Premium
PCR‐based assay for the detection of Curtobacterium flaccumfaciens pv. flaccumfaciens in bean seeds
Author(s) -
Tegli S.,
Sereni A.,
Surico G.
Publication year - 2002
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2002.01187.x
Subject(s) - biology , genomic dna , polymerase chain reaction , microbiology and biotechnology , dna , pcr test , nat , virology , genetics , gene , physics , quantum mechanics
Aims: To develop a PCR‐based protocol for the rapid, sensitive and specific detection of Curtobacterium flaccumfaciens pv. flaccumfaciens ( Cff ) in bean seeds. Methods and Results: A pair of PCR primers ( Cff FOR2‐ Cff REV4), targeting the sequence of a cloned DNA fragment of 550 bp amplified in Repetitive‐sequence‐based‐PCR (Rep‐PCR) experiments, were designed and shown to specifically amplify a 306‐bp DNA fragment using Cff DNA as template. Moreover, this PCR protocol was demonstrated to successfully detect Cff in naturally infected bean seeds in 36 h. Conclusions: A specific, highly sensitive and rapid PCR assay for the detection of Cff was achieved. Significance and Impact of the Study: Cff is a seed‐borne bacterium on the EPPO A2 quarantine list; this procedure may be useful for routine diagnosis of Cff , overcoming the problems of conventional techniques.