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Title: DIFFERENTIATION OF CLOSELY RELATED BUT BIOLOGICALLY DISTINCT ISOLATES OF PRUNUS NECROTIC RINGSPOT VIRUS BY POLYMERASE CHAIN REACTION

Author
item Hammond, Rosemarie
item Crosslin, James
item PASINI, RITA - DEPT OF MICROBIOL., UMCP
item HOWELL, WILLIAM - WASH. STATE UNIV, PROSSER
item MINK, GAYLORD - WASH. STATE UNIV, PROSSER

Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/30/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary: Prunus necrotic ringspot virus (PNRSV) occurs worldwide wherever stone fruits are grown and is a serious pathogen of many woody species, including sweet and sour cherry, peach, apple, and plum. All species and cultivars of Prunus appear to be susceptible to one or more strains of the virus. The virus can cause serious diseases in nurseries and orchards with reduced dtree growth and yield. Highly virulent strains cannot be readily distinguished from mild strains by rapid, non-biological methods. We have developed reverse transcriptase-polymerase chain reaction (RT-PCR) assays that can identify isolates in sweet cherry displaying different symptoms. The method provides a rapid, valuable tool for identification of severe isolates and will aid scientists and extension services to reduce field spread of the disease. The assays will also aid quarantine and action agencies in indexing of incoming and outgoing germplasm for the presence of fthis virus.

Technical Abstract: Prunus necrotic ringspot ilarvirus (PNRSV) exists as a number of biologically distinct variants which differ in host specificity, serology, and pathology. Previous nucleotide sequence alignment and phylogenetic analysis of cloned reverse transcription-polymerase chain reaction (RT-PCR) products amplified from genomic RNA 3 of several biologically distinct PNRSV isolates obtained from sweet cherry revealed correlations between symptom type and the nucleotide and amino acid sequences of the 3a (putative movement protein) and 3b (coat protein) open reading frames (Hammond and Crosslin, 1998). Based upon this analysis, RT-PCR assays have been developed that can identify isolates displaying different symptoms and serotypes. The incorporation of primers in a multiplex PCR protocol permits rapid detection and discrimination among the strains. The results of PCR amplification using type-specific primers that amplify a portion of the coat protein gene demonstrate that the primer-selection procedure developed for PNRSV constitutes a reliable method of viral strain discrimination in cherry for disease control and will also be useful in examining biological diversity within the PNRSV virus group.