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  • 1
    Electronic Resource
    Electronic Resource
    Detection of squash mosaic virus in seeds of melon (Cucumis melo) by enzyme-linked immunosorbent assay (ELISA) (1990)
    Springer
    European journal of plant pathology 96 (1990), S. 91-102 
    Details
    ISSN: 1573-8469
    Keywords: serology ; identification ; virus purification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Samenvatting Het pompoenemozaïekvirus gaat over met het zaad van verscheideneCucurbitaceae. Het gebruik van virusvrij zaad is belangrijk om te voorkomen dat het virus zijn intrede doet in Nederland en zich naar andere landen verspreidt. Een antiserum werd geproduceerd tegen een serotype 1 isolaat van meloen. Met behulp van dit antiserum werd een ELISA ontwikkeld om pompoenemozaïekvirus in zaden van meloen aan te tonen. Twee varianten van ELISA werden vergeleken, namelijk een variant waarbij monster en enzymconjugaat gelijktijdig geïncubeerd werden (ELISA 1) en een variant waarbij monster en enzymconjugaat na elkaar geïncubeerd werden (ELISA 2). De gevoeligheid van de ELISA varianten werd uitgetoetst door meel van zieke zaden in verschillende verhoudingen te mengen met meel van gezonde zaden. Het pompoenemozaïekvirus werd met beide ELISA varianten aangetoond in verdunningen van 1 ∶ 160 (1 deel meel van zieke zaden gemengd met 159 delen meel van gezonde zaden) of hoger na 4 uur incubatie met substraat. ELISA 1 gaf doorgaans hogere extinctiewaarden dan ELISA 2 voor bijna alle verdunningen. Omdat ELISA 1 ook nog sneller is dan ELISA 2, wordt ELISA 1 aanbevolen voor routinematig gebruik. Wanneer voor routinematig gebruik 100 meloenezaden per submonster getoetst worden, kan het pompoenemozaïek virus betrouwbaar worden aangetoond. In Nederland worden momenteel per zaadpartij 20 submonsters van 100 zaden getoetst.
    Notes: Abstract Squash mosaic virus (SqMV, comovirus) is seed-transmitted in severalCucurbitaceae. Therefore, the use of virus-free seed is important to prevent establishment of this virus in the Netherlands and to avoid spread to other countries. This study was undertaken to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of SqMV in melon seeds. An antiserum was produced to a serotype 1 isolate from melon. Two ELISA variants were investigated viz. an ELISA variant with simultaneous incubation of sample and enzyme conjugate (ELISA 1) and an ELISA variant with successive incubation of sample and enzyme conjugate (ELISA 2). The sensitivity of ELISA was tested by mixing fluor of ground infected and non-infected seeds in different proportions. SqMV was detected by both ELISA variants at dilutions of 1 ∶ 160 (1 part of infected flour mixed with 159 parts of non-infected flour) or higher after a substrate incubation period of 4 h. However, ELISA 1 gave relatively higher absorbance values than ELISA 2 for nearly all dilutions. Since ELISA 1 is also faster than ELISA 2, ELISA 1 is advised for routine testing. In these test, using subsamples of 100 melon seeds SqMV is detected reliably. ELISA 1 is now used in the Netherlands for routine-indexing of melon seed lots for SqMV.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02005133
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  • 2
    Electronic Resource
    Electronic Resource
    Characteristics of immunofluorescence microscopy and of dilution-plating to detect Pseudomonas syringae pv. phaseolicola in bean seed lots and for risk assessment of field incidence of halo blight (1991)
    Springer
    European journal of plant pathology 97 (1991), S. 233-244 
    Details
    ISSN: 1573-8469
    Keywords: Phaseolus vulgaris ; Phaseolus coccineus ; bush bean ; runner bean ; method evaluation ; grey test area ; detection level ; saprophytic interference ; predictive value ; diagnostic sensitivity ; diagnostic specificity ; analytical sensitivity ; analytical specificity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Routine laboratory testing of 710 bean seed lots from various origins forPseudomonas syringae pv. phaseolicola (Psp) with immunofluorescence microscopy (IF) showed that 27.5% of the seed lots (five subsamples of 1000 seeds tested per sample) contained two or more IF-positive cells in a total of 500 microscope fields (magnification 500×). Simultaneously performed dilution-platings of IF-positive subsamples on King's medium B confirmed presence of Psp for one-third of these IF-positive seed lots. The ‘grey area’ of disagreement between both laboratory tests was studied by comparison of test data and by field trials. The number of IF-positive cells per subsample was positively correlated with isolation and identification of Psp (R=0.85). The detection level of IF was ca. 102 Psp cells per ml of undiluted subsample extract. The detection level of Psp by isolation on King's medium B was variable, being inversely related with the saprophyte to Psp ratio. The high sensitivity of IF was in part due to high percentages of dormant or dead IF-positive cells in the sample extract. Field trials over two years with 10 000 seeds per seed lot, showed disease incidence for 9 of the 22 seed lots. Of ten IF-positive lots with five positive subsamples per sample, nine were positive in the field test plot (the negative lot gave primary infection spots of Psp when used for commercial growing). By isolation, seven of these ten IF-positive lots were positive. Of the five IF-positive lots with two or less positive subsamples, isolation and field trial were both negative. Based on data on seed transmission from literature, field incidence was unlikely for these five samples in a 10 000 seeds field trial. All seven IF-negative lots were negative in the field trial. The value of IF and isolation for indexing bean seed lots for Psp is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01989820
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  • 3
    Electronic Resource
    Electronic Resource
    Specificity of polycllonal and monoclonal antibodies for the identification of Xanthomonas campestris pv. campestris (1992)
    Springer
    European journal of plant pathology 98 (1992), S. 81-94 
    Details
    ISSN: 1573-8469
    Keywords: black rot ; serology ; sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) ; proteinase K
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Polyclonal and monoclonal antibodies (PCAs and MCAs), produced to whole cells and flagellar extracts ofXanthomonas campestris pv.campestris (Xcc), respectively, were tested for specificity. In immunofluorescence microscopy (IF) the three PCAs tested, reacted at low dilutions with all Xcc strains, some other xanthomonads and non-xanthomonads. At higher dilutions most cross-reactivity with non-xanthomonad strains disappeared. However, the cross-reactivity with strains ofX. c. pv.vesicatoria (Xcv),X. c. pv.amoraciae (Xca) andX. c. pv.phaseoli var. fuscans (Xcpf) remained. Six MCA-producing cell clones viz. 20H6, 2F4, 18G12, 10C5, 17C12 and 16B5 were selected for specificity tests with an enzyme immunoassay (EIA), IF and a dot-blot immunoassay (DBI). None of the MCAs reacted with all Xcc strains in IF and EIA. In DBI, only MCAs 17C12 and 16B5 reacted with all Xcc strains. All six MCAs tested, cross-reacted in one of either tests with other pathovars ofX. campestris, such as Xcv or Xca. The MCAs were also tested in immunoblotting experiments using total bacterial extracts, cell envelope and flagellar extracts. MCAs 20H6, 2F4, 18G12 and 10C5 reacted with the lipopolysaccharide (LPS) of Xcc. MCAs 16B5 and 17C12 reacted with a 39 kilodalton and a 29 kilodalton protein, respectively. It is concluded that the PCAs and MCAs discussed in this study may be used for routine identification and differentiation of (a group of) Xcc strains. The significance of the cross-reactions with other pathovars ofX. campestris needs to be determined by testing seed lots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01996321
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  • 4
    Electronic Resource
    Electronic Resource
    Application of polyclonal and monoclonal antibodies for the detection of Xanthomonas campestris pv. campestris in crucifer seeds using immunofluorescence microscopy (1992)
    Springer
    European journal of plant pathology 98 (1992), S. 95-106 
    Details
    ISSN: 1573-8469
    Keywords: pathogenicity testing ; virulence ; dilution-plating ; YDC-agar ; NSCA ; NSCAA ; nutrient starch medium ; antibiotics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Polyclonal and monoclonal antibodies (PCAs and MCAs) were tested for the detection ofXanthomonas campestris pv.campestris (Xcc) in cabbage seeds using immunofluorescence microscopy (IF). It was concluded that PCA 94, MCAs 20H6, 2F4, 18G12 and a mixture of MCAs 20H6, 18G12, 2F4 and 16B5 could be used to detect Xcc in seed extracts when 5 min and 2.5 h shaking of seeds are used as extraction methods. The reliability of confirming suspect colonies with MCAs and PCA 94 in IF depended in part on the seed lot tested and the antibody used. Some virulent Xcc strains derived from seed lots, did not react with MCAs 10C5, 2F4, 18G12, 17C12 and 16B5. On the other hand, saprophytic isolates obtained from one seed lot cross-reacted with MCA 17C12 and to a lesser extent with MCAs 2F4, 18G12 and PCA 94. No relationship was found between IF-reactions of Xcc strains using MCAs and reactions of Xcc strains in pathogenicity testing. Xcc andX. c. pv.amoraciae (Xca) could in general not be distinguished on the basis of reactions with MCAs and PCAs. Also in pathogenicity tests Xcc and Xca were hard to distinguish.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01996322
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  • 5
    Electronic Resource
    Electronic Resource
    Comparison of immunofluorescence microscopy and dilution-plating for the detection of Xanthomonas campestris pv. campestris in crucifer seeds (1992)
    Springer
    European journal of plant pathology 98 (1992), S. 169-178 
    Details
    ISSN: 1573-8469
    Keywords: correlation ; test evaluation ; extraction method ; media ; serology ; seed-borne bacteria ; Pseudomonas syringae pv.phaseolicola
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The correlation between immunofluorescence microscopy (IF) and dilution-plating on nutrient starch cycloheximide agar (NSCA) or NSCA with the addition of nitrofurantoin and vancomycin (NSCAA) was studied for the detection ofXanthomonas campestris pv.campestris (Xcc) in crucifer seeds. When checking 50 μl of the seed extract in IF, IF and dilution-plating gave corresponding results (both positive or negative) for 45.4–56.4% of the samples tested. No differences were observed in this respect between tests using a polyclonal antiserum (PCA 94) and replicate tests using monoclonal antibodies (MCA 20H6). When 20 μl of the seed extract was checked in IF, 67.3–71.3% of the samples tested were both positive or negative with dilutionplating and IF. IF negative and dilution-plating positive samples were found for 0.0–7.3% of all samples tested. The percentage of IF positive and dilution-plating negative samples ranged from 26.7–29.2 (20 μl seed extract checked) to 41.8–47.3% (50 μl seed extract checked). Generally, the probability of isolating Xcc increased with increasing numbers of fluorescent cells found in IF. Above 10 000 cells per ml the probability of isolating Xcc ranged from 57.1–81.8%. Increasing the extraction time from 5 min to 2.5 h shaking showed no significant increase of the number of samples found positive in IF and dilution-plating. However, when using both 5 min and 2.5 h shaking as compared to 5 min shaking only, more samples can be found positive in IF (1.0–14.5%) and dilution-plating (3.0–18.5%). Examining 1 μl instead of 50 μl of the sample smear, would increase the correspondence between IF and dilution-plating results up to minimally 69.1% (MCA 20H6). However, the risk of false-negative results in IF as compared to dilution-plating would also increase.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01974380
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  • 6
    Electronic Resource
    Electronic Resource
    Colours of florets of several gerbera (Gerbera jamesonii Bolis ex Adlam) cultivars measured with a colorimeter (1995)
    Springer
    Euphytica 84 (1995), S. 49-55 
    Details
    ISSN: 1573-5060
    Keywords: colorimeter ; colour variation ; flower colour ; Gerbera jamesonii ; seasonal effects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The colour variation between several gerbera cultivars were analyzed with a tristimulus colorimeter. A pilot study with a few cultivars showed that the flower colour variation between cultivars and colour effects during the growing season can be calculated quantitatively on the basis of data measured by the colorimeter. On the basis of these results the colour differences between a larger group of gerbera cultivars were measured. A consistent number of cultivars was distinct on the basis of colorimetric data and visual colour assessments.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01677556
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  • 7
    Electronic Resource
    Electronic Resource
    Detection of Clavibacter michiganensis ssp. michiganensis in tomato seeds by immunofluorescence microscopy and dilution plating (1993)
    Springer
    European journal of plant pathology 99 (1993), S. 125-137 
    Details
    ISSN: 1573-8469
    Keywords: bacterium ; detection ; identification ; media ; pathogenicity-testing ; serology ; seed-borne
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A method for detectingClavibacter michiganensis ssp.michiganensis in tomato seeds was evaluated. The method is based on rapid screening of tomato seed lots using indirect immunofluorescence staining (IF), followed by dilution plating of IF positive seed lots. Different polyclonal antisera, prepared againstC. michiganensis ssp.michiganensis were tested for their specificity using IF. All strains ofC. michiganensis ssp.michiganensis tested reacted with the polyclonal antisera. Two of nine saprophytic isolates from tomato seeds were positive with the antisera as well as with the control normal serum, but cells of these isolates were distinct in shape from cells ofC. michiganensis ssp.michiganensis. For extraction of the pathogen from the seed, seeds were either blended with a stomacher or soaked at 4–6 °C. The stomacher method yielded more fluorescent cells in IF than 24 h soaking of seed samples. However, soaking of seeds for 48 h generally yielded less saprophytes and overall higher numbers ofC. michiganensis ssp.michiganensis colonies in dilution plating when compared to blending by a stomacher. SCM medium was generally more selective than KBT and modified CNS medium. However, the efficacy of the medium was dependent on the seed lot and/or extraction method used. Confirmation of suspected colonies with YDC (yeast-dextrose-carbonate medium), IF and a pathogenicity test on tomato seedlings proved to be highly reliable (P〉0.95). For routine testing of seed lots it is recommended to screen tomato seed lost after soaking seeds for 24 h at 4–6 °C with IF, followed by plating of IF-positive seed lots on modified CNS and SCM after soaking seeds for an additional 24 h.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01974265
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  • 8
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    The development of a conductimetric assay for automated detection of metabolically active soft rot Erwinia spp. in potato tuber peel extracts (1996)
    Wiley
    Details
    Publication Date: 1996-10-01
    Print ISSN: 0021-8847
    Electronic ISSN: 2056-5232
    Topics: Biology
    Published by Wiley
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