• Title/Summary/Keyword: spore germination

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Purification and Characterization of a Major Extracellular Chitinase from a Biocontrol Bacterium, Paenibacillus elgii HOA73

  • Kim, Yong Hwan;Park, Seur Kee;Hur, Jin Young;Kim, Young Cheol
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.318-328
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    • 2017
  • Chitinase-producing Paenibacillus elgii strain HOA73 has been used to control plant diseases. However, the antimicrobial activity of its extracellular chitinase has not been fully elucidated. The major extracellular chitinase gene (PeChi68) from strain HOA73 was cloned and expressed in Escherichia coli in this study. This gene had an open reading frame of 2,028 bp, encoding a protein of 675 amino acid residues containing a secretion signal peptide, a chitin-binding domain, two fibronectin type III domains, and a catalytic hydrolase domain. The chitinase (PeChi68) purified from recombinant E. coli exhibited a molecular mass of approximately 68 kDa on SDS-PAGE. Biochemical analysis indicated that optimum temperature for the actitvity of purified chitinase was $50^{\circ}C$. However, it was inactivated with time when it was incubated at $40^{\circ}C$ and $50^{\circ}C$. Its optimum activity was found at pH 7, although its activity was stable when incubated between pH 3 and pH 11. Heavy metals inhibited this chitinase. This purified chitinase completely inhibited spore germination of two Cladosporium isolates and partially inhibited germination of Botrytis cinerea spores. However, it had no effect on the spores of a Colletotricum isolate. These results indicate that the extracellular chitinase produced by P. elgii HOA73 might have function in limiting spore germination of certain fungal pathogens.

Postharvest Disease Control of Colletotrichum gloeosporioides and Penicillium expansum on Stored Apples by Gamma Irradiation Combined with Fumigation

  • Cheon, Wonsu;Kim, Young Soo;Balaraju, Kotnala;Kim, Bong-Su;Lee, Byeong-Ho;Jeon, Yongho
    • The Plant Pathology Journal
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    • v.32 no.5
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    • pp.460-468
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    • 2016
  • To study the control of postharvest decay caused by Colletotrichum gloeosporioides and Penicillium expansum, gamma irradiation alone or in combination with fumigation was evaluated to extend the shelf life of apples in South Korea. An irradiation dose of 2.0 kGy resulted in the maximum inhibition of C. gloeosporioides and P. expansum spore germination. The gamma irradiation dose required to reduce the spore germination by 90% was 0.22 and 0.35 kGy for C. gloeosporioides and P. expansum, respectively. Microscopic observations revealed that when the fungal spores were treated with gamma irradiation (4.0 kGy), conidial germination was stopped completely resulting in no germ tube formation in C. gloeosporioides. Treatment with the eco-friendly fumigant ethanedinitrile had a greater antifungal activity against C. gloeosporioides and P. expansum in comparison with the non-treated control under in vitro conditions. The in vitro antifungal effects of the gamma irradiation and fumigation treatments allowed us to further study the effects of the combined treatments to control postharvest decay on stored apples. Interestingly, when apples were treated with gamma irradiation in combined with fumigation, disease inhibition increased more at lower (< 0.4 kGy) than at higher doses of irradiation, suggesting that combined treatments reduced the necessary irradiation dose in phytosanitary irradiation processing under storage conditions.

Inactivation of Bacterial Spores by High Pressure and Food Additive Combination (초고압과 식품첨가물 병용을 이용한 세균 포자의 살균)

  • Chung, Yoon-Kyung
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1094-1099
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    • 2011
  • Antimicrobial efficacy of high pressure (HP) can be enhanced by the application of additional hurdles. The objective of this study was to assess the enhancement in pressure lethality by tert-butylhydroquinone (TBHQ) treatment, against bacterial spores that are considered significant in the food industry. Spores of Clostridium sporogenes, Bacillus cereus and B. subtilis were prepared. Spore suspensions containing TBHQ (200 ppm, dissolved in dimethyl sulfoxide, DMSO) were pressurized at 650 or 700 MPa at 54-72$^{\circ}C$ for 5 min. Inactivation of bacterial spores resulted only with HP treatment. The population of B. subtilis spores was more inactivated by HP than those of B. cereus and C. sporogenes spores. Inactivation of C. sporogenes spores using pressure was more affected by the germinated population, compared to Bacillus spores. The inactivation of Bacillus spores increased when pressurized at 70$^{\circ}C$, compared to 54$^{\circ}C$. On the other hand, the degree of germination-induced lethality for Bacillus spores decreased at 70$^{\circ}C$. When spores were treated with a combination of DMSO-HP and TBHQ-HP, these treatments seemed to protect the spores against HP, especially at 54$^{\circ}C$. Further mechanistic studies involved in inducing germination by HP and using a subsequent sporicidal agent will be needed for a better understanding of bacterial spore inactivation.

Spore Associated Bacteria (SAB) of Arbuscular Mycorrhizal Fungi (AMF) and Plant Growth Promoting Rhizobacteria (PGPR) Increase Nutrient Uptake and Plant Growth Under Stress Conditions

  • Gopal, Selvakumar;Chandrasekaran, Murugesan;Shagol, Charlotte;Kim, Ki-Yoon;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.4
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    • pp.582-592
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    • 2012
  • Microorganisms present in the rhizosphere soil plays a vital role in improving the plant growth and soil fertility. Many kinds of fertilizers including chemical and organic has been approached to improve the productivity. Though some of them showed significant improvement in yield, they failed to maintain the soil properties. Rather they negatively affected soil eventually, the land became unsuitable for agricultural. To overcome these problems, microorganisms have been used as effective alternative. For past few decades, plant growth promoting rhizobacteria (PGPR) and arbuscular mycorrhizal fungi (AMF) have been used as effective inoculants to enhance the plant growth and productivity. PGPR improves the plant growth and helps the plant to withstand biotic and abiotic stresses. AM fungi are known to colonize roots of plants and they increase the plant nutrient uptake. Spore associated bacteria (SAB) are attached to spore wall or hyphae and known to increase the AMF germination and root colonization but their mechanism of interaction is poorly known. Better understanding the interactions among AMF, SAB and PGPR are necessary to enhance the quality of inoculants as a biofertilizers. In this paper, current knowledge about the interactions between fungi and bacteria are reviewed and discussed about AMF spore associated bacteria.

Inhibitory Effect of Flusilazole on the Spore Formation of Aspergillus niger Causing the Onion Black Mold in Vapour Phase (Flusilazole의 훈증 효과에 의한 양파검은곰팡이병균(Aspergillus niger)의 포자 형성 억제)

  • Kim, Heung-Tae;Park, Se-Won;Choi, Gyung-Ja;Kim, Jin-Cheol;Cho, Kwang-Yun
    • Research in Plant Disease
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    • v.8 no.2
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    • pp.124-130
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    • 2002
  • In 1998, a pathogen isolated from infected onions was identified as Aspergillus niger. At 3$0^{\circ}C$, A. niger AnYD-1 showed the best mycelial growth, spore germination, and high pathogenicity to onions. In spite of the weak inhibitory effect of flusilazole and hexaconazole on the mycelial growth on PDA, they showed the specific inhibitory activity against the formation of spores in the vapour phase. With flusilazole and hexaconazole, the effects of the solvent, the applied concentration and the incubating temperature on the activities inhibiting the spore formation were confirmed. Their inhibitory effect on the spore formation in vapour phase was excellent by solving them with dimethylsulfoxide and dimethylformamide among tested solvents, and applying them at high temperature such as 30~35$^{\circ}C$.

Bacterial Sporulation and germination of Biocontrol agent Bacilus subtilis YBL-7 (항진균성 길항세균 Bacillus subtilis YBL-7의 종자피막용 포자체의 생산과 발아조건)

  • 장종원;김상달
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.236-242
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    • 1995
  • Biological control of soilborne plant pathogens by the addition of antagonistic microorganisms to the soil may offer a practical supplement or alternative to existing disease management strategies that depend heavily on chemical pesticides. Soil amendment with antagonistic microbes was non-effective because of high cost, low efficacy, and inconvenient usage on the treatment course. Therefore, seed coating formulation for the application of biological seed treatments has been being to apply successful disease suppression for many important crops. The objectives of this study were to investigate the optimal condition for the spore production of biocontrol agent Bacillus subtilis YBL-7 and the liquid coating formulation that contained a suspension of a proper aqueous binder, as well as a ground fine solid particulate material. The maximum yield has been obtained from 60 hrs-old culture at 30$\circ$C in spore forming (SF) medium containing 0.8% nutrient broth, 0.05% yeast extract, 10$^{-1}$ M MgCl$^{2}$, 10$^{-4}$ M MnCl$^{2}$, 10$^{-5}$ M dipicolinic acid, and pH 6.5. The optimal condition of dried spore preparation was achieved when cells of B. subtilis YBL-7 was heat-dried with 50$\circ$C for 2 hrs.

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Antifouling effects of the periostracum on algal spore settlement in the mussel Mytilus edulis

  • Kang, Ji-Young;Bangoura, Issa;Cho, Ji-Young;Joo, Jin;Choi, Yoo Seong;Hwang, Dong Soo;Hong, Yong-Ki
    • Fisheries and Aquatic Sciences
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    • v.19 no.1
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    • pp.7.1-7.6
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    • 2016
  • In nature, marine mussels (Mytilus edulis) suffer less fouling colonization on the newly formed sides of their shells. Using settlement assays with algal spores of Porphyra suborbiculata, we determined that spore attachment and germination on the periostracum decreased to 36.8 and 3.3 %, respectively. Additionally, the spore settlement was considerably diminished by periostracum dichloromethane extracts containing 19 % oleamide, a major antifouling compound. A scanning electron micrograph of the surface revealed a regular ripple structure with approximately $1.4{\mu}m$ between ripples. Based on these results, mussel periostraca or their associated biomimetic materials may become environmentally friendly, antifouling agents for preventing the settlement of soft foulants.

Effects of Several Fungicides on the Spore Growth Period of Alternaria dauci, a Carrot Black Leaf Blight Fungus, Using a Rezasulin-based Spore Survival Assay (Rezasulin 기반 포자 생존 검정법을 이용한 당근검은잎마름병균 Alternaria dauci의 포자 생장 시기에 따른 몇 가지 살균제의 효과)

  • Jiwon Do;Heung Tae Kim
    • Research in Plant Disease
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    • v.30 no.2
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    • pp.131-138
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    • 2024
  • The effects of five fungicides on the spore growth phase of Alternaria dauci, which causes carrot leaf blight, were tested using the spore viability assay (SVA) and agar dilution method (ADM). The average EC50 values for chlorothalonil against seven isolates of A. dauci examined by SVA and ADM were 14.21 ㎍/ml and more than 100 ㎍/ml. Dithianon and folpet also had lower EC50 values in SVA than in ADM, while iminoctadine trisalbesilate had lower EC50 value in ADM. For fluazinam, the EC50 values of SVA and ADM were 1.63 and 2.40 ㎍/ml, respectively. As EC50 values of five fungicides according to the spore growth phase of A. dauci KACC 42997, the efficacy of each fungicide as chlorothalonil, dithianon, and folpet decreased when treated after spore germination rather than when treated with spores before germination. However, iminoctadine tris-albesilate was more effective when treated after spores germinated than when treated before treatment. The excellent effect of fluazinam on the pathogen was maintained until A. dauci KACC 42997 was cultured in potato dextrose broth for 6 hr and the germ tube grew beyond the size of the spore. However, when treated with iminoctadine tris-albesilate and fluazinam after culturing for 12 hr, as the EC50 values of the two fungicides increased to 8.87 and 20.65 ㎍/ml, their efficacies decreased. The results of this study show that the treatment time of the fungicide should be determined by considering the effect of the fungicide on the spore growth phase of pathogens.

Physiological Responses of Bacillus amyloliquefaciens Spores to High Pressure

  • Ahn, Ju-Hee;Balasubramaniam, V.M.
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.524-529
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    • 2007
  • Pressure inactivation behavior of Bacillus amyloliquefaciens spores was investigated in deionized water. The spores of B. amyloliquefaciens were subjected to $105^{\circ}C$ and 700 MPa. The magnitude of the decrease in viability after pressure treatment was similar to that after pressure treatment followed by heat shock. The increase of dipicolinic acid (DPA) release was correlated with the spore inactivation, and the hydrophobicity did not significantly change during the pressure-assisted thermal processing (PATP). Lag phase duration increased with increasing pressure process time. The mechanisms of spore germination and inactivation during the PATP were related to a complex physiological process.

Studies on the Long-Term Preservable Meat Products Based on the Water Activity Control (수분활성도 조절에 의한 육제품의 장기저장에 관한 연구)

  • 신현길;김행하;이원창
    • Journal of Food Hygiene and Safety
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    • v.1 no.2
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    • pp.115-120
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    • 1986
  • This experiment was carried out to develop long-term preservable meat products by controlling a water activity which has a potential inhibitory effects on the growth of microorganism and germination of spore. For this purpose the accuracy of instrument for the measurement of water activity, electric hygrometer was used for this study as well as water activity of final products were investigated according to the various amounts of different kinds of additives. Spore of Bacillus spp, Clostridium sporogenes and Cl. botulinum were inoculated to the meat products with different aw and their growth states in $25^{\circ}C$ were observed to find out the lowest aw which inhibit their growth (aw 0.950).

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