Abstract:

Specific primer pairs were designed according to the gene sequence of cucumber mosaic virus(CMV)and tobacco mosaic virus(TMV)coat protein region. The two viruses could be detected by simple RT-PCR with the primers and the suitable primers were selected for multiplex RT-PCR. Using 18S rRNA of gladiolus as an internal control, a multiplex RT-PCR program was established to simultaneously detect the two viruses of Gladiolus hybridus. Two specific bands,CMV(629 bp)and TMV(423 bp)were amplified from all the samples infected with CMV and TMV by the multiplex RT-PCR. Sequence analysis of the amplified products showed that the nucleotide sequences homology of CMV and TMV was 93%~97% and 99%, respectively, compared with the sequences of other isolates.

Key words: Gladiolus hybridus, Multiplex RT-PCR, CMV, TMV, 18S rRNA