One hundred and seventeen colonies were screened for the detection of the production of exodextranase on the dextran-mineral salts medium. Ten colonies out of them produced the dextranase. Flavobacterium multivorum greatly producing the enzyme was isolated from soil, identified and then studied for various biochemical characteristics. The activity of the dextranase in the cultured medium was high between pH8 and 9 at $35^{\circ}C$, and between $45^{\circ}C$ and $55^{\circ}C$ at pH8. By the growth curves the generation times of the bacterium were approximately 52 minutes in the LB broth, 38 minutes in the LB plus 1% dextran and 660 minutes in the dextran-salts. The strain did not have ant plasmid, and was susceptible to genramicin, cotrimoxazole and cefoperazone, and moderately susceptible to chloramphenicol, cefamandole and cefotaxime, but resistant to ampicillin, cephalothin, tetracycline, amikacin and tobramycin.
Fusarium and Rhizoctonia genera are important pathogens of many field crops worldwide. They are constantly evolving and expanding their host range. Selecting resistant cultivars is an effective strategy to break their infection cycles. To this end, we screened a collection of Medicago truncatula accessions against Fusarium oxysporum, Fusarium solani, and Rhizoctonia solani strains isolated from different plant species. Despite the small collection, a biodiversity in the disease response of M. truncatula accessions ranging from resistant phenotypes to highly susceptible ones was observed. A17 showed relative resistance to all fungal strains with the lowest disease incidence and ratings while TN1.11 was among the susceptible accessions. As an initiation of the characterization of resistance mechanisms, the antioxidant enzymes' activities, at the early stages of infections, were compared between these contrasting accessions. Our results showed an increment of the antioxidant activities within A17 plants in leaves and roots. We also analyzed the responses of a population of recombinant inbred lines derived from the crossing of A17 and TN1.11 to the infection with the same fungal strains. The broad-sense heritability of measured traits ranged from 0.87 to 0.95, from 0.72 to 0.96, and from 0.14 to 0.85 under control, F. oxysporum, and R. solani conditions, respectively. This high estimated heritability underlines the importance of further molecular analysis of the observed resistance to identify selection markers that could be incorporated into a breeding program and thus improving soil-borne pathogens resistance in crops.
BACKGROUND: Methane is a major greenhouse gas attributed to global warming partly contributed by agricultural activities from ruminant fermentation and rice paddy fields. Methanotrophs are microorganisms that utilize methane. Their unique metabolic lifestyle is enabled by enzymes known as methane monooxygenases (MMOs) catalyzing the oxidation of methane to methanol. Rice absorbs, transports, and releases methane directly from soil water to its stems and the micropores and stomata of the plant epidermis. Methylobacterium species associated with rice are dependent on their host for metabolic substrates including methane. METHODS AND RESULTS: Methylobacterium spp. isolated from rice were evaluated for methane oxidation activities and screened for the presence of sMMO mmoC genes. Qualitatively, the soluble methane monooxygenase (sMMO) activities of the selected strains of Methylobacterium spp. were confirmed by the naphthalene oxidation assay. Quantitatively, the sMMO activity ranged from 41.3 to 159.4 nmol min-1 mg of protein-1. PCR-based amplification and sequencing confirmed the presence and identity of 314 bp size fragment of the mmoC gene showing over 97% similarity to the CBMB27 mmoC gene indicating that Methylobacterium strains belong to a similar group. CONCLUSION(S): Selected Methylobacterium spp. contained the sMMO mmoC gene and possessed methane oxidation activity. As the putative methane oxidizing strains were isolated from rice and have PGP properties, they could be used to simultaneously reduce paddy field methane emission and promote rice growth.
Mohamed H. El-Sayed;Fahdah A. Alshammari;Mohammed H. Sharaf
Journal of Microbiology and Biotechnology
/
v.33
no.1
/
pp.61-74
/
2023
The global increase in multidrug-resistant (MDR) bacteria has inspired researchers to develop new strategies to overcome this problem. In this study, 23 morphologically different, soil-isolated actinomycete cultures were screened for their antibacterial ability against MDR isolates of ESKAPE pathogens. Among them, isolate BOGE18 exhibited a broad antibacterial spectrum, so it was selected and identified based on cultural, morphological, physiological, and biochemical characteristics. Chemotaxonomic analysis was also performed together with nucleotide sequencing of the 16S rRNA gene, which showed this strain to have identity with Streptomyces lienomycini. The ethyl acetate extract of the cell-free filtrate (CFF) of strain BOGE18 was evaluated for its antibacterial spectrum, and the minimum inhibitory concentration (MIC) ranged from 62.5 to 250 ㎍/ml. The recorded results from the in vitro anti-biofilm microtiter assay and confocal laser scanning microscopy (CLSM) of sub-MIC concentrations revealed a significant reduction in biofilm formation in a concentration-dependent manner. The extract also displayed significant scavenging activity, reaching 91.61 ± 4.1% and 85.06 ± 3.14% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), respectively. A promising cytotoxic ability against breast (MCF-7) and hepatocellular (HePG2) cancer cell lines was obtained from the extract with IC50 values of 47.15 ± 13.10 and 122.69 ± 9.12 ㎍/ml, respectively. Moreover, based on gas chromatography-mass spectrometry (GC-MS) analysis, nine known compounds were detected in the BOGE18 extract, suggesting their contribution to the multitude of biological activities recorded in this study. Overall, Streptomyces lienomycini BOGE18-derived extract is a good candidate for use in a natural combating strategy to prevent bacterial infection, especially by MDR pathogens.
Proceedings of the Korean Society of Crop Science Conference
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2022.10a
/
pp.289-289
/
2022
Waterlogging tolerance of corn is one of the important factor for cultivate in paddy soil condition to increase cultivation area and self-sufficiency of corn in Korea. In order to develop elite waterlogging tolerance corn, the new corn lines bred by crossing wild corn, Teosinte, and cultivated corn inbred lines. Five accessions among the 2 species, Zea mays sub spp. mexicana and Zea mays spp. parviglumis, of 81 Teosinte were selected through the waterlogging treatment. The waterlogging treatments were implemented for 7 days at the seedling(V3) stage. The inbred lines were developed by crossing 5 teosinte accessions and cultivated corn lines and they were estimated waterlogging tolerance. It was screened and analyzed the metabolites extracted from roots of 19KT-32(KS141 × teosinte) that was treated waterlogging. We selected 8 of 180 metabolites like as γ-aminobutyric acid(GABA), putrescine, citrulline, Gly, and Ala that expression was remarkably changed over 2.5-times, 7 metabolites increased and 1 metabolite decreased in waterlogging, respectively. Glutamate decarboxylase(GAD) catalyzing GABA accumulation gene have 10 haplotypes, and exon1 was highly conserved, but identified to 135 SNPs after the first intron. Among the 135 SNPs, the number of transversion mutations (52) surpassed the number of transition mutations (38). Most of metabolites were related to abiotic stress in plant that it regulated to pH, osmotic pressure K+/Ca++ and ATPase activity. We are analyzing the association using these results for increase breeding efficiency.
We screened biotic and abiotic preventative,i(preventers) from natural resources to prevent the rusty phenomenon in ginseng roots. To select preventatives(preventers), soil microbes such as Agrobacterium and certain microbes isolated from the rusty ginsengs and the soil in which the rusty ginsengs were planted and used. It is also performed with germination tests of the seeds of Latuca Sativa L. We identified that how selected preventatives(preventers) effect the germination of ginseng seeds. Furthermore, how these influence on the rusty phenomenon and the growth of 1 -year-old ginsengs treated in the pavement. The final preventatives; ICPE-C1$\sub$05/, ICPE-P$\^$107/ were effective in not only the growth of ginseng, but also inhibition of the rusty phenomenon. Moreover, we selected abiotic soil improvers; called P, R, and W, respectively; to promote the effects of preventatives. R and W was excellented among choring improvers. The germination rate of 2-year-old ginsengs treated with ICPE-C$\sub$105/P, and ICPE-P$\sub$107/P was the highest under the effects of naturally selected preventatives mixing with abiotic soil improvers. All treat which was compounding preventers & improvers were so excellented of growth ginseng. Especially treats of ICPE-C$\sub$105/R and ICPE-P$\sub$107/R showed growth increased of each 67.3% and 52.7% As well, the growth of ginseng was the highest in the treatment of ICPE-C$\sub$105/R, and ICPE-P$\sub$105/R. Though rusty of rate was emerged 35% in control, preventers ICPE-C$\sub$105/R and ICPE-P$\sub$107/R were emerged 5.3%. It was affirmed effective of preventer. On the other hands, amounts of ginsenoside treated with preventatives showed to be changed. The ginsenoside was increased to 14.2% with treatment with ICPE-P$\sub$107/R which is highest among groups compared to control, and ICPE-C$\sub$105/P was increased to 5.0%. To sum up with total results, it is judged that biotic preventatives (ICPE-C$\sub$105/R, and ICPE-P$\sub$107/R) which we created improve both a high yield of ginseng and the inhibition of the rusty phenomenon. phenomenon.
Journal of the Korean Society of Food Science and Nutrition
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v.37
no.11
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pp.1507-1514
/
2008
Cholesterol oxidase catalyses the conversion of cholesterol to 4-cholesten-3-one. This enzyme has been used for clinical assay of human serum cholesterol and for reduction of cholesterol level in foods and feeds. In order to search the microorganism which has a high extracellular and stable activity of cholesterol oxidase, soil microorganisms were screened. As a result, the one with the highest extracellular cholesterol oxidase activity was obtained and named as the BEN 115. The BEN 115 strain was identified as one of the Nocardia species based on our taxonomic studies. The cholesterol oxidase from this strain was shown to have two bands of extracellular proteins on SDS-PAGE and Western blot. Their molecular masses were estimated to be about 55 and 57 kDa, respectively. In addition, this cholesterol oxidase was considerably stable at the broad range of pH $3.5{\sim}9.5$ and at the temperature of $25{\sim}55^{\circ}C$. The optimum pH and temperature of this cholesterol oxidase were pH 5.5 and $35^{\circ}C$, respectively. The activity of extracellular cholesterol oxidase could be enhanced 1.6 to 2.0 folds by the addition of nonionic detergent such as Triton X-114, Triton X-100, or Tween-80 into the culturing broth. The substrate specificities against campesterol, sitosterol and stigmasterol were measured to be 50%, 50%, and 27%, respectively, compared to the cholesterol. These results suggest that Nocardia sp. BEN 115 may be useful as a microbial source of cholesterol oxidase production.
Application of rhizospheric fungi is an effective and environmentally friendly method of improving plant growth and controlling many plant diseases. The current study was aimed to identify phytohormone-producing fungi from soil, to understand their roles in sesame plant growth, and to control Fusarium disease. Three predominant fungi (PNF1, PNF2, and PNF3) isolated from the rhizospheric soil of peanut plants were screened for their growth-promoting efficiency on sesame seedlings. Among these isolates, PNF2 significantly increased the shoot length and fresh weight of seedlings compared with controls. Analysis of the fungal culture filtrate showed a higher concentration of indole acetic acid in PNF2 than in the other isolates. PNF2 was identified as Penicillium sp. on the basis of phylogenetic analysis of ITS sequence similarity. The in vitro biocontrol activity of Penicillium sp. against Fusarium sp. was exhibited by a 49% inhibition of mycelial growth in a dual culture bioassay and by hyphal injuries as observed by scanning electron microscopy. In addition, greenhouse experiments revealed that Fusarium inhibited growth in sesame plants by damaging lipid membranes and reducing protein content. Co-cultivation with Penicillium sp. mitigated Fusarium-induced oxidative stress in sesame plants by limiting membrane lipid peroxidation, and by increasing the protein concentration, levels of antioxidants such as total polyphenols, and peroxidase and polyphenoloxidase activities. Thus, our findings suggest that Penicillium sp. is a potent plant growth-promoting fungus that has the ability to ameliorate damage caused by Fusarium infection in sesame cultivation.
This study was conducted to evaluate the degree of acid tolerance of indigenous Rhizobium in natural soils, and to select the superior Rhizobia for legume forage on hilly land. 153 Rhizobial strains isolated from two host genera, Medicago and Trifolium in the pastures of Yeongnam area were screened on acidic agar plates. The results obtained are summarized as follows: 1. The degrees of acid tolerance of R. meliloti and R. trifolii were ranged from 4.3 to pH 5.5 and from 4.0 to pH 5.5 respectively. 2. Among acid tolerant Rhizobia isolates, two strains of R. meliloti at pH 4.3 and three strains of R. trifolii at pH 4.0 were shown to have the activities of nodulation and nitrogenase. 3. R. trifolii strains were shown to survive about 80 to 99% at the pH range 4.8 to 5.0, while R. meliloti were about 35 to 47% at that pH range. Therefore, more acid tolerant R. meliloti should be developed for legume forage in Korean hilly land.
Kim, Ho-Sang;Seong, Ki-Young;Eun, Moo-Young;Hwang, Cher-Won
Applied Biological Chemistry
/
v.41
no.2
/
pp.125-129
/
1998
Xanthomonas sp. isolated from soil exhibited cell wall lytic activity of Candida albicans and secreted chitinase in chitin media. Especially, the chitinase activity was induced by chitin and reached a maximum level at 3 days culture in chitin media. We constructed genomic library of Xanthomonas sp. using cosmid vector in E. coli. Oligonucleotide probe was synthesized from the consensus sequence corresponding to chitinase active site, which was derived from the comparison of amino acid sequences of bacterial chitinase genes. Using this oligonucleotide probe, we screened the genomic library. By restriction enzyme mapping of the positive clones, we identified 4 independent clones which may contain the chitinase gene. One of the clones, named pXCH1 (1.2 kb insert), was further analyzed. Northern blot analysis indicated that is transcripts, 1 kb and 0.8 kb, were induced by chitin. When the cloned gene was induced by IPTG in E.coli cell, chitinase activity which was secreted onto culture media was not observed. However, when the cell was disrupted by using sonicator and then centrifuged, the supernatant exhibited chitinase activity. SDS-PAGE of the supernatant indicated that about 35 kDa protein was induced by IPTG. From these results, it was concluded that the cloned DNA was one of the chitinase genes of Xanthomonas sp.
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