• Title/Summary/Keyword: Environmental spore

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Monoclonal Antibody-Based Indirect-ELISA for Early Detection and Diagnosis of Epiphytic Didymella bryoniae in Cucurbits

  • Lee, Sun-Cheol;Han, Ki-Soo;Lee, Jung-Han;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.19 no.5
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    • pp.260-265
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    • 2003
  • Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.

Environmental Factors Favoring the Disease Development and Chemical Control of Clubroot(Plasmodiophora brassicae) in Chinese Cabbage (배추 무사마귀병(Plasmodiophora brassicae)의 발병유인 및 약제방제)

  • Oh, Jeung-Haing;Cho, Jang-Hwan;Kim, Bong-Gu;Chae, Je-Chun;Chung, Gil-Ung;Hwang, Chul-Ho;Kim, Doo-Wook
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.244-247
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    • 1997
  • The experiment was conducted to obtain a basic information on the enviroumental factors favoring the disease development and chemical control of clubroot in chinese cabbage. The inoculation by insertion of infested soil was the most effective for the disease development as compared to the other inoculation methods such as pouring the spore suspension into soil and dipping roots into the spore suspension. On the environmental factors favoring the clubroot development, optimum slil pH and soil temperature were pH 5-6 and $20{\sim}30^{\circ}C$, respectvely. The combination of soil treatment and seed coating treatment was more effective than each single treatment in the chemical control of clubroot by 0.5% of fluazinam dust.

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Diversity of Bacillus thuringiensis Strains Isolated from Citrus Orchards in Spain and Evaluation of Their Insecticidal Activity Against Ceratitis capitata

  • J.C., Vidal-Quist;Castanera, P.;Gonzalez-Cabrera, J.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.749-759
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    • 2009
  • A survey of Bacillus thuringiensis (Berliner) strains isolated from Spanish citrus orchards has been performed, and the strains were tested for insecticidal activity against the Mediterranean fruit fly Ceratitis capitata (Wiedemann), a key citrus pest in Spain. From a total of 150 environmental samples, 376 isolates were selected, recording a total B. thuringiensis index of 0.52. The collection was characterized by means of phase-contrast microscopy, SDS-PAGE, and PCR analysis with primer pairs detecting toxin genes cry1, cry2, cry3, cry4, cry5, cry7, cry8, cry9, cry10, cry11, cry12, cry14, cry17, cry19, cry21, cry27, cry39, cry44, cyt1, and cyt2. Diverse crystal inclusion morphologies were identified: bipyramidal (45%), round (40%), adhered to the spore (7%), small (5%), and irregular (3%). SDS-PAGE of spore-crystal preparations revealed 39 different electrophoresis patterns. All primer pairs used in PCR tests gave positive amplifications in strains of our collection, except for primers for detection of cry3, cry19, cry39, or cry44 genes. Strains containing cry1, cry2, cry4, and cry27 genes were the most abundant (48.7%, 46%, 11.2%, and 8.2% of the strains, respectively). Ten different genetic profiles were found, although a total of 109 strains did not amplify with the set of primers used. Screening for toxicity against C. capitata adults was performed using both spore-crystal and soluble fractions. Mortality levels were less than 30%. We have developed a large and diverse B. thuringiensis strain collection with huge potential to control several agricultural pests; however, further research is needed to find out Bt strains active against C. capitata.

Secondary Contamination is the Main Source for Spread of Nosema bombycis Resulting in Outbreak of Pebrine Disease in Bombyx mori L.

  • Chakrabarty, Satadal;Saha, A.K.;Manna, B.;Kumar, S. Nirmal
    • International Journal of Industrial Entomology and Biomaterials
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    • v.27 no.2
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    • pp.282-288
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    • 2013
  • In nature, the population of Nosema bombycis (Microsporidia) causing pebrine disease is small and their development is extremely slow and only few ultimately producing spores. Pebrine infected silkworm, Bombyx mori larvae collected from sericulture field were alive till $3^{rd}$ generation though the concentration of N.bombycis spore was very high ($2.4-3.0{\times}10^8$ spores. $mL^{-1}$). All larvae were died during $4^{th}$ generation with extremely high concentration of pebrine spores ($3.0-4.0{\times}10^9$ spores. $mL^{-1}$) and mostly contain long polar tube (LT). Alternately, all larvae were died immediately (at $3^{rd}$ stage of $1^{st}$ generation) when it was artificially inoculated with same concentration of N.bombycis spores harvested from field ($2.4-3.0{\times}10^8$ spores. $mL^{-1}$) though concentration of spores harvest was very less ($3.0-4.0{\times}10^6$ spores. $mL^{-1}$) and mostly contain short polar tube (ST). Artificially pebrine infected male moth when mated with healthy female moth took six generations to develop pebrine disease and all larvae were died at the $2^{nd}$ stage with very less spore harvest ($3.0-10.0{\times}10^6$ spores. $mL^{-1}$). Survival percentage was increased in all generations (~92.0% at $4^{th}$ generation) when silkworm rearing was conducted under new integrated disease management system.

Proteomic Reference Map and Comparative Analysis between Streptomyces griseus S4-7 and wbiE2 Transcription Factor-Mutant Strain

  • Kim, Jisu;Kwon, Young Sang;Bae, Dong-Won;Kwak, Youn-Sig
    • The Plant Pathology Journal
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    • v.36 no.2
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    • pp.185-191
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    • 2020
  • Streptomyces griseus S4-7, a well-characterized keystone taxon among strawberry microbial communities, shows exceptional disease-preventing ability. The whole-genome sequence, functional genes, and bioactive secondary metabolites of the strain have been described in previous studies. However, proteomics studies of not only the S4-7 strain, but also the Streptomyces genus as a whole, remain limited to date. Therefore, in the present study, we created a proteomics reference map for S. griseus S4-7. Additionally, analysis of differentially expressed proteins was performed against a wblE2 mutant, which was deficient in spore chain development and did not express an antifungal activity-regulatory transcription factor. We believe that our data provide a foundation for further in-depth studies of functional keystone taxa of the phytobiome and elucidation of the mechanisms underlying plant-microbe interactions, especially those involving the Streptomyces genus.

Particulate Matter and Spores of Fungi Imperfecti in the Ambient Air of Seosan in Spring (서산지역 춘계의 대기 분진과 불완전균류 포자)

  • 여환구;김종호
    • Korean Journal of Environmental Biology
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    • v.19 no.2
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    • pp.153-157
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    • 2001
  • The relationship between SPM (Suspended Particulate Matter) and fungal spore in the air was investigated in Seosan, a rural county of Korea, in spring of 2000. SPM concentrations in the air were $199.8\mu{g}\;m^{-3}$, in the 1st Yellow Sand Period (March, 23-24), $249.4\mu{g}\;m^{-3}$ in the 2nd Yellow Sand Period (April, 7-9) and $98.9\mu{g}\;m^{-3}$ in the Non Yellow Sand Period (May, 12-16), respectively. Although there was somewhat difference in total SPM concentration between the two Yellow Sand Periods, majority of the total SPM were composed of $5\mu{m}$ sized coarse particles over the two periods. However, fine particles sized about $1\mu{m}$and coarse particles sized about $5-6\mu{m}$ ultimately showed peaks, which was within typical bimodal pattern at the graph of SPM size distribution in the Non Yellow Sand Period. Four mold genera grown from airborne fungal spores were finally identified in full-grown colonies at the SPM samples during the Yellow Sand Periods. These genera were Fusarium, Aspergillus, Penicillium and Basipetospora.

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A Study About Analysis Results for Kudoa septempunctata (Myxosporea: Multivalvulida) in Tissue at Olive Flounder, using PCR (polymerase chain reaction) and Histological Methods (PCR (polymerase chain reaction)법과 조직학적 방법을 통한 넙치 조직에서의 Kudoa septempunctata (Myxosporea: Multivalvulida)의 분석에 관한 연구)

  • Do, Jeong Wan;Cho, Miyoung;Jung, Sung Hee;Lee, Nam-Sil
    • Korean Journal of Environmental Biology
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    • v.35 no.4
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    • pp.468-475
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    • 2017
  • This study is for the consideration of the existence tendency of Kudoa septempunctata in olive flounder. In general, muscle has shown a strong PCR positive reaction in spores containing tissues rather than non-containing tissues. However, blood PCR results showed opposed tendency. In various organs of the tested fish containing spores in muscle tissue, heart had shown positive reaction along with muscle at PCR analysis. Muscle fiber necrosis was observed at the histological observation, and this degeneration was common in both samples. The one sample was the PCR positive muscle containing spore and the other was the PCR positive muscle non-containing spore. Both of muscle tissues indicated a positive reaction at ISH (in-situ hybridization) against K. septempunctata.

Modelling Algae Transport in Coastal Areas with Marine Afforestation (바다숲 조성해역의 해조류 포자 확산모델링)

  • Cho, Jae-Kweon;Lim, Young-Soo;Hong, Do-Ung;Kim, Jong-Kyu
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.15 no.1
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    • pp.1-8
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    • 2012
  • To arrange effectively artificial reefs for marine afforestation, tidal currents were analyzed by numerical experiments, and particle tracking based upon tidal currents were carried out to clarify the path of algae spore. The experiments were carried out by EFDC (Environmental Fluid Dynamics Code), and water column was vertically divided 10 layers. Tidal current patterns showed to be affected by main current at outside of study area, and circle currents of two were observed from analysis of residual currents. Particle tracking were experimented for 15 days at 2 installation places in which artificial reefs for marine afforestation would be deployed. According to the results of particle tracking experiment, particle movement at St.1 showed belt type along coastal line, and St.2 showed ellipse type at 300~500 m distant from coastal line. It suggest that artificial reefs for marine afforestation should be installed belt zone at station of St.1 and ellipse zone at St.2. Modelling algae transport was also tested to account for local dispersion of algae spore due to the suspended materials.

Culture Method of Spore for Entomopathogenic Fungus Using Natural Zeolite Ceramic Ball (천연제오라이트 세라믹볼을 이용한 곤충병원성 곰팡이 포자 생산 방법)

  • Lee, Jung-Bok;Kim, Beaum-Soo;Joo, Woo-Hong;Kwon, Gi-Seok
    • Korean Journal of Environmental Agriculture
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    • v.35 no.1
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    • pp.72-78
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    • 2016
  • BACKGROUND: Entomopathogenic fungi have been studied to develop for biological control agents as an alternative to chemical control agents in insect pest management. This investigated to determine the optimal culture conditions in ceramic balls for maximal sporulation of entomopathogenic fungi Beauveria bassiana M130 by use rice bran extract.METHODS AND RESULTS: METHODS AND RESULTS: A culture of entomopathogenic fungi for 12day on rice bran extract(1:8, w/v) incubated in ceramic matrix at 28℃. Natural zeolite ceramic ball was high production of 4.2×108 conidial/mL. The culture condition optimized initial pH, temperature, rice bran extract concentration, adhesives substance and concentration of NaCl, respectively. The high production of spore optimal conditions were temperature 28℃, initial pH 3, rice bran extract 3 mL, starch 33 g, 5 % NaCl and sopre suspension 7 mL, respectively.CONCLUSION: This study was carried out for the mass production of entomopathogenic fungi conidia recover rate 65% in matrix of natural zeolite ceramic ball, and to develop ingredient-used formulation of Beauveria bassiana M130 conidia for biological control agents.

A Biostimulant Preparation of Brown Seaweed Ascophyllum nodosum Suppresses Powdery Mildew of Strawberry

  • Bajpai, Sruti;Shukla, Pushp Sheel;Asiedu, Samuel;Pruski, Kris;Prithiviraj, Balakrishnan
    • The Plant Pathology Journal
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    • v.35 no.5
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    • pp.406-416
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    • 2019
  • Strawberry, an important fruit crop, is susceptible to a large number of pathogens that reduce fruit quality and productivity. In this study, the effect of a biostimulant prepared from Ascophyllum nodosum extract (ANE) (0.1%, 0.2%, and 0.3%) was evaluated on powdery mildew progression under greenhouse and field conditions. In the greenhouse, application of 0.2% ANE showed maximum reduction in powdery mildew progression as compared to the control. Forty-eight hour post-inoculation, foliar spray of 0.2% ANE reduced spore germination by 75%. Strawberry leaves sprayed with ANE showed higher total phenolic and flavonoid content in response to powdery mildew infection. Furthermore, application of ANE elicited defense response in strawberry plants by induction of defense-related enzymes, such as phenylalanine ammonia lyase, polyphenol oxidase, and peroxidase activity. In field conditions, foliar spray of 0.2% ANE showed a reduction of 37.2% of natural incidence of powdery mildew infection as compared to the control. ANE sprayed plant also reduces the severity of powdery mildew infection under natural conditions. These results indicate that application of ANE induces the strawberry plant's active defense against powdery mildew infection by induction of secondary metabolism and regulating the activities of defense-related enzymes.