A bacterium producing acidic cellulase was isolated from pig feces. The isolate, ATC6 strain, was found to be Gram-positive, non-motile, catalase-positive, and spore-forming stain. Under an electron microscope, the cells were observed to be rod-shaped. The isolate was identified as Bacillus amyloliquefaciens ATC6 on the basis of morphological and biochemical properties as well as 16S rRNA gene sequences. Optimum pH and temperature for the cellulase activity of the culture supernatant of B. amyloliquefaciens ATC6 were found to be pH 4.5 and $55^{\circ}C$, respectively. More than 80% of its maximum activity was maintained at pH 4.0. The cellulase activity was maintained at temperatures ranging from 35 to $55^{\circ}C$ after 2 h incubation at pH 4.5, whereas it's activity decreased rapidly at $65^{\circ}C$.
The gene for the Bdi I modification enzyme, which is one of Bdi I restriction-modification system, fromBrevibacterium divaricatum FERM 5948 was cloned and expressed in E. coli. For cloning of the Bdi I methylase gene, we have initially used three cloning site(EcoRI, BamHI and Sal I) of plasmid vector pBR 322 and adopted the retransformation method after Bdi I restriction endonuclease cleavage. Selection of transformants carrying the gene was based on the resistance of the modified plasmid encoding the enzyme to cleavage by Bdi I restriction enzyme, and the recombinant plasmid pBDIM 116 containing 5.6kb EcoRI insery was proved to carry the gene. Crude cell extracts prepared from strains carrying the plasmid pBDIM 116 contained an S-adenosylmethionine-dependent methyltransferase activity specific for the Bdi I recognition site, ATCGAT. The restriction map was constructed with 11 restriction enzyme, and the Bdi I restriction-modification system was also discussed.
The distribution and abundance of fish eggs and larvae were investigated from February to December 2020 along the coastal waters of Korea. The eggs and larvae were identified using the mitochondrial DNA cytochrome c oxidase subunit I (mtDNA COI) and 16s rRNA gene. During the study period, eggs of overall 45 taxa belonging to 26 families were collected and larvae of overall 39 taxa belonging to 23 families were collected. In Yeongil Bay, eggs of Engraulis japonicus, which accounted for 83.9% of the total population, was the most dominant species, followed by Sardinops sagax (4.0%), Repomucenus valenciennei (3.8%) and E. japonicus larvae, which accounted for 34.9% of the total population. These were followed by Sebastiscus marmoratus (31.0%). In Gomso Bay, E. japonicus eggs accounted for 61.7% of the total population, followed by Sillago japonica (14.0%), Johnius grypotus (8.8%) and Pholis fangi larvae, which accounted for 53.5% of the total population, followed by Ammodytes personatus (34.1%). In Jinhae Bay, E. japonicus eggs accounted for 86.0% of the total population, followed by Leiognathus nuchalis (4.1%), Konosirus punctatus (3.7%) and E. japonicus larvae, which accounted for 48.7% of the total population, followed by Parablennius yatabei (21.6%).
Sohn, Soo-In;Ahn, Byung-Ohg;Chi, Hee-Youn;Cho, Byung-Kwan;Cho, Min-Seok;Shin, Kong Sik
Korean Journal of Soil Science and Fertilizer
/
v.45
no.5
/
pp.829-835
/
2012
The cultivation of genetically modified (GM) crops has increased due to their economic and agronomic advantages. Before commercialization of GM crops, however, we must assess the potential risks of GM crops on human health and environment. The aim of this study was to investigate the possible impact of Bt rice on the soil microbial community. Microbial communities were isolated from the rhizosphere soil cultivated with Bt rice and Nakdong, parental cultivar and were subjected to be analyzed using both culture-dependent and molecular methods. The total counts of bacteria, fungi, and actinomycetes in the rhizosphere of transgenic and conventional rice were not significantly different. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that the bacterial community structures during cultural periods were very similar each other. Analysis of dominant isolates in the rhizosphere cultivated with Bt and Nakdong rice showed that the dominant isolates from the soil of Bt rice and Nakdong belonged to the Proteobacteria, Cloroflexi, Actinobacteria, Firmicutes, and Acidobacteria. These results indicate that the Bt rice has no significant impact on the soil microbial communities during cultivation period. Further study remains to be investigated whether the residue of Bt rice effect on the soil environment.
Park, Seong-Joo;Yoon, Jerng-Chang;Shin, Kwang-Soo;Kim, Eung-Ho;Yim, Soo-Bin;Cho, Yeon-Je;Sung, Gi-Moon;Lee, Dong-Geun;Kim, Seung-Bum;Lee, Dong-Uk;Woo, Sung-Hoon;Koopman, Ben
Journal of Microbiology
/
v.45
no.2
/
pp.113-121
/
2007
The bacterial diversity inherent to the biofilm community structure of a modified rotating biological contactor wastewater treatment process, referred to as the Rotating Activated Bacillus Contactor (RABC) process, was characterized in this study, via both culture-dependent and culture-independent methods. On the basis of culture-dependent methods, Bacillus sp. were found to exist in large numbers on the biofilm (6.5% of the heterotrophic bacteria) and the microbial composition of the biofilms was quite simple. Only three phyla were identified-namely, the Proteobacteria, the Actinobacteria (High G+C Gram-positive bacteria), and the Firmicutes (Low G+C Gram-positive bacteria). The culture-independent partial 16S rDNA sequence analysis revealed a considerably more diverse microbial composition within the biofilms. A total of eight phyla were recovered in this case, three of which were major groups: the Firmicutes (43.9%), the Proteobacteria (28.6%), and the Bacteroidetes (17.6%). The remaining five phyla were minor groups: the Planctomycetes (4.4%), the Chlorobi (2.2%), the Actinobacteria (1.1%), the Nitrospirae (1.1%), and the Verrucomicrobia (1.1%). The two most abundant genera detected were the endospore-forming bacteria (31.8%), Clostridium and Bacillus, both of which are members of the Firmicutes phylum. This finding indicates that these endospore-forming bacteria successfully colonized and dominated the RABC process biofilms. Many of the colonies or clones recovered from the biofilms evidenced significantly high homology in the 16S rDNA sequences of bacteria stored in databases associated with advanced wastewater treatment capabilities, including nitrification and denitrification, phosphorus accumulation, the removal of volatile odors, and the removal of chlorohydrocarbons or heavy metals. The microbial community structures observed in the biofilms were found to correlate nicely with the enhanced performance of advanced wastewater treatment protocols.
Chung, Eu Jin;Hossain, Mohammad Tofajjal;Khan, Ajmal;Kim, Kyung Hyun;Jeon, Che Ok;Chung, Young Ryun
The Plant Pathology Journal
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v.31
no.2
/
pp.152-164
/
2015
Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.
It is urgently required to construct safety data on agricultural by-products imported for use as medium materials for domestic mushroom production. However, research on microorganisms is insufficient. This study was conducted to investigate the presence of bacteria that have the possibility of harmful effects on human, plants and mushroom in wheat straw, peatmoss, cottonseed hull, cottonseed meal, and beet pulp imported from Australia, Canada, China, Egypt, Germany. Bacteria were found in the range of $1.35{\times}10^2$ to $8.34{\times}10^6CFU/g$. As a result of 16S rDNA sequence analysis, total of 19 genera and 45 species of bacteria were identified. Bacillus genus was dominant, followed by Paenibacillus genus. At the species level, diverse species was in the order of Firmicute, Proteobacteria and Actinobacteria. Regarding the agricultural by-products, straw and peat moss had more diverse bacteria than other agricultural by-products. Among the indentified bacteria, 6 species of 5 genera (Enterobacter asburiae, Enterobacter ludwigii, Stenotrophomonas maltophilia, Pseudomonas monteilii, Bacillus anthracis, and Cellulosimicrobium funkei) were present as potent harmful bacteria to human. Surprisingly, both the human and plant pathogenic Klebsiella pneumoniae subsp. pneumonia was present. Bacillus altitudinis was present as a plant pathogen. Lysinibacillus sphaericus, an insect pathogen, and Ochrobactrum pseudogrignonense, a mushroom pathogen, were also present. The results of this study confirmed that several kinds of pathogenic bacteria were present in the agricultural by-products for the mushroom cultivation medium imported into Korea. Our work suggests that hygiene inspection and management is urgently needed for imported agricultural by-products to be safely used for mushroom production.
A thermophilic nonspore-forming rod isolated from hay compost in Korea was subjected to a taxonomic study. The microorganism, designated as $SC-1^T$, was identified as a nitrate-reducing and nonmotile bacterium. Although the strain was negatively Gram-stained, a KOH test showed that the strain $SC-1^T$ belonged to a Gram-positive species. Growth was observed between 45 and $70^{\circ}C$. The optimal growth temperature and pH were $60^{\circ}C$ and pH 7.5, respectively. The G+C content of the genomic DNA was 65 mol% and the major quinone types were MK-6 and MK-7. A phylogenetic analysis based on 16S rDNA sequences revealed that the strain $SC-1^T$ was most closely related to Symbiobacterium thermophilum. However, the level of DNA-DNA relatedness between strain $SC-1^T$ and the type strain for Symbiobacterium thermophilum was approximately 30%. Accordingly, on the basis of the phenotypic traits and molecular systematic data, the strain $SC-1^T$ would appear to represent a new species within the genus Symbiobacterium. The type strain for the new species is named $SC-1^T$ ($=KCTC\;0307BP^T;\;DSM15906^T$).
Yang, Sung Jun;Kim, Chang Yong;Lee, Jo Byoung;Kang, Sung Sun;Lee, Jong Jin
Journal of Sericultural and Entomological Science
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v.51
no.2
/
pp.99-106
/
2013
The purpose of this study is to investigate the effects of supplementation of mulberry powder, mulberry extract and silkworm powder during the 8 weeks of resistance exercise on Androgen receptor(AR) mRNA and Myogenic regulatory factors(MRFs) expression of rats muscle. Fifty males, Sprague-Dawley rat, were randomly divided into 5 groups: CON(control group, n = 10), REG(resistance exercise group, n = 10), MP REG(mulberry powder intake and resistance exercise group, n = 10), ME REG(mulberry extract intake and resistance exercise group, n = 10) and SP REG(silkworm powder intake and resistance exercise group, n = 10). After climbing the ladder without weights during the 1 week of adaptation period, the rats in the resistance exercise group were trained to climb a 0.98-m vertical(80 degree incline) ladder with weights in their tail during 7 weeks(10 times each day, 2 days per week). After exercise, the skeletal muscle was extracted from the flexor hallucis longus. After separating the total ribonucleic acid (RNA) of each group, quantitative polymerase chain reaction was used to analyze RNA quantitatively. AR mRNA and MRFs expression revealed that all of the treated groups had significantly difference. AR mRNA expression increased in ME REG $6.24{\pm}1.85$ and SP REG $9.68{\pm}0.28$ fold compared to CON. Myod mRNA expression increased in MP REG $6.04{\pm}0.47$, ME REG $4.31{\pm}1.58$ and SP REG $8.11{\pm}0.57$ fold compared to CON. And myogenin mRNA expression increased in MP REG $4.11{\pm}0.42$, ME REG $4.12{\pm}0.45$ and SP REG $6.50{\pm}0.61$ fold compared to CON. In conclusion, during the resistance exercise, providing mulberry and silkworm gives positive effect on AR mRNA and MRFs expression increase.
Objective: The present study employed 5-aza-2'-deoxycytidine (5-Aza-CdR) to treat non-small cell lung cancer (NSCLC) cell line A549 to investigate the effects on proliferation and expression of the TFPI-2 gene. Methods: Proliferation was assessed by MTT assay after A549 cells were treated with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR, a specific demethylating agent, for 24, 48 and 72h. At the last time point cells were also analyzed by flow cytometry (FCM) to identify any change in their cell cycle profiles. Methylation-specific polymerase chain reaction (MSPCR), real time polymerase chain reaction(real-time PCR) and western blotting were carried out to determine TFPI-2 gene methylation status, mRNA expression and protein expression. Results: MTT assay showed that the growth of A549 cells which were treated with 5-Aza-CdR was significantly suppressed as compared with the control group (0 ${\mu}mol/L$ 5-Aza-CdR). After treatment with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h, FCM showed their proportion in G0/G1 was $69.7{\pm}0.99%$, $76.1{\pm}0.83%$, $83.8{\pm}0.35%$, $95.5{\pm}0.55%$ respectively (P<0.05), and the proportion in S was $29.8{\pm}0.43%$, $23.7{\pm}0.96%$, $15.7{\pm}0.75%$, $1.73{\pm}0.45%$, respectively (P<0.05), suggesting 5-Aza-CdR treatment induced G0/G1 phase arrest. MSPCR showed that hypermethylation in the promoter region of TFPI-2 gene was detected in control group (0 ${\mu}mol/L$ 5-Aza-CdR), and demethylation appeared after treatment with 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h. Real-time PCR showed that the expression levels of TFPI-2 gene mRNA were $1{\pm}0$, $1.49{\pm}0.14$, $1.86{\pm}0.09$ and $5.80{\pm}0.15$ (P<0.05) respectively. Western blotting analysis showed the relative expression levels of TFPI-2 protein were $0.12{\pm}0.01$, $0.23{\pm}0.02$, $0.31{\pm}0.02$, $0.62{\pm}0.03$ (P<0.05). TFPI-2 protein expression in A549 cells was gradually increased significantly with increase in the 5-Aza-CdR concentration. Conclusions: TFPI-2 gene promoter methylation results in the loss of TFPI-2 mRNA and protein expression in the non-small cell lung cancer cell line A549, and 5-Aza-CdR treatment could induce the demethylation of TFPI-2 gene promoter and restore TFPI-2 gene expression. These findings provide theoretic evidence for clinical treatment of advanced non-small cell lung cancer with the demethylation agent 5-Aza-CdR. TFPI-2 may be one molecular marker for effective treatment of advanced non-small cell lung cancer with 5-Aza-CdR.
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