Development of PCR for the detection of Polymyxa betae in sugar beet roots and its application in field studies

• Published in: Physiological and molecular plant pathology Vol. 47; no. 5; pp. 303 - 313
• Main Authors: Mutasa, E.S., Chwarszczynska, D.M., Adams, M.J., Ward, E., Asher, M.J.C.
• Format: Journal Article
• Language: English
• Published: London Elsevier India Pvt Ltd 01.11.1995; Elsevier
• Subjects: BBES, Broom's Barn; beet nectrotic yellow vein virus; Biological and medical sciences; Biotechnology; BNYVV; CTAB, hexadecyltrimethyl-ammonium bromide; dNTPs, deoxyribonucleoside triphospates; EMBL, European Molecular Biology Library; EtBr, ethidium bromide; Fundamental and applied biological sciences. Psychology; Fungal plant pathogens; General aspects and techniques; Generalities. Techniques; Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control; PCR, polymerase chain reaction; Phytopathology. Animal pests. Plant and forest protection; Plant viruses and viroids; Protozoa; RES, Rothamsted; SDS, sodium dodecylsulphate; SSPE, 0·001 M phosphate buffer pH7·4 containing 0·15 M NaCl, 1· M EDTA; England; United Kingdom; Great Britain; Europe; BBES, Broom's Barn; beet nectrotic yellow vein virus; EtBr, ethidium bromide; CTAB, hexadecyltrimethyl-ammonium bromide; SDS, sodium dodecylsulphate; PCR, polymerase chain reaction; RES, Rothamsted; BNYVV; dNTPs, deoxyribonucleoside triphospates; EMBL, European Molecular Biology Library; SSPE, 0·001 M phosphate buffer pH7·4 containing 0·15 M NaCl, 1· M EDTA; Cultivated field; Plant pathogen; Plant juvenile growth stage; Organic soil; Sandy soil; Beta vulgaris var. saccharata; Epidemiology; Incidence; Dicotyledones; Angiospermae; Loam soil; Diagnosis; Southern blotting; Vector; Nucleic acid; Protozoa; Silty soil; Root; Polymyxa betae; Mineral soils; Sugar plant; Chenopodiaceae; Polymerase chain reaction; Screening; Dot blotting; Analysis method; DNA; Peat soil; Spermatophyta; Technique; Molecular biology; Detection; Mycetozoa
• ISSN: 0885-5765; 1096-1178
• DOI: 10.1006/pmpp.1995.1060

A previously cloned 1·8 kb PCR Polymyxa betae DNA fragment has been fully sequenced and used to design PCR primers for the specific detection of P. betae in sugar beet seedling roots. The primers did not amplify sequences from fungi closely related to P. betae, or from sugar beet or other microorganisms commonly associated with sugar beet roots. Protocols which combine PCR with Southern/dot blot analysis of products were developed to detect P. betae in sugar beet roots. A nested PCR protocol was also developed and shown to provide levels of sensitivity that allow more rapid screening for P. betae infection. Subsequently, PCR was used to detect P. betae in a survey of infection incidence in 65 sugar beet fields. Seventy one percent of fields and 33% of plants sampled were infected by late June, with a lower incidence on peat as compared to mineral soils.