A sensitive method for detecting bamboo mosaic virus (BaMV) and establishment of BaMV-free meristem-tip cultures

Citation
Yh. Hsu et al., A sensitive method for detecting bamboo mosaic virus (BaMV) and establishment of BaMV-free meristem-tip cultures, PLANT PATH, 49(1), 2000, pp. 101-107
Citations number
38
Categorie Soggetti
Plant Sciences
Journal title
PLANT PATHOLOGY
ISSN journal
00320862 → ACNP
Volume
49
Issue
1
Year of publication
2000
Pages
101 - 107
Database
ISI
SICI code
0032-0862(200002)49:1<101:ASMFDB>2.0.ZU;2-B
Abstract
A sensitive method was used to detect bamboo mosaic virus (BaMV) and its as sociated satellite RNA (satBaMV) by P-32- and digoxigenin (Dig)-labelled pr obes synthesized from cDNA clones of BaMV genomic (L probe) and satBaMV (S probe) RNA. Both the P-32- and Dig-labelled L and S probes could detect as little as 490 pg of BaMV viral RNA by slot- and dot-blot hybridization. In infected leaf extracts, P-32-labelled L and S probes detected virus at 25-f old higher dilutions than Dig-labelled probes, which were also successfully used to detect BaMV infection in plants derived from meristem-tip culture. However, immunoassays failed to detect BaMV in meristem culture. By dot-bl ot hybridization assays, 25% of the seedlings were shown to be virus-free. These results suggest that a highly sensitive method for the detection of B aMV infection is required for the establishment of BaMV-free cultures. Meri stem-tip culture also provides an efficient method for obtaining virus-free bamboo plants.