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A Technique Combining Immunoprecipitation and RT ‐ PCR for RNA Plant Virus Detection
Author(s) -
Lima José Albersio Araujo,
Nascimento Aline Kelly Queiroz,
Radaelli Paula,
Silva Ana Kelly Firmino,
Silva Fabiana Rodrigues
Publication year - 2014
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/jph.12208
Subject(s) - immunoprecipitation , rna , biology , virus , reverse transcription polymerase chain reaction , real time polymerase chain reaction , reverse transcriptase , microbiology and biotechnology , virology , polymerase chain reaction , messenger rna , biochemistry , gene
Plant virus identification and characterization can be accomplished by several methods involving their morphological, physical, biological, cytological, serological and molecular properties. The use of molecular techniques is increasing worldwide, and some have been developed for identification and characterization of plant viruses. Reverse transcription polymerase chain reaction ( RT ‐ PCR ) has been shown to be a suitable method for research with RNA plant viruses. In this study, a new approach of RT ‐ PCR involving previous virus immunoprecipitation ( IP ) was used for RNA amplification of five virus species of the genera C omovirus , C ucumovirus , P otyvirus and S obemovirus from infected plant tissues. IP ‐ RT ‐ PCR was practical, sensitive and minimized problems with total RNA extractions from infected tissues. The technique provides partial virus particle purification by its specific immunoprecipitation, and it should be especially useful for RNA amplification of viruses that occur in low or variable concentrations in plant tissues or when the tissues contain various forms of RT ‐ PCR amplification inhibitors.