title: Detection of Chrysanthemum stunt viroid (CSV) using nick translated probes in a dot-blot hybridization assay.
abstract: We developed a dot-blot hydridization assay for the detection of Chrysanthemum stunt viroid (CSV) in Chrysanthemum plant samples. The probe, a recombinant plasmid containing a full-length monomeric cDNA copy of CSV, is labelled with (32P) by nick-translation. The influence of the hybridization conditions, of the sample denaturation technique and of the plant sap components on the final sensitivity has been studied. The optimized system, involving a formaldehyde denaturation step, allows the detection of as little as 5 pg of purified viroid. Under these conditions, 100 pg of pure viroid diluted in plant sap, or infected plant extract diluted 1:25 in healthy extract can be detected, showing the potential of this method for indexing of Chrysanthemum for CSV infection.
authors.affiliation: Station de pathologie végétale, INRA, Centre de Recherches de Bordeaux, Pont de la Maye, France.
authors.forename: T
authors.initials: T
authors.lastname: Candresse
authors.forename: G
authors.initials: G
authors.lastname: Macquaire
authors.forename: M
authors.initials: M
authors.lastname: Monsion
authors.forename: J
authors.initials: J
authors.lastname: Dunez
mesh_terms: D011723:Chrysanthemum cinerariifolium / Q000382:microbiology*; D004274:DNA, Recombinant; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D010935:Plant Diseases; D010942:Plant Viruses / Q000302:isolation & purification*; D010944:Plants / Q000382:microbiology*; D014772:Viroids / Q000302:isolation & purification*
chemical_list: D004274:DNA, Recombinant