• Title/Summary/Keyword: Wild strain

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Genotype distribution of infectious haematopoietic necrosis virus (IHNV) in Korea

  • Cha, Seung Joo;Jung, Yo Han;Lee, Hyun Young;Jung, Ji Yoon;Cho, Hee Jung;Park, Mi Seon
    • Journal of fish pathology
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    • v.25 no.3
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    • pp.143-150
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    • 2012
  • Infectious haematopoietic necrosis virus (IHNV) is an important fish pathogen that infects both wild and cultured salmonids. Since the first isolation of IHNV from rainbow trout and masu salmon in 1991, a series of IHN disease outbreak has been reported in Korea. In 2011, we isolated two IHNV isolates from rainbow trout cultured in Korea. The full open-reading frame (ORF) encoding the glycoprotein (G) of them were sequenced and the amino acid sequences were phylogenetically analyzed. Phylogenetic analysis of the G revealed that both IHNV isolates were grouped into an Asian genogroup containing Korean IHNV isolates and Japanese IHNV isolates. However, based on their sequence variation, they were divided into different subgroup. While one isolate was similar to other Korean isolates, the other isolate showed a high level of similarity with Japanese isolates, suggesting the possibility of influx of new IHNV strain into Korea.

Survival Factor Gene FgSvf1 Is Required for Normal Growth and Stress Resistance in Fusarium graminearum

  • Li, Taiying;Jung, Boknam;Park, Sook-Young;Lee, Jungkwan
    • The Plant Pathology Journal
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    • v.35 no.5
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    • pp.393-405
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    • 2019
  • Survival factor 1 (Svf1) is a protein involved in cell survival pathways. In Saccharomyces cerevisiae, Svf1 is required for the diauxic growth shift and survival under stress conditions. In this study, we characterized the role of FgSvf1, the Svf1 homolog in the homothallic ascomycete fungus Fusarium graminearum. In the FgSvf1 deletion mutant, conidial germination was delayed, vegetative growth was reduced, and pathogenicity was completely abolished. Although the FgSvf1 deletion mutant produced perithecia, the normal maturation of ascospore was dismissed in deletion mutant. The FgSvf1 deletion mutant also showed reduced resistance to osmotic, fungicide, and cold stress and reduced sensitivity to oxidative stress when compared to the wild-type strain. In addition, we showed that FgSvf1 affects glycolysis, which results in the abnormal vegetative growth in the FgSvf1 deletion mutant. Further, intracellular reactive oxygen species (ROS) accumulated in the FgSvf1 deletion mutant, and this accumulated ROS might be related to the reduced sensitivity to oxidative stress and the reduced resistance to cold stress and fungicide stress. Overall, understanding the role of FgSvf1 in F. graminearum provides a new target to control F. graminearum infections in fields.

A Mutation of a Putative NDP-Sugar Epimerase Gene in Ralstonia pseudosolanacearum Attenuates Exopolysaccharide Production and Bacterial Virulence in Tomato Plant

  • Hyoung Ju Lee;Sang-Moo Lee;Minseo Choi;Joo Hwan Kwon;Seon-Woo Lee
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.417-429
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    • 2023
  • Ralstonia solanacearum species complex (RSSC) is a soil borne plant pathogen causing bacterial wilt on various important crops, including Solanaceae plants. The bacterial pathogens within the RSSC produce exopolysaccharide (EPS), a highly complicated nitrogencontaining heteropolymeric polysaccharide, as a major virulence factor. However, the biosynthetic pathway of the EPS in the RSSC has not been fully characterized. To identify genes in EPS production beyond the EPS biosynthetic gene operon, we selected the EPS-defective mutants of R. pseudosolanacearum strain SL341 from Tn5-inserted mutant pool. Among several EPSdefective mutants, we identified a mutant, SL341P4, with a Tn5-insertion in a gene encoding a putative NDP-sugar epimerase, a putative membrane protein with sugar-modifying moiety, in a reverse orientation to EPS biosynthesis gene cluster. This protein showed similar to other NDP-sugar epimerases involved in EPS biosynthesis in many phytopathogens. Mutation of the NDP-sugar epimerase gene reduced EPS production and biofilm formation in R. pseudosolanacearum. Additionally, the SL341P4 mutant exhibited reduced disease severity and incidence of bacterial wilt in tomato plants compared to the wild-type SL341 without alteration of bacterial multiplication. These results indicate that the NDP-sugar epimerase gene is required for EPS production and bacterial virulence in R. pseudosolanacearum.

Overexpression of S-Adenosylmethionine Synthetase in Recombinant Chlamydomonas for Enhanced Lipid Production

  • Jeong Hyeon Kim;Joon Woo Ahn;Eun-Jeong Park;Jong-il Choi
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.310-318
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    • 2023
  • Microalgae are attracting much attention as promising, eco-friendly producers of bioenergy due to their fast growth, absorption of carbon dioxide from the atmosphere, and production capacity in wastewater and salt water. However, microalgae can only accumulate large quantities of lipid in abiotic stress, which reduces productivity by decreasing cell growth. In this study, the strategy was investigated to increase cell viability and lipid production by overexpressing S-adenosylmethionine (SAM) synthetase (SAMS) in the microalga Chlamydomonas reinhardtii. SAM is a substance that plays an important role in various intracellular biochemical reactions, such as cell proliferation and stress response, and the overexpression of SAMS could allow cells to ithstand the abiotic stress and increase productivity. Compared to wild-type C. reinhardtii, recombinant cells overexpressing SAMS grew 1.56-fold faster and produced 1.51-fold more lipids in a nitrogen-depleted medium. Furthermore, under saline-stress conditions, the survival rate and lipid accumulation were 1.56 and 2.04 times higher in the SAMS-overexpressing strain, respectively. These results suggest that the overexpression of SAMS in recombinant C. reinhardtii has high potential in the industrial-scale production of biofuels and various other high-value-added materials.

Proteomic and Phenotypic Analyses of a Putative YggS Family Pyridoxal Phosphate-Dependent Enzyme in Acidovorax citrulli

  • Lynn Heo;Yongmin Cho;Junhyeok Choi;Jeongwook Lee;Yoobin Han;Sang-Wook Han
    • The Plant Pathology Journal
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    • v.39 no.3
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    • pp.235-244
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    • 2023
  • Acidovorax citrulli (Ac) is a phytopathogenic bacterium that causes bacterial fruit blotch (BFB) in cucurbit crops, including watermelon. However, there are no effective methods to control this disease. YggS family pyridoxal phosphate-dependent enzyme acts as a coenzyme in all transamination reactions, but its function in Ac is poorly understood. Therefore, this study uses proteomic and phenotypic analyses to characterize the functions. The Ac strain lacking the YggS family pyridoxal phosphate-dependent enzyme, AcΔyppAc(EV), virulence was wholly eradicated in geminated seed inoculation and leaf infiltration. AcΔyppAc(EV) propagation was inhibited when exposed to L-homoserine but not pyridoxine. Wild-type and mutant growth were comparable in the liquid media but not in the solid media in the minimal condition. The comparative proteomic analysis revealed that YppAc is primarily involved in cell motility and wall/membrane/envelop biogenesis. In addition, AcΔyppAc(EV) reduced biofilm formation and twitching halo production, indicating that YppAc is involved in various cellular mechanisms and possesses pleiotropic effects. Therefore, this identified protein is a potential target for developing an efficient anti-virulence reagent to control BFB.

Unraveling the Role of Cytochrome P450 as a Key Regulator Lantipeptide Production in Streptomyces globisporus

  • Da-Ran Kim;Su In Lee;Youn-Sig Kwak
    • The Plant Pathology Journal
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    • v.39 no.6
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    • pp.566-574
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    • 2023
  • The aim of this study was to investigate the regulation of lantipeptide production in Streptomyces globisporus SP6C4, which produces the novel antifungal lantipeptides conprimycin and grisin, and to identify the role of cytochrome P450 (P450) in tis regulation. To investigate the regulation of lantipeptide production, we created gene deletion mutants, including ΔP450, ΔtsrD, ΔlanM, ΔP450ΔtsrD, and ΔP450ΔlanM. These mutants were characterized in terms of their morphology, sporulation, attachment, and antifungal activity against Fusarium oxysporum. The gene deletion mutants showed distinct characteristics compared to the wild-type strain. Among them, the ΔP450ΔlanM double mutant exhibited a recovery of antifungal activity against F. oxysporum, indicating that P450 plays a significant role in regulating lantipeptide production in S. globisporus SP6C4. Our findings highlight the significant role of P450 in the regulation of lantipeptide production and morphological processes in S. globisporus. The results suggest a potential link between P450-mediated metabolic pathways and the regulation of growth and secondary metabolism in SP6C4, thereby highlighting P450 as a putative target for the development of new antifungal agents.

Cold Shock Response and Low Temperature Stable Transcript of DEAD-box RNA Helicase in Bacillus subtilis (DEAD-box RNA Helicase 유전자가 결핍된 Bacillus subtilis의 저온 충격 반응성과 저온 안정성 전사물)

  • Oh, Eun-Ha;Lee, Sang-Soo
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.289-294
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    • 2011
  • We investigated the cold shock sensitivity of DEAD-box RNA helicase gene deleted strains of in Bacillus subtilis CU1065. To understand cold shock effects, cells were cultivated at $37^{\circ}C$ to log phase ($O.D_{600}$=0.5-0.6) and then temperature was shifted to $15^{\circ}C$. Cold shock slow down the growth rate of wild type and deleted strains of DEAD-box RNA helicase gene (ydbR, yfmL, yqfR, deaD). The growth rate of ydbR deleted strain is 5 times severely reduced compared to that of wild type strain (CU1065). But the growth rate of other three (yfmL, yqfR, deaD) deleted strains is nearly equal to the growth rate of wild type. Compared to $37^{\circ}C$, the amount of ydbR and yqfR mRNA transcripts are increased at the growth temperature of $15^{\circ}C$. On the other hands the mRNA transcripts of yfmL and deaD are not changed at both conditions of $37^{\circ}C$ and $15^{\circ}C$. Upon cold shock treatment ydbR mRNA transcript is clearly increased. After treatment of rifampicin (bacteria transcription inhibitor) the amount of ydbR mRNA was measured. Temperature shift from $37^{\circ}C$ to $15^{\circ}C$ and rifampicin treatment showed slowly decay of ydbR mRNA. But at $37^{\circ}C$ and rifampicin treatment ydbR mRNA is rapidly reduced. These results showed that cold shock induction of ydbR mRNA resulted from the stability of ydbR mRNA and not from the transcription induction of ydbR. In relation to these results, we found the cold box element of csp (cold shock protein gene) in 5' untranslated region of ydbR gene. Cold shock induction of ydbR is caused by the stability of ydbR mRNA like the stability of csp mRNA.

Effect of Manganese Sulfate Concentration in Media on Production Speed of Insecticidal Crystal by Bacillus thuringiensis (배지 중 Manganese sulfate 농도가 Bacillus thuringiensis의 곤충독소 생성 시간에 미치는 영향)

  • Ro-Un Lee;Do Gyung Oh;Eun-Sun Jeong;Jung-Beom Kim
    • Journal of Food Hygiene and Safety
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    • v.38 no.3
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    • pp.170-175
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    • 2023
  • In this study, the effect of MnSO4 on the insecticidal crystal (IC) produced by Bacillus thuringiensis for a rapid detection medium was analyzed. The strains used included one B. thuringiensis reference (KCTC 1511) and nine wild-type strains. The IC in B. thuringiensis was detected following the method published by the Ministry of Food and Drug Safety in Korea. In the nutrient agar to which 0.005% MnSO4 was added, IC was observed on two of the three plates after 48 hours of incubation and on all three plates after 120 hours. In AK agar, IC was observed on one and two of the three plates after 48 and 96 hours of incubation, respectively. These results indicated that 0.005% MnSO4 nutrient agar is more appropriate than AK agar for production of IC in B. thuringiensis. The effect of various MnSO4 concentrations on IC production was studied after 24 hours of incubation. IC was produced on 1 of the 10 plates with 0.000% MnSO4 nutrient agar, 2 of the 10 plates with 0.001% MnSO4 nutrient agar, and 3 of the 10 plates with 0.002% MnSO4 nutrient agar. IC was not observed for the other nutrient agars containing 0.003%-0.009% MnSO4. These results indicated that nutrient agar with 0.002% MnSO4 led to the most rapid production of IC by B. thuringiensis after 24 hours of incubation. However, the conditions for IC production by B. thuringiensis depended on the incubation conditions and strain activity. Therefore, further studies are needed to verify the effects of 0.002% MnSO4 on the production of IC by various Bacillus thuringiensis strains.

Mycelial Production and Amylase Activity of Fungi for Brewing in Different Submerged Culture Conditions (액체배양에서의 양조용 곰팡이의 균체 생산 및 전분분해효소 활성)

  • Noh, Jong-Min;Choi, Ji-Ho;Jung, Seok-Tae;Yeo, Soo-Hwan;Park, Jang-Woo;Lee, Jin-Won;Choi, Han-Seok
    • Journal of the East Asian Society of Dietary Life
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    • v.23 no.6
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    • pp.833-838
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    • 2013
  • In this study, twelve strains of brewing fungi were individually cultivated on wheat extract broth (WEB), potato dextrose broth (PDB) and malt extract broth (MEB) in order to determine the microorganism with good culture characteristics as well as with amylolytic activity. The strain cultured in PDB exhibited mycelia production from 12.6 g/L (Rhizopus oryzae KACC 45714) to a maximum of 48.0 g/L (Aspergillus oryzae KACC 46959), which was 2.3~9.2 times lower than that of the strain cultured in WEB and 1.7~14.6 times lower than that of the strain cultured in MEB. Accorfing the results, We found that the commercial strains of A. oryzae Suwon, CF1001 and CF1003 had a higher dry cell mass than the wild-type strains KACC 46421, 46423, 46424 and 46959. For Rhizopus sp., the acidity levels in WEB, PDB and MEB were 0.12~0.47%, 0.22~1.0% and 0.16~0.68% (equivalent lactic acid concentration) respectively. For A. oryzae, the acidity levels were 0.06~0.11%, 0.03~0.04% and 0.06~0.08% (equivalent lactic acid concentration), respectively. Amylase enzyme from Rhizopus delemar KACC 46419 exhibited an enzyme activity of 0.013 U and 0.019 U in WEB and MEB cultures, respectively. The enzyme activity of the amylase enzyme from A. oryzae was 0.019~0.037, 0.017~0.033 and 0.028~0.046 U in WEB, PDB and MEB cultures, respectively.

Studies on the safety of Brucella abortus RB51 vaccine I. Comparison of the biochemical and genetic characteristics of Brucella abortus RB51 vaccine strains (부루세라백신(RB51)의 안전성에 관한 연구 I. Brucella abortus RB51 백신균주의 생화학적 및 유전학적 성상비교)

  • Kim, Jong-man;Woo, Sung-ryong;Lee, Ji-youn;Jung, Suk-chan;Kang, Seung-won;Kim, Jong-yeom;Yoon, Yong-dhuk;Cho, Sang-nae;Yoo, Han-sang;Olsen, Steven C.
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.533-541
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    • 2000
  • Biochemical and genetic analysis were carried out to investigate the potential recovery of pathogenecity or related mutations of Brucella abortus RB51 vaccine strains. RB51 strains were recovered from commercial vaccines, including related seed stocks from private companies in Republic of Korea, strain from USA, a reference strain from C university and a field isolate (Daehungjin) from aborted dairy cow after RB51 vaccination were compared with two identified virulent wild strains (S2308 and a field strain isolated from dairy cow in Korea) at the same conditions. All the strains examined, except identified pathogenic strains, revealed the identical characteristics to the original RB51 in biochemical properties, antigen and bacteriophage typing. Outer membrane protein (OMP) profiles from strains of RB51 showed the same patterns with standard RB51 in SDS-PAGE. In addition, Western blotting with the brucella specific monoclonal antibody also indicated that all the vaccine strains were completely deficient in their LPS compared to the pathogenic Br abortus strains. The differences in DNA structures among strains were also possible to detect after PCR. All vaccine strains, except S19, S1119-3, S1075, S544 and Br suis, were amplified a 178bp DNA fragment of eri-gene, and 364bp of IS711 elements. In contrast, 498bp DNA product was only found with Br abortus. Overall evidences in the present study confirmed that the RB51 strains for vaccine production in Korea did not originated from the phenomena of possible recovery of pathogenicity or related to any potential mutation event at all.

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