ISHS Acta Horticulturae 550: XVIII International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Top Fruit Diseases
DETECTION OF VIROID-LIKE RNAS IN POMEGRANATE (PUNICA GRANATUM L.) |
| Authors: | G. Gómez, V. Pallás |
| Keywords: | pomegranate (Punica granatum L.); Viroids; Small circular RNAs |
| 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. | |
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