• Horticultural Science & Technology
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• v.31 no.6
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• pp.657-665
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• 2013

A large patch disease caused by Rhizoctonia solani AG2-2 (IV) is a serious problem in Korean lawngrass (Zoysia japonica) sites including golf courses and sports fields in Korea. Antagonistic microorganisms against R. solani AG2-2 (IV) were isolated from various forest and crop soil sources in Southern Korea. Among the 61 isolates, I-009, FRIN-001-1, and YPIN-022 strains showing dramatic inhibition of the mycelial growth of R. solani AG2-2 (IV) in the pairing culture were selected as the most potential antagonistic microorganisms for this study. Based on the 16s RNA sequence comparison, I-009 and FRIN-001-1 isolates were identified as Bacillus spp., while YPIN-022 isolate belongs to the genus Pseudomonas. The greater inhibition (clear) zone between two edges of the selected and pathogenic microbes ranged from 11 to 15 mm in three selections, but the others averaged to 7 mm out of 30 mm distance. In another antifungal test using culture filtrate, those three isolates represented a range of 51.7 to 63.5% suppression potential. The selected isolates also inhibited significantly the stem-segment colonization by R. solani AG2-2 (IV) in vivo test by 28.1%, 43.0%, and 23.7% when inoculated with I-009, FRIN-001-1, and YPIN-022, respectively. The highest antagonistic activity for the large patch disease was demonstrated by the isolate FRIN-001-1, which will be useful for developing a bio-pesticide against Rhizoctonia.

• Journal of Applied Biological Chemistry
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• v.60 no.3
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• pp.213-218
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• 2017

The objective was to examine the antifungal activity of Pinus densiflora extract for the control of turfgrass fungal diseases. Antifungal activities of the various fractions of n-hexane, methylene chloride (Ch), ethyl acetate (EtOAc), and n-butanol from P. densiflora were evaluated against Rhizoctonia solani AG1-1B, R. solani AG2-2IV, Sclerotinia homoeocarpa, R. cerealis, Pythium spp., and Colletotrichum graminicola. The Ch and EtOAc fractions showed antifungal activity against Pythium sp. and C. graminicola in paper disc assay. The effective concentration to produce 50% mycelial inhibition ($EC_{50}$) using five discriminatory concentrations of pinosylvin (1) from the Ch fraction of P. densiflora was evaluated on R. solani AG1-1B, R. solani AG2-2IV, R. cerealis, and S. homoeocarpa. S. homoeocarpa showed the highest sensitivity with the lowest mean $EC_{50}$ value ($8.426{\mu}g/mL$) among the four pathogens. Among the three Rhizoctonia pathogens, R. cerealis had the highest mean $EC_{50}$ value ($99.832{\mu}g/mL$) and R. solani AG2-2IV, with the lowest sensitivity, had the lowest $EC_{50}$ value ($39.696{\mu}g/mL$). These results suggested that pinosylvin (1) from P. densiflora could be a valuable lead compound in the improvement of a novel antifungal agent.

• The Korean Journal of Mycology
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• v.26 no.3 s.86
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• pp.373-379
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• 1998

Rhizoctonia solani [Thanatephorus cucumeris (frank) Donk], one of the major soil-borne plant pathogens with world-wide distribution, can cause great damages on various crops. In Korea, sheath blight on rice caused by this pathogen is the major concern, and active studies on this pathogen have been performed. However, most of these studies were concerned with pathogenicity of the isolates instead of molecular analyses of different AGs of R. solani. Therefore, in this study, thirty isolates of Rhizoctonia solani collected from various sources were used for the analyses of genetic relationships among themselves and for the rapid anastomosis grouping with RAPD method. As a result, thirty isolates of known and unknown AGs were grouped into five subgroups and each group included AG-1, AG-2, AG-3, AG-4, and AG-5. RS-1 isolate was found to be closely related to AG-5. Isolates RS-4, RS-14, RS-17, and RS-16 were found to be closely related to AG-2-2(III B). Isolate RS-13 was closely related to AG-4, isolates RS-8 and RS-10 were closely related to AG-1(I B), and isolates RS-7 and RS-21 were closely related to AG-2-2(IV). Isolate RS-19 was closely related to AG-1(I C), and isolates RS-3, RS-5, RS-18, RS-6, and RS-15 were found to be closely related to AG-1.

• Asian Journal of Turfgrass Science
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• v.25 no.1
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• pp.48-51
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• 2011

This study was performed to functional improvement by fermenting of SCB (Slurry Composting and Biofiltering) liquid fertilizer. After screening of cellulose decomposing test using microorganisms originated from composts produced with turf grass clipping, five kinds of Cellulose Decomposing Microorganisms (CDMs) were selected. The optimum medium for CDMs culture was Tryptic soy broth. The optimum fermenting time of CDMs in SCB liquid fertilizer was 36 hour. In addition, CDMs showed antimicrobial activity in vitro. Sclerotinia homoeocarpa, Rhizoctonia solani AG2-2(IV) and Pythium spp. were inhibited by CDMs, but Rhizoctonia solani AG2-2(IIIB) in vitro uneffected by CDMs we isolated.

• The Korean Journal of Mycology
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• v.22 no.4
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• pp.309-324
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• 1994

A total of 2,276 isolates of Rhizoctonia solani obtained from diseased crops of 68 species was classified into anastomosis groups AG-1, AG-2-1, AG-2-2, AG-3, AG-4 and AG-5 by anastomosis test. Among the isolates, 1,091 isolates were identified as AG-1, 326 isolates as AG-2-1, 191 isolates as AG-2-2, 71 isolates as AG-3, 505 isolates as AG-4, and 92 isolates as AG-5. Among the isolates of AG-1, 791 isolates were grouped as cultural type IA, 280 isolates as cultural type IB, and the others as cultural type IC. Among the isolates of AG-2-2, 112 isolates were grouped as cultural type IIIB, and the others as cultural type IV. Cultural types IA, IB and IC of AG-1 were isolated from 7, 26 and 2 species of crops, respectively. AG-2-1 was isolated from 10 species of crops. Cultural types IIIB and IV of AG-2-2 were isolated from 7 and 3 species of crops, respectively. AG-3 was only isolated from Solanum tuberosum. AG-4 was isolated from 43 species of crops, and AG-5 from 13 species of crops. A single anastomosis group was isolated from each of 45 species of crops, but two or more than two anastomosis groups were isolated from each of the other crops. Cultural appearance of the isolates belonging to an anastomosis group or a cultural type was mostly distinct from that belonging to others, although cultural appearances of some anastomosis groups or cultural types were similar to one another. Optimum temperature for mycelial growth of AG-1, AG-2-2, AG-4 and AG-5 ranged from 26 to $30^{\circ}C$, and that of AG-2-1 and AG-3 from 22 to $26^{\circ}C$. Minimum temperature for mycelial growth of AG-2-1 was the lowest as $2{\sim}3^{\circ}C$, that of AG-1(IA) and AG-4 was the highest as $10{\sim}11^{\circ}C$, and that of the others ranged from 5 to $10^{\circ}C$. Maximum temperature for mycelial growth of AG-2-2(IIIB) was the highest as $36{\sim}37^{\circ}C$, that of AG-2-1 was the lowest as $29{\sim}30^{\circ}C$, and that of the others ranged from 31 to $36^{\circ}C$. When the mycelial growth rates at $26^{\circ}C$ were compared, AG-1(IC) grew most rapidly, followed by AG-1(IA) and AG-1(IB), and AG-2-1 grew most slowly.

• Weed & Turfgrass Science
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• v.4 no.2
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• pp.124-128
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• 2015

Large patch, caused by Rhizoctonia solani AG2-2 IV, is a soil-born disease that is the most important of warm season turfgrass such as zoysia and Bermuda grass. This study was conducted to analysis of the soil microbial community structure on large patch. Center of the large patch (CLC), edge (CLE) and healthy (CLH) part of microbial communities were examined using metagenomics in Phylum level. Distribution trends of the rhizosphere microorganisms were similar to the order Proteobacteria, Acidobacteria, Chloroflexi, Firmicutes, Planctomycetes, Gemmatimonadetes, Nitrospira, Cyanobactria and Verrucomicrobia in soil collections. Contrastively Actinobacteria was more 56% abundant in healthy part soil (16%) than in the center (9.28%) or edge (10.84%) parts. Taxonomic distributions were compared among the CLC, CLE and CLH, total 6,948 OTUs were detected in the CLC, 6,505 OTUs for the CLE and 5,537 OTUs were detected in the CLE. Distributions of Actinobacteria OTUs were appeared 615 OTUs in the CLC, 709 OTUs in the CLE and 891 OTUs in the CLH. Among Actinobacteria, 382 OTUs were overlapped in the all soils. Not matched OTUs of CLH (286 OTUs) was detected 23 times higher than CLC (91 OTUs) and CLE (126 OTUs).

• Horticultural Science & Technology
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• v.32 no.6
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• pp.879-886
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• 2014

This study was conducted to test in vitro the antagonistic effect of composted liquid manure (CLM) against soil-borne turfgrass pathogenic fungi, Rhizoctonia solani AG-2-2 (IIIB) (brown patch), R. solani AG-2-2 (IV) (large patch), and Sclerotinia homoeocarpa (dollar spot) for environmentally friendly turfgrass management. CLMs were collected from 9 livestock excretion treatment facilities around the country including Gunwi (GW), Hapcheon (HC), Hoengseong (HS), Icheon (IC), Iksan (IS), Muan (MA), Nonsan (NS), and Yeoju (YJ). CLMs of IC, GW, and IS showed s ignificant (p < 0.05) mycelium growth inhibition that was 17.8%, 20.4%, and 48.0% against R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. A t otal of 110 bacterial isolates were obtained from the CLMs that showed antagonistic effects. Among them, 5, 4, and 10 microbe isolates showed promising antifungal activity against mycelium growth of R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. The bacterial isolates ICIIIB60, GWIV70, and ISSH20 effectively inhibited the mycelial growth of three soil-borne turfgrass pathogens. Selected bacterial isolates were identified as Alcaligenes sp., Bacillus licheniformis Ab2, and B. subtilis C7-3 through 16s rDNA gene sequence analysis. Among 5 fungicides, the most compatible fungicide with ICIIIB60, GWIV70, and ISSH20 was tebuconazol, toclofos-methyl and toclofos-methyl, respectively. These findings suggested that CLMs could be effectively used not only as organic liquid fertilizer sources but also as biological control agents for soil-borne turfgrass diseases such as brown patch, large patch, and dollar spot.

• Weed & Turfgrass Science
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• v.2 no.1
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• pp.70-75
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• 2013

Seven medicinal plant extracts were tested for antifungal activities against six species of the major turfgrass pathogenic fungi (Colletotrichum graminicola, Pythium spp., Rhizoctonia cerealis, Rhizoctonia solani AG1-1, Rhizoctonia solani AG2-2, and Sclerotinia homoeocarpa) using paper disk diffusion method. Three medicinal plant extracts, including Pinus densiflora showed antifungal activities. In suppression of mycelium growth test, on medium adding P. densiflora extract showed that inhibition rate of mycelium growth were above 80% in 10 mg/10 ml concentration of the extract. The inhibition rate of Pythium spp. was 100% and C. graminicola was 84.3% in 10 mg/10 ml concentrations of P. densiflora extract, respectively. In particularly, the inhibition rate of Pythium spp. was 89.5% in 2 mg/10 ml concentrations of P. densiflora extract. As a result, P. densiflora extract showed high antifungal activity to Pythium spp. and C. graminicola of the turfgrass pathogen in in vitro test.

• Asian Journal of Turfgrass Science
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• v.26 no.1
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• pp.17-23
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• 2012

Large patch caused by Rhizoctonia solani AG2-2 (IV) is one of the most serious diseases in zoysiagrass. The objectives of this research were to evaluate the in vitro fungicidal effect and in field control of large patch disease of Paraffin oil. In the field experiments, paraffin oil was applied with 1 L/$m^2$ after diluted at 5 ml to 20 ml of oil/1 L of water for the control of large patch on Zoysia japonica in golf course. The same material was tested for inhibition of mycelial growth in vitro. Paraffin oil at 0.5% and 2.0% had a fungicidal effect by 43 to 67% on R. solani AG2-2 (IV) in vitro. In two locations of the field experiments, the paraffin oil provided moderate protection of large patch. It was effective in suppressing large patch development by 48.0% in comparison with a water check after one time application on mid-September in golf course. Paraffin oil may be used as an alternative control agent for environment friendly management of large patch on Zoysiagrass in golf course.

• Journal of Plant Biotechnology
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• v.43 no.4
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• pp.450-456
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• 2016

Rhizoctonia leaf blight (large patch) has become a serious problem in Korean lawn grass, which is extremely hard to treat and develops mostly from the roots of lawn grass to wither it away. Rhizoctonia leaf blight (large patch) is caused by Rhizoctonia solani AG2-2 (IV). To develop zoysia japonica with strong disease tolerance against this pathogenic bacterium, ${\beta}-1,3-glucanase$ was cloned from zoysia japonica, which is one of the PR-Proteins known to play a critical role in plant defense reaction. ${\beta}-1,3-glucanase$ is known to be generated within the cells when plant tissues have a hypersensitive reaction due to virus or bacterium infection and secreted outside the cells to play mainly the function of resistance against pathogenic bacteria in the space between the cells. This study utilized the commonly preserved part in the sequence of corn, wheat, barley, and rice which had been researched for their disease tolerance among the ${\beta}-1,3-glucanase$ monocotyledonous plants. Based on the part, degenerate PCR was performed to find out the sequence and full-length cDNA was cloned. E.coli over-expression was conducted in this study to mass purify target protein and implement in vitro activation measurement and antibacterial test. In addition, to interpret the functions of ZjGlu1 gene, each gene-incorporating plant transformation vectors were produced to make lawn grass transformant. Based on ZjGlu1 protein, antibacterial activity test was conducted on 9 strains. As a result, R. cerealis, F. culmorum, R.solani AG-1 (1B), and T. atroviride were found to have antibacterial activity. The gene-specific expression amount in each organ showed no huge difference in the organs based upon the transformant and against 18s gene expression amount.