Detection of complementary RNA intermediates of viroid replication by Northern blot hybridization.
Molecular hybridization by the Northern blot technique in combination with 125I-labeled PSTV (+) RNA and 32P-labeled PSTV cDNA as probes has been applied to detect viroid-specific sequences in healthy and viroid(PSTV)-infected tomato plants. Conditions are described which allow differentiation of (+) and (-) viroid sequences on the basis of the different thermostabilities of the corresponding hybrid molecules. By this experimental approach, it is documented that no viroid-specific DNA sequences can be detected and that viroid replication proceeds via complementary RNA intermediates. Out of the seven (-) RNA species found, six are apparently larger than the circular viroid (+) RNA and one is about the same size as the linear (+) RNA molecule.
D001483:Base Sequence; D009693:Nucleic Acid Hybridization; D010942:Plant Viruses / Q000254:growth & development / Q000378:metabolism*; D010944:Plants / Q000382:microbiology*; D012367:RNA, Viral / Q000096:biosynthesis*; D014772:Viroids / Q000254:growth & development / Q000378:metabolism*; D014779:Virus Replication
D012367:RNA, Viral