[Construction of infectious cDNA clone of cucumber mosaic virus satellite RNA XJs1 and preliminary study on its...

Cucumber mosaic virus (CMV) sugar beet isolate caused yellow mosaic, leaf distortion, crinkle and stunt symptoms on sugar beet in nature. It exhibited some special biological properties with narrower host range and had no symptom on Nicotiana glutinosa L. and Nicotiana tobacum L. cv. NC-89. A new satellite RNA, XJs1 was found to be associated with the helper virus. In order to know the cause of the special pathogenicity of the CMV isolate. Full-length infectious cDNA clone of CMV satellite RNA XJs1, pMSC20, was constructed by reverse transcriptase-polymerase chain reaction (RT-PCR). Sequence analysis showed that the satellite RNA consists of 384 nucleotides (nt) (GenBank accession number: D0070748). Compared the nucleotide sequence of satellite RNA XJsl with those of other representative CMV satellite RNAs displayed that it contains typical necrogenic consensus sequence block from positions 325 to 350, and shared 73.27% - 91.93% nucleotide sequence identity with some published CMV satellite RNAs. By in vitro transcription, satellite RNA XJsl was inoculated on Nicotiana tabacum and Nicotiana glutinosa together with CMV-AH, a CMV isolate without satellite RNA. The results showed that satellite RNA XJsl could attenuate symptoms on Nicotiana tabacum and Nicotiana glutinosa induced by CMV-AH. Detection by RT-PCR and Northern blot hybridization revealed XJs1 obtained replication in the above two host plants, showing the pathogenicity changes of CMV-AH on Nicotiana tabacum and Nicotiana glutinosa were induced by co-infecting with satellite RNA XJsl. These results indicated that XJsl is probably an attenuate satellite RNA. The relationship between helper virus, satellite RNA and host plants is discussed.