DETECTION OF VIABLE CELLS OF RALSTONIA-SOLANACEARUM IN SOIL USING A SEMISELECTIVE MEDIUM AND A PCR TECHNIQUE

Citation
S. Ito et al., DETECTION OF VIABLE CELLS OF RALSTONIA-SOLANACEARUM IN SOIL USING A SEMISELECTIVE MEDIUM AND A PCR TECHNIQUE, Journal of phytopathology, 146(8-9), 1998, pp. 379-384
Citations number
19
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Plant Sciences
Journal title
ISSN journal
0931-1785
Volume
146
Issue
8-9
Year of publication
1998
Pages
379 - 384
Database
ISI
SICI code
0931-1785(1998)146:8-9<379:DOVCOR>2.0.ZU;2-3
Abstract
A semiselective medium (PCCG) and a polymerase chain reaction (PCR) te chnique were combined to detect viable cells of Ralstonia (Pseudomonas ) solanacearum E. F. Smith (synonym Burkholderia solanacearum) in soil . DNA was extracted from 92 strains of soil bacteria including R. sola nacearum that grew on PCCG and then used as template for PCR with a pa ir of primers designed to amplify a single fragment (281 bp) of R. sol anacearum DNA. The 281-bp fragment was amplified only from DNA of R. s alanacearum (12 strains). For DNA from soil bacteria other than R. sol anacearum, the PCR amplification generated no products for 66 strains, a single DNA band with different sizes from 281 bp for 2 strains, and several DNA bands for 12 strains. Southern analysis showed that any o f those products other than the 281-bp fragment had no homology with t he 281-bp fragment of R. solanacearum, indicating the specificity of t he primers to generate the 281-bp fragment from R. solanacearum. A sim ple method consisting of a plating step using PCCG and succeeding PCR for amplifying the 281-bp fragment from colonies on PCCG was described .