Discrimination between tobamoviruses and their pathotypes for L-gene-mediated resistance in green pepper (Capsicum annuum L.) by reverse transcription-polymerase chain reaction

• Published in: Journal of general plant pathology : JGPP Vol. 71; no. 1; pp. 60 - 67
• Main Authors: Takeuchi, S. (Kochi-ken. Agricultural Research Center, Nankoku (Japan)), Hamada, H, Toyoda, K, Suzuki, K, Kiba, A, Hikichi, Y, Okuno, T
• Format: Journal Article
• Language: English
• Published: Tokyo Springer Nature B.V 01.02.2005
• Subjects: BIOLOGICAL DIFFERENCES; CAPSICUM ANNUUM; Deoxyribonucleic acid; DISEASE RESISTANCE; DNA; GENETIC RESISTANCE; MOLECULAR BIOLOGY; PATHOTYPES; PCR; Plant pathology; TOBAMOVIRUSES; Tomatoes; Vegetables; Viruses
• ISSN: 1345-2630; 1610-739X
• DOI: 10.1007/s10327-004-0161-4

A new method to discriminate between tobamoviruses and their pathotypes that infect green pepper (Capsicum annuum L.) was developed using reverse transcription-polymerase chain reaction (RT-PCR). The P sub(0) pathotype (Tobacco mosaic virus. Tomato mosaic virus, and Tobacco mild green mosaic virus) and the P sub(1) pathotype (Paprika mild mottle virus) were distinguished by RT-PCR using primers specific to each pathotype. However, the P sub(12) and P sub(1,2,3) pathotypes of Pepper mild mottle virus (PMMoV) could not be distinguished from each other using this procedure. The P sub(1,2) and P sub(1,2,3) pathotypes were differentiated by RT-nested PCR, in which a DNA fragment was first produced by RT-PCR using primers containing conserved sequences of PMMoV. The product was then used as the DNA template in a second PCR using primers specific to each pathotype. An immunocapture (IC) RT-PCR method was developed based on results from this study, which facilitated detection of tobamoviruses in pepper plants, seeds, and field soils and allowed the identification of their pathotypes.