Mitochondrial plasmids of sugar beet amplified via rolling circle method detected during curtovirus screening.
Crops of sugar beet have been considerably impaired by infection with Beet curly top virus (BCTV) during the past decades. Quick and reliable diagnostic techniques are therefore desirable to detect this circular single-stranded DNA-containing geminivirus. Techniques combining either tissue printing or blot hybridization, or rolling circle amplification (RCA) and restriction fragment length polymorphism (RFLP) were compared. Although they easily detected BCTV with certainty, both exhibited apparent false positive results which have been scrutinized in closer detail. Uninfected control plants revealed unspecific signals due to probe attachment on tissue blots, and dominant fragment patterns upon RCA/RFLP which did not hybridize with BCTV-specific probes. Cloning and sequencing of these DNA fragments showed that they were amplified from mitochondrial plasmids. Examination of their genome structure revealed no relationship with geminiviruses or their satellites.
Institute of Biology, Department of Molecular Biology and Plant Virology, University of Stuttgart, Pfaffenwaldring 57, 70550 Stuttgart, Germany.
Maria
M
Homs
Sigrid
S
Kober
Gabi
G
Kepp
Holger
H
Jeske
D027461:Beta vulgaris / Q000235:genetics*; D003001:Cloning, Molecular; D016172:DNA Fingerprinting; D004261:DNA Replication; D004272:DNA, Mitochondrial / Q000378:metabolism*; D005189:False Positive Reactions; D017791:Geminiviridae / Q000235:genetics / Q000302:isolation & purification*; D008928:Mitochondria / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization / Q000379:methods; D010957:Plasmids / Q000378:metabolism*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA
D004272:DNA, Mitochondrial