We have examined the correlation between the physicochemical and microbiological environment variables for the different layers of oak forest soil in Mt. Gyeryong, Korea. The result shows that there is a high correlation in the environment variables between the soil parameters of the fermented (F) layer and humus (H) layer. In particular, the pH level in the F layer shows a high correlation with C and N, while the various organic acids of the H layer turns out to be closely correlated with soil bacteria density. As we evaluated phylogenetic characteristics of bacterial populations by DGGE analysis with DNA extracted. Total of 175 bands including 43 bands from litter (L) layer, 42 bands from F layer, 43 bands from H layer and 47 bands from rhizosphere (A) layer were selected as the major DGGE band of oak forest soil. Based on the 16S rRNA gene sequences, 175 DGGE bands were classified into 32 orders in 7 phylum. The heat map was analyzed in order to compare the quantity of the base sequences of each order and based on the clustering of the different layers of oak forest soil, the result confirms that the F layer and H layer belong to a different cluster from that of L layer and A layer. Furthermore, it also showed that approximately 50% of the total microbial population in different layers is ${\alpha}$-proteobacteria, which indicates that they belong to the dominant system group. In particular, Rhizobiales, Burkholderiales and Actinobacteriales were observed in all the seasons and layers of oak forest soil, which confirms that they are the indigenous soil bacterial community in oak forest soil.
Relationship of soil properties and seasonal variation on microbilogical population to-continuous culture and first-time culture of ginseng was investigated by bimonthly from May 1976 to January 1977. pH and P contents of 2 years continuous culture of soil were higher than other culture plot of soil, and contraty to the above, 2 years first-time culture of ginseng soil was conplot of soil, and contraty to the above, 2 years first-time culture of ginseng soil was contained more potassium contents than other culture plot of soil. In microbiological fluctuation with seasonr in various soil conditions, the population, trends of Fusarium spp., Erwiniaspp., and flourescent Psedudomonas spp. were increased in May and July in general, but decreased in the other month. It was observed that in all type of soil, Fusarium spp. was distributed in abundance in and on rihizosphere, and decreased the propagules numbers as soil depth increase. The numbers of Erwinia spp. and fluorescent Pseudo-monas spp. were distributed greater in numbers on the surface zone of soil depth decreasing the numbers along the soil layer increase, and also in 2years continuous culture of soil especially, a great numbers of Erwinia spp. and fluorescent Pseudomonas were evenly distributed in surface zone and rhizosphere. Ginseng disease with a high incidence of bacterial disease in continuous culture of 2 and 4 years was seemed to be associated with soil bacteria that was high in numbers of Erwinia spp. and fluorescent Pseudomonas spp. in May and July.
The soil of the reclaimed tidal land, located in Chogi-ri, Is. Kanghwa, Korea was used in this experiment. The experimented soil samples were collected from 18 sites with its time elapsed after the shore-protection works, soil-depth and the vegetation of saline plants, and at each site samplings were conducted monthly from March through October, 1968, for the purposes of examining the changes of microbial populations for the microbes such as bacteria, actinomycetes and fungi, by using the dilution plate method. The numbers of the microbes in these soils generally showed lower levels comparing with those of other soils. The more time elapsed after the reclamation, the higher numbers of the microbes inhibited the soils. Higher populations were there in the surface soils than in the lower part of the area. The surface soils included comparatively better conditions in aeration and contents of organic matter than in the lower part, and this fact was. same as in general soils. However, not so was this in the case of March, April and October due to the higher soil temperatures in the lows. At the experimental sites where the halophytes such as Salicorniu were grown vigourously, the more densly the plants grew, the higher populations of actinomycetes and fungi were, but not in the case of bacterial population. This means, in this soil with dense Salicornia, it is difficult to obtain good-natured soils in short time without a higher population of bacteria. For the rapid utilization of the land soil, in this view of point, the methods increasing the number of bacteria in the soil are needed as well as the cultivation and harvesting Salicorniu which indicated in the privious paper(Hong, et al., 1969a). According to the results of this experiment, the changes of soil-microbial populations in the reclaimed tidal land soil containing high salinity depend deeply upon the interrelations of many environmental factors such as soil-salinity, soil-components and contents, concentration of organic matters, pH, aeration, and air and soil temperatures, as in the general soils.
Total bacterial community DNA, which was extracted from microcosm soil and field soil after 2,4-D amendments, was analyzed on Southern blots, using the tfdA gene probe derived from plasmid pJP4 and the Spa probe from Sphingomonas paucimobilis. Southern blot analyses with total bacterial DNA extracted from soils Inoculated with Pseudomonas cepacia/pJP4 revealed that DNA probe method could detect the 2,4-D degrading bacteria down to $10^5\;cells/g$ dry soil. In the microcosm experiment, there was a good correlation between 2,4-D degradation and banding patterns in hybridization analyses performed after each 2,4-D treatment using the two probes. When bacterial DNA extracted from microcosm soil was hybridized with the Spa probe, a change in the position of hybrid bands was observed over time in a Southern blot, suggesting that population change or possibly genetic rearrangement in 2,4-D degrading microbial populations occurred in this soil. With the Spa probe, one hybrid DNA band was persistently observed throughout the five 2,4-D additions. When bacterial DNA isolated from the field soil was probed with the tfdA and Spa, strong hybridization signal was observed in the 100 ppm-treated subplot, weak signal In the 10 ppm-treated subplot, and no significant signal in the 1 ppm-treated and control subplots. The data show that DNA probe analyses were capable of detecting and discriminating the indigenous 2,4-D degrading microbial populations in soil amended with 2,4-D under laboratory and field conditions.
The production of Korean ginseng, one of the most important medicinal root crops, is limited by many factors including soil sickness, root rots in fields as well as during storage prior to consumption. Although much research has been conducted on the diseases in field condition, little information is available on the control of postharvest roots rots. To obtain better management strategy of postharvest root rots in ginseng, biological control using antagonistic bacteria was attempted. Of 208 bacteria obtained form suppressive soil samples, 4 were selected based on the inhibitory effect on mycelial growth of two major causal fungi for postharvest root rots in ginseng, Botrytis cinerea and Fusarium solani. The culture filtrates of these bacterial antagonists greatly inhibited the conidial germination of both pathogenic fungi and produced abnormal morphology such as swollen germ tubes in F. solani and vacuolation of nongerminated conidia in B. cinerea. The population levels of bacterial antagonists on the ginseng roots were gradually increased up to 8 days of incubation. Postharvest root rots of ginseng caused by f. solani and B. cinerea were controlled in dipping tests in the ranges of 60∼80% by antagonistic Bacillus spp. obtained from suppressive soil. These results suggest that biological control using these antagonistic bacteria would be an alternative strategy to control postharvest root rots in ginseng.
Fusarium wilt of radish (Raphanus sativus L.) is caused by the Fusarium oxysporum f. sp. raphani (FOR) which mainly attacks Raphanus spp. The pathogen is a soil-borne and forms chlamydospores in infected plant residues in soil. Infected pathogen colonizes the vascular tissue, leading to necrosis of the vascular tissue. Growth promoting beneficial organisms such as Pseudomonas fluorescens WCS374 (strain WCS374), P. putida RE10 (strain RE10) and Pseudomonas sp. EN415 (strain EN415) were used for microorganisms-mediated induction of systemic resistance in radish against Fusarium wilt. In this bioassy, the pathogens and bacteria were treated into soil separately or concurrently, and mixed the bacteria with the different level of combination. Significant suppression of the disease by bacterial treatments was generally observed in pot bioassy. The disease incidence of the control recorded 46.5% in the internal observation and 21.1% in the external observation, respectively. The disease incidence of P. putida RE10 recorded 12.2% in the internal observation and 7.8% in the external observation, respectively. However, the disease incidence of P. fluorescens WCS374 which was proved to be highly suppressive to Fusarium wilt indicated 45.6% in the internal observation and 27.8% in the external observation, respectively. The disease incidence of P. putida RE10 mixed with P. fluorescens WCS374 or Pseudomonas sp. EN415 was in the range of 10.0-22.1%. On the other hand, the disease incidence of P. putida RE10 mixed with Pseudomonas sp. EN415 was in the range of 7.8-20.2%. The colonization by FOR was observed in the range of $2.4-5.1{\times}10^3/g$ on the root surface and $0.7-1.3{\times}10^3/g$ in the soil, but the numbers were not statistically different. As compared with $3.8{\times}10^3/g$ root of the control, the colonization of infested ROR indicated $2.9{\times}10^3/g$ root in separate treatments of P. putida RE10, and less than $3.8{\times}10^3/g$ root of the control. Also, the colonization of FOR recorded $5.1{\times}10^3/g$ root in mixed treatments of 3 bacterial strains such as P. putida RE10, P. fluorescens WCS374 and Pseudomonas sp. EN415. The colonization of FOR in soil was less than that of FOR in root part. Based on soil or root part, the colonization of ROR didn't indicate a significant difference. The colonization of introduced 3 fluorescent pseudomonads was observed in the range of $2.3-4.0{\times}10^7/g$ in the root surface and $0.9-1.8{\times}10^7/g$ in soil, but the bacterial densities were significantly different. When growth promoting organisms were introduced into the soil, the population of Pseudomonas sp. in the root part treated with P. putida RE10 was similar in number to the control and recorded the low numerical value as compared with any other treatments. The population density of Pseudomonas sp. in the treatment of P. putida RE10 indicated significant differences in the root part, but didn't show significant differences in soil. The population densities of infested FOR and introduced bacteria on the root were high in contrast to those of soil. P. putida RE10 and Pseudomonas sp. EN415 used in this experiment appeared to induce the resistance of the host against Fusarium wilt.
The CFDA (6-carboxyfluorescein diacetate) direct viable count method and plate count (PC) method using conventional nutrient broth (NB) medium and $10^{-2}$ diluted NB (DNB) medium were applied to samples collected from Mt. Yongdoo In Andong, in an effect to determine the number of living bacteria pine mushroom forest soil. The number of living bacteria determined via plate count in NB medium comprised $5{\sim}8%$ of the CFDA direct viable count, and the bacteria in the DNB medium comprised $40{\sim}47%$. This result indicated that viable but nonculturable (VBNC) bacteria existed in the pine mushroom forest soil at a high percentage. The phylogenetic characteristics of the VBNC bacterial populations in the samples of pine mushroom (Tricholoma matsutake) forest soil were analyzed via the direct extraction of DNA and 16S rDNA-ARDRA. The 115 clones from pine mushroom forest soil were clustered into 31 different RFLP phylotypes by ARDRA. Based on the 16S rDNA sequences, the 31 ARDRA clusters were classified into 6 phylogenetic groups: ${\alpha}-,\;{\beta}-,\;{\gamma}-Proteobacteria$, Acidobacteria, Actinobacteria and Firmicutes. Among these bacterial populations, approximately 85% were classified as members of phylum Acidobacteria. The Acidobacteria phylum was shown to exist abundantly in the pine mushroom forest soil.
Seo, Sang-Tae;Park, Jong-Han;Kim, Kyung-Hee;Lee, Sang-Hyun;Oh, Eun-Sung;Shin, Sang-Chul
Research in Plant Disease
/
제14권1호
/
pp.32-36
/
2008
Bacterial wilt caused by Ralstonia solanacearum has become a severe problem on tomato in Korea and no effective control measures are available yet. Pseudomonas species play key roles for the biocontrol of many plant diseases especially in soil. A rhizobacterial population of 150 Pseudomonas strains, isolated from the rhizosphere soil of various plants grown at different sites, was screened for 2,4-diacetylphloroglucinol producing gene (PhlD) by PCR. Two strains (P83 and P84) among them were found to be phlD positive. When the isolates were analysed by 16S rDNA (Sensu Stricto), all isolates yielded amplified products of 1,018bp. Of the 150 isolates of Pseudomonas spp., a bacterial strain P. putida P84 isolated from tomato rhizosphere showed to suppress a wide range of phytopathogenic bacteria in vitro. The best source of carbon for P84 strain were glucose, arabinose, inositol and melibiose. In greenhouse experiments, P84 strain suppressed the development of bacterial wilt in tomato with a control value of 60%.
The effects of diesel oil on the microbial community in sandy loam soil were investigated, and the effects of bioremediation which was performed to enhance the removal of diesel oil from soil were also measured. The residual percentage of diesel oil was about 50% after 16 week incubation period. The bioremediation treatment increased the removal rate at 60~95%. When the soil was contaminated with diesel oil, the direct bacterial count, length of fungal hyphae, aerobic heterotroph and hydrocarbon degrader were increased by 2~3 orders of magnitude. The bioremediation further increased these numbers 10 to 100-fold. There were no difinite patterns of change in fluorescein diacetate hydrolysis activity in bioremediation-untreated soil, but about 10 times of increase of activity was observed in bioremediation-treated soil. Similar change was occurred in soil dehydrogenase activity.
The population diversity and seasonal changes of bacterial communities in rice soils were monitored using both culture-dependent approaches and molecular methods. The rice field plot consisted of twelve subplots planted with two genetically-modified (GM) rice and two non-GM rice plants in three replicates. The DGGE analysis revealed that the bacterial community structures of the twelve subplot soils were quite similar to each other in a given month, indicating that there were no significant differences in the structure of the soil microbial populations between GM rice and non-GM rice during the experiment. However, the DGGE profiles of June soil after a sudden flooding were quite different from those of the other months. The June profiles exhibited a few intense DNA bands, compared with the others, indicating that flooding of rice field stimulated selective growth of some indigenous microorganisms. Phylogenetic analysis of l6S rDNA sequences from cultivated isolates showed that, while the isolates obtained from April soil before flooding were relatively evenly distributed among diverse genera such as Arthrobacter, Streptomyces, Terrabacter, and Bacillus/Paenibacillus, those from June soil after flooding mostly belonged to the Arthrobacter species. Phylogenetic analysis of 16S rDNA sequences obtained from the soil by cloning showed that April, August, and October had more diverse microorganisms than June. The results of this study indicated that flooding of rice fields gave a significant impact on the indigenous microbial community structure; however, the initial structure was gradually recovered over time after a sudden flooding.
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