Fernando M. Alves-Santos

A DNA-Based Procedure for In Planta Detection of Fusarium oxysporum f. sp. phaseoli.

ABSTRACT We have characterized strains of Fusarium oxysporum from common bean fields in Spain that were nonpathogenic on common bean, as well as F. oxysporum strains (F. oxysporum f. sp. phaseoli) pathogenic to common bean by random amplified polymorphic DNA (RAPD) analysis. We identified a RAPD marker (RAPD 4.12) specific for the highly virulent pathogenic strains of the seven races of F. oxysporum f. sp. phaseoli. Sequence analysis of RAPD 4.12 allowed the design of oligonucleotides that amplify a 609-bp sequence characterized amplified region (SCAR) marker (SCAR-B310A280). Under controlled environmental and greenhouse conditions, detection of the pathogen by polymerase chain reaction was 100% successful in root samples of infected but still symptomless plants and in stem samples of plants with disease severity of >/=4 in the Centro Internacional de Agricultura Tropical (CIAT; Cali, Colombia) scale. The diagnostic procedure can be completed in 5 h and allows the detection of all known races of the pathogen in plant samples at early stages of the disease with no visible symptoms.

Influence of the ectomycorrhizal fungus Laccaria laccata on pre-emergence, post-emergence and late damping-off by Fusarium oxysporum and F. verticillioides on Stone pine seedlings.

In greenhouse experiments, the ectomycorrhizal fungus Laccaria laccata was evaluated for biological control of preemergence, post-emergence and late damping-off of Pinus pinea caused by Fusarium verticillioides and F. oxysporum. In pre-emergence damping-off assays, preinoculation with Laccaria laccata did not significantly improve germination of seeds and no statistical significant differences were found in Fusarium treatments when compared with controls. At 18 weeks after sowing, inoculation with L. laccata reduced the incidence of post-emergence damping-off but differences were significant only in F. oxysporum treatments. Pinus pinea transplanted plants were used in late damping off assays, and only F. oxysporum produced significant damage. Inoculation with L. laccata did not attenuate significantly the virulence of F. oxysporum. However, the percentage of mycorrhization did not reached significant level, so the amount of mycorrhizal fungus was insufficient for effective protection. Although very low percentages of mycorrhization were recorded in all mycorrhized treatments, and Fusarium occurrence significantly reduced mycorrhization, those levels have been efficient to reduce damage in F. oxysporum post-emergence damping-off assays. In short, pre-emergence damping-off was not found; only F. oxysporum produced significant damage on P. pinea seedlings and L. laccata reduced damage when the percentage of mycorrhization reached a significant level. These results have been compared with previous work on P. sylvestris inoculated with the same mycorrhizae isolate and Fusarium pathogens.