• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.108.1-108
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• 2003

Soil metagenome is untapped total microbial genome including that of the majority of unculturable bacteria present in soil. We constructed soil metagenomic library in Escherichia coli using DNA directly extracted from two different soils, pine tree rhizosphere soil and forest topsoil. Metagenomic libraries constructed from pine tree rhizosphere soil and forest topsoil consisted of approximately 33,700 clones and 112,000 clones with average insert DNA size of 35-kb, respectively. Subsequently, we screened the libraries to select clones with antimicrobial activities against Saccharomyces cerevisiae and Agrobacterium tumefaciens using double agar layer method. So far, we have a clone active against S. cerevisiae and a clone active against A. tumefaciens from the forest topsoil library. In vitro mutagenesis and DNA sequence analysis of the antifungal clone revealed the genes involved in the biosynthesis of antimicrobial secondary metabolite. Metagenomic libraries constructed in this study would be subject to search for diverse genetic resources related with useful microbial products.

• Journal of The Korean Society of Grassland and Forage Science
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• v.18 no.2
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• pp.133-142
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• 1998

This study was conducted to investigate the effects of antagonistic bacteria and pathogenic fungi on the growth of tall fescue(Festuca arundinacea Schreb.) in continuous cropping soil(CCS) and non-continuous cropping soil(NCCS). Tall fescue was established by seeding into pots of 11 cm in diameter and 9 cm in depth containing 1 : 1 mixture of soil and vermiculite, and cultivated at pots with antagonistic bacteria and pathogenic fungi in a vinyl house. The bacteria used in this study were Bacillus subtilis and hsants. B. subtilis was isolated and identified kern forage rhizosphere soil and fusants were isolated through cell hsion from B. subtilis and B. thwingiensis. B. subtilis was named as B. subtilis 101 and hsants were named as F-3, F-7 and F-8. In dark culture experiment, tall fescue inoculated with the antagonistic bacteria lived longer than that of control in both CCS and NCCS. However, tall fescue of CCS lived shorter than that of NCCS. Dry weight of tall fescue inoculated with the antagonistic bacteria was higher than that of tall fescue inoculated with pathogenic hngi in both CCS and NCCS(P< 0.05), and the antagonistic bacteria showed positive effects on the growth of tall fescue. However, Dry weight of tall fescue was decreased by the inoculation of the pathogenic b g i in both CCS and NCCS(P< 0.05).

• Korean Journal of Environmental Agriculture
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• v.40 no.4
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• pp.345-352
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• 2021

BACKGROUND: Salinity is one of the major limiting factors in agriculture that affect the growth and productivity of crops. It is economically difficult to artificially purify the soil affected by salt. Therefore, the use of plant growth-promoting bacteria (PGPB) in an effort to reduce stress caused by salt is emerging as a cost-effective and environment-friendly method. In this study, the purpose was to isolate the salt-tolerant bacteria from the rhizosphere soil and identify their ability to promote plant growth under salt stress condition. METHODS AND RESULTS: The isolates KST-1, KST-2, AST-3, and AST-4 that showed plant growth-promoting activity for barley in salt conditions were close to Bacillus cereus (KST-1, KST-2, and AST-4) and Bacillus thuringiensis (AST-3) and showed high salt tolerance up to 7% of additional NaCl to the media. When inoculated to barley, the strains had only minor effect on the length of the barley. However, the concentrations of chlorophyll in the barley leaves were found to be higher from the bacteria-inoculated pots than those from the uninoculated control. In particular, the chlorophyll concentration in Bacillus cereus AST-4 experiment was 5.45 times higher than that of the uninoculated control under the same experimental condition. CONCLUSION(S): The isolated salt-tolerant bacteria were found to influence on chlorophyll concentration of the barley. As represented by the strain AST-4, microbes may suggest a cost-effective and environmentally benign method to alleviate salt stress of crops cultivated in salt-accumulated soils such as reclaimed lands.

• The Plant Pathology Journal
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• v.18 no.2
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• pp.57-62
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• 2002

In vivo expression technology (IVET) has been developed to study bacterial gene expression in Salmonella typhimurium during host infection. The expression of selected genes by IVET has been elevated in vivo but not in vitro. The selected genes turned out to be important for bacterial virulence and/or pathogenicity. IVET depends on a synthetic operon with a promoterless transcriptional fusion between a selection marker gene and a reporter gene. The IVET approach has been successfully adapted in other bacterial pathogens and plant-associated bacteria using different selection markers. Pseudomonas putida suppresses citrus root rot caused by Phytophthora parasitica and enhances citrus seedling growth. The WET strategy was adapted based on a transcriptional fusion, pyrBC'-lacZ, in P. putida to study the bacterial traits important far biocontrol activities. Several genes appeared to be induced on P. parasitica hyphae and were found to be related with metabolism and regulation of gene expression. It is likely that the biocontrol strain took a metabolic advantage from the plant pathogenic fungus and then suppressed citrus root rot effectively. The result was parallel with those from the adaptation of IVET in P. fluorescens, a plant growth promoting rhizobacteria (PGPR). Interestingly, genes encoding components for type III secretion system have been identified as rhizosphere-induced genes in the PGPR strain. The type III secretion system may play a certain role during interaction with its counterpart plants. Application of IVET has been demonstrated in a wide range of bacteria. It is an important strategy to genetically understand complicated bacterial traits in the environment.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1994.06a
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• pp.50-61
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• 1994

Biological control of important fungal diseases such as Phytophthora blight of red pepper, gary mold rot of vegetables, and powdery mildew of many crops was attempted using an antagonistic bacterium, Bacillus sp. AC-1 in greenhouses and fields. The antagonistic bacterium isolated from the rhizosphere soils of healthy red pepper plant was very effective in the inhibition of mycelial growth of plant pathogenic fungi in vitro including Phytophthora capsici, Rhizoctonia solani, Pyricularia oryzae, Botrytis cinerea, Valsa mali, Fusarium oxysporum, Pythium ultimum, Alternari mali, Helminthosporium oryzae, and Colletotrichum gloeosporioides. Culture filtrate of antagonistic Bacillus sp. AC-1 applied to pot soils infested with Phytophthora capsici suppressed the disease occurrence better than metalaxyl application did until 37 days after treatment in greenhouse tests. Treatments of the bacterial suspension on red pepper plants also reduced the incidence of Phytophthora blight in greenhouse tests. In farmers' commercial production fields, however, the controlling efficacy of the antagonistic bacteria was variable depending on field locations. Gray mold rot of chinese chives and lettuce caused by Botrytis cinerea was also controlled effectively in field tests by the application of Bacillus sp. AC-1 with control values of 79.7% and 72.8%, respectively. Spraying of the bacterial suspension inhibited development of powdery mildew of many crops such as cucumber, tobacco, melon, and rose effectively in greenhouse and field tests. The control efficacy of the bacterial suspension was almost same as that of Fenarimol used as a chemical standard. Further experiments for developing a commercial product from the antagonistic bacteria and for elucidating antagonistic mechanism against plant pathogenic fungi are in progress.

• Journal of Applied Biological Chemistry
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• v.60 no.2
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• pp.125-131
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• 2017

Nine strains of high concentrations of salt-tolerant bacteria were isolated from the rhizosphere and rhizoplane of the halophyte plant Suaeda japonica grown in Gochang Buan tidal flat. The isolated bacteria were classified as genera Vibrio (strains JRS-1, -2, -3, -4, and -5, and JRL-1 and -4) and Bacillus (strains JRL-2 and -3) based on the 16S rRNA gene sequence similarity. The optical growth condition for salt concentration was examined on the selected, representative strains. Strain JRS-1 with the closest relative of Vibrio neocaledonicus showed the highest growth rate at the total salt concentration of 6% among the incubation conditions of 3-8% salt concentrations. Strain JRL-2 with the closest relative of Bacillus thuringiensis showed the tendency that growth rate increased with increasing salt concentrations and the maximum growth rate at 7% of the total salt concentration. The isolated bacteria showed salt-resistances to higher salt concentrations than their habitat soils with 3%. In addition, we identified evidences of potentially plant interaction-relevant enzymatic activities, from utilization of some substrates rich in plants, such as triglyceride, ${\rho}$-nitrophenyl-${\alpha}$,$\text\tiny{D}$-glucoside, and ${\rho}$-nitrophenyl-${\beta}$,$\text\tiny{D}$-glucoside.

• Korean Journal of Soil Science and Fertilizer
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• v.33 no.4
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• pp.266-274
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• 2000

Understanding of microbial community structure in soil-root system is necessary to use beneficial soil and rhizosphere microbes for improvement of crop production and biocontrol. The knowledge of behavior and function of microbes in soil-root system plays a key role for the application of beneficial inocula. Because the majority of the intact bacteria in soil are unable to grow on nutrient media, both culturable and nonculturable bacteria have to be studied together. In our study, culture-independent survey of bacterial community in the soil-root system of red pepper fields was conducted by the sequence analysis of three universal clone libraries of genes which code for small-subunit rRNA (rDNA). Universal small subunit rRNA primers were used to amplify DNA extracted from each sample and PCR products were cloned into pGEM-T. Out of 27 clones sequenced, 25 clones were from domain bacteria. Two of the rDNA sequences were derived from eukaryotic organelles. Within the domain bacteria, several kingdoms were represented : the Proteobacteria (16 clones). Cytophyga-Flexibacter-Bacteroides group (2 clones). the high G+C content gram-positive group(1 clone) and 4 unknown clones.

• Journal of Life Science
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• v.24 no.4
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• pp.461-466
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• 2014

Three plant species, Aster sphathulifolius, Sedum oryzifolium, and Lysimachia mauritiana, native to the Dokdo Islands in South Korea, were examined for rhizosphere microorganisms by using 16S rDNA sequences. Nine species of rhizosphere microorganisms were isolated from the three native plant species, respectively. Phylogenetic analysis showed that the microorganisms could be classified into 19 species belonging to four phyla (Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria), and the characteristics of the microbes were confirmed. Rhizosphere microorganisms from the six orders (Bacillales, Corynebacteriales, Flavobacteriales, Micrococcales, Oceanospirillales, and Rhodobacterales) were isolated from S. oryzifolium. From L. mauritiana, microbes belonging to the seven orders (Bacillales, Flavobacteriales, Micrococcales, Oceanospirillales, Rhizobiales, and Rhodobacterales) were isolated. From A. sphathulifolius, the six orders of rhizosphere microorganisms (Alteromonadales, Bacillales, Corynebacteriales, Flavobacteriales, Micrococcales, and Rhizobiales) were isolated. These data showed that Actinobacteria and Proteobacteria were the dominant phyla for the rhizosphere of all three plants. To confirm the bacterial diversity in rhizospheres, Shannon's diversity index (H') was used at the genus level. In these data, the rhizosphere from S. oryzifolium and L. mauritiana had more diverse bacteria compared to that from A. sphathulifolius.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.4
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• pp.582-592
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• 2012

Microorganisms present in the rhizosphere soil plays a vital role in improving the plant growth and soil fertility. Many kinds of fertilizers including chemical and organic has been approached to improve the productivity. Though some of them showed significant improvement in yield, they failed to maintain the soil properties. Rather they negatively affected soil eventually, the land became unsuitable for agricultural. To overcome these problems, microorganisms have been used as effective alternative. For past few decades, plant growth promoting rhizobacteria (PGPR) and arbuscular mycorrhizal fungi (AMF) have been used as effective inoculants to enhance the plant growth and productivity. PGPR improves the plant growth and helps the plant to withstand biotic and abiotic stresses. AM fungi are known to colonize roots of plants and they increase the plant nutrient uptake. Spore associated bacteria (SAB) are attached to spore wall or hyphae and known to increase the AMF germination and root colonization but their mechanism of interaction is poorly known. Better understanding the interactions among AMF, SAB and PGPR are necessary to enhance the quality of inoculants as a biofertilizers. In this paper, current knowledge about the interactions between fungi and bacteria are reviewed and discussed about AMF spore associated bacteria.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.437-447
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• 2012

Natural and beneficial associations between plants and bacteria have demonstrated potential commercial application for several agricultural crops. The sunflower has acquired increasing importance in Brazilian agribusiness owing to its agronomic characteristics such as the tolerance to edaphoclimatic variations, resistance to pests and diseases, and adaptation to the implements commonly used for maize and soybean, as well as the versatility of the products and by-products obtained from its cultivation. A study of the cultivable bacteria associated with two sunflower cultivars, using classical microbiological methods, successfully obtained isolates from different plant tissues (roots, stems, florets, and rhizosphere). Out of 57 plant-growth-promoting isolates obtained, 45 were identified at the genus level and phylogenetically positioned based on 16S rRNA gene sequencing: 42 Bacillus (B. subtilis, B. cereus, B. thuringiensis, B. pumilus, B. megaterium, and Bacillus sp.) and 3 Methylobacterium komagatae. Random amplified polymorphic DNA (RAPD) analysis showed a broad diversity among the Bacillus isolates, which clustered into 2 groups with 75% similarity and 13 subgroups with 85% similarity, suggesting that the genetic distance correlated with the source of isolation. The isolates were also analyzed for certain growth-promoting activities. Auxin synthesis was widely distributed among the isolates, with values ranging from 93.34 to 1653.37 ${\mu}M$ auxin per ${\mu}g$ of protein. The phosphate solubilization index ranged from 1.25 to 3.89, and siderophore index varied from 1.15 to 5.25. From a total of 57 isolates, 3 showed an ability to biologically fix atmospheric nitrogen, and 7 showed antagonism against the pathogen Sclerotinia sclerotiorum. The results of biochemical characterization allowed identification of potential candidates for the development of biofertilizers targeted to the sunflower crop.