Studies were conducted during 2 months from May of 1997 to evaluate the effects of pig manure compost(PMC) on soil microbial flora. To do so, a field experiment of Chinese cabbage(Brassica campestris L.) was conducted in a randomized block design on a sandy loam soil and microbial floral characteristics in soils were analyzed. Treatments to control included the application of PMC at (A) $8Mg\;ha^{-1}$CM-8), (B) $29Mg\;ha^{-1}$(CM-2,9), and (C) $57Mg\;ha^{-1}$(CM-57), and of chemical fertilizer(D) at $320N-80P_2O_5-200K_2O\;kg\;ha^{-1}$(NPK). In each treatment, the rhizosphere and non-rhizosphere soils were tested for the analysis of microbial populations. The populations of bacteria, actinomycetes, and fungi increased in soils with the applications of PMC and chemical fertilizer, but that of Bacillus sp. decreased. However, the population of fluorescent Pseudomonas sp. was reduced in NPK plots only. With increasing application rates of PMC, the number of colony forming units(cfu) of bacteria (Pseudomonas sp. and actinomycetes) and fungi increased. in all PMC-treated plots, the population density peaked at early growth stage for bacteria(including Bacillus sp.), at late growth for fluorscent Pseudomonas sp., and at harvest for fungi and actinomycetes. The rhizosphere effect was greatest for fluorscent Pseudomonas sp. As the application rates of PMC increased, Total N, organic matter, available phosphate, and exchangeable -K, -Ca, and -Mg increased compared to control, but soil pH was lowered. In NPK plots, EC was 3.4-fold and exchangeable K was 5-fold higher than control.
To detect whole ammonia-oxidizing bacteria in the activated sludge, group-specific primers targeting the 16S-rRNA gene of ammonia-oxidizing bacteria were used. The electrophoresis pattern of the PCR products seemed to produce a single band of approximately 1.0 k bp for the bacteria in activated sludge and Nitrosomonas europaea. No band was observed for nitrite-oxidizer Nitrobacter winogradskyi and heterotrophs such as Pseudomonas putida. Then direct measurement of the PCR product was made by fluorometry using the reagent Hoechist 33258, so that the fluorescent intensity was in proportional to the cell number of the sample up to 240. Total time required for the test was about 4 h including DNA extraction. The DNA fragments produced were cloned and their sequences showed high similarity to those of Nitrosomonas spp. This study showed the feasibility to detect ammonia-oxidizing bacteria and to esti-mate their population rapidly for the control of the nitrogen elimination process.
Although biological metabolism in soil is very important for evaluating the soil properties, most of researches have concerned mainly about physical and chemical sides. In this study, biological characteristics were examined to demonstrate the biota in the plastic film house soils. Contents of organic matter and phosphate in soil were increased with cultivation period. ECs of soil cultivated spinach and melon were $3.59dS\;m^{-1}$ and $3.46dS\;m^{-1}$ respectively: these values were higher than that of rose and flower, which were $1.23dS\;m^{-1}$ and $1.32dS\;m^{-1}$ respectively. The population of fluorescent Pseudomonas strains of the soil cultivated flowers: $113.8{\times}10^4{\sim}129.7{\times}10^4cfu\;g^{-1}$ was higher than that of leafy vegetables: $40.7{\times}10^4{\sim}97.9{\times}10^4cfu\;g^{-1}$ and fruiting vegetables: $25.0{\times}10^4{\sim}91.7{\times}10^4cfu\;g^{-1}$. However the number of Fusarium strains of the soil cultivated with flowers: $3.8{\times}10^2{\sim}4.0{\times}10^2cfu\;g^{-1}$ was lower than that of leafy vegetables: $4.3{\times}10^2{\sim}16.3{\times}10^2cfu\;g^{-1}$ and fruiting vegetables: $7.6{\times}10^2{\sim}30.0{\times}10^2cfu\;g^{-1}$. In relation to the cultivation period, the habitation density of aerobic bacteria, mesophilic Bacillus, thermophilic Bacillus, and fluorescent Pseudomonas strains was the highest in the soil cultivated over 11 years, but diversity index showed negative correlation with cultivation period. Microbial biomass C in these soils had positive correlation with each number of microorganisms including aerobic bacteria, actinomycetes, and strains of mesophilic Bacillus as well as the total number of these microorganisms.
The effects of metalaxyl(granule), carbofuran(granule) and simazine(water soluble powder) on the soil microflora were conducted at field soil between Iksan and Chonju province. Pesticides were divided into 0.5, 1, 1.5 and 2.0 times of normal of field, respectively. The number of fluorescent Pseudomonas was ranged from $10^3$ to $10^6/g$ in both field soil treated with cabofuran. Pseudomonas concentration of Chonju field soil slowly increased and approached the maximum level at 56 day after treatment(DAT). It showed the higher at 14DAT than other DAT in Iksan field soil treated with metalaxyl or simazine, whereas it increased again at 112 DAT in metalaxyl treatment. Cabofuran treatment of both field soil showed maximum Pseudomonas number at 28 DAT compared to that of other treatments. In Chonju field soil, those Pseudomonads of metalaxyl and simazine treatment increased the highest level at 7 DAT. Simazine treatment decreased it's number from the beginning of experiment. In both soil, metalaxyl treatment decrease the general fungi number at 7 DAT, but increase at 14 and 56 DAT in Iksan field soil. However it increased at 56 DAT in Chonju field soil. Cabofuran treatment of Iksan field soil tended to decrease general fungi number at 28 DAT, but was ranged from 1.0 to $8.6{\times}104/g$ for the rest of experimental period. It started to increase at 56 DAT simazine treatment of Iksan. General bacterial concentration both soil treated with cabofuran was belong to $26.6{\sim}29.6{\times}106$. It was the highest at 56 DAT, but was not significantly different. General actinomyces number was highly increased at 7 and 112 DAT compared to that of other DAT. Pseudomonas putida or P. fluorescens from both field soil was separated and identified 10 to 30 of all 104 Pseudomonas, respectively. All isolated microorganisms showed chemical resistance of 100ppm metalaxyl, cabofuran and simazine treatment.
Elsinoe fawcettii is one of major pathogenic fungi which cause citrus scab diseases, resulting in fruit blemishes that reduce the economic value of fruit. By increasing interest to safe products of crops, the alternative methods of disease control is highly required. We investigated whether the 215 bacterial strains isolated from Jeju Island possess antifungal effect or suppression effect on the symptom development by Elsinoe fawcettii on citrus. Among them, three bacterial strains THJ 609-3, MRL408-3, and TRH423-3 that exhibited antifungal capacity against Elsinoe fawcettii were selected. To illustrate the disease suppression mechanism, pre-inoculation with the selected bacterial strains was carried out whether could suppress the citrus crab on the leaves. The observation with a fluorescence microscope revealed that the selected bacteria could decrease the number of fungal spores. The ratio of germ tube formation was also decreased by the selected bacterial strains at one day after fungus challenge. The strain THJ 609-3 was identified as Pseudomonas putida as a result of analyzing the internal transcript spaces of the rhizobacterial rDNA. The strains MRL 408-3 and TRH 423-3 were identified as Burkholderia gladioli. Our results may be valuable when the selected rhizobacterial strains used as the environment-friendly microbe for biological control on citrus scab caused by Elsinoe fawcettii.
Journal of Korean Society of Environmental Engineers
/
v.32
no.12
/
pp.1094-1101
/
2010
Microbial fuel cells (MFC) were enriched using sediment Nakdong river, Hoidong river and protected water area in Gijang. The microbial community of sediment and enriched MFC was analyzed by FISH (fluorescent in situ hybridization) and 16S rDNA sequencing. ${\alpha}$-Proteobacteria, Acidobacter and Cyanobactia group were dominant in sediment by FISH. The coulombs of the final 10 peak of the 3 MFC (Nakdong, Hoidong, Gijang) were 0.64 C, 0.50 C, 0.61 C, respectively. When MFCs were enriched by sediment, ${\beta}$-, ${\gamma}$-Proteobacteria, Acidobacter and Firmicutes group increased 45~90%, 50~90%, 40~80% and 45~125%, respectively. In results of 16S rDNA sequencing, Roseomonas sp., Azospillium sp., Frateuria sp., Dyella sp., Enterobacter sp. and Deinocossus were isolated from Nakdong river and Azospillium sp., Delftia sp., Ralstonia sp., Klebsiella sp. and Deinococcus sp. were isolated from protected water area in Gijang and Pseudomonas sp., Klebsiella sp., Deinococcus sp., Leifsonia sp. and Bacillus sp. were isolated from Hoidong river.
This study was conducted to investigate the characteristics of the medium used on the composting step, comparing the excavator agitator with the self-propelled turner. The temperature of the outdoor composting medium tended to increase rapidly after flipping in the turner. The late composting medium temperature was maintained at the excavator treatment area (farm practice), and the late composting effect progressed. During the field composting stage, various microorganisms such as Bacillus spp., Actinomycetes, fluorescent Pseudomonas spp., and filamentous fungi were distributed in the medium, and the density of aerobic bacteria involved in the decomposition of the medium was increased. Under high-temperature composting conditions, blue fungi, and mesophilic actinomycetes were inhibited or killed. Thermophilic actinomycetes, which play an important role in decomposing organic matter, showed higher densities than those observed in farm practices in the self-propelled turner process. The length of rice straw was slightly shorter when the self-propelled turner was used, and the water content did not show any significant difference between treatments. The a and b values tended to increase as the inverter was turned over. The CN ratio of the composting broth was lowered from 23.1 to 16.2 for the 5th turnover in the context of farming practices, and from 23.3 to 16.9 in the context of the self-propelled turner. The yield of each treatment was increased by 20% in 1 period, 28% in 2 periods, and 26% in 3 periods; the overall yield was 23%.
Kim, Jong-Soo;Kim, Ji-Hyun;Ryu, Eun-Kyeong;Kim, Jin-Kyoo;Kim, Chi-Kyung;Hwang, In-Gyu;Lee, Kyoung
Journal of Microbiology and Biotechnology
/
v.14
no.2
/
pp.302-311
/
2004
Comamonas testosteroni (formerly Pseudomonas sp.) NCIMB 10643 can grow on biphenyl and alkylbenzenes $(C_2-C_7)$ via 3-substituted catechols. Thus, to identify the genes encoding the degradation, transposon-mutagenesis was carried out using pAG408, a promoter-probe mini-transposon with a green fluorescent protein (GFP), as a reporter. A mutant, NT-1, which was unable to grow on alkylbenzenes and biphenyl, accumulated catechols and exhibited an enhanced expression of GFP upon exposure to these substrates, indicating that the gfp had been inserted in a gene encoding a broad substrate range catechol 2,3-dioxygenase. The genes (2,826 bp) flanking the gfp cloned from an SphI-digested fragment contained three complete open reading frames that were designated bphCDorfl. The deduced amino acid sequences of bphCDorfl were identical to 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC), 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase (BphD), and OrfI, respectively, that are all involved in the degradation of biphenyl/4-chlorobiphenyl (bph) by Ralstonia oxalatica A5. The deduced amino acid sequence of the orfl revealed a similarity to those of outer membrane proteins belonging to the OmpW family. The introduction of the bphCDorfl genes enabled the NT-l mutant to grow on aromatic hydrocarbons. In addition, PCR analysis indicated that the DNA sequence and gene organization of the bph operon were closely related to those in the bph operon from Tn4371 identified in strain A5. Furthermore, strain A5 was also able to grow on a similar set of alkylbenzenes as strain NCIMB 10643, demonstrating that, among the identified aromatic hydrocarbon degradation pathways, the bph degradation pathway related to Tn4371 was the most versatile in catabolizing a variety of aromatic hydrocarbons of mono- and bicyclic benzenes.
BACKGROUND: The urgency of feeding the world's growing population while combating soil pollution, salinization and desertification requires suitable biotechnology not only to improve crop productivity but also to improve soil health through interactions of soil nutrient and soil microorganism. Interest in the utilization of microbial fertilizer has increased. A principle of nature farming is to produce abundant and healthy crops without using chemical fertilizer and pesticides, and without interrupting the natural ecosystem. Beneficial microorganisms may provide supplemental nutrients in the soil, promote crop growth, and enhance plant resistance against pathogenic microorganisms. We mixed beneficial microorganisms such as Bacillus sp. Han-5 with anti-fungal activities, Trichoderma harziaum, Trichoderma longibrachiatum with organic material degrading activity, Actinomycetes bovis with antibiotic production and Pseudomonas sp. with nitrogen fixation. This study was carried out to investigate the mixtures on the soil microflora and soil chemical properties and the effect on the growth of lettuce and cucumber under greenhouse conditions. METHODS AND RESULTS: The microbial mixtures were used with each of organic fertilizer, swine manure and organic+swine manure and compared in regard to changes in soil chemical properties, soil microflora properties and crop growth. At 50 days after the treatment of microorganism mixtures, the pH improved from 5.8 to 6.3, and the EC, $NO_3$-Na and K decreased by 52.4%, 60.5% and 29.3%, respectively. The available $P_2O_5$ and $SiO_2$ increased by 25.9% and 21.2%, respectively. Otherwise, the population density of fluorescent Pseudomonas sp. was accelerated and the growth of vegetables increased. Moreover, the population density of E. coli and Fusarium sp., decreased remarkably. The ratio of bacteria to fungi (B/F) and the ratio of Actinomycetes bovis to fungi (A/F) increased 2.3 (from 272.2 to 624.4) and 1.7 times (from 38.3 to 64), respectively. Furthermore, the growth and yield of cucumber and lettuce significantly increased by the treatment of microorganism mixtures. CONCLUSION(S): These results suggest that the treatment of microorganism mixtures improved the chemical properties and the microflora of soil and the crop growth. Therefore, it is concluded that the microorganism mixtures could be good alternative soil amendments to restore soil nutrients and soil microflora.
Levulinic acid (LA) is a valuable chemical used in fuel additives, fragrances, and polymers. In this study, we proposed possible biosynthetic pathways for LA production from lignin and poly(ethylene terephthalate). We also created a genetically encoded biosensor responsive to LA, which can be used for screening and evolving the LA biosynthesis pathway genes, by employing an LvaR transcriptional regulator of Pseudomonas putida KT2440 to express a fluorescent reporter gene. The LvaR regulator senses LA as a cognate ligand. The LA biosensor was first examined in an Escherichia coli strain and was found to be non-functional. When the host of the LA biosensor was switched from E. coli to P. putida KT2440, the LA biosensor showed a linear correlation between fluorescence intensity and LA concentration in the range of 0.156-10 mM LA. In addition, we determined that 0.156 mM LA was the limit of LA detection in P. putida KT2440 harboring an LA-responsive biosensor. The maximal fluorescence increase was 12.3-fold in the presence of 10 mM LA compared to that in the absence of LA. The individual cell responses to LA concentrations reflected the population-averaged responses, which enabled high-throughput screening of enzymes and metabolic pathways involved in LA biosynthesis and sustainable production of LA in engineered microbes.
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