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Diagnotics of the important apple viruses
Winkowska, Lucie ; Ryšánek, Pavel (advisor) ; Petr, Petr (referee)
Apple mosaic virus (ApMV), Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV) and Apple chlorotic leaf spot virus (ACLSV) are economically the most important viruses in pome fruit trees, which are distributed worldwide and can caused significantly yield reduction. The major control strategies (namely pathogen detection, exclusion by crop certification or quarantine, control in infected orchards by eradication from infected cultivars and rootstocks, etc.) rely heavily on accurate and sensitive detection methods and on perfect knowledge of pathogens.
In the doctoral thesis the diagnostic method quantitative RT-PCR (qRT-PCR) was optimized for detection and quantification of four studied viruses. The results suggested that qRT-PCR method was the most reliable technique in comparison with conventional diagnostic methods DAS/I-ELISA and RT-PCR.
In our study the concentration of ASGV, ASPV and ACLSV, measured by qRT-PCR, were stable during vegetation and in different plant tissue. Only the concentration of ApMV changed during vegetation in leaves and inner bark. This result indicates that changes of virus concentration observed by DAS/I-ELISA and RT-PCR in plant tissues are caused by other way (inhibitors, plant senescence, lower sensitivity, ect.) than by changes of virus concentrations in plant.
Under the monitoring (at all 351 trees were tested) it was showed, that studied viruses were more spread in orchards and gardens then in wild apple trees.
Selected virus isolates from wild apple trees and apples from orchards and gardens were sequenced and molecular variability was studied also with already published isolates. However individual isolates of studied viruses were similar. The variability associated with geographic origin or with type of planting has not been confirmed.
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