mu}g/l$), but at a much shorter time (12.65 h) compared with resting cells (23.62 mg/l, 91.5 h). The specific PDG production levels were 1.92 and $24{\mu}g$ per gram cells per gram glucose for freeze-dried cells and resting cells, respectively. Both systems indicated a new and potentially efficient way to produce PDG independent of microbial cell growth.

• Research in Plant Disease
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• v.14 no.2
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• pp.112-116
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• 2008

Powdery mildew of Paprika caused by Leveillula taurica has been a serious problem in greenhouse. It is an unusual endophytic powdery mildew because the mycelia grow inside the leaf, such that the pale yellow lesions on adaxial surfaces appear first and the white powdery lesion/signs develop later on the corresponding to the spots of the abaxial leaf surface, where the conidiophores are typically emerge through the stomatal opening. Although one foliar application of cooking oil and yolk mixture(COY) to the foliage was not practically effective enough, two or three, weekly application of COY to the foliage at either 0.3 or 0.5% concentration resulted in excellent control against powdery mildew with disease index less/lower than 1, respectively. This treatment could provide protection for three weeks, which, we believe, is not only cost-effective, but also environment-friendly. Powdery mildew fungus was affected by COY treatment quickly which is recognizable in three days. Net photosynthesis and evapotranspiration was remarkably reduced by powdery mildew infection compared to healthy leaves, suggesting that prevention and early protection is the most critical strategy for peak paprika fruit yield. Moreover, COY treatment did not adversely affect the photosynthesis and evapotranspiration of foliages.

• The Plant Pathology Journal
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• v.34 no.5
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• pp.381-392
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• 2018

Clavibacter michiganensis subsp. michiganesis (Cmm) is a quarantine-worthy pest in $M{\acute{e}}xico$. The implementation and validation of new technologies is necessary to reduce the time for bacterial detection in laboratory conditions and Raman spectroscopy is an ambitious technology that has all of the features needed to characterize and identify bacteria. Under controlled conditions a contagion process was induced with Cmm, the disease epidemiology was monitored. Micro-Raman spectroscopy ($532nm\;{\lambda}$ laser) technique was evaluated its performance at assisting on Cmm detection through its characteristic Raman spectrum fingerprint. Our experiment was conducted with tomato plants in a completely randomized block experimental design (13 plants ${\times}$ 4 rows). The Cmm infection was confirmed by 16S rDNA and plants showed symptoms from 48 to 72 h after inoculation, the evolution of the incidence and severity on plant population varied over time and it kept an aggregated spatial pattern. The contagion process reached 79% just 24 days after the epidemic was induced. Micro-Raman spectroscopy proved its speed, efficiency and usefulness as a non-destructive method for the preliminary detection of Cmm. Carotenoid specific bands with wavelengths at 1146 and $1510cm^{-1}$ were the distinguishable markers. Chemometric analyses showed the best performance by the implementation of PCA-LDA supervised classification algorithms applied over Raman spectrum data with 100% of performance in metrics of classifiers (sensitivity, specificity, accuracy, negative and positive predictive value) that allowed us to differentiate Cmm from other endophytic bacteria (Bacillus and Pantoea). The unsupervised KMeans algorithm showed good performance (100, 96, 98, 91 y 100%, respectively).

• 한국균학회소식:학술대회논문집
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• 2016.05a
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• pp.25-25
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• 2016

Biological control has many advantages as a disease control method, particularly when compared with pesticides. One of the most important benefits is that biological control is an environmental friendly method and does not introduce pollutants into the environment. Another great advantage of this method is its selectivity. Selectivity is the important factor regarding the balance of agricultural ecosystems because a great damage to non target species can lead to the restriction of natural enemies' populations. The objective of this research was to evaluate the effects of several different bacterial isolates on the efficacy of biological control of soil borne diseases. White rot caused by Sclerotium cepivorum was reported to be severe disease of garlic and chive. The antifungal bacteria Burkholderia pyrrocinia CAB08106-4 was tested in field bioassays for its ability to suppress white rot disease. In field tests, B. pyrrocinia CAB08106-4 isolates suppressed white rot in garlic and chive, with the average control efficacies of 69.6% and 58.9%, respectively. In addition, when a culture filtrate of B. pyrrocinia CAB08106-4 was sprayed onto wounded garlic bulbs after inoculation with a Penicillium hirstum spore suspension in a cold storage room ($-2^{\circ}C$), blue mold disease on garlic bulbs was suppressed, with a control efficacy of 79.2%. These results suggested that B. pyrrocinia CAB08106-4 isolates could be used as effective biological control agents against both soil-borne and post-harvest diseases of Liliaceae. Chinese cabbage clubroot caused by Plasmodiophora brassicae was found to be highly virulent in Chinese cabbage, turnips, and cabbage. In this study, the endophytic bacterium Flavobacterium hercynium EPB-C313, which was isolated from Chinese cabbage tissues, was investigated for its antimicrobial activity by inactivating resting spores and its control effects on clubroot disease using bioassays. The bacterial cells, culture solutions, and culture filtrates of F. hercynium EPB-C313 inactivated the resting spores of P. brassicae, with the control efficacies of 90.4%, 36.8%, and 26.0%, respectively. Complex treatments greatly enhanced the control efficacy by 63.7% in a field of 50% diseased plants by incorporating pellets containing organic matter and F. hercynium EPB-C313 in soil, drenching seedlings with a culture solution of F. hercynium EPB-C313, and drenching soil for 10 days after planting. Soft rot caused by Pectobacterium carotovorum subsp. carotovorum was reported to be severe disease to Chinese cabbage in spring seasons. The antifungal bacterium, Bacillus sp. CAB12243-2 suppresses the soft rot disease on Chinese cabbage with 73.0% control efficacy in greenhouse assay. This isolate will increase the utilization of rhizobacteria species as biocontrol agents against soft rot disease of vegetable crops. Sclerotinia rot caused by Sclerotinia sclerotiorum has been reported on lettuce during winter. An antifungal isolate of Pseudomonas corrugata CAB07024-3 was tested in field bioassays for its ability to suppress scleritinia rot. This antagonistic microorganism showed four-year average effects of 63.1% of the control in the same field. Furthermore, P. corrugata CAB07024-3 has a wide antifungal spectrum against plant pathogens, including Sclerotinia sclerotiorum, Sclerotium cepivorum, Botrytis cinerea, Colletotrichum gloeosporioides, Phytophotra capsici, and Pythium myriotylum.