• BMB Reports
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• v.40 no.2
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• pp.172-179
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• 2007

PCR-mediated recombination describes the process of in vitro chimera formation from related template sequences present in a single PCR amplification. The high levels of genetic redundancy in eukaryotic genomes should make recombination artifacts occur readily. However, few evolutionary biologists adequately consider this phenomenon when studying gene lineages. The cytosolic glyceraldehyde-3-phosphate dehydrogenase gene (GapC), which encodes a NADP-dependent nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase in the cytosol, is a classical lowcopy nuclear gene marker and is commonly used in molecular evolutionary studies. Here, we report on the occurrence of PCR-mediated recombination in the GapC gene family of Hibiscus tiliaceus. The study suggests that recombinant areas appear to be correlated with DNA template secondary structures. Our observations highlight that recombination artifacts should be considered when studying specific and allelic phylogenies. The authors suggest that nested PCR be used to suppress PCRmediated recombination.

• The Plant Pathology Journal
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• v.18 no.1
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• pp.30-35
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• 2002

The hyphal growth and biocontrol efficacy of Trichodemo harzianum in soil may depend on its interactions with biotic components of the soil environment. The effect of soil microbial biomass on growth and biocontrol efficacy of T. hanianum isolate ThzIDl-M3 (green fluorescent protein transformant) was investigated using artificially prepared different levels of soil microbial biomass (153,328, or 517ug biomass carbon per g of dry soil; BC). The hyphal growth of T. harzanum was significantly inhibited in the soil with 328 or 517 $\mu$g BC compared with 153 ug BC. When ThzIDl-M3 was added to the soils as an alginate pellet formulation, the recoverable population of ThzIDl-M3 varied, but the highest population occurred in 517ug BC. Addition of alginate pellets of ThzIDl-M3 to the soils (10 per 50 g) resulted in increased indigenous microbial populations (total fungi, bacterial fluorescent Pseudomonas app., and actinomycetes). Furthermore, colonizing ability of ThzIDl-M3 on sclerotia of Sclerotinia sclerotiorum was significantly reduced in the soil with high revel of BC. These results suggest that increased soil microbial biomass contributes to increased interactions between introduced T. harzianum and soil microorganisms, consequently reducing the biocontrol efficacy of 1T. harzianum.

• The Plant Pathology Journal
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• v.32 no.3
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• pp.273-280
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• 2016

Most biocontrol agents for plant diseases have been isolated from sources such as soils and plants. As an alternative source, we examined the feces of tertiary larvae of the herbivorous rhino beetle, Allomyrina dichotoma for presence of biocontrol-active microbes. The initial screen was performed to detect antifungal activity against two common fungal plant pathogens. The strain with strongest antifungal activity was identified as Bacillus amyloliquefaciens KB3. The inhibitory activity of this strain correlated with lipopeptide productions, including iturin A and surfactin. Production of these surfactants in the KB3 isolate varied with the culture phase and growth medium used. In planta biocontrol activities of cell-free culture filtrates of KB3 were similar to those of the commercial biocontrol agent, B. subtilis QST-713. These results support the presence of microbes with the potential to inhibit fungal growth, such as plant pathogens, in diverse ecological niches.

• Research in Plant Disease
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• v.20 no.3
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• pp.182-188
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• 2014

Because bacterial isolates from only a few genera have been developed commercially as biopesticides, discovery and characterization of novel bacterial strains will be a key to market expansion. Our previous screen using plant bioassays identified 24 novel biocontrol isolates representing 12 different genera. In this study, we characterized the 3 isolates showing the best biocontrol activities. The isolates were Pantoea dispersa WCU35, Proteus myxofaciens WCU244, and Exiguobacterium acetylicum WCU292 based on 16S rRNA sequence analysis. The isolates showed differential production of extracellular enzymes, antimicrobial activity against various fungal or bacterial plant pathogens, and induced systemic resistance activity against tomato gray mold disease caused by Botrytis cinerea. E. acetylicum WCU292 lacked strong in vitro antimicrobial activity against plant pathogens, but induced systemic resistance against tomato gray mold disease. These results confirm that the trait of biological control is found in a wide variety of bacterial genera.

• Mycobiology
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• v.46 no.1
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• pp.33-46
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• 2018

Gray mold (Botrytis cinerea) is one of the most common diseases of strawberries (Fragaria${\times}$ananassa Duchesne) worldwide. Although many chemical fungicides are used for controlling the growth of B. cinerea, the risk of the fungus developing chemical resistance together with consumer demand for reducing the use of chemical fungicides have necessitated an alternative method to control this pathogen. Various naturally occurring microbes aggressively attack plant pathogens and benefit plants by suppressing diseases; these microbes are referred to as biocontrol agents. However, screening of potent biocontrol agents is essential for their further development and commercialization. In this study, 24 strains of yeast with antagonistic ability against gray mold were isolated, and the antifungal activity of the volatile and diffusible metabolites was evaluated. Putative mechanisms of action associated with the biocontrol capacity of yeast strains against B. cinerea were studied through in vitro and in vivo assays. The volatile organic compounds produced by the Galactomyces candidum JYC1146 could be useful in the biological control of plant pathogens and therefore are potential alternative fungicides with low environmental impact.

• Journal of Ginseng Research
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• v.43 no.3
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• pp.408-420
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• 2019

Background: Ginseng (Panax ginseng Meyer) is an invaluable medicinal plant containing various bioactive metabolites (e.g., ginsenosides). Owing to its long cultivation period, ginseng is vulnerable to various biotic constraints. Biological control using endophytes is an important alternative to chemical control. Methods: In this study, endophytic Trichoderma citrinoviride PG87, isolated from mountain-cultivated ginseng, was evaluated for biocontrol activity against six major ginseng pathogens. T. citrinoviride exhibited antagonistic activity with mycoparasitism against all ginseng pathogens, with high endo-1,4-${\beta}$-D-glucanase activity. Results: T. citrinoviride inoculation significantly reduced the disease symptoms caused by Botrytis cinerea and Cylindrocarpon destructans and induced ginsenoside biosynthesis in ginseng plants. T. citrinoviride was formulated as dustable powder and granules. The formulated agents also exhibited significant biocontrol activity and induced ginsenosides production in the controlled environment and mountain area. Conclusion: Our results revealed that T. citrinoviride has great potential as a biological control agent and elicitor of ginsenoside production.

• The Plant Pathology Journal
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• v.22 no.1
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• pp.103-106
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• 2006

In this biocontrol research, we evaluated disease suppressive effects of antagonistic bacterial strains ISE13 and KJ1R5 against Korean ginseng root rot caused by P. eaetorum. We also examined the effects of nutrient solution in the hydroponic culture system for Korean ginseng on biological activity of the bacterial strains. As results of dual culture tests of the bacterial strains on $V_{8}$ juice agar, the strain ISE13 showed antifungal activity against P. eaetorum and other plant pathogenic fungi, but the strain KJ1R5 did not. When their inhibitory effects against infection of P. eaetorum on the roots grown in either nutrient solution or water were tested, the strains ISE13 and KJ1R5 inhibited the disease severity of Korean ginseng roots only grown with water, compared to buffer-treated, inoculated controls. However, the nutrient solution used for hydroponic cultures of ginseng in pots caused higher levels of disease severity by the strains ISE13 and KJ1R5 from 418.8\%$ to 40.0\%$ and from 24.3\%$ to 45.0\%$, respectively. In this study, the bacterial strains ISE13 and KJ1R5 could be potentially biocontrol agents to suppress Korean ginseng root rot caused by P. eaetorum. However, more attention using nutrient solution in hydroponic cultures for Korean ginseng production should be applied in biocontrol of plant diseases using the antagonistic microorganisms.

• The Plant Pathology Journal
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• v.25 no.1
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• pp.62-69
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• 2009

In this study, we characterized the bacterial strain KJ2C12 in relation with its biocontrol activity against Phytophthora capsici on pepper, and identified this strain using morphological, physiological, biochemical, fatty acid methyl ester, and 16S rRNA gene sequence analyses. Strain KJ2C12 significantly (P=0.05) reduced both final disease severity and areas under the disease progress curves of 5-week-old pepper plants inoculated with P. capsici compared to buffer-treated controls. As for the production of antibiotics, biofilms, biosurfactant, extracellular enzyme, HCN, and swarming activity, strain KJ2C12 produced an extracellular enzyme with protease activity, but no other productions or swarming activity. However, Escherichia coli produced weak biofilm only. Strain KJ2C12 could colonize pepper roots more effectively in a gnotobiotic system using sterile quartz sand compared to E. coli over 4 weeks after treatments. However, no bacterial populations were detected in 10 mM $MgSO_4$ buffer-treated controls. Strain KJ2C12 produced significantly higher microbial activity than the $MgSO_4$-treated control or E. coli over 4 weeks after treatments. Bacterial strain KJ2C12 was identified as Bacillus luciferensis based on morphological, physiological, and biochemical characteristics as well as FAME and 16S rRNA gene sequence analyses. In addition, these results suggested that B. luciferensis strain KJ2C12 could reduce Phytophthora blight of pepper by protecting infection courts through enhanced effective root colonization with protease production and an increase of soil microbial activity.

• The Plant Pathology Journal
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• v.22 no.4
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• pp.364-368
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• 2006

Survival of biocontrol agents and their effective colonization of rhizhosphere are the essential components for successful disease suppression. The effects of hydrogel supplement on bacterial survival and disease control were evaluated in pot and in the field. Addition of 2% hydrogel material to potting soil resulted in significant enhancement of colonization of biocontrol agent Serratia plymuthica A21-4 both in soil and rhizosphere of pepper plants. Rhizosphere colonization of S. plymuthica A21-4 retrieved from 40 days old pepper seedlings indicated 100 times higher bacterial population in hydrogel treated soil than in ordinary pot soil. The pepper plants sown in hydrogelated potting soil showed higher seed germination rate and the better growth of pepper plant than those in ordinary commercial pot soil. Although the suppression of Phytophthora capsid density in the potting soil by treatment of biocontrol agent A21-4 was not significantly different between in hydrogelated soil and ordinary potting soil, the suppression of Phytophthora blight between two treatments was significantly different. A21-4 treatment in hydrogelated potting soil was completely disease-free while same treatment in ordinary potting soil revealed 36% disease incidence. Our field study under natural disease occurrence also showed significantly less disease incidence(12.3%) in the A21-4 treatment in the hydrogelated soil compared to other treatments. Yield promotion of pepper by the A21-4 treatment in the hydrogelated potting soil was also recognized. Our results indicated that hydrogel amendment with biocontrol agent in pot soil would be a good alternative to protect pepper seedlings and increase plant yield.

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.831-838
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• 2018

Trophic interactions of introduced biocontrol fungi with soil animals can be a key determinant in the fungal proliferation and activity. This study investigated the trophic interaction of an introduced biocontrol fungus with soil nematodes. The biocontrol fungus Trichoderma harzianum ThzID1-M3 and the fungivorous nematode Aphelenchoides sp. (10 per gram of soil) were added to nonsterile soil, and microbial populations were monitored for 40 days. Similar results were obtained when the experiment was duplicated. ThzID1-M3 stimulated the population growth of indigenous nematodes (p < 0.05), regardless of whether Aphelenchoides sp. was added. Without ThzID1-M3, indigenous nematodes did not increase in number and the added Aphelenchoides sp. nematodes almost disappeared by day 10. With ThzID1-M3, population growth of nematodes was rapid between 5 and 10 days after treatment. ThzID1-M3 biomass peaked on day 5, dropped at day 10, and then almost disappeared at day 20, which was not influenced by the addition of nematodes. In contrast, a large quantity of ThzID1-M3 hyphae were present in a heat-treated soil in which nematodes were eliminated. Total fungal biomass in all treatments peaked on day 5 and subsequently decreased. Addition of nematodes increased the total fungal biomass (p < 0.05), but ThzID1-M3 addition did not affect the fungal biomass. Hyphae of total fungi when homogenously distributed did not support the nematode population growth; however, hyphae of the introduced fungus did when densely localized. The results suggest that soil fungivorous nematodes are an important constraint on the hyphal proliferation of fungal agents introduced into natural soils.