A New Method for the Detection of Minor Populations of Citrus Tristeza Virus Strains Infecting Single Citrus Trees

Authors : Hanife GENÇ

Pages : 449-459

View : 13 | Download : 4

Publication Date : 2005-12-01

Article Type : Research Paper

Abstract :Fifteen different biologically important citrus tristeza virus insert ignore into journalissuearticles values(CTV); isolates insert ignore into journalissuearticles values(B14, B15, B23, B31, B52, B57, T36, D9, D10, D11, D12, D15, D16, D20 and D32); were tested using strain-specific biotin-labeled oligonucleotide probes. Coat protein genes insert ignore into journalissuearticles values(CPGs); of the samples were amplified by PCR, and the products were hybridized with probes insert ignore into journalissuearticles values(direct PCR hybridization insert ignore into journalissuearticles values(DPH);); and sequenced. In the DPH, all samples having probe 0 reactions were infected with CTV. Some isolates giving more than one specific probe reaction showed that they may be infected by mixtures of different CTV strains. In order to investigate this, CPGs from individual bacterial colonies were used as PCR templates and the PCR products were tested. This method developed in this study was called colony PCR hybridization insert ignore into journalissuearticles values(CPH);. It was found that samples such as B57, which were apparently mixtures of strains, were indeed mixtures and other samples, which apparently contained a single strain insert ignore into journalissuearticles values(B23, D9, D11 and D16);, actually contained one or more additional strains not detectable by DPH or other methods. CPH is a sensitive method for the detection and differentiation of CTV strains infecting single citrus trees.
Keywords : CTV, Probes, PCR, hybridization, Colony PCR hybridization