• Weed & Turfgrass Science
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• v.2 no.3
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• pp.306-311
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• 2013

Pythium blight occurred by Pythium aphanidermatum on chewing fescue cv. "Jamestowm II" from early June, 2010 and 2011 at the test field in Daegu University in Gyeongbuk Province, Korea. Disease symptoms on the turfgrass were leaf blights dying from the leaf tip and root rot, which appeared patches of brown to dark brown color or gray brown color in the field. The pathogens (40-1 isolate) of Pythium blight was isolated from the diseased leaf and crown tissue and cultured on potato-dextrose agar (PDA) for identification. Lobulate sporangia were inflated, complex structures, and filamentous sporangia were usually indistinguishable from vegetative hyphae. Sequences of ribosomal RNA gene of the fungus were homologous with similarity of 100% to those of P. aphanidermatum isolates in GenBank database. Pathogenicity was also confirmed on the chewing fescue, creeping betgrass and Kentucky bluegrass by Koch's postulates. This is the first report of Pythium blight on chewing fescue caused by P. aphanidermatum in Korea.

• Research in Plant Disease
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• v.8 no.4
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• pp.234-238
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• 2002

In vitro and in vivo activities of a biocontrol agent, Bacillus stearothermophilus strain YC4194 was evaluated for the control of Pythium damping-off of cucumber. B. stearothermophilus YC4194 inhibited germination of cystospores and formation of zoosporangia of Pythium aphanidermatum in vitro. Incorporation of a bentonite and talc based formulation(10$^{9}$ cfu/g) of B. stearothermophilus YC4194 to the nursery soils (10 g/ι soil) resulted In a significant (p=0.01) reduction in the disease severity of cucumber damping-off after inoculation with P. aphanidermatum. The control efficacy of B. stearothermophilus YC4194 formulation was not different from that of the fungicides, dimethomorph, metalaxyl, ethaboxam. When the cucumber plants were transplanted to the soil inoculated with P. aphanidermatum zoospores, the B. stearothermophilus YC4194 maintained the high population density in rhizosphere soil upto 10$^{7}$ cfu/g until 15 days after treatment.

• Mycobiology
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• v.32 no.3
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• pp.139-147
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• 2004

A severe damping-off disease of bush okra caused by Pythium aphanidermatum, was diagnosed in plastic houses in Der Attia village, 15 km southwest of El-Minia city, Egypt, during the winter of 2001. Bush okra seedlings showed low emergence with bare patches inside the plastic houses. Seedlings that escaped pre-emergence damping-off showed poor growth, stunting and eventually collapsed. Examination of the infected tissues confirmed only Pythium aphanidermatum, showing its typical intercalary antheridia, and lobulate zoosporangia. P. aphanidermatum was shown to be pathogenic on bush okra under pot and field experiments. Bacteria making inhibition zones against the damping-off fungus P. aphanidermatum were selected. Agar discs from rhizosphere soil of bush okra containing colonies were transferred onto agar plate culture of P. aphanidermatum. After 2 days of incubation, colonies producing clear zones of non-Pythium growth were readily detected. The two bacteria with the largest inhibition zones were identified as Pseudomonas fluorescens. Bush okra emergence(%) in both pot and plastic houses experiments indicated that disease control could be obtained by applying P. fluorescens to the soil or coating the bacteria to the bush okra seeds before sowing. In the plastic houses, application of the bacteria onto Pythium-infested soil and sowing bush okra seeds dressed with bacteria gave 100% emergence. In addition, This was the first reported disease of bush okra by this oomycete in Egypt.

• The Plant Pathology Journal
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• v.15 no.2
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• pp.112-118
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• 1999

Eleven species of Pythium were isolated from leaf blight symptoms on creeping bentgrass (Agrostis palustirs Huds.), Kentucky bluegrass (Poa pratenisis L.) and zoysiagrasses (Zoysia japonica Steud., and Z. matrella (L.) Merr.) planted on golf courses in Korea. Mycelial growth on potato carrot agar medium under various temperature conditions indicated that Pythium species obtained in this study could be divided into four groups based on their responses to temperature conditions. P. vanterpoolii was found to favor low temperature conditions with the optimum temperature of $25^{\circ}$, whereas P. aphanidermatum and P. myriotylum favored relatively high temperature conditions with the optimum temperature of $35^{\circ}$. Other species including P. arrhenomanes, P. catenulatum, P. graminicola, P. oligandrum, P. rostratum, P. torulosum, and P. ultimum were the intermediate group with the optimum temperature of 25~$35^{\circ}$. P. periplocum was similar to the intermediate group but the minimum temperature for its mycelial growth was $15^{\circ}$, which was approximately $5^{\circ}$ above that for the intermediate Pythium spp.group. In the pathogenicity tests conducted in the lab using potted plants, P. aphanidermatum, P.a arrhenomanes, P. catenulatum, P. graminicola, P. myriotylum, P. periplocum, P. rostratum, P. torulosum, P. ultimum, and P. vanterpoolii were found to be pathogenic to creeping bentgrass and Kentucky bluegrass. P. aphanidermatum, P. catenulatum, and P. graminicola were frequently isolated from leaf blight symptoms of creeping bentgrass and Kentucky bluegrass in golf courses during the warm and humid periods in July-August. On the other hand, P. vanterpoolii and P. torulosum were frequently isolated during the cool and humid periods in March-May, suggesting both species might be the major causes of leaf blight occurring in the spring time. Zoysiagrass was susceptible to P. arrhenomanes and the heterothallic Pythium sp. (Ht-F), showing stem and crown rot of turf-grasses at poorly drained areas under coool and humid or rainy conditions. P. oligandrum and the heterothallic Pythium sp. (Ht-L) isolated from creeping bentgrass were avirulent to all species of turfgrasses tested in this study.

• The Plant Pathology Journal
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• v.34 no.3
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• pp.218-235
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• 2018

Plant growth promoting rhizobacteria and endophytic bacteria were isolated from different varieties of turmeric (Curcuma longa L.) from South India. Totally 50 strains representing, 30 PGPR and 20 endophytic bacteria were identified based on biochemical assays and 16S rDNA sequence analysis. The isolates were screened for antagonistic activity against Pythium aphanidermatum (Edson) Fitzp., and Rhizoctonia solani Kuhn., causing rhizome rot and leaf blight diseases in turmeric, by dual culture and liquid culture assays. Results revealed that only five isolates of PGPR and four endophytic bacteria showed more than 70% suppression of test pathogens in both assays. The SEM studies of interaction zone showed significant ultrastructural changes of the hyphae like shriveling, breakage and desication of the pathogens by PGPR B. cereus (RBacDOB-S24) and endophyte P. aeruginosa (BacDOB-E19). Selected isolates showed multiple Plant growth promoting traits. The rhizome bacterization followed by soil application of B. cereus (RBacDOB-S24) showed lowest Percent Disease Incidence (PDI) of rhizome rot and leaf blight, 16.4% and 15.5% respectively. Similarly, P. aeruginosa (BacDOB-E19) recorded PDI of rhizome rot (17.5%) and leaf blight (17.7%). The treatment of these promising isolates exhibited significant increase in plant height and fresh rhizome yield/plant in comparison with untreated control under greenhouse condition. Thereby, these isolates can be exploited as a potential biocontrol agent for suppressing rhizome rot and leaf blight diseases in turmeric.

• The Korean Journal of Mycology
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• v.25 no.4 s.83
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• pp.276-290
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• 1997

Eleven species of Pythium were identified from 125 isolates collected from leaf blight lesions on creeping bentgrass (Agrostis palustris Huds.), Kentucky bluegrass (Poa pratensis L.), and zoysiagrasses (Zoysia japonica Steud., and Z. matrella (L.) Merr.) at 35 golf courses in Korea in $1990{\sim}1996$. The identified species included P. aphanidermatum, P. arrhenomanes, P. catenulatum, P. graminicola, P. myriotylum, P. oligandrum, P. periplocum, P. rostratum, P. torulosum, P. ultimum var. ultimum, and P. vanterpoolii. Mycological characteristics of sporangia, oogonia, antheridia, and oospores observed on the sucrose-asparagine bentgrass leaf culture medium were described for each species. Of the species, P. arrhenomanes, P. catenulatum, P. gmminicola, P. oligandrum, P. periplocum, P. rostratum, P. torulosum and P. vanterpoolii were reported for the first time in Korea. P. myriotylum, P. rostratum, P. torulosum and P. vanterpoolii showed characteristic colony patterns on the potato-carrot agar medium, which can be used as criteria for species identification of Pythium.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.40-44
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• 2003

Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.386-389
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• 2002

Antagonistic Bacillus sp. #16 to phytopathogenic fungi were selected based on the growth rate, inhibition rate and surface tension reduction. Based on the 16S rRNA sequences, Bacillus sp. #16 is closely related to the B. subtilis DSM10. In the pot test, Bacillus sp. #16 show the most effective growth inhibition against damping-off disease of cucumber seeding.

• Asian Journal of Turfgrass Science
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• v.13 no.4
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• pp.223-234
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• 1999

Antibiotic activity of selected biocontrol agent Trichoderma harzianum ABGS-95 showed 59% to P. graminicola, 65% to P. aphanidermatum and 57% to Rhizoctonia solani compare to non-treated control. ABGC-95 showed resistant to major agrochemicals such as metalaxyl+mancozeb, etridiazole, propamocarb, toclofos methly, terbuconazole, pencycuron and flutolanil. The biocontrol agent T. harzianum ABGC-95 grew vigorously in low nutrient media and water agar. And sand mixture with wheat bran or mowing debris of zoysia grass also provided good growth of the organism. Application of sand mixture of Trichoderma spp. into aeration cores in golf showed most effective biocontrol of pythium blight. Top dressing application of T. harzianum ABGC-95 reached 83% control efficient while spray application of same biocontrol agent showed only 69% control. The biocontrol agent ABGC-95 successfully suppressed the population density of Pythium spp. in soil. The population density of total Pythium spp. in ABGC-95 treated soil was sustained almost same population at beginning(early May) up to end of August, while the population in untreated control plot was increased 5 times that of beginning and even 10 times in pathogen accumulated soil.

• The Plant Pathology Journal
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• v.21 no.4
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• pp.328-333
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• 2005

Paenibacillus elgii SD17 (KCTC $10016BP^T$=NBRC $100335^T$) was recently reported as a new species. Based on its inhibitory activity to Thanatephorus cucumeris AG1-1, strain SD17 was further evaluated for its potential as a biocontrol agent against soil-borne diseases of turf grasses in Korea. P. elgii SD17 showed a broad spectrum of antimicrobial activity in vitro test and suppressed development of turf grass diseases; Pythium blight caused by Pythium aphanidermatum and brown patch caused by T. cucumeris AG1-1 on creeping bentgrass (Agrostis palustris) in the growth chamber tests. Under a condition for massive culture in a 5,000 L fermenter, P. elgii SD17 reached $6.4{\times}10^8$ spores/ml that resulted in approximately $1.0{\times}10^7$ cfu/g when formulated into a granule formulation (GR) using the whole culture broth instead of water. Using the GR formulation, biocontrol activity of P. elgii SD17 was confirmed. In the growth chamber tests, the GR formulation was effective against brown patch and Pythium blight with similar level of disease severity compared to each of the standard fungicides at the application rates of 10 g/$m^2$ or above. In the field tests, compared to each untreated control, the GR formulation also effectively controlled Pythium blight, brown patch and large patch at all the application rates of 5, 10 and 20 g/$m^2$, respectively, without significant response by the application rates. However its performance was inferior to each of the standard chemical fungicides. Based on these results, we consider this GR formulation of P. elgii SD17 as an effective biocontol agent to suppress Pythium blight, brown patch and large patch of turf grasses in Korea.