• Journal of Life Science
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• v.31 no.1
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• pp.28-36
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• 2021

Edwardsiella piscicida is a significant cause of hemorrhagic septicemia in fish and gastrointestinal infections in humans. Survival bacteria require specialized mechanisms to adapt to environmental fluctuations. Hence, to understand the mechanism through which E. piscicida senses and responds to environmental osmolarity changes, we determined the protein expression profile and physiological properties under various salinity conditions in this study. The OmpR protein is a part of the Env-ZOmpR two-component system that has been implicated in sensing salt stress in bacteria. However, the physiological role played by this protein in E. piscicida remains to be elucidated. Therefore, in this work, the function of the OmpR protein in response to salt stress was investigated. Phenotypic analysis revealed that, in the mutant, three of the biochemical phenotypes were different from the wild type, including, citrate utilization, hydrogen sulfide, and indole production. Introduction of the plasmid containing the entire ompR gene to the mutant strain returned it to its parental phenotype. The retarded growth rate also partially recovered. Furthermore, in our studies, OmpR was not found to be related to cell motility. Taken together, our results from the mutational analysis, the growth assay, MALDI-TOF MS, qRT-PCR, and the phenotype studies suggest that the OmpR of E. piscicida is implicated in osmoregulation, growth, expression of porins (ETAE_1826), virulence-related genes (EseC, EseD and EvpC), and certain genes of unknown function (ETAE_1540 and ETAE_2706).

• Journal of fish pathology
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• v.20 no.3
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• pp.201-209
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• 2007

Viral hemorrhagic septicemia (VHS) is one of the most serious viral disease of farmed rainbow trout and some marine fishes in Europe and North America. It has been reported in various marine fish species of Asian countries and induced cause mass mortality in Japanese flounder (Paralichthys olivaceus) culturing in Korea. The aims of this study were to monitor VHSV in wild marine fishes and to give critical information for controling the disease through prophylactic methods. Prevalence of the viral disease, geological distribution and reservoir of the virus were investigated using wild marine fishes captured in southern coast and east china sea for two years. (Reverse Transcriptase Polymerase Chain Reaction) RT-PCR results showed that VHSV were detected in 17 (10.6%) out of 160 fish. G gene sequences of viral strains isolated in this study were closely related to that of a reference strain, KVHS01-1, belonging to VHSV genotype Ⅰ. The results suggest that some of wild marine fishes are VHSV carriers and may spread the pathogen directly to fish farmed in coastal area.

• Development and Reproduction
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• v.20 no.4
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• pp.297-304
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• 2016

Lectins belong to the pattern-recognition receptors (PRRs) class and play important roles in the recognition and elimination of pathogens via the innate immune system. Recently, it was reported that lily-type lectin-1 is involved when a pathogen attacks in the early immune response of fish. However, this study is limited to information that the lectin is involved in the innate immune response against viral infection. In the present study, the lily-type lectin-2 and -3 of Oplegnathus fasciatus (OfLTL-2 and 3) have been presented to be included B-lectin domain and two D-mannose binding sites in the amino acid sequence that an important feature for the fundamental structure. To investigate the functional properties of OfLTLs, the tissue distribution in the healthy rock bream and temporal expression during early developmental stage analysis are performed using quantitative real-time PCR. OfLTL-2 and 3 are predominantly expressed in the liver and skin, but rarely expressed in other organ. Also, the transcripts of OfLTLs are not expressed during the early developmental stage but its transcripts are increased after immune-related organs which are fully formed. In the challenge experiment with RBIV (rock bream iridovirus), the expression of OfLTLs was increased much more strongly in the late response than the early, unlike previously known. These results suggest that OfLTLs are specifically expressed in the immune-related tissues when those organs are fully formed and it can be inferred that the more intensively involved in the second half to the virus infection.

• Journal of Life Science
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• v.18 no.8
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• pp.1023-1030
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• 2008

NtMEK2, which is the tobacco MAPK kinase that is upstream of SIPK and WIPK, was identified using the dexamethasone (DEX)-inducible gain-of-function transgenic system. Expression of $NtNEK2^{DD}$, a constitutively active mutant of NtNEK2, leads to HR-like cell death, which indicates that the NtMEK2-SIPK/WIPK cascade controls defense responses in tobacco. However, little is known about the downstream target substrates or defense-related genes that are regulated by the NtMEK2-SIPK/ WIPK cascade. In this study, ACP-based differential display RT-PCR was used to isolate the downstream effectors mediated by the NtMEK2-SIPK/WIPK cascade in $NtNEK2^{DD}$ transgenic plants. The results identified 6 novel differentially expressed genes (DEGs). These included pathogen induced protein 2-4 (pI2-4), monoterpene synthase 2 (MTS2), seven in absentia protein (SINA), cell death marker protein 1 (CDM1), hydroxyproline-rich glycoprotein (HRGP) and unknown genes (DEG45). The induction of these genes was confirmed by RT-PCR of samples obtained from $NtNEK2^{DD}$ plants. Additionally, when compared with other isolated DEGs, the pI2-4, CDM1 and HRGP genes were significantly up-regulated in response to treatment with salicylic acid and tobacco mosaic virus. Taken together, these results suggest that three novel DEGs were regulated by the NtMEK2-SIPK/WIPK cascade involved in disease resistance in tobacco.

• Journal of fish pathology
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• v.20 no.3
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• pp.211-220
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• 2007

Red sea bream iridovirus disease (RSIVD) cause massive economic losses in marine aquaculture industry in Korea. The causative agent of this disease (RSIV) infects a wide range of fish species. The aims of this study were to monitor RSIV in wild marine fishes and to give critical information for controling the disease through prophylactic methods. Prevalence of the viral disease, geological distribution and reservoir of the virus were investigated using wild marine fishes captured in southern coast and east china sea for two years. (Polymerase Chain Reaction) PCR results showed that RSIV were detected in 39 (24.3%) out of 160 fish. MCP gene sequences of viral strains isolated in this study were closely related to that of a reference strain, red seabream-K, belonging to Megalocytivirus subgroup Ⅲ. The results suggest that some of wild marine fishes are RSIV carriers and may spread the pathogen directly to fish farmed in coastal area.

• Research in Plant Disease
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• v.10 no.4
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• pp.331-336
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• 2004

The fungus genus Sclerotinia contains a number of important plant pathogens. Vegetable growers in our country are probably most familiar with Sclerotinia sclerotiorum, the causes of sclerotinia rot on crisphead lettuce. S. sclerotiorum has a wide host range which can include lettuce as well as crops such as broccoli, cabbage, carrots, celery, beans, peppers, potatoes, stocks, and tomato. Some fungicides, including benomyl, are effective in some crops, but not all. So, we isolated a antagonistic bacteria that are active on sclerotinia rot caused by S. sclerotiorum and that can be used to control it. About 702 strains had been isolated from soil around plant roots in the field. Ten strains showed strong antifungal activity against S. sclerotiorum. In pot test for antagonistic activity, A-7 strain showed high control value against the pathogen when compared with others. The strain was, therefore, selected as a biocontrol candidate against sclerotinia rot and its biochemical properties and 16S rDNA sequence was analyzed. The A-7 strain was highly related to Bacillus subtilis and B. amyloliquefaciens. To confirm precise identification, we had performed gyr A gene sequences analysis. Its sequence had 96% similarity with B. amyloliquefaciens. Consequently, the isolate was identified as B. amyloliquefaciens A-7.

• Plant Biotechnology Reports
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• v.3 no.2
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• pp.147-155
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• 2009

Oryza grandiglumis Chitinase IVa (OgChitIVa) cDNA encoding a class IV chitinase was cloned from wild rice (Oryza grandiglumis). OgChitIVa cDNA contains an open reading frame of 867 nucleotides encoding 288 amino acid residues with a predicted molecular weight of 30.4 kDa and isoelectric point of 8.48. Deduced amino acid sequences of OgChitIVa include the signal peptide and chitin-binding domain in the N-terminal domain and conserved catalytic domain. OgChitIVa showed significant similarity at the amino acid level with related monocotyledonous rice and maize chitinase, but low similarity with dicotyledoneous chitinase. Southern blot analysis showed that OgChitIVa genes are present as two copies in the wild rice genome. It was shown that RNA expression of OgChitIVa was induced by defense/stress signaling chemicals, such as jasmonic acid, salicylic acid, and ethephon or cantharidin and endothall or wounding, and yeast extract. It was demonstrated that overexpression of OgChitIVa in Arabidopsis resulted in mild resistance against the fungal pathogen, Botrytis cinerea, by lowering disease rate and necrosis size. RT-PCR analysis showed that PR-1 and PR-2 RNA expression was induced in the transgenic lines. Here, we suggest that a novel OgChitIVa gene may play a role in signal transduction process in defense response against B. cinerea in plants.

• BMB Reports
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• v.54 no.2
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• pp.118-123
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• 2021

The bacterial effector protein RavZ from a pathogen can impair autophagy in the host by delipidating the mammalian autophagy-related gene 8 (mATG8)-phosphatidylethanolamine (PE) on autophagic membranes. In RavZ, the membrane-targeting (MT) domain is an essential function. However, the molecular mechanism of this domain in regulating the intracellular localization of RavZ in cells is unclear. In this study, we found that the fusion of the green fluorescent protein (GFP) to the MT domain of RavZ (GFP-MT) resulted in localization primarily to the cytosol and nucleus, whereas the GFP-fused duplicated-MT domain (GFP-2xMT) localized to Rab5- or Rab7-positive endosomes. Similarly, GFP fusion to the catalytic domain (CA) of RavZ (GFP-CA) resulted in localization primarily to the cytosol and nucleus, even in autophagy-induced cells. However, by adding the MT domain to GFP-CA (GFP-CA-MT), the cooperation of MT and CA led to localization on the Rab5-positive endosomal membranes in a wortmannin-sensitive manner under nutrient-rich conditions, and to autophagic membranes in autophagy-induced cells. In autophagic membranes, GFP-CA-MT delipidated overexpressed or endogenous mATG8-PE. Furthermore, GFP-CA△α3-MT, an α3 helix deletion within the CA domain, failed to localize to the endosomal or autophagic membranes and could not delipidate overexpressed mATG8-PE. Thus, the CA or MT domain alone is insufficient for stable membrane localization in cells, but the cooperation of MT and CA leads to localization to the endosomal and autophagic membranes. In autophagic membranes, the CA domain can delipidate mATG8-PE without requiring substrate recognition mediated by LC3-interacting region (LIR) motifs.

• Korean Journal of Breeding Science
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• v.51 no.4
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• pp.395-403
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• 2019

Rice blast caused by the fungus Magnaporthe grisea (anamorphic: Pyricularia oryzae) is an important disease in rice and development of resistant varieties to blast is one of the most important goals in rice breeding programs. A japonica rice variety, Palgong, has shown resistance to the Korean blast pathogen since it was developed in 1996. Nine blast resistance quantitative trait loci (QTLs) in Palgong alleles were identified on chromosomes 2, 4, 7, and 11. Four QTLs of qBn2.3, qBn4.2, qBn11.1, and qBn11.2 explained 28-56.7% of total phenotypic variation, while five QTLs of qBn2.2, qBn2.4, qBn4.1, qBn7.1, and qBn7.2 explained 9.7-18.8%. In a previous study, one to four resistance genes were located on the loci qBn2.2, qBn2.3, qBn4.2, qBn11.1, and qBn11.2, however, resistance genes were not located on the loci qBn2.4, qBn4.1, and qBn7.1. A major QTL, qBn11.2, explaining 56.7% of total phenotypic variation was related to the durable resistance of Palgong. Additionally, rice stripe virus resistance of Palgong was assumed to be based on the Stvb-i gene, which is located on a major QTL qBn11.2.

• Korean Journal of Environment and Ecology
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• v.31 no.5
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• pp.444-454
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• 2017

There are many ongoing studies of infectious diseases as the major factor responsible for global declining of the amphibian population. Although some point out the amphibian rearing facilities like frog farms as one of the important sources of harboring and spreading amphibian infectious pathogens in the wild, there have been few related studies in South Korea. In this study, we investigated the bacterial and fungal colonies on the skin and in the internal organs of frogs and tadpoles collected inside and outside of Dybowski's brown frog farms in Inje, Goesan, and Gongju to compare the difference according to the region and between inside and outside the farm. We also intended to classify the bacteria collected from the tadpoles into species by analyzing 16s rDNA gene sequences. The result showed that the number of bacterial colonies found in the skin and gut of frogs and the number of fungal colonies found in the skin and liver of frogs collected in Goesan was significantly greater than those in the frogs in Inje. However, there was no difference between the frogs collected inside and outside of farms in both regions. In the case of tadpoles, the number of fungal colonies in the tadpoles collected from Gongju was greater than that in the tadpoles collected from Inje. The comparison of inside and outside frog farms showed that there were more bacterial colonies on the skin of the tadpoles collected from inside than outside the frog farm in Inje and more bacterial colonies in the organs of the tadpoles collected from outside than inside the farm in Gongju. The frogs with higher condition factor (body weight/snout-vent length*100) showed fewer bacterial colonies on the skin and fewer fungal colonies in the heart, but there were no significant relationships in tadpoles. We identified the total of 15 genera and four phyla of bacteria, but the difference according to regions and between inside and outside farm was not evident. The result of this study indicates that the different conditions according to the locality of farm and between inside and outside farm cause the difference in the population sizes of bacterial and fungal colonies and that it can affect the overall health condition of Dybowski's brown frogs in the farm. Moreover, the result suggests that effective disease control in the facility is greatly necessary to ensure successful operation of amphibian rearing facility and to prevent the possible spread of diseases from the facility to the wild.