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Title: PCR-DETECTION OF APPLE STEM PITTING VIRUS FROM POME FRUIT HOSTS AND SEQUENCE VARIABILITY AMONG VIRAL ISOLATES

Author
item NEMICHINOV, L - CONTRACT EMPLOYEE
item Hadidi, Ahmed

Submitted to: Acta Horticulturae
Publication Type: Proceedings
Publication Acceptance Date: 9/14/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary: Apple stem pitting virus is an economically important virus which infects apple, pear, and quince worldwide and is of quarantine importance. Currently this virus is detected by biological indexing which may be time consuming, labor intensive, and/or not virus specific. Serological methods have not been used for detection because of the lack of commercially available viral antiserum. We have developed a sensitive and rapid assay method for apple stem pitting virus from infected pome fruit tissue. This method is based on polymerase chain reaction (PCR) technology. We have also demonstrated a potential molecular variability among virus isolates examined. The PCR assays developed have the potential to be utilized in quarantine and certification programs for early and opportune detection of the pathogen.

Technical Abstract: A relatively simple and rapid PCR assay has been developed for the detection of apple stem pitting virus (ASPV) from apples and its' pear strain known as pear vein yellows virus (PVYV), from pears. The assay allows accurate diagnosis of ASPV and PVYV from leaf, bark and bud tissues of stem pitting or pear vein yellows diseased apple and pear trees, respectively. Nucleotide sequences of the cloned PCR products, derived from different ASPV isolates, have been obtained and analyzed; inserts were used to generate ASPV-and PVYV-specific DIG-labeled cRNA probes. Sequencing and hybridization analyses revealed a potentially high level of molecular variability among the isolates of the virus.