• Korean Journal of Soil Science and Fertilizer
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• v.40 no.6
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• pp.447-453
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• 2007

With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.

• Journal of Bio-Environment Control
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• v.32 no.2
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• pp.165-171
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• 2023

Potato dry rot is one of the potato storage diseases caused by Fusarium species and is a representative pathological disorder that induced post-harvest loss during storage. Chlorpropham treatment for sprouting inhibition is mainly used for room temperature storage of potatoes for processing. In this study, the inhibitory effect of chlorpropham on Fusarium-induced dry rot of potato 'Dano'. To investigate the mycelial growth rate of the dry rot fungus (Fusarium solani Appel & Wollenw), mycelial growth was investigated in a chlorpropham (5.0, 50.4, 503.8, and 5,038 ppm) and prochloraz (0.1, 1.0, 10.0, and 100.0 ppm) medium containing F. oxysporum mycelia. Mycelia were more inhibited as the concentration of chlorpropham and prochloraz increased during incubation at 20℃, and the inhibition rate was 98.2% and 100% when treated with 503.8 ppm of chlorpropham and 10ppm of prochloraz in 14 days, respectively. Potato Dano tubers inoculated with F. oxysporum were dipped in chlorpropham (5.0, 50.4, and 503.8 ppm) and prochloraz (100 ppm) to investigate the effect of preventing dry rot during cold storage at 20℃ and 4℃ in vivo. The disease diameter of potatoes stored at room temperature (about 20℃) was reduced to 13.0 mm in the prochloraz 100 ppm teatment, and 10.7 mm in the chlorpropham 50.4 ppm treatment compared to 13.7 mm in the control tuber at 70 days of storage. The disease progression in all treatments including control was similar with no statistically significant difference at 4℃ air temperature. From the results of this study, it is considered that treatment with 50.4 ppm of chlorpropham after harvest will be useful for suppressing dry rot of stored potatoes.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.58 no.4
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• pp.376-382
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• 2013

Rice sheath blight (ShB), caused by Rhizoctonia solani K$\ddot{u}$hn, is one of the serious fungal disease resulting in severe yield loss in rice field worldwide. There are limited sources of genetic resistance and no commercial cultivar with high level of ShB resistance is currently available in Korea. In order to seek available resources with high level of resistance to ShB, 40 rice germplasms were used to evaluate disease reactions including ShB, leaf blast and bacterial leaf blight and these germplasms also examined agronomic traits such as days to heading, culm length, panicle length, No. of panicles, No. of spikelets per panicle and so on. There is wide variation in agronomic characters and disease reactions. Rice germplasms also showed considerably different ShB reaction caused by inoculation at seedling stage. Areumbyeo, Gayabyeo, IR579-Es44 and IR64 showed more strong reaction to ShB than the others. Especially, Gayabyeo is considerably available to develop a new variety with resistance to ShB in Korea.

• Research in Plant Disease
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• v.18 no.3
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• pp.201-209
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• 2012

The multi-function of 18 Bacillus subtilis isolates collected from agricultural extension centers of local government and National Academy of Agricultural Science was investigated by measuring their antifungal activities against five plant pathogens, such as Rhizoctonia solani, Colletotrichum acutatum, Fusarium oxysporum, Magnaporthe oryzae and Phytophthora capsici, phosphorus solubilization ability, production of indole acetic acid (IAA) and siderophore, and nitrogen fixation. The B. subtilis isolates showed antifungal activity against several plant pathogens and nitrogen fixation activity, and produced siderophore and IAA. They could control pepper powdery mildew (Leveillula taurica), but there was no difference in control efficacy among the B. subtilis isolates. In fields, the control efficacy of B. subtilis R2-1 ($10^8$ cells/ml) was compared with two microbial fungicides, Q-pect and Topsid. In 2009, the control efficacy of B. subtilis R2-1 (37.7%) was lower than that of Topsid (47.6%), but higher than that of Q-pect (25.7%). In 2010, the control efficacy of B. subtilis R2-1 (83.3%) was higher than that of Topsid (67.9%). In order to elucidate mode of action of B. subtilis R2-1 for controlling pepper powdery mildew, spore germination rates of pepper powdery mildew pathogen collected on treated leaves was investigated when suspensions of B. subtilis R2-1 and two microbial fungicides (Q-pect and Topsid) were foliar-sprayed. They highly suppressed spore germination of the pathogen with inhibition values of 84.2% for B. subtilis R2-1, 97.9% for Q-pect and 94.7% for Topsid. Further study on the mass-culturing method and formulation is needed for development of a microbial fungicide.

• Microbiology and Biotechnology Letters
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• v.37 no.3
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• pp.280-286
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• 2009

To investigate anti-phytopathogenic fungal activity of Coptis chinensis, the methanol extract and its organic solvent fractions were prepared. Using the extract and the fractions, in-vitro spore-germination inhibition and mycelial-growth inhibition activities were evaluated against Colletotrichum gloeosporioides, Phytohpthora capsici, Pyricularia grisea, Rhizoctonia solani, Botryosphaeri dothidea, Glomerella cingulata, respectively. Treatment of the methanol extract (500 mg/mL) into the spore of phytopathogenic fungi completely inhibited germinations for 5 days, except B. dothidea, and showed strong antifungal activities against P. grisea and B. cinerea, and antioomycetes activity against P. capsici. The minimal growth inhibition concentrations of the methanol extract against P. grisea, B. cinerea and P. capsici were 300, 300, and 500 mg/mL, respectively. For practical application of C. chinensis in red-pepper field, the hot-water extract (1,000 mg/mL) was prepared in commercial facility, after evaluation of heat stability and solvent-extraction yields of antifungal substances. The 3-times leaf-spray of the extract from June to August, 2008 did not show any deleterious effect to red-pepper. In fact, the leaf-spray promoted plant growth including leaf, root and fruit. The average weight and rind of each fruit were increased to 119% and 117% comparison to those of without treatments. Our results suggest that C. chinensis is a useful source for control of red-pepper diseases and plant growth.

• Research in Plant Disease
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• v.25 no.3
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• pp.114-123
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• 2019

QD LED has an ideal light source for growing crops and can also be used to control plant pathogenic microorganisms. The mycelial growth inhibition effect of QD LED light on Rhizoctonia solani, Phytophthora drechsleri, Sclerotinia sclerotiorum, Sclerotinia minor, Botrytis cinerea, Fusarium oxysporum, Pectobacterium carotovorum, and Xanthomonas campestris were investigated. According to the results, BLUE (450 nm) light, suppressed S. sclerotiorum by 16.7% at 50 cm height from the light source, and 94.1% mycelial growth at 30 cm height. Mycelial growth of Sclerotinia minor was inhibited by 80.4% at 50 cm height and 36.3% at 50 cm height in B. cinerea. S. minor, and B. cinerea was inhibited by 100% mycelial growth at a height of 30 cm from the light source. At 15 cm height, all three pathogens (B. cinerea, S. minor, and S. sclerotiorum) was inhibited by 100%. QD RED (M1) and QD RED (M2) light suppressed mycelial growth of S. minor and B. cinerea by 100% at 30 cm and 15 cm height from the light source. For S. sclerotiorum, QD RED (M1) and QD RED (M2) showed 75.2% and 100% inhibition, respectively. Further experiment was conducted to know the suppression effect of lights after inoculating the fungal pathogens on lettuce crop. According to the results, QD RED (M2) suppressed the S. sclerotiorum by 59.9%. In addition, Blue (450 nm), QD RED (M1), and QD RED (M2) light reduce the infestation by 59.9%. In case of B. cinerea, disease reduction was found 84% by BLUE (450 nm) light. Results suggest that the growth inhibition of mycelium increases by Quantum dot LED light.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.301-310
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• 2010

Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1223-1229
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• 2009

To construct a new recombinant strain of Bacillus velezensis that has antifungal and insecticidal activity via the expression of the insecticidal Bacillus thuringiensis crystal protein, a B. thuringiensis expression vector (pHT1K-1Ac) was generated that contained the B. thuringiensis cry1Ac gene under the control of its endogenous promoter in a minimal E. coli-B. thuringiensis shuttle vector (pHT1K). This vector was introduced into a B. velezensis isolate that showed high antifungal activities against several plant diseases, including rice blast (Magnaporthe grisea), rice sheath blight (Rhizotonia solani), tomato gray mold (Botrytis cinerea), tomato late blight (Phytophthora infestans), and wheat leaf rust (Puccinia recondita), by electroporation. The recombinant B. velezensis strain was confirmed by PCR using cry1Ac-specific primers. Additionally, the recombinant strain produced a protein approximately 130 kDa in size and parasporal inclusion bodies similar to B. thuringiensis. The in vivo antifungal activity assay demonstrated that the activity of the recombinant B. velezensis strain was maintained at the same level as that of wild-type B. velezensis. Furthermore, it exhibited high insecticidal activity against a lepidopteran pest, Plutella xylostella, although its activity was lower than that of a recombinant B. thuringiensis strain, whereas wild-type B. velezensis strain did not show any insecticidal activity. These results suggest that this recombinant B. velezensis strain can be used to control harmful insect pests and fungal diseases simultaneously in one crop.

• Research in Plant Disease
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• v.28 no.1
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• pp.19-25
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• 2022

Ginseng is an important medicinal plant cultivated in East Asia for thousands of years. It is typically cultivated in the same field for 4 to 6 years and is exposed to a variety of pathogens. Among them, ginseng root rot is the main reason that leads to the most severe losses. In this study, endophytic bacteria were isolated from healthy ginseng, and endophytes with antagonistic effect against ginseng root rot pathogens were screened out. Among the 17 strains, three carried antagonistic effect, and were resistant to radicicol that is a mycotoxin produced by ginseng root rot pathogens. Finally, Bacillus velezensis CH-15 was selected due to excellent antagonistic effect and radicicol resistance. When CH-15 was inoculated on ginseng root, it not only inhibited the mycelial growth of the pathogen, but also inhibited the progression of disease. CH-15 also carried biosynthetic genes for bacillomycin D, iturin A, bacilysin, and surfactin. In addition, CH-15 culture filtrate significantly inhibited the growth and conidial germination of pathogens. This study shows that endophytic bacterium CH-15 had antagonistic effect on ginseng root rot pathogens and inhibited the progression of ginseng root rot. We expected that this strain can be a microbial agent to suppress ginseng root rot.

• The Plant Pathology Journal
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• v.33 no.5
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• pp.499-507
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• 2017

In an attempt to develop a biological control agent against mycotoxigenic Fusarium species, we isolated Bacillus amyloliquefaciens strain DA12 from soil and explored its antimicrobial activities. DA12 was active against the growth of mycotoxigenic F. asiaticum, F. graminearum, F. proliferatum, and F. verticillioides both in vitro and in planta (maize). Further screening using dual culture extended the activity range of strain DA12 against other fungal pathogens including Botrytis cinerea, Colletotrichum coccodes, Endothia parasitica, Fusarium oxysporum, Raffaelea quercus-mongolicae, and Rhizoctonia solani. The butanol extract of the culture filtrate of B. amyloliquefaciens DA12 highly inhibited the germination of F. graminearum macroconidia with inhibition rate 83% at a concentration of $31.3{\mu}g/ml$ and 100% at a concentration of $250{\mu}g/ml$. The antifungal metabolite from the butanol extract was identified as iturin A by thin layer chromatography-bioautography. In addition, volatile organic compounds produced by DA12 were able to inhibit mycelial growth of various phytopathogenic fungi. The volatile compounds were identified as 2-heptanone, 5-methyl heptanone and 6-methyl heptanone by gas chromatography-mass spectrometry (GC-MS) analysis. These results indicate that the antagonistic activity of Bacillus amyloliquefaciens DA12 was attributable to iturin A and volatile heptanones, and the strain could be used as a biocontrol agent to reduce the development of Fusarium diseases and mycotoxin contamination of crops.