PCR-based specific detection of Ralstonia solanacearum by amplification of cytochrome c1 signal peptide sequences.
A polymerase chain reaction (PCR)-based method was developed to detect the DNA of Ralstonia solanacearum, the causal agent of bacterial wilt in various crop plants. One pair of primers (RALSF and RALSR), designed using cytochrome c1 signal peptide sequences specific to R. solanacearum, produced a PCR product of 932 bp from 13 isolates of R. solanacearum from several countries. The primer specificity was then tested using DNA from 21 isolates of Ralstonia, Pseudomonas, Burkholderia, Xanthomonas, and Fusarium oxysporum f. sp. dianthi. The specificity of the cytochrome c1 signal peptide sequences in R. solanacearum was further confirmed by a DNA-dot blot analysis. Moreover, the primer pair was able to detect the pathogen in artificially inoculated soil and tomato plants. Therefore, the present results indicate that the primer pair can be effectively used for the detection of R. solanacearum in soil and host plants.
Kang
Man Jung
MJ
National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon 441-707, Korea.
Lee
Mi Hee
MH
Shim
Jae Kyung
JK
Seo
Sang Tae
ST
Shrestha
Rosemary
R
Cho
Min Seok
MS
Hahn
Jang Ho
JH
Park
Dong Suk
DS
D003575:Cytochromes c1 / Q000235:genetics*; D018551:Solanum lycopersicum / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D021382:Protein Sorting Signals / Q000235:genetics*; D043368:Ralstonia solanacearum / Q000235:genetics / Q000302:isolation & purification*; D012988:Soil Microbiology
D021382:Protein Sorting Signals; D003575:Cytochromes c1