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PCR-based assay for the detection of Curtobacterium flaccumfaciens pv. flaccumfaciens in bean seeds
1Dipartimento di Biotecnologie Agrarie (DiBA) - Sezione di Patologia Vegetale, Università degli Studi di Firenze, Firenze, Italy. stefania.tegli@unifi.it
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 (CffFOR2-CffREV4), 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.
