• Mycobiology
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• v.32 no.4
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• pp.197-198
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• 2004

A new technique for single spore isolation was developed for predacious fungi forming constricting rings directly on the spores using Dactylaria brochopaga and Arthrobotrys dactyloides. Constricting rings were induced directly on the spores by transferring the spores in 25 ppm solution of DL-Valine in sterile distilled water. Freshly hatched and thoroughly washed second stage juveniles of Meloidogyne incognita were transferred into cavity blocks containing induced rings for trapping and killing of nematodes. The killed nematodes were surface sterilized with streptomycin and inoculated into petri dishes containing maize meal agar media with 100 ppm streptomycin. The petri dishes were incubated at $29{\pm}1^{\circ}C$ for few days which yielded axenic culture of these fungi.

• Korean Journal of Soil Science and Fertilizer
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• v.20 no.3
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• pp.269-274
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• 1987

A laboratory experiment was conducted to find out the effect of inoculation with plant growth of associative $N_2$-fixing bacteria on the combination of eight different rice seedling with seven different associative $N_2$-fixing bacteria in which were isolated and identified from grasses and rice in saline and reclaimed saline paddy soil. Those were examined in N-free axenic culture medium. The result of this experiment exemplified the contribution of associative $N_2$-fixing bacteria to their host plant were varied much by the combination of rice cultivars and bacterial strains. Acetylene reduction ($N_2$-fixation) activity seems to be more variable with respect to the combination of rice cultivars and bacterial strains. The relationship between acetylene reduction activity and rice performance such as dry matter weight, plant height, root length, and number of root were statistically insignificant. However, in comparision with the control, the dry matter yield of Annapuruna was increased by inoculation of all the seven bacteria strains. Among the seven rice cultivars, the average acetylene reducing activity was the highest in Annapuruna, and Shingwang, and the lowest in IR-8 inoculated with Pseudomonas sp H8. Among the seven bacterial strains the highest acetylene reducing activity was obtained by the combination of Shingwang with Pseudomonas sp H8.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.429-433
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• 2000

Giardia lamblia, a human pathogen causing outbreaks of diarrhea, recently became a focus of great concerns in the fields of both medical and environmental microbioloty. To develop the experimental tools to study giardiasis, encystation, one of the major processes in its life cycle, was reconstituted by inducing an axenic culture of a flagellated form of G. lamblia into a cyst from under high concentration of bile and alkaline pH condition. The successful induction was confirmed by Northern analysis of resulting increased expression of the CWPl gene encoding the cyst wall protein 1. An examination of the encystation process with SEM (scanning electron microscopy) and TEM (transmission electron microscopy) revealed that the trophozoite, a flagellate with a bilateral symmetry, was transformed to a cyst form with an oval-shape and defined filamentous wall. The encystation was found to cause a disappearance of the flagella and an invagination of the adhesive disc. An extensive formation of rER (rough endoplasmic reticulum) was observed after 24h of induction, indication an active synthesis and export of proteins during this process. The vital staining of the invitro-induced systs showed that most cysts maintained their viability.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.60-60
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• 2015

Hybrid histidine kinase is a part of two-component system that is required for various stress responses and pathogenesis of pathogenic fungi. In the present study, Tco1, a homologue of human pathogen Cryptococcus neoformans Tco1 encoding a hybrid histidine kinase, was identified in corn smut pathogen Ustilago maydis by bioinformatic analysis. To explore the role of Tco1 in the virulence of U. maydis, mutants in which the tco1 gene was partially deleted were constructed by allelic exchange. The U. maydis tco1 mutants did show unaltered growth rate on axenic medium but were unable to produce conjugation tubes and develop fuzzy filaments, resulting in impaired mating of compatible strains. The expression levels of prf1, pra1, and mfa1 which are involved in the pheromone pathway significantly decreased in the tco1 mutants. In inoculation tests to host, the tco1 mutants showed significantly reduced ability in the production of anthocyanin pigments and tumor development on maize leaves. Overall, the combined results indicated that Tco1 plays important roles in sexual development and virulence of U. maydis by regulating the expression of the genes involved in the pheromone pathway.

• Parasites, Hosts and Diseases
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• v.55 no.1
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• pp.81-84
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• 2017

Trypanosoma cruzi is the etiological agent of Chagas disease. Epimastigote forms of T. cruzi can be readily cultured in axenic conditions. Ethanol and dimethyl sulfoxide (DMSO) are commonly used solvents employed as vehicles for hydrophobic compounds. In order to produce a reference plot of solvent dependent growth inhibition for T. cruzi research, the growth of epimastigotes was analyzed in the presence of different concentrations of ethanol (0.1-4.0%) and DMSO (0.5-7.5%). The ability of the parasites to resume growth after removal of these solvents was also examined. As expected, both ethanol and DMSO produced a dose-dependent inhibition of cellular growth. Parasites could recover normal growth after 9 days in up to 2% ethanol or 5% DMSO. Since DMSO was better tolerated than ethanol, it is thus recommended to prefer DMSO over ethanol in the case of a similar solubility of a given compound.

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1805-1810
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• 2007

Purple nonsulfur bacteria were isolated from river sediments and their growth promoting capabilities on tomato were examined. Isolated strains KL9 and BL6 were identified as Rhodopseudomonas spp. by 16S rDNA sequence analysis. Rhodopseudomonas strain KL9 maximally produced 5.56 mM/min/mg protein and $67.2\;{\mu}M/min/mg$ protein of indole-3-acetic acid (IAA) and 5-aminolevulinic acid (ALA), respectively, which may be one of the mechanisms of plant growth enhancement. The germination percentage of tomato seed, total length, and dry mass of germinated tomato seedling increased by 30.2%, 71.1%, and 270.8%, respectively, compared with those of the uninoculated control 7 days after inoculation of strain KL9. The lengths of the root and shoot of germinated seedling treated with 3 mM tryptophan, a precursor of IAA, increased by 104.4% and 156.5%, respectively, 7 days after inoculation of strain KL9. Rhodopseudomonas KL9 increased 123.5% and 54% of the root and shoot lengths of germinated seedling, respectively, treated with 15 mM glycine and succinate, precursors of ALA. This plant growth promoting capability of purple nonsulfur bacteria may be a candidate for a biofertilizer in agriculture.

• Journal of Microbiology
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• v.35 no.2
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• pp.117-122
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• 1997

Several dominant 4-CPA-degrading bacteria were isoalted from agricultural soils. Most of the isolates were identified as Burkholderia species by fatty acid methyl ester (FAME) analysis, but they were idstinct in chromosomal patterns obtained by PCR amplification of repetitive extragenic palindromic (REP) sequences. These strains were generally restricted in their substrate utilization capabilities. The 4-CPA degradative enzymes were idnducible by 4-CPA and some isolates appeared to mineralize 4-CPA via formation of 4-chlorophenol and 4-chlorocatechol as intermediates during its biodegradation pathway. Plasmid DNAs were not detected from most of the isoaltes and their 4-CPA genes wer on the chromosomal DAN. The 4-CPA degradation patterns in axenic cultures and natural soils varied depending on the strains and soils. The inoculation of 4-CPA degraders much improved the removal of 4-CPA from the 4-CPA treated soils.

• The Plant Pathology Journal
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• v.19 no.3
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• pp.152-158
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• 2003

Stromata of the white root rot fungus, Rosellinia necatrix, were produced on diseased roots although they were reported to develop rarely in nature. Forty-two (42) out of 47 samples produced synnemata while 23 developed stromata. Forty-seven (47) isolates obtained from diseased root samples were divided into 24 mycelium compatibility groups (MCGs). Sixteen (16) out of 24 MCGs produced stromata. Single ascospore isolates from 10 stroma samples produced dsRNA-containing isolates from diseased tissue beneath stromata. The frequency of synnema production on axenic culture varied among isolates with different origin. The dsRNA was not transmitted vertically to the ascospore offspring despite the infection of various dsRNA in the parental isolates. The dsRNA was absent in 35 ascospore isolates in two stroma samples that originated from the isolates, in which dsRNA was not eliminated by hyphal tip isolation. Consequently, sexual reproduction in the white root rot fungus was suggested to produce propagules as a new infection source and to have the function to eliminate infectious factors such as mycoviruses.

• ALGAE
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• v.24 no.3
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• pp.163-168
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• 2009

Grateloupia filicina (J.V. Lamouroux) C. Agardh (Halymeniaceae, Cryptonemiales, Rhodophyta) is an edible seaweed as well as an important source of carrageenan. In the present study, attempt has been made to develop a suitable protocol for effective regeneration of the seaweed and the rapid multiplication of the desired varieties. The young upright thallus of G. filicina was grown in axenic culture using both solid and liquid media. The various media tested were f/2, Provasoli’s Enriched Seawater (PES) and Enriched Seawater (ESW). The effect of glycerol (as a carbon source) and various plant growth regulators i.e. auxin (NAA) and cytokinins (Kinetin and BA) were tested. Although, regeneration of young thalli was observed from the cut ends in all the media, better growth was found in f/2, PES, f/2 (0.5% Glycerol), f/2 (NAA ${10^{-5}}_M)\;and\;f/2\;(BA\;{10^{-6}}_M$). On the other hand callusing was observed only in solid media supplemented with low concentration of Glycerol (0.5%) in f/2, NAA ${10^{-5}}_M\;in\;f/2,\;PES\;and\;BA\;{10^{-5}}_M$ in f/2. Young thalli were developed from the callus sub culture after 40 days of inoculation.

• Parasites, Hosts and Diseases
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• v.52 no.4
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• pp.429-433
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• 2014

To identify sequences of Entamoeba histolytica associated with the development of amebic liver abscess (ALA) in hamsters, subtractive hybridization of cDNA from E. histolytica HM-1:IMSS under 2 growth conditions was performed: 1) cultured in axenic medium and 2) isolated from experimental ALA in hamsters. For this procedure, 6 sequences were obtained. Of these sequences, the mak16 gene was selected for amplification in 29 cultures of E. histolytica isolated from the feces of 10 patients with intestinal symptoms and 19 asymptomatic patients. Only 5 of the 10 isolates obtained from symptomatic patients developed ALA and amplified the mak16 gene, whereas the 19 isolates from asymptomatic patients did not amplify the mak16 gene nor did they develop ALA. Based on the results of Fisher's exact test (P<0.001), an association was inferred between the presence of the mak16 gene of E. histolytica and the ability to develop ALA in hamsters and with the patient's symptoms (P=0.02). The amplification of the mak16 gene suggests that it is an important gene in E. histolytica because it was present in the isolates from hamsters that developed liver damage.