Articles
DETECTION OF VIROID-LIKE RNAS IN POMEGRANATE (PUNICA GRANATUM L.)
Article number
550_46
Pages
321 – 326
Language
English
Abstract
Leaves of pomegranate (of six different varieties) exhibiting some kind of alterations were analysed for the presence of viroids or viroid-like RNAs.
The standard RNA extraction procedure was applied and RNAs were analysed by bi-dimensional polyacrylamide gel electrophoresis.
Two sets of three different bands (estimated sizes between 270 and 400 nt) were clearly observed in the diagonal of the second dimension gel, indicating a possible circular nature for these RNAs.
A 300 nt band was not recognised by a hop stunt viroid riboprobe.
These RNAs were pooled, RT-PCR amplified and cloned.
One of the recombinant plasmid (called 5/24) was able to detect two of the six putative viroid-like RNAs in Northern blot assays.
The sequence of the 5/24 clone showed 100% of homology with chloroplastic 16S rRNA. These results, and the observation that the putative viroid-like RNAs were detected in all varieties tested, permit to propose that at least two of the these six RNAs were from a host origin.
The standard RNA extraction procedure was applied and RNAs were analysed by bi-dimensional polyacrylamide gel electrophoresis.
Two sets of three different bands (estimated sizes between 270 and 400 nt) were clearly observed in the diagonal of the second dimension gel, indicating a possible circular nature for these RNAs.
A 300 nt band was not recognised by a hop stunt viroid riboprobe.
These RNAs were pooled, RT-PCR amplified and cloned.
One of the recombinant plasmid (called 5/24) was able to detect two of the six putative viroid-like RNAs in Northern blot assays.
The sequence of the 5/24 clone showed 100% of homology with chloroplastic 16S rRNA. These results, and the observation that the putative viroid-like RNAs were detected in all varieties tested, permit to propose that at least two of the these six RNAs were from a host origin.
Publication
Authors
G. Gómez, V. Pallás
Keywords
pomegranate (Punica granatum L.); Viroids; Small circular RNAs
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