Functional analysis of beet necrotic yellow vein virus (BNYVV) RNA4 in fungal transmission

Full-length RNA4 of beet necrotic yellow vein virus (BNYVV) and its mutants, including a frame-shift and deletions within the coding region, were inserted into a transcription plasmid. Transcripts derived from these plasmids and RNAs extracted from a BNYVV isolate containing only RNAs 1, 2 and 3 were coinoculated to Tetragona expansa. Then these recombinant isolates were inoculated to sugar beet plants grown in a sand culture system containing Po-lymyxa betae that had been freed of virus particles to test efficiency of BNYVV transmission. The result showed that the efficiency of BNYVV transmission among the plants by Polymyxa betae was highly decreased by different deletions in the coding region of RNA4, but not affected by the frame-shift mutant with four nucleotides insertion. The observations suggest that at least some of the RNA4 sequence encompassing the deleted region of 580 nucleotide acids (525 -1105 nt) are necessary for high efficient transmission of the virus by the fungus.