Current studentsExisting staffContact us
 

Detection of Coconut cadang-cadang viroid sequences in oil and coconut palm by ribonuclease protection assay

Simple item page
dc.contributor.authorVadamalai, G.
dc.contributor.authorPerera, A.
dc.contributor.authorHanold, D.
dc.contributor.authorRezaian, M.
dc.contributor.authorRandles, J.
dc.date.issued2009
dc.descriptionJournal compilation © 2008 Association of Applied Biologists
dc.description.abstractA ribonuclease protection assay (RPA) has been developed for detecting Coconut cadang-cadang viroid (CCCVd) sequences. An RNA probe complementary to full-length CCCVd₂₄₆ was used, terminating at nucleotide 65 in the upper conserved region, and linked to a non-viroid 5' sequence, which acted as an internal control for ribonuclease activity. Extracts from CCCVd-infected coconut (Cocos nucifera) and African oil (Elaeis guineensis) palms protected three major fragments of approximately 250, 125 and 50 nt and a variable number of minor fragments. Extracts of healthy coconut palms, Potato spindle tuber viroid-infected tomato and transfer RNA did not protect the probe. The approximately 250 nt fragment is predicted to indicate the presence of monomers and dimers of circular CCCVd₂₄₆, linear CCCVd₂₄₆ with the same termini as the probe and point mutants of these forms. The origin of smaller protected fragments is discussed. RPA-detected CCCVd sequences in 13 of 18 oil palms surveyed in a commercial plantation in Malaysia. Signal intensity varied between the positive oil palms and was generally lower than in coconut palms infected with CCCVd. An infection phenotype was implied but not confirmed by the observation that in a group of 10 oil palms with orange leaf spotting, 9 contained CCCVd, whereas in a group of 8 palms without orange spotting, the viroid was detected in 4. Of four coconut palms in Sri Lanka shown by dot-blot assay to contain CCCVd-related RNA, one was shown by RPA to be positive for the CCCVd₂₄₆ sequence. RPA is therefore a robust and sensitive test for CCCVd sequences, and our results show that sequences closely related to CCCVd₂₄₆ are not confined to the Philippines.
dc.description.statementofresponsibilityG. Vadamalai, A.A.F.L.K. Perera, D. Hanold, M.A. Rezaian & J.W. Randles
dc.identifier.citationAnnals of Applied Biology, 2009; 154(1):117-125
dc.identifier.doi10.1111/j.1744-7348.2008.00278.x
dc.identifier.issn0003-4746
dc.identifier.issn1744-7348
dc.identifier.orcidRandles, J. [0000-0003-2231-816X]
dc.identifier.urihttp://hdl.handle.net/2440/50858
dc.language.isoen
dc.publisherAssoc Applied Biologists
dc.source.urihttps://doi.org/10.1111/j.1744-7348.2008.00278.x
dc.subjectAfrican oil palm
dc.subjectCCCVd-related RNA
dc.subjectCoconut cadang-cadang viroid
dc.subjectcoconut palm
dc.subjectcocadviroid
dc.subjectMalaysia
dc.subjectribonuclease protection assay
dc.subjectSri Lanka.
dc.titleDetection of Coconut cadang-cadang viroid sequences in oil and coconut palm by ribonuclease protection assay
dc.typeJournal article
pubs.publication-statusPublished

Files

Collections

Agriculture, Food and Wine publications