Premium
Detection of plant viruses by molecular hybridization using non‐radioactive probes 1
Author(s) -
KUMMERT J.,
COLINET D.,
LEPOIVRE P.
Publication year - 1995
Publication title -
eppo bulletin
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.327
H-Index - 36
eISSN - 1365-2338
pISSN - 0250-8052
DOI - 10.1111/j.1365-2338.1995.tb01471.x
Subject(s) - complementary dna , nucleic acid , biology , rna , plant virus , nucleic acid thermodynamics , dot blot , hybridization probe , microbiology and biotechnology , molecular probe , context (archaeology) , dna , virus , virology , biochemistry , gene , paleontology
In the context of the phytosanitary legislation governing the trade of plants, accurate highly sensitive and rapid techniques of virus detection need to be developed. Such objectives can be achieved by molecular hybridization and PCR. Specific nucleic acid sequences of RNA viruses can be directly recognized by hybridization in the presence of labelled complementary DNA (cDNA) probes, synthesized either from purified viral RNA, from ds‐RNA from infected plants, or from total RNA preparations from infected plants by RT‐PCR. After characterization and selection, specific cDNA are cloned in bacteria by means of recombinant plasmids, allowing continuous availability of identical and homologous probes. Techniques of non‐radioactive labelling, which can reach similar levels of sensitivity and specificity to the previous exclusive use of radioactivity, are used. Specific probes are generally hybridized to viral nucleic acid sequences from plant samples immobilized on solid support (dot‐blot hybridization). Several examples for different viruses and host plants are presented, demonstrating the possibilities of the use of molecular hybridization for diagnosis of virus infection in plants.