Identification and characterization of a calla lily-infecting isolate of Lisianthus necrosis virus

• Main Authors: Yi-Shan Chang, 張怡珊
• Other Authors: 陳煜焜
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/16559367572375422549

碩士 === 國立中興大學 === 植物病理學系所 === 95 === Calla lily (Zantedeschia spp.) showing symptoms of systemic necrosis was observed in the fields of central Taiwan in the 2003. Electron microscopic observation revealed the presence of isometric virus particles of 32-34 nm in diameter in the preparations of crude extracts, purified virus suspension and ultrathin sections of diseased tissues. The relative molecular weight of the subunit of viral capsid protein, similar to that of Lisianthus necrosis virus (LNV), was estimated as 39 kDa by SDS-polyacrylamide gel electrophoresis. The isolated virus reacted positively to LNV antiserum in indirect-ELISA and western blotting tests. The cDNA fragments were amplified from the total RNA extracted from LNV-Z infected Nicotiana benthamiana by reverse transcription-polymerase chain reaction (RT-PCR). Full length sequence of the virus genome has been compiled from cDNA fragments amplified by RT-PCR with LNV-specific primer sets. Sequence analysis showed that the complete genome of LNV-Z consists of 4785 nucleotides with five open reading frames (Acc. No. AM71119). The amino acid identity of the coat proteins between tested strain (LNV-Z) and the reported lisianthus strain (LNV-L) was 97.7％. The results shown in this study indicate that LNV is the causal agent of the systemic necrosis of calla lily in the fields.