To investigate the effects of elevated $CO_2$ on the denitrifying bacterial community structure in a wetland soil, dynamics of bacterial community structure was explored in an artificial wetland ecosystem with one of three plant species (T. latifolia, S. lacustris, and 1. effusus) under two levels of $CO_2$(370 ppm or 740 ppm) after 110day incubation. For the analysis of bacterial community structure, functional genes such as nitrite reductase genes (nirS) were PCR-amplified followed by cloning of PCR products and screening by restriction fragment length polymorphism (RFLP). nirS gene fragments were amplified in all analyzed soil samples. Species richness estimated by the number of distinct phylotypes were 83 and 95 in the ambient $CO_2$ treatment and the elevated treatment, respectively. Two phylotypes (type 1 and type 2) were dominant in both of the treatments. Elevated $CO_2$ treatment increased species richness of denitrifying as well as changed a large proportion of denitrifier phylotypes compared to those of the ambient treatment. Overall, the data in this study suggested that the denitrifying communities in the wetland soil are diverse and that the richness of denitrifying bacterial community might be affected by elevated $CO_2$ treatment.
Yoo, Sung-Je;Lee, Shin Ae;Weon, Hang-Yeon;Song, Jaekyeong;Sang, Mee Kyung
Korean Journal of Environmental Agriculture
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v.40
no.1
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pp.49-59
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2021
BACKGROUND: Soil salinity causes reduction of crop productivity. Rhizosphere microbes have metabolic capabilities and ability to adaptation of plants to biotic and abiotic stresses. Plant growth-promoting bacteria (PGPB) could play a role as elicitors for inducing tolerance to stresses in plants by affecting resident microorganism in soil. This study was conducted to demonstrate the effect of selected strains on rhizosphere microbial community under salinity stress. METHODS AND RESULTS: The experiments were conducted in tomato plants in pots containing field soil. Bacterial suspension was inoculated into three-week-old tomato plants, one week after inoculation, and -1,000 kPa-balanced salinity stress was imposed. The physiological and biochemical attributes of plant under salt stress were monitored by evaluating pigment, malondialdehyde (MDA), proline, soil pH, electrical conductivity (EC) and ion concentrations. To demonstrate the effect of selected Bacillus strains on rhizosphere microbial community, soil microbial diversity and abundance were evaluated with Illumina MiSeq sequencing, and primer sets of 341F/805R and ITS3/ITS4 were used for bacterial and fungal communities, respectively. As a result, when the bacterial strains were inoculated and then salinity stress was imposed, the inoculation decreases the stress susceptibility including reduction in lipid peroxidation, enhanced pigmentation and proline accumulation which subsequently resulted in better plant growth. However, bacterial inoculations did not affect diversity (observed OTUs, ACE, Chao1 and Shannon) and structure (principle coordinate analysis) of microbial communities under salinity stress. Furthermore, relative abundance in microbial communities had no significant difference between bacterial treated- and untreated-soils under salinity stress. CONCLUSION: Inoculation of Bacillus strains could affect plant responses and soil pH of tomato plants under salinity stress, whereas microbial diversity and abundance had no significant difference by the bacterial treatments. These findings demonstrated that Bacillus strains could alleviate plant's salinity damages by regulating pigments, proline, and MDA contents without significant changes of microbial community in tomato plants, and can be used as effective biostimulators against salinity stress for sustainable agriculture.
Michelle Miguel;Seon-Ho Kim;Sang-Suk Lee;Yong-Il Cho
Animal Bioscience
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v.36
no.9
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pp.1453-1464
/
2023
Objective: This study investigated the changes in bacterial communities within decomposing swine microcosms, comparing soil with or without intact microbial communities, and under aerobic and anaerobic conditions. Methods: The experimental microcosms consisted of four conditions: UA, unsterilized soil-aerobic condition; SA, sterilized soil-aerobic condition; UAn, unsterilized soil-anaerobic condition; and San, sterilized soil-anaerobic condition. The microcosms were prepared by mixing 112.5 g of soil and 37.5 g of ground carcass, which were then placed in sterile containers. The carcass-soil mixture was sampled at day 0, 5, 10, 30, and 60 of decomposition, and the bacterial communities that formed during carcass decomposition were assessed using Illumina MiSeq sequencing of the 16S rRNA gene. Results: A total of 1,687 amplicon sequence variants representing 22 phyla and 805 genera were identified in the microcosms. The Chao1 and Shannon diversity indices varied in between microcosms at each period (p<0.05). Metagenomic analysis showed variation in the taxa composition across the burial microcosms during decomposition, with Firmicutes being the dominant phylum, followed by Proteobacteria. At the genus level, Bacillus and Clostridium were the main genera within Firmicutes. Functional prediction revealed that the most abundant Kyoto encyclopedia of genes and genomes metabolic functions were carbohydrate and amino acid metabolisms. Conclusion: This study demonstrated a higher bacteria diversity in UA and UAn microcosms than in SA and SAn microcosms. In addition, the taxonomic composition of the microbial community also exhibited changes, highlighting the impact of soil sterilization and oxygen on carcass decomposition. Furthermore, this study provided insights into the microbial communities associated with decomposing swine carcasses in microcosm.
Removal of methane, benzene and toluene was evaluated in a lab-scale biocover packed with a soil mixture of forest soil and earthworm cast (75:25 weight ratio). The bacterial community in the biocover was characterized using quantitative real-time PCR and terminal restriction fragment length polymorphism. Methane was removed at the upper layer of the biocover (-0.1 ~ -0.4 m), where the oxygen concentration was remarkably lower. The average removal efficiencies for methane and benzene/toluene were 90% and 99%, respectively. The pmoA gene copy numbers, responsible for methane oxidation, in the upper layer were higher than those in the lower layer. While type I methanotrohs dominated the lower layer, type II methanotrophs, such as Methylocystis and Methylosinus, were noted to be predominant in the upper layer. Benzene and toluene were removed from the lower layer (-0.6 ~ -0.9 m) as well as the upper layer. Moreover, the tmoA gene copy number, responsible for benzene/toluene oxidation, seen in the upper layer was not significantly different from those seen in the lower layer. These results suggest that a biocover packed with a soil and earthworm cast mixture is a promising method which could be utilized for the control of methane and volatile organic compounds such as benzene and toluene.
Soybean is well known to be originated from Korea and far-east Asian countries, and studies of many root nodule bacteria associated with soybean have mainly-focused on nitrogen fixation, but much less study was carried out on bacterial community in the rhizosphere of soybean. In this study, we analyzed the bacterial community in rhizosphere of Korean soybean, Daepungkong using the pyrosequencing method based on the 16S rRNA gene to characterize the change of the rhizosphere community structure according to the growth stages of soybeans and to elucidate bacterial core community in rhizosphere of soybean. Our results revealed that bacterial community of rhizosphere soil differed from that of bulk soil and was composed of a total of 21 bacterial phyla. The predominant phylum in the rhizosphere of soybean was Proteobacteria (36.6-42.5%) and followed by Acidobacteria (8.6-9.4%), Bacteroidetes (6.1-10.9%), Actinobacteria (6.4-9.8%), and Firmicutes (5.7-6.3%). The bacterial core community in soybean rhizosphere was mainly composed of the operational taxonomic units (OTUs) belonging to the phylum Proteobacteria throughout all growth stages. The OTU00006 belonged to the genus Bradyrhizobium had the highest abundance and Steroidobacter, Streptomyces, Devosia were followed. These results show that bacterial core community in soybean rhizosphere was mainly composed of OTUs associated with plant growth promotion and nutrient cycles.
In this study, microorganism community characteristics of organic managed soil which applied rye (Secale cereal L.) as green manure for 25 years, were determined. The chemical properties of organic soil showed high level of organic matter and available $P_2O_5$, while the level of exchangeable cation was low. The analysis of dehydrogenase activity and carbon source utilization indicated that the values in on organic soil were significantly higher than those of the control. It suggested that the microorganism community of organic soil had high microorganism activity, compared to the control. In addition, when the 16S rRNA gene-targeted NGS (Next generation sequencing) analysis was conducted to estimate the class of bacterial community, the class level of bacterial taxon composition on organic soil showed higher portion of Sphingobacteriia, Acidobacteriia, Gammaproteobacteria, Solibacteres and Planctomycetia. By base on the results of various reports in which organic managed soil had high portion of Acidobacteriia and Planctomycetia, the characteristic of taxon composition in organic soil, which showed the high percentages of Ktedonobacteria, Sphingobacteriia, Acidobacteriia and Gammaproteobacteria, was resulted from the application of rye as a green manure for the long term. However, further researches were needed because the crop effect was not considered in this study.
Light hydrocarbons accumulated in subsurface soil by long-term microseepage could favor the anomalous growth of indigenous hydrocarbon-oxidizing microorganisms, which could be crucial indicators of underlying petroleum reservoirs. Here, Illumina MiSeq sequencing of the 16S rRNA gene was conducted to determine the bacterial community structures in soil samples collected from three typical oil and gas fields at different locations in China. Incubation with n-butane at the laboratory scale was performed to confirm the presence of "universal microbes" in light-hydrocarbon microseepage ecosystems. The results indicated significantly higher bacterial diversity in next-to-well samples compared with background samples at two of the three sites, which were notably different to oil-contaminated environments. Variation partitioning analysis showed that the bacterial community structures above the oil and gas fields at the scale of the present study were shaped mainly by environmental parameters, and geographic location was able to explain only 7.05% of the variation independently. The linear discriminant analysis effect size method revealed that the oil and gas fields significantly favored the growth of Mycobacterium, Flavobacterium, and Pseudomonas, as well as other related bacteria. The relative abundance of Mycobacterium and Pseudomonas increased notably after n-butane cultivation, which highlighted their potential as biomarkers of underlying oil deposits. This work contributes to a broader perspective on the bacterial community structures shaped by long-term light-hydrocarbon microseepage and proposes relatively universal indicators, providing an additional resource for the improvement of microbial prospecting of oil and gas.
Song Hong-Gyu;Kim Ok-Sun;Yoo Jae-Jun;Jeon Sun-Ok;Hong Sun-Hee;Lee Dong-Hun;Ahn Tae-Seok
Journal of Microbiology
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v.42
no.4
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pp.285-291
/
2004
The soil bacterial community and some inoculated bacteria were monitored to assess the microbial responses to prescribed fire in their microcosm. An acridine orange direct count of the bacteria in the unburned control soil were maintained at a relatively stable level $(2.0\~2.7\times10^9\;cells/g^{-1}{\cdot}soil)$ during the 180 day study period. The number of bacteria in the surface soil was decreased by fire, but was restored after 3 months. Inoculation of some bacteria increased the number of inoculated bacteria several times and these elevated levels lasted several months. The ratios of eubacteria detected by a fluorescent in situ hybridization (FISH) method to direct bacterial count were in the range of $60\~80\%$ during the study period, with the exception of some lower values at the beginning, but there were no definite differences between the burned and unburned soils or the inoculated and uninoculated soils. In the unburned control soil, the ratios of $\alpha-,\beta-\;and\;\gamma-subgroups$ of the proteobacteria, Cytophaga-Flavobacterium and other eubacteria groups to that of the entire eubacteria were 13.7, 31.7, 17.1, 16.8 and $20.8\%,$ respectively, at time 0. The overall change on the patterns of the ratios of the 5 subgroups of eubacteria in the uninoculated burned and inoculated soils were similar to those of the unburned control soil, with the exception of some minor variations during the initial period. The proportions of each group of eubacteria became similar in the different microcosms after 6 months, which may indicate the recovery of the original soil microbial community structure after fire or the inoculation of some bacteria. The populations of Azotobacter vinelandii, Bacillus megaterium and Pseudomonas fluorescens, which had been inoculated to enhance the microbial activities, and monitored by FISH method, showed similar changes in the microcosms, and maintained high levels for several months.
Bacterial diversity based on the denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA gene sequences was determined for soil samples from two abandoned mine sites and the corresponding enrichment cultures using soil sample as key inoculum. Sequencing analysis of DGGE bands obtained from both the soil samples matched mostly with sequences of uncultured and newly described organisms, or organisms recently associated with the acid mine drainage environment. However, the enrichment of soil samples in ferrous sulfate and elemental sulfur media yielded sequences that were consistent with well-known iron- and sulfur-oxidizing acidophilic bacteria. Analysis of enrichment cultures of soil samples from Dalsung mine revealed abundant ${\gamma}$-$Proteobacteria$, whereas that of Gubong mine sample displayed acidophilic groups of ${\gamma}$-$Proteobacteria$, ${\alpha}$-$Proteobacteria$, $Actinobacteria$ and $Firmicutes$. Chemical elemental analysis of the mine samples indicated that the Dalsung site contained more iron and sulfate along with other toxic components as compared with those of the Gubong site. Biogeochemistry was believed to be the primary control on the acidophilic bacterial group in the enrichment samples.
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.342-342
/
2017
Rice seeds are a home to endophytic bacterial communities which serve as a source of the plant's endophytes. As rice undergo physiological and adaptive modifications through cross breeding in the process of attaining salinity tolerance, this may also lead to changes in the endophytic bacterial community especially those residing in the seeds. This study explores the community structure of seed bacterial endophytes as influenced by rice parental lineage, genotype and physiological adaptation to salinity stress. Endophytic bacterial diversity was studied through culture dependent technique, cloning and Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. Results revealed considerably diverse communities of bacterial endophytes in the interior of rice seeds. The richness of ribotypes ranges from 5-14 T-RFs corresponding to major groups of bacterial endophytes in the seeds. Endophytic bacterial diversity of the salt-sensitive IR29 is significantly more diverse compared to those of salt-tolerant cultivars. Proteobacteria followed by Actinobacteria and Firmicutes dominated the overall endophytic bacterial communities of the indica rice seeds based on 16S rDNA analysis of clones and isolates. Community profiles show common ribotypes found in all cultivars of the indica subspecies representing potential core microbiota belonging to Curtobacterium, Flavobacterium, Enterobacter, Xanthomonas, Herbaspirillum, Microbacterium and Stenotrophomonas. Multivariate analysis showed that the bacterial endophytic community and diversity of rice seeds are mainly influenced by their host's genotype, physiological adaptation to salt stress and parental lineage.
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