Improved immunological detection of S pongospora subterranea
Article Accepted: 07 November 2004 Abstract
Spongospora has two members which are important pathogens of vegetables, S. subterranea f.sp. subterranea ( Sss) and S. subterranea f.sp. nasturtii ( Ssn). The close taxonomic relationship of these formae speciales is based on similar cystosori morphology. The potato disease powdery scab, caused by Sss, is difficult to control. The key control measure is avoidance, aimed at planting clean seed in clean soil. For the development of routine tests for the presence of the pathogen on tubers and in soil, a monoclonal antibody (MAb) was developed using Sss cystosori as immunogen. It detected less than one Sss cystosorus and recognised Sss material from many parts of the world. No cross-reactions with other Plasmodiophoromycetes including Plasmodiophora brassicae, Polymyxa betae, Polymyxa graminis and different Streptomyces species causing common and netted scab of potatoes were observed. A novel tuber sample test method was developed using a kitchen peeling machine. This detected two tubers with one powdery scab lesion each in a sample including eighteen uninfected tubers. When soil samples spiked with cystosori were tested with the MAb, different Sss infestation levels could be discriminated. Ssn cystosori gave absorbance values in ELISA as high as Sss cystosori, whereas fresh crook roots of watercress containing Ssnzoosporangia and plasmodia or mud from an Ssninfected watercress bed gave low absorbance values or no reaction. The potential of these findings for the development of a disease control management are discussed. Keywords Inoculum monoclonal antibody potato quantification watercress References Bell, KS, Roberts, J, Verrall, S, Cullen, DW, Williams, NA, Harrison, JG, Toth, LK, Cooke, DEL, Duncan, JM, Claxton, JR 1999 Detection and quantification of Spongospora subterranea f.sp. subterranea in soils and on tubers using specific PCR primers European Journal of Plant Pathology 105 905 915 Google Scholar Bouchek-Mechiche, K, Gardan, L, Normand, P, Jouan, B 2000 DNA relatedness among strains of Streptomyces pathogenic to potato in France: Description of three new species, S. europaeiscabiei sp. nov. and S. stelliscabiei sp. nov. associated with common scab, and S. reticuliscabiei sp. nov. associated with netted scab International Journal of Systematic and Evolutionary Microbiology 50 91 99 Google Scholar Braselton, JP 1995 Current status of the Plasmodiophorids Critical Reviews in Microbiology 21 263 275 PubMed Google Scholar Bulman, SR, Marshall, JW 1998 Detection of Spongospora subterranea in potato tuber lesions using the polymerase chain reaction (PCR) Plant Pathology 47 759 766 Google Scholar Clark, MF, Adams, AN 1977 Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses Journal of General Virology 34 475 483 PubMed Google Scholar St. Groth, S, Scheidegger, D 1980 Production of monoclonal antibodies: Strategy and tactics Journal of Immunological Methods 35 1 21 PubMed Google Scholar Down, GJ, Clarkson, JM 2002 Development of a PCR-based diagnostic test for Spongospora subterranea f.sp. nasturtii, the causal agent of crook root of watercress ( Rorippa nasturtium-aquaticum) Plant Pathology 51 275 280 Google Scholar Falloon, R, Genet, R, Wallace, A, Nott, H 1999 IPM use for powdery scab Commercial Grower 54 33 34 Google Scholar Harrison, JG, Rees, EA, Barker, H, Lowe, R 1993 Detection of spore balls of Spongospora subterranea on potato tubers by enzyme-linked immunosorbent assay Plant Pathology 42 181 186 Google Scholar Lambert, DH, Loria, R 1989a Streptomyces scabies sp. nov. nom International Journal of Systematic Bacteriology 39 387 392 Google Scholar Lambert, DH, Loria, R 1989b Streptomyces acidiscabies sp. nov. nom International Journal of Systematic Bacteriology 39 393 396 Google Scholar Merz, U 1989 Infectivity, inoculum density and germination of Spongospora subterranea resting spores: A solution-culture test system EPPO Bulletin 19 585 592 Google Scholar
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