Fire blight disease, caused by Erwinia amylovora, is a devastating affliction in apple cultivation worldwide. Chemical pesticides have exhibited limited effectiveness in controlling the disease, and biological control options for treating fruit trees are limited. Therefore, a relatively large-scale survey is necessary to develop microbial agents for apple trees. Here we collected healthy apple trees from across the country to identify common and core bacterial taxa. We analyzed the endophytic bacterial communities in leaves and twigs and discovered that the twig bacterial communities were more conserved than those in the leaves, regardless of the origin of the sample. This finding indicates that specific endophytic taxa are consistently present in healthy apple trees and may be involved in vital functions such as disease prevention and growth. Furthermore, we compared the community metabolite pathway expression rates of these endophyte communities with those of E. amylovora infected apple trees and discovered that the endophyte communities in healthy apple trees not only had similar community structures but also similar metabolite pathway expression rates. Additionally, Pseudomonas and Methylobacterium-Methylorobrum were the dominant taxa in all healthy apple trees. Our findings provide valuable insights into the potential roles of endophytes in healthy apple trees and inform the development of strategies for enhancing apple growth and resilience. Moreover, the similarity in cluster structure and pathway analysis between healthy orchards was mutually reinforcing, demonstrating the power of microbiome analysis as a tool for identifying factors that contribute to plant health.
Kim, Kiyoon;Samaddar, Sandipan;Ahmed, Shamim;Roy, Choudhury Aritra;Sa, Tongmin
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.364-364
/
2017
Saemangeum reclaimed land is a part of Saemangeum Development Project. Most of the persistent problems of Saemangeum reclaimed land remain to be related to soil salinity. Soil salinity is a major abiotic factor related to microbial community structure and also fungi have been reported to be more sensitive to salinity stress than bacteria. The aim of this study was conducted to investigate the effect of soil salinity levels on the microbial communities in Saemangeum reclaimed land using 454 pyrosequencing analysis. Soil samples was collected from 12 sites of in Saemangeum reclaimed land. For pyrosequencing, 27F/518R (bacteria) and ITS3/ITS4 (fungi) primers were used containing the Roche 454 pyrosequencing adaptor-key-linker (underlined) and unique barcodes (X). Pyrosequencing was performed by Chun's Lab (Seoul, Korea) using the standard shotgun sequencing reagents and a 454 GS FLX Titanium sequencing System (Roche, Inc.). In the soil samples, Proteobacteria (bacteria) and Ascomycota (fungi) shows the highest relative abundance in all the soil sample sites. Proteobacteria, Bacteroidetes, Plantomycetes, Gemmatimonadetes and Parcubacteria were shown to have significantly higher abundance in high salinity level soils than low salinity level soils, while Acidobacteria and Nitrospirae has significantly higher relative abundance in low salinity level soils. The abundance of fungal, Ascomycota has the highest relative abundance in soil samples, followed by Basidiomycota, Chlorophyta, Zygomycota and Chytridiomycota. Basidiomycota, Zygomycota, Glomeromycota and Cerozoa were show significantly higher relative abundance in low salinity level soils. The principal coordinate analysis (PCoA) and correlation analysis shown to salinity-related soil parameters such as ECe, Na+, SAR and EPS were affected to bacterial and fungal community structure. Proteobacteria, Bacteroidetes, Plantomycetes exhibited significantly positive correlation with soil salinity, while Acidobacteria exhibited significantly negative correlation. In the case of fungal community, Basidiomycota and Zygomycota were seen show significantly negative correlation with salinity related soil parameters. These results suggest that provide understanding effect of soil salinity on microbial community structure and correlation of microbial community with soil parameters in Saemangeum reclaimed land.
Journal of Korean Society of Environmental Engineers
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v.33
no.11
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pp.847-854
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2011
To deal with issues from global climate changes, renewable bioenergy has become important. Algae have been regarded as a good resource for biorefinery and bioenergy, and also have potential capability to remove nutrient and non-decompositional pollutants for wastewater advanced treatment. Although algal-bacterial ecological interaction would be a crucially important factor in using algae for wastewater advanced treatment and resource recovery from wastewater, very little is known about ecological interaction between algae and bacteria in a real wastewater environment. In this study, under a real municipal wastewater condition, we characterized wastewater pollutant treatability and bacterial communities in response to growth of Ankistrodesmus gracilis SAG278-2, which can grow in wastewater and has a high lipid contents. The growth of algal population using the wastewater was inhibited by increase in wastewater bacteria while bacterial survival and cellular decay rate were not influenced by the algal growth. Removals of recalcitrant organic matters and total nitrogen were improved in the presence of algal growth. According to T-RFLP and statistical analysis, algal growth affected time-course changes in bacterial community structures. The following 16S rRNA gene amplicon, cloning results showed that the algal growth changes in bacterial community structure, and that bacterial populations belonging to Sediminibacterium, Sphingobacterium, Mucilaginibacter genera were identified as cooperative with the algal growth in the wastewater.
To understand the ecological function of heterotrophic bacterial community in water column of large freshwater lakes in the permafrost zone, we investigated the structure and function of bacterial community in Lake Khuvsgul, Mongolia. Species composition of overall bacterial community was analyzed by denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments, and bacteria that can be cultured at 10oC were isolated and characterized. Based on the depth profile of environmental parameters, thermocline and chemocline were recognized at the 5~10 m zone of the water column. The stratified DGGE profile indicated that the discontinuity of water properties might influence the structure of bacterial community: band profiles in the 0~5 m zone were diverse with large change by depth, but the profile was relatively stable at the $\geq$10 m zone, with predominance of the band identified as Acidovorax facilis. Bacterial cultures were screened for protease, cellulase, amylase and lipase activity, and 23 isolates were selected for high activity of the hydrolytic enzymes. The isolates were identified based on their 16S rRNA gene sequences. In the surface water (zero meter depth), Acidovorax defluvii and Sphingobacterium faecium with high cellulase activity were present. Flavobacterium succinicans, Mycoplana bullata and A. facilis were stably predominant isolates at 2 m, 5 m, and $\geq$10 m depths, respectively. F. succinicans isolates showed high protease activity while M. bullata isolates showed moderate levels of protease and celluase activity. A. facilis isolates showed either cellulase or lipase activity, exclusively to each other. According to the profile of growth rates of the isolates in the temperature range of $0\sim42^{\circ}C$, the surface-zone (0~5 m) isolates were facultative psychrophiles while isolates from $\geq$10 m depth were typical mesophiles. This stratification is believed to be due to stratified availability of organic materials to the bacterial decomposers. In the water column below the chemoline, the environment is extremely oligotrophic so that the trait of rapid growth in low temperature might not be demanded by deep-lake decomposers. The stratified distribution of community composition and decomposer activity in Lake Khuvsgul implies that ecological functions of bacterial community in lakes of cold region are sharply divided by water column stratification.
Culture-dependent methods, such as heterotrophic plate counting (HPC), are usually applied to evaluate the bacteriological quality of hemodialysis water. However, these methods cannot detect the uncultured or viable but non-culturable (VBNC) bacteria, both of which may be quantitatively predominant throughout the hemodialysis water treatment system. Therefore, propidium monoazide (PMA)-qPCR associated with HPC was used together to profile the distribution of the total viable bacteria in such a system. Moreover, high-throughput sequencing of 16S rRNA gene amplicons was utilized to analyze the microbial community structure and diversity. The HPC results indicated that the total bacterial counts conformed to the standards, yet the bacteria amounts were abruptly enhanced after carbon filter treatment. Nevertheless, the bacterial counts detected by PMA-qPCR, with the highest levels of $2.14{\times}10^7copies/100ml$ in softener water, were much higher than the corresponding HPC results, which demonstrated the occurrence of numerous uncultured or VBNC bacteria among the entire system before reverse osmosis (RO). In addition, the microbial community structure was very different and the diversity was enhanced after the carbon filter. Although the diversity was minimized after RO treatment, pathogens such as Escherichia could still be detected in the RO effluent. In general, both the amounts of bacteria and the complexity of microbial community in the hemodialysis water treatment system revealed by molecular approaches were much higher than by traditional method. These results suggested the higher health risk potential for hemodialysis patients from the up-to-standard water. The treatment process could also be optimized, based on the results of this study.
Kimchi is a traditional Korean fermented vegetable food, the production of which involves brining of Korean cabbage, blending with various other ingredients (red pepper powder, garlic, ginger, salt-pickled seafood, etc.), and fermentation. Recently, kimchi has also become popular in the Western world because of its unique taste and beneficial properties such as antioxidant and antimutagenic activities, which are derived from the various raw materials and secondary metabolites of the fermentative microorganisms used during production. Despite these useful activities, analysis of the microbial community present in kimchi has received relatively little attention. The objective of this study was to evaluate the bacterial community structure from the raw materials, additives, and final kimchi product using the culture-independent method. Specifically, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the 16S rRNA partial sequences of the microflora. One primer set for bacteria, 341FGC-518R, reliably produced amplicons from kimchi and its raw materials, and these bands were clearly separated on a 35-65% denaturing gradient gel. Overall, 117 16S rRNA fragments were identified by PCR-DGGE analysis. Pediococcus pentosaceus, Leuconostoc citreum, Leuconostoc gelidum, and Leuconostoc mesenteroides were the dominant bacteria in kimchi. The other strains identified were Tetragenococcus, Pseudomonas, Weissella, and uncultured bacterium. Comprehensive analysis of these microorganisms could provide a more detailed understanding of the biologically active components of kimchi and help improve its quality. PCR-DGGE analysis can be successfully applied to a fermented food to detect unculturable or other species.
Do, Thi Huyen;Dao, Trong Khoa;Nguyen, Khanh Hoang Viet;Le, Ngoc Giang;Nguyen, Thi Mai Phuong;Le, Tung Lam;Phung, Thu Nguyet;Straalen, Nico M. van;Roelofs, Dick;Truong, Nam Hai
Asian-Australasian Journal of Animal Sciences
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v.31
no.5
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pp.738-747
/
2018
Objective: In a previous study, analysis of Illumina sequenced metagenomic DNA data of bacteria in Vietnamese goats' rumen showed a high diversity of putative lignocellulolytic genes. In this study, taxonomy speculation of microbial community and lignocellulolytic bacteria population in the rumen was conducted to elucidate a role of bacterial structure for effective degradation of plant materials. Methods: The metagenomic data had been subjected into Basic Local Alignment Search Tool (BLASTX) algorithm and the National Center for Biotechnology Information non-redundant sequence database. Here the BLASTX hits were further processed by the Metagenome Analyzer program to statistically analyze the abundance of taxa. Results: Microbial community in the rumen is defined by dominance of Bacteroidetes compared to Firmicutes. The ratio of Firmicutes versus Bacteroidetes was 0.36:1. An abundance of Synergistetes was uniquely identified in the goat microbiome may be formed by host genotype. With regard to bacterial lignocellulose degraders, the ratio of lignocellulolytic genes affiliated with Firmicutes compared to the genes linked to Bacteroidetes was 0.11:1, in which the genes encoding putative hemicellulases, carbohydrate esterases, polysaccharide lyases originated from Bacteroidetes were 14 to 20 times higher than from Firmicutes. Firmicutes seem to possess more cellulose hydrolysis capacity showing a Firmicutes/Bacteroidetes ratio of 0.35:1. Analysis of lignocellulolytic potential degraders shows that four species belonged to Bacteroidetes phylum, while two species belonged to Firmicutes phylum harbouring at least 12 different catalytic domains for all lignocellulose pretreatment, cellulose, as well as hemicellulose saccharification. Conclusion: Based on these findings, we speculate that increasing the members of Bacteroidetes to keep a low ratio of Firmicutes versus Bacteroidetes in goat rumen has resulted most likely in an increased lignocellulose digestion.
The bacterial community structure in biological activated carbon (BAC) process in drinking water treatment plant was investigated by Fluorescent in situ Hybridization (FISH) with rRNA-targeted oligonucleotide probe. Samples were collected at different three points in BAC process every month for one year. They were hybridized with a probe specific for the alpha, beta, gamma subclass of the class Proteobacteria, Cytophaga-Flavobacteria group and Gram-positive high G+C content (HGC) group. Total numbers of bacteria in BAC process counted by 4',6-diamidino-2-phenylindole (DAPI) staining were $5.4{\times}10^{10}$ (top), $4.0{\times}10^{10}$ (middle) and $2.8{\times}10^{10}$ cells/ml (bottom). The number of the culturable bacteria was from $1.0{\times}10^7$ to $3.6{\times}10^7$ cells/ml and the culturability was about 0.05%. The faction of bacteria detectable by FISH with the probe EUB338 was about 83% of DAPI counts. Gamma and alpha subclass of the class Proteobacteria were predominant in BAC process and their ratios were over 20% respectively. In top and middle, alpha, beta and gamma subclass of the class Proteobacteria competed with each other and their percentages was changed according to the season. In bottom, gamma subclass of the class Proteobacteria was predominant all through the year. It could be successfully observed the seasonal distribution of bacterial community in biological activated carbon process using FISH.
For elucidating the correlation between the eubacterial community structure and environmental parameters in Nammae Reservoir located in Kyungsan, Kyungbuk, the bacterial community structure and their structure affecting environmental parameters were analyzed using Fluorescent In Situ Hybridization (FISH) monthly over year. $\alpha$ . $\beta$ . $\gamma$-subclasses of Proteobacteria and Cytophaga-Flavobacterium (CF) group known as dominant bacterial group in freshwater were detected in 3 stations over year. The ratio of each subclass to total bacteria was determined; $\alpha$.$\beta$ . $\gamma$-subclasses and CF group varied in the range of 4.0~29.2%, 1.7~25.8%, 1.8~12.8%, 4.9~36.3%, respectively and there was no substantial differences between stations. In terms of the correlation between each group specific bacteria and environmental parameters such as temperature, SS, pH, DOC, NH$_4$-N, NO$_3$-N, PO$_4$-P, standing crops of algae, the results were as follows: 1) total bacterial numbers correlated positively with temperature, SS and DOC, 2) Eubacteria positively with DOC and Chl-$\alpha$, 3)${\gamma}$-subclass positively with DOC, and 4) CF group positively with standing crops of chlorophyceae, 5) whereas $\beta$-subclass bacteria correlated negatively with standing crop of cyanobacteria and that of total algae.
We compared the composition of phospholipid fatty acids (PLFA) to assess the microbial community structure in the soil and rhizosphere community of non-transgenic watermelons and transgenic watermelons in Miryang farmlands in Korea during the spring and summer of 2005. The PLFA data were seasonally examined for the number of PLFA to determine whether there is any difference in the microbial community in soils from two types of watermelons, non-transgenic and transgenic. We identified 78 PLFAs from the rhizosphere samples of the two types of watermelons. We found eight different PLFAs for the type of plants and sixteen PLFAs for the interaction of plant type and season. The PLFA data were analyzed by analysis of variance separated by plant type (P<0.0085), season (P<0.0154), and the plant type${\times}$season interaction (P<0.1595). Non-parametric multidimensional scaling (NMS showed a small apparent difference but multi-response permutation procedures (MRPP) confirmed that there was no difference in microbial community structure for soils of both plant types. Conclusively, there was no significant adverse effect of transgenic watermelon on bacterial and fungal relative abundance as measured by PLFA. We could reject our hypothesis that there might be an adverse effect from transgenic watermelon with our statistical results. Therefore, we can suggest the use of this PLFA methodology to examine the adverse effects of transgenic plants on the soil microbial community.
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