• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1577-1584
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• 2010

In total, 140 halotolerant bacterial strains were isolated from both the soil of barren fields and the rhizosphere of six naturally growing halophytic plants in the vicinity of the Yellow Sea, near the city of Incheon in the Republic of Korea. All of these strains were characterized for multiple plant growth promoting traits, such as the production of indole acetic acid (IAA), nitrogen fixation, phosphorus (P) and zinc (Zn) solubilization, thiosulfate ($S_2O_3$) oxidation, the production of ammonia ($NH_3$), and the production of extracellular hydrolytic enzymes such as protease, chitinase, pectinase, cellulase, and lipase under in vitro conditions. From the original 140 strains tested, on the basis of the latter tests for plant growth promotional activity, 36 were selected for further examination. These 36 halotolerant bacterial strains were then tested for 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Twenty-five of these were found to be positive, and to be exhibiting significantly varying levels of activity. 16S rRNA gene sequencing analyses of the 36 halotolerant strains showed that they belong to 10 different bacterial genera: Bacillus, Brevibacterium, Planococcus, Zhihengliuella, Halomonas, Exiguobacterium, Oceanimonas, Corynebacterium, Arthrobacter, and Micrococcus. Inoculation of the 14 halotolerant bacterial strains to ameliorate salt stress (150 mM NaCl) in canola plants produced an increase in root length of between 5.2% and 47.8%, and dry weight of between 16.2% and 43%, in comparison with the uninoculated positive controls. In particular, three of the bacteria, Brevibacterium epidermidis RS15, Micrococcus yunnanensis RS222, and Bacillus aryabhattai RS341, all showed more than 40% increase in root elongation and dry weight when compared with uninoculated salt-stressed canola seedlings. These results indicate that certain halotolerant bacteria, isolated from coastal soils, have a real potential to enhance plant growth under saline stress, through the reduction of ethylene production via ACC deaminase activity.

• The Plant Pathology Journal
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• v.34 no.2
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• pp.126-138
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• 2018

Rice blast caused by Magnaporthe oryzae is a major disease. In the present study, we aimed to identify and evaluate the novel bacterial isolates from rice rhizosphere for biocontrol of M. oryzae pathogen. Sixty bacterial strains from the rice plant's rhizosphere were tested for their biocontrol activity against M. oryzae under in vitro and in vivo. Among them, B. amyloliquefaciens had significant high activity against the pathogen. The least disease severity and highest germination were recorded in seeds treated with B. amyloliquefaciens UASBR9 (0.96 and 98.00%) compared to untreated control (3.43 and 95.00%, respectively) under in vivo condition. These isolates had high activity of enzymes in relation to growth promoting activity upon challenge inoculation of the pathogen. The potential strains were identified based on 16S rRNA gene sequencing and dominance of these particular genes were associated in Bacillus strains. These strains were also confirmed for the presence of antimicrobial peptide biosynthetic genes viz., srfAA (surfactin), fenD (fengycin), spaS (subtilin), and ituC (iturin) related to secondary metabolite production (e.g., AMPs). Overall, the results suggested that application of potential bacterial strains like B. amyloliquefaciens UASBR9 not only helps in control of the biological suppression of one of the most devastating rice pathogens, M. grisea but also increases plant growth along with a reduction in application of toxic chemical pesticides.

• The Plant Pathology Journal
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• v.29 no.2
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• pp.168-173
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• 2013

Apple ring rot disease, caused by Botryosphaeria dothidea (Moug. ex. Fr) Ces. et de Not., is one of the most important diseases on apple fruits. In this study, strain 9001 isolated from healthy apple fruits from an infested orchard was evaluated for its biocontrol activity against apple ring rot in vitro and in vivo. Strain 9001 showed obvious antagonistic activity to B. dothidea YL-1 when plated on potato dextrose agar. Soaking healthy apples in the bacterial suspensions of strain 9001 prior to artificial inoculation of fungal pathogen resulted in a dramatic decrease in disease incidence when compared to the control. Moreover, either field application in the growth season or postharvest treatment of apples from infected orchards with bacterial suspensions of strain 9001 resulted in significantly reduced disease incidence within the storage period for 4 months at room temperature. Based on the phylogenetic analysis of 16S rRNA and the gyrA gene, strain 9001 was identified as Bacillus amyloliquefaciens. These results indicated that B. amyloliquefaciens 9001 could be a promising agent in biocontrol of apple ring rot on fruit, which might help to minimize the yield loss of apple fruit during the long postharvest period.

• Journal of The Korean Society of Grassland and Forage Science
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• v.35 no.3
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• pp.201-206
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• 2015

An experiment was carried out to determine the homofermentative activity of Lactobacillus plantarum KCC-10 and KCC-19 on the ensiling of whole crop barley (WCB). The crude protein in the silages was slightly higher in the KCC-10 and KCC-19 treatments compared to the control, but there was no significant difference between the two inoculant-treated silages. Nutrient parameters such as acid detergent fiber, neutral detergent fiber and in vitro dry matter digestibility in L. plantarum KCC-10 and KCC-19 treated silages did not differ from those in the control silage. The lactic acid content increased in KCC-10 and KCC-19 treated silage when compared with the control silage but the contents of acetic acid and butyric acid produced in KCC-10 and KCC-19 treated silages were similar with the control silage. Further, the number of lactic acid bacteria (LAB) in KCC-10 treated silage demonstrated a significant increase when compared to the control. Especially, KCC-19 treated silage showed greater lactic acid bacterial growth potential. Other microbes such as yeast and fungi were not detected in KCC-10 and KCC-19 treated WCB silages. Hence, this study suggests that the addition of L. Plantarum KCC-10 and KCC-19 to the WCB silage can improve fermentation quality for the production of high-quality silage.

• Korean Journal of Poultry Science
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• v.27 no.2
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• pp.155-161
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• 2000

Unfortunately, there is no technique which is stable and repetitive to produce transgenic chicken, although various ways of gene transfer including PGC-and embryonic cell-mediated gene transfer, DNA microinjection, virus inoculation and sperm cells have been employed. The aims of this study were 세 develop and establish such a stable, repetitive and efficient way of gene transfer giving a faithful gene expression during development after the reconstruction of embryo in an UV-irradiated egg. A dual reporter plasmid (pJJ9), a fusion gene containing lacZ and GFP driven by a CMV promoter was used to exploit either merits of both reporting markers. lacZ with strong signal or GFP with vital marking. Electroporated embryonic blastodermal cells (EBCs) in the presence of the pJJ9 DNA faithfully showed 377 bp PCR product and lacZ or GFP expressions in the identical cells in vitro of in vivo. Furthermore, analyses of expression pattern of the foreign DNA demonstrated that microinjected EBCs cells into the UV-irradiated recipient egg should participate in normal developmental process, for example, proliferation and differentiation into various tissues. Thirty percentages of the manipulated eggs showed lacZ expression in their tissues. These results together with the specific procedures used in this study should facilitate avian transgenesis.

• Korean Journal of Soil Science and Fertilizer
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• v.37 no.1
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• pp.36-40
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• 2004

It is now widely accepted that immobilized microbial cells can overcome some of the problems associated with microbial survival stability, efficacy, storage, transportation and ease of application in agricultural environments. Pantoea agglomerans, a phosphate solubilizing bacterium, was immobilized in alginate, agar and gelatin carriers. All the three immobilfized carriers with bacterial cells of P. agglomerans were compared for solubilization of tricalcium phosphate in pure liquid cultures. While alginate beads were tested for phosphate solubilization on alternate days up to five days, agar beads and gelatin cubes were subjected for one time phosphate solubilization analysis after seven days. Both alginate and agar immobilized cells of P. agglomerans exhibited higher efficiency in increasing the solubilizaliun of tricalcium phosphate than gelatin immobilized cells. The culture filtrate of alginate bead inoculation treatment registered a rapid increase in soluble phosphate concentration upon incubation. A corresponding decrease in the pH of the medium was also observed in all the treatments.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.352-359
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• 2012

One hundred thirty-four putative Bacillus isolates were recovered from soybean rhizosphere soils of Nimar region to select effective zinc solubilizers for increased assimilation of zinc (Zn) in soybean seeds. These isolates were screened in vitro for zinc-solubilization ability on Tris-minimal agar medium supplemented separately with 0.1% zinc in the form of zinc oxide, zinc phosphate, and zinc carbonate. Of all, 9 isolates and a reference Bacillus cereus ATCC 13061 were characterized and identified as Bacillus species based on Gram-positive reaction, endospore-forming cells, and the presence of iso-$C_{15:0}$ and anteiso-$C_{15:0}$ as predominant fatty acids. On plate assay, two isolates KHBD-6 and KHBAR-1 showed a greater diameter of solubilization halo and colony diameter on all the three zinc compounds. The isolates KHBD-6, KHBAR-1, BDSD-2-2C, and KHTH-4-1 and the reference strain ATCC 13061 had higher soluble zinc concentration in liquid medium supplemented with zinc phosphate and zinc carbonate compounds as compared with the other isolates and uninoculated control. Evaluation under microcosm conditions showed that inoculation of isolates KHBD-6 (57.34 ${\mu}g/g$), KHBAR-1 (55.67 ${\mu}g/g$), and strain ATCC 13061 (53.10 ${\mu}g/g$) significantly increased the Zn concentration in soybean seeds as compared with the other isolates and uninoculated control (47.14 ${\mu}g/g$). This study suggests the occurrence of zinc-solubilizing Bacillus in soils of Nimar region and isolates KHBD-6 and KHBAR-1 were found to be promising zinc solubilizers for increased assimilation of Zn in soybean seeds.

• Journal of Microbiology
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• v.40 no.2
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• pp.91-97
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• 2002

Enteroaggregative E. coil (EAEC) is an important aethiological causal agent of diarrhea in people of developed and undeveloped countries. Different in vitro and in vivo models have been proposed to study the pathdgenic and immune mechanisms of EAEC infaction. The aim of this study was to analyze whether BALB/c mice could be used as an animal model to study EAEC pathogenesis Six-week-old BALB/c mice were inoculated with EAEC strain 042 (044:H88) nalidixic acid resistant, and re-inoc-ulated ten days after. Mice feces were monitored for the presence of the EAEC strain over a period of 20 days . Bacteria were enumerated on MacConkey agar containing 100$\mu$g of nalidixic acid per ml. Results showed that 35% of the animals were colonized for 3 days, 15% for 5 and 10% for more than 7 days . After re-inoculation only 16% of the animals remained colonized for more than 3 days. During the necropsy, the intestinal fluid of same of the infected animals presented mucus and blood. Six of these fluids showed the presence of IgA antibodies againset Pet toxin and IgG natibodies raised against the toxin were also detected in the animal serum. Histopathologic evidence confirms the stimulation of mucus hypersecretion, an increased amount of goblet cells and the presence of bacterial aggregates in the apical surfaces of intestinal epithelial cells. Edema was present in the submucosa. These results suggest that BALB/c mice could be used as an animal model for in vivo study of EAEC infection.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.5
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• pp.522-529
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• 1998

This experiment was conducted to study the effects of lactic acid bacteria (LAB) inoculation either alone or in combination with cell wall degrading enzymes on the fermentation characteristics and chemical compositions of Rhodesgrass silage. Over to 1 kg of fresh Rhodesgrass sample a treatment of inoculant LAB with or without addition of an enzyme of Acremoniumcellulase (A) or Meicelase (M) or a mixture of both enzymes (AM) was applied. The treatments were control untreated, LAB-treated (application rate $1.0{\times}10^5cfu/g$ fresh sample), LAB+A 0.005%, LAB+A 0.01%, LAB+A 0.02%, LAB+M 0.005%, LAB+M 0.01%, LAB+M 0.02 %, LAB+AM 0.005%, LAB+AM 0.01%, and LAB+AM 0.02%. The sample was ensiled into 2-L vinyl bottle silo, with 9 silages of each treatment were made. Three silages of each treatment were incubated at 20, 30 and $40^{\circ}C$ for 2-months of storage period. All silages were well preserved with their fermentation quality has low pH values (3.91-4.26) and high lactic acid concentrations (4.11-9.89 %DM). No differences were found in fermentation quality and chemical composition of the control untreated silage as compared to the LAB-treated silage. Combined treatment of LAB+cellulases improved the fermentation quality of silages measured in terms of lower (p < 0.01) pH values and higher (p < 0.05) lactic concentrations than those of LAB-treated silages. Increasing amount of cellulase addition resulted in decrease (p < 0.05) of pH value and increase (p < 0.05) of lactic acid concentration. LAB + cellulase treatments (all cellulase types) reduced (p < 0.01) NDF, ADF and in vitro dry matter digestibility of silages compared with the control untreated silages. The fermentation quality and the rate of cell wall reduction were higher (p < 0.01) in the silages treated with LAB + cellulase A than in the silages treated with either LAB+cellulase M or LAB + cellulase AM. Incubation temperature of $40^{\circ}C$ was likely to be more appropriate environment for stimulating the fermentation of Rhodesgrass silages than those of 20 and $30^{\circ}C$.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.4
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• pp.511-517
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• 2014

This study examined the effects of bacterial inoculants on chemical composition and fermentation indices of barley silage. Barley forage (Youngyang) was harvested at 24% dry matter (DM) and wilted to 47.9% DM. The wilted barley forage was chopped to 3-5 cm length and applied with no inoculant (CON), L. plantarum ($1{\times}10^{10}cfu/g$, LP) or Effective Microorganisms ($0.5{\times}10^9cfu/g$, EM). Then the forages were ensiled in four replications for each treatment in 20 L mini silos and stored for 100 days. The contents of crude protein and ether extract were higher in CON silage ensiled for 100-d, while the contents of DM and crude ash were higher in EM silage (p<0.05). The contents of ADF, NDF and hemicellulose as well as the in vitro DM digestibility were not affected by microbial inoculation (p>0.05). The pH, ammonia-N concentration and lactate to acetate ratio were higher (p<0.05) in CON silage, while lactate concentrations were higher (p<0.05) in CON and LP silage. Acetate concentration and lactic acid bacteria was increased (p<0.05) by both inoculants (LP and EM), but propionate concentration and yeast was increased (p<0.05) by EM and LP, respectively. These results indicated that the fermentation quality of barley silage was improved by the application of bacterial inoculants.