Detection and Concentration of Coconut Cadang-Cadang Viroid in Coconut Leaf Extracts

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Abstract
Procedures for the detection of coconut cadang-cadang viroid (CCCV) by polyacrylamide gel electrophoresis (PAGE) and molecular hybridization with 3H- or 32P-label complementary DNA (cDNA) are compared. The lower limits of detection per assay for the 3 techniques were the following: PAGE, 50 ng; 3H-cDNA hybridization in solution, 0.5 ng; 32P-cDNA spot hybridization on nitrocellulose membranes, 0.25 ng. Spot hybridization was considered the best for routine diagnosis of CCCV because of its sensitivity, ease of operation, and reliability. Infection in inoculated seedlings could be detected by this method at least 4 mo before the viroid band was detectable by PAGE. Concentrations of CCCV in partially purified nucleic acid extracts were estimated by molecular hybridization and measurements of yields of purified viroid. In the same palm, young fronds contained lower amounts of CCCV than mature fronds. Palms in the early and medium stages of disease contained similar amounts of CCCV while late stage palms contained lower amounts. Palms at the same stage of disease from varied geographical locations with different levels of disease incidence contained comparable levels of CCCV.

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