• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.25-25
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• 2014

Three species of Fusarium, F. fujikuroi, F. verticillioides and F. proliferatum, are known to be associated with bakanae disease of rice [1, 2]. F. fujikuroi infects rice flowers and survive in endosperm and embryo of the seeds. Infected seed is an important source of primary inoculum of pathogens [3]. Seeds of rice (Oryza sativa cv. Boramchan) collected from bakanae-infected field were found to be 96% infected with Fusarium sp., 52% with F. fujikuroi, 42% with F. verticillioides, and 12% with F. proliferatum as determined by incubation method and species-specific PCR assays. F. fujikuroi was detected at lemma/palea, endosperm and embryo whereas F. verticillioides and F. proliferatum were recovered only from lemma/palea by means of component plating test. Seed disinfection methods have been developed to control bakanae disease and prochloraz has been most widely used for rice seeds. Two chemicals formulated with prochloraz (PC 1) and prochloraz + hexaconazole (PC 2) that inhibit biosynthesis of ergosterol strongly reduced the incidence of Fusarium spp. on selective media to 4.7% and 2.0%, respectively. Disease symptoms of rice seedlings in nursery soil were alleviated by chemical treatment; seedlings with elongated leaves or wide angle between leaf and stem were strikingly reduced from 15.6 to 3.2% (PC 1) and 0 (PC 2), stem rots were reduced from 56.9 to 26.2% (PC 1) and 32.1% (PC 2), and normal seedling increased from 0.4 to 13.3% (PC 2). Prochloraz has some disadvantages and risks such as the occurrence of tolerant pathogens [4] and effects on the sterol synthesis in animals and humans [5]. For these reasons, it is necessary to develop new disinfection method that do not induce fungal tolerance and are safe to humans and animals. Chlorine dioxide ($ClO_2$), that is less toxic, produces no harmful byproducts, and has high oxidizing power, has been reported to be effective at disinfection of several phytopathogenic fungi including Colletotrichum spp. and Alternaria spp. [6]. Gaseous $ClO_2$ applied to rice seeds at a concentration of 20 ppm strongly suppressed mycelial growth of Fusarium fujikuroi, F. verticillioides and F. proliferatum. The incidence of Fusarium spp. in dry seed with 8.7% seed moisture content (SMC) tended to decrease as the concentration of $ClO_2$ increased from 20 to 40 ppm. Applying 40 ppm $ClO_2$ at 90% relative humidity, incidence was reduced to 5.3% and resulted in significant reduction of disease symptoms on MS media. In nursery soil, stem rot was reduced from 56.9 to 15.4% and the number of normal seedlings increased from 0.4 to 25.5%. With water-soaked seeds (33.1% SMC) holding moisture in the endosperm and embryo, the effectiveness of disinfection using $ClO_2$ increased, even when treated with only 20 ppm for four hours. This suggests that moisture was a key element for action of $ClO_2$. Removal of the palea and lemma from seeds significantly decreased the incidence of Fusarium spp. to 3.0%. Seed germination appeared to decrease slightly by water-soaking at $30^{\circ}C$ because of increased SMC and by physical damage of embryos from hulling. These results indicate that the use of gaseous $ClO_2$ was effective as a means to disinfect rice seeds infected with Fusarium spp. and that moisture around the pathogens in the seed was an important factor for the action of $ClO_2$. Further investigations should be conducted to ascertain the best conditions for complete disinfection of Fusarium spp. that infect deep site of rice seeds.

• Journal of Microbiology and Biotechnology
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• v.31 no.9
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• pp.1231-1240
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• 2021

Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.

• Journal of Bio-Environment Control
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• v.21 no.2
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• pp.127-133
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• 2012

Inhibition effects on spore germination and mycelia growth for pepper anthracnose fungi (Collectricum acutatum) were investigated in vitro using eco-friendly agricultural materials as well as ecofriendly pesticides. The inhibition effect on mycelia growth of anthracnose fungi is the highest when the anthracnose mycelia were treated with a pesticide (commercial name: Koreayeok) that contains a mixture of Bacillus subtilis and Panibacillus polymyxa, resulting in 100% inhibition of the mycelia growth. Meanwhile, the range of 20~40% inhibition effects on the growth of anthracnose mycelia was observed with other commercial agricultural materials. The significant inhibition effects on spore formation of anthracnose fungus were shown in vitro with two water dispersible pesticides containing sulfur [BTB (100%) and SulfurStar (95.1%)], Koreayeok (95.0%), Borstar (99.0%) containing Bordeaux mixture, and Jihabudea-KM containing Psedomonas spp. (96.1%), respectively. Taken from these in vitro results of inhibiting of the spore germination and mycelia growth together, Koreayeok is the most effective on control of pepper anthracnose disease in vitro. In addition, two water dispersible pesticides containing sulfur (BTB and SulfurStar) and Borstar (99.0%) containing Bordeaux mixture are also significantly applicable to prevent pepper plants from anthracnose disease in vitro. It remains to be determined whether the selected eco-friendly agricultural materials in effective control of anthracnose in vitro can be used to control pepper anthracnose in field.

• The Korean Journal of Pesticide Science
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• v.20 no.4
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• pp.369-374
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• 2016

Bacillus velezensis YP2 inhibited the mycelial growth of several plant pathogens including Cercespora spp., Septoria sp., Phoma sp., Botrytis cinerea and Sclerotinia scleotiorum occurring in leafy vegetables. Control efficacy for powdery mildew caused by Erysiphe cruciferarm on red leaf mustard and cheong mustard by treatment of spraying with 10-fold diluted Luria-Bertani (LB) broth of B. velezensis YP2 was 91.8% and 80.9%, respectively. When B. velezensis YP2 was treated four times with five-day interval, three times at seven-day interval and two times at ten day interval in the greenhouse test, the control effect of red leaf mustard powdery mildew was 70.6%, 65.0% and 40.9%, respectively. Also B. velezensis YP2 could promote the seed germination and plant growth of led leaf mustard. The results showed that the culture broth of B. velezensis YP2 was very effective to control the powdery mildew of leaf mustard.

• 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.

• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1614-1623
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• 2010

Bacteria were isolated from roots of sugarcane varieties grown in the fields of Punjab. They were identified by using API20E/NE bacterial identification kits and from sequences of 16S rRNA and amplicons of the cpn60 gene. The majority of bacteria were found to belong to the genera of Enterobacter, Pseudomonas, and Klebsiella, but members of genera Azospirillum, Rhizobium, Rahnella, Delftia, Caulobacter, Pannonibacter, Xanthomonas, and Stenotrophomonas were also found. The community, however, was dominated by members of the Pseudomonadaceae and Enterobacteriaceae, as representatives of these genera were found in samples from every variety and location examined. All isolates were tested for the presence of five enzymes and seven factors known to be associated with plant growth promotion. Ten isolates showed lipase activity and eight were positive for protease activity. Cellulase, chitinase, and pectinase were not detected in any strain. Nine strains showed nitrogen fixing ability (acetylene reduction assay) and 26 were capable of solubilizing phosphate. In the presence of 100 mg/l tryptophan, all strains except one produced indole acetic acid in the growth medium. All isolates were positive for ACC deaminase activity. Six strains produced homoserine lactones and three produced HCN and hexamate type siderophores. One isolate was capable of inhibiting the growth of 24 pathogenic fungal strains of Colletotrichum, Fusarium, Pythium, and Rhizoctonia spp. In tests of their abilities to grow under a range of temperature, pH, and NaCl concentrations, all isolates grew well on plates with 3% NaCl and most of them grew well at 4 to $41^{\circ}C$ and at pH 11.

• The Plant Pathology Journal
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• v.39 no.5
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• pp.430-448
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• 2023

Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.

• Research in Plant Disease
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• v.22 no.2
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• pp.81-93
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• 2016

Out of plant-associated bacteria, certain plant growth-promoting bacteria (PGPB) have been reported to increase plant growth and productivity and to elicit induced resistance against plant pathogens. In this study, our objective was to broaden the range of applications of leaf-colonizing PGPB for foliar parts of road tress and pepper. Total 1,056 isolates of endospore-forming bacteria from tree phylloplanes were collected and evaluated for the enzymatic activities including protease, lipase, and chitinase and antifungal capacities against two fungal pathogens, Colletotrichum graminicola and Botrytis cinerea. Fourteen isolates classified as members of the bacilli group displayed the capacity to colonize pepper leaves after spraying inoculation. Three strains, 5B6, 8D4, and 8G12, and the mixtures were employed to evaluate growth promotion, yield increase and defence responses under field condition. Additionally, foliar application of bacterial preparation was applied to the road tress in Yuseong, Daejeon, South Korea, resulted in increase of chlorophyll contents and leaf thickness, compared with non-treated control. The foliar application of microbial preparation reduced brown shot-hole disease of Prunus serrulata L. and advanced leaf abscission in Ginkgo biloba L. Collectively, our results suggest that leaf-colonizing bacteria provide potential microbial agents to increase the performance of woody plants such as tree and pepper through spray application.

• Korean Journal of Environmental Agriculture
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• v.37 no.1
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• pp.34-40
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• 2018

BACKGROUND: In order to select a fungicide that can effectively control anthracnose disease in Japanese plum fruit, mycelial growth inhibition effect and spore germination inhibition effect of six fungicides were tested in vitro against six isolates of Colletotrichum acutatum and five isolates of C. gloeosporioides that were isolated from diseased Japanese plum fruit. METHODS AND RESULTS: Inhibitory effects of fungicides on mycelial growth were investigated after inoculating each isolate on potato dextrose agar amended with four discriminatory concentrations of each fungicide for 7 days at $25^{\circ}C$. For spore germination inhibitory effect, each isolate of the Colletotrichum spp. was cultured in potato dextrose agar for 7-14 days at $25^{\circ}C$. After adjusting the concentration of spores of each isolate to $1{\times}10^6mL^{-1}$ by diluting with 0.025% PDB, the spore suspension was mixed with each fungicide (1:4, v/v), and $60{\mu}L$ aliquots were dispensed to sterile hole slide glass. Hole slide glasses were placed in a humidified box and incubated for 15 hours at $25^{\circ}C$. Then, spore germination was observed under an optical microscope. At recommended concentration of fungicide prochloraz manganese showed the highest mycelial growth inhibitory effect and dithianon showed the lowest mycelial growth inhibition. The $EC_{50}$ values for the inhibition of spore germination by dithianon and pyraclostrobin were $0.069-0.126{\mu}g/mL$ and $0.37-1.59{\mu}g/mL$, respectively. Although benomyl, prochloraz manganese, azoxystrobin, and tebuconazole did not inhibit the spore germination, they appeared to restrain mycelial growth by abnormal growth of germ tube and mycelium after germination. CONCLUSION: Dithianon seemed to have preventive effect. Prochloraz manganese, azoxystrobin, and tebuconazole were likely to have control effect. Pyraclostrobin is considered to have both preventive and control effect against anthracnose disease of Japanese plum fruit.

• Korean Journal of Agricultural Science
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• v.13 no.1
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• pp.52-61
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• 1986

Mycelial growth of Colletotrichun dematium and C. gloeosporioides isolated from anthracnose of Lycium chinense was favorable on V-8 Juice Agar media and Oatmeal Agar media of the four different media tested. Sporulation of C. dematium was favorable on PDA media and V-8 Juice Agar media and that of C. gloeosporioides was favorable on all media tested. The optimum temperature for mycelial growth of C. gloeosporioides and C. dematium was $25^{\circ}C$ and $28^{\circ}C$, respectively and sporulation of the fungi was stimulated under alternating NUV light and darkness. Of the seven nitrogen sources tested, gelatin was the most favorable for mycelial growth of C. gloeosporioides and glycin and gelatin were favorable for that of C. dematium. Of the eleven carbon sources tested, dextrin and sorbitol were favorable for mycelial growth of C. gloeosporioides and dextrin was the most favorable for that of C. dematium. Among the eleven fugicides tested, Benomyl, Folpet, Dithianon, Carbendazim and S-3308L were found superior for the inhibition of mycelial growth and spore germination of the fungi. They were, also, found superior for the control of anthracnose disease of L. chinense when they were applied to the plants in the field.