Jaekyeong Song

Pyrosequencing analysis of the bacterial communities in the guts of honey bees Apis cerana and Apis mellifera in Korea.

The bacterial communities in the guts of the adults and larvae of the Asian honey bee Apis cerana and the European honey bee Apis mellifera were surveyed by pyrosequencing the 16S rRNA genes. Most of the gut bacterial 16S rRNA gene sequences were highly similar to the known honey bee-specific ones and affiliated with Pasteurellaceae or lactic acid bacteria (LAB). The numbers of operational taxonomic units (OTUs, defined at 97% similarity) were lower in the larval guts (6 or 9) than in the adult guts (18 or 20), and the frequencies of Pasteurellaceae-related OTUs were higher in the larval guts while those of LAB-related OTUs in the adult guts. The frequencies of Lactococcus, Bartonella, Spiroplasma, Enterobacteriaceae, and Flavobacteriaceae-related OTUs were much higher in A. cerana guts while Bifidobacterium and Lachnospiraceae-related OTUs were more abundant in A. mellfera guts. The bacterial community structures in the midguts and hindguts of the adult honey bees were not different for A. cerana, but significantly different for A. mellifera. The above results substantiated the previous observation that honey bee guts are dominated by several specific bacterial groups, and also showed that the relative abundances of OTUs could be markedly changed depending on the developmental stage, the location within the gut, and the honey bee species. The possibility of using the gut bacterial community as an indicator of honey bee health was discussed.

Effects of PCR cycle number and DNA polymerase type on the 16S rRNA gene pyrosequencing analysis of bacterial communities

The effects of PCR cycle number and DNA polymerase type on 16S rRNA gene pyrosequencing analysis were investigated using an artificially prepared bacterial community (mock community). The bacterial richness was overestimated at increased PCR cycle number mostly due to the occurence of chimeric sequences, and this was more serious with a DNA polymerase having proofreading activity than with Taq DNA polymerase. These results suggest that PCR cycle number must be kept as low as possible for accurate estimation of bacterial richness and that particular care must be taken when a DNA polymerase having proofreading activity is used.

Characterization of the bacterial and archaeal communities in rice field soils subjected to long-term fertilization practices

The bacterial and archaeal communities in rice field soils subjected to different fertilization regimes for 57 years were investigated in two different seasons, a non-planted, drained season (April) and a rice-growing, flooded season (August), by performing soil dehydrogenase assay, real-time PCR assay and pyrosequencing analysis. All fertilization regimes increased the soil dehydrogenase activity while the abundances of bacteria and archaea increased in the plots receiving inorganic fertilizers plus compost and not in those receiving inorganic fertilizers only. Rice-growing and flooding decreased the soil dehydrogenase activity while they increased the bacterial diversity in rice field soils. The bacterial communities were dominated by Chloroflexi, Proteobacteria, and Actinobacteria and the archaeal communities by Crenarchaeota at the phylum level. In principal coordinates analysis based on the weighted Fast UniFrac metric, the bacterial and archaeal communities were separated primarily by season, and generally distributed along with soil pH, the variation of which had been caused by long-term fertilization. Variations in the relative abundance according to the season or soil pH were observed for many bacterial and archaeal groups. In conclusion, the microbial activity, prokaryotic abundance and diversity, and prokaryotic community structure in the rice field soils were changed by season and long-term fertilization.

Draft Genome Sequence of the Biocontrol Bacterium Bacillus amyloliquefaciens Strain M27

Bacillus amyloliquefaciens strain M27 is a biocontrol agent with antagonistic activities against a wide range of fungal pathogens. Here we present the 3.86-Mb draft genome sequence of the bacterium with the aims of providing insights into the genomic basis of its antifungal mechanism and facilitating its application in the biocontrol of plant diseases.

Soil pH and electrical conductivity are key edaphic factors shaping bacterial communities of greenhouse soils in Korea

Soil microorganisms play an essential role in soil ecosystem processes such as organic matter decomposition, nutrient cycling, and plant nutrient availability. The land use for greenhouse cultivation has been increasing continuously, which involves an intensive input of agricultural materials to enhance productivity; however, relatively little is known about bacterial communities in greenhouse soils. To assess the effects of environmental factors on the soil bacterial diversity and community composition, a total of 187 greenhouse soil samples collected across Korea were subjected to bacterial 16S rRNA gene pyrosequencing analysis. A total of 11,865 operational taxonomic units at a 97% similarity cutoff level were detected from 847,560 sequences. Among nine soil factors evaluated; pH, electrical conductivity (EC), exchangeable cations (Ca2+, Mg2+, Na+, and K+), available P2O5, organic matter, and NO3-N, soil pH was most strongly correlated with bacterial richness (polynomial regression, pH: R2 = 0.1683, P < 0.001) and diversity (pH: R2 = 0.1765, P < 0.001). Community dissimilarities (Bray-Curtis distance) were positively correlated with Euclidean distance for pH and EC (Mantel test, pH: r = 0.2672, P < 0.001; EC: r = 0.1473, P < 0.001). Among dominant phyla (> 1%), the relative abundances of Proteobacteria, Gemmatimonadetes, Acidobacteria, Bacteroidetes, Chloroflexi, and Planctomycetes were also more strongly correlated with pH and EC values, compared with other soil cation contents, such as Ca2+, Mg2+, Na+, and K+. Our results suggest that, despite the heterogeneity of various environmental variables, the bacterial communities of the intensively cultivated greenhouse soils were particularly influenced by soil pH and EC. These findings therefore shed light on the soil microbial ecology of greenhouse cultivation, which should be helpful for devising effective management strategies to enhance soil microbial diversity and improving crop productivity.

Complete genome sequence of Bacillus velezensis M75, a biocontrol agent against fungal plant pathogens, isolated from cotton waste

Bacillus species have been widely used as biological control agents in agricultural fields due to their ability to suppress plant pathogens. Bacillus velezensis M75 was isolated from cotton waste used for mushroom cultivation in Korea, and was found to be antagonistic to fungal plant pathogens. Here, we report the complete genome sequence of the M75 strain, which has a 4,007,450-bp single circular chromosome with 3921 genes and a G+C content of 46.60%. The genome contained operons encoding various non-ribosomal peptide synthetases and polyketide synthases, which are responsible for the biosynthesis of secondary metabolites. Our results will provide a better understanding of the genome of B. velezensis strains for their application as biocontrol agents against fungal plant pathogens in agricultural fields.

Microbial Diversity and Community Analysis in Lettuce or Cucumber Cultivated Greenhouse Soil in Korea

The soil chemical properties, microbial community structures and biochemical properties of lettuce or cucumber-cultivated greenhouse soil samples were analyzed to assess soil health and characterize microbial distribution in 8 locations in Korea. Although most of chemical properties were within the soil management guidelines, the available phosphate, and the contents of exchangeable potassium and calcium were higher than those of recommended levels. In the culture-dependent analysis, 841 bacterial strains were isolated from the greenhouse soils and were identified at the genus level by 16S rRNA gene sequences analysis. The dominant bacterial genera were Bacillus (35.7%), Microbacterium (9.3%), Arthrobacter (5.7%) and Lysobacter (5.1%). The abundance of pseudomonads was highly variable depending on the soil samples. In the culture-independent analysis, soil microbial community was investigated by using phospholipid fatty acid (PLFA) method. Principal component analysis (PCA) showed that a specific grouping for microbial community structure in the greenhouse soils was not observed based on cultivated crops and investigated sites. The results revealed that the greenhouses soils examined are relatively sound managed in terms of soil chemical contents and microbial properties.

Terrabacter aerolatus sp. nov., isolated from an air sample

A novel actinomycete, designated strain 5414T-18(T), was isolated from an air sample collected from the Taean region, Korea. The strain contained oxidase and grew in the presence of 7 % NaCl. A neighbour-joining tree constructed on the basis of the 16S rRNA gene sequence showed that strain 5414T-18(T) is a member of the genus Terrabacter, sharing 97.8-98.3 % 16S rRNA gene sequence similarities to type strains of species of the genus Terrabacter (98.3 % sequence similarity with Terrabacter lapilli LR-26(T)). It contained peptidoglycan containing ll-diaminopimelic acid of A3gamma type, with three glycine residues as the interpeptide bridge. Whole-cell sugars were glucose, mannose and ribose. Mycolic acids were absent. The predominant menaquinone was MK-8(H(4)). The major fatty acids (>7 % of total fatty acids) were iso-C(15 : 0), iso-C(16 : 0), C(17 : 1)omega8c and iso-C(14 : 0). The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol and an unidentified phosphoglycolipid. The DNA G+C content of the type strain was 73 mol%. Strain 5414T-18(T) exhibited DNA-DNA relatedness levels of 44, 43, 39, 34 and 34 % to the type strains of Terrabacter lapilli, Terrabacter aerolatus, Terrabacter terrae, Terrabacter tumescens and Terracoccus luteus, respectively. These findings suggest that strain 5414T-18(T) represents a novel species within the genus Terrabacter. The name Terrabacter aeriphilus sp. nov. is proposed for this novel species, with the type strain 5414T-18(T) (=KACC 20693(T)=DSM 18563(T)).

Effects of diet type, developmental stage, and gut compartment in the gut bacterial communities of two Cerambycidae species (Coleoptera)

The gut bacterial community of wood-feeding beetles has been examined for its role on plant digestion and biocontrol method development. Monochamus alternatus and Psacothea hilaris, both belonging to the subfamily Lamiinae, are woodfeeding beetles found in eastern Asia and Europe and generally considered as destructive pests for pine and mulberry trees, respectively. However, limited reports exist on the gut bacterial communities in these species. Here, we characterized gut bacterial community compositions in larva and imago of each insect species reared with host tree logs and artificial diets as food sources. High-throughput 454 pyrosequencing of bacterial 16S rRNA gene revealed 225 operational taxonomic units (OTUs) based on a 97% sequences similarity cutoff from 138,279 sequence reads, the majority of which were derived from Proteobacteria (48.2%), Firmicutes (45.5%), and Actinobacteria (5.2%). The OTU network analysis revealed 7 modules with densely connected OTUs in specific gut samples, in which the distributions of Lactococcus-, Kluyvera-, Serratia-, and Enterococcus-related OTUs were distinct between diet types or developmental stages of the host insects. The gut bacterial communities were separated on a detrended correspondence analysis (DCA) plot and by c-means fuzzy clustering analysis, according to diet type. The results from this study suggest that diet was the main determinant for gut bacterial community composition in the two beetles.

Nocardioides paucivorans sp. nov. Isolated from Soil

One strain, designated KIS31-44T, was isolated from a soil sample collected from Dokdo Island, South Korea. The strain is Gram-stain-positive, aerobic, non-spore-forming and non-motile. It grows optimally at 28–30°C, at pH 7.0 and 0% NaCl. 16S rRNA gene sequence analysis showed that strain KIS31-44T belonged to the genus Nocardioides and shared the highest sequence similarities with Nocardioides aestuarii JC2056T (95.5%) and Nocardioides terrae VA15T (95.0%). The major fatty acids of strain KIS31-44T were C17:1ω6c, C18:1ω9c, summed feature 3 (C16:1ω6c and/or C16:1ω7c), iso-C16:0, C18:0 10-methyl (TBSA), C16:0 2-OH, C17:0 10-methyl, and iso-C16:1 H. The major isoprenoid quinone was MK-8 (H4). The strain contained diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol as the major polar lipids. The peptidoglycan structure was A3γ-type with ll-diaminopimelic acid. Based on these data, the isolate represents one novel species in the genus Nocardioides, for which the name Nocardioides paucivorans sp. nov. (type strain KIS31-44T =DSM 27142T =KACC 17309T) is proposed.