Origin of Race 3 of Fusarium oxysporum f. sp. lycopersici at a Single Site in California.

ABSTRACT Thirty-nine isolates of Fusarium oxysporum were collected from tomato plants displaying wilt symptoms in a field in California 2 years after F. oxysporum f. sp. lycopersici race 3 was first observed at that location. These and other isolates of F. oxysporum f. sp. lycopersici were characterized by pathogenicity, race, and vegetative compatibility group (VCG). Of the 39 California isolates, 22 were in VCG 0030, 11 in VCG 0031, and six in the newly described VCG 0035. Among the isolates in VCG 0030, 13 were race 3, and nine were race 2. Of the isolates in VCG 0031, seven were race 2, one was race 1, and three were nonpathogenic to tomato. All six isolates in VCG 0035 were race 2. Restriction fragment length polymorphisms (RFLPs) and sequencing of the intergenic spacer (IGS) region of rDNA identified five IGS RFLP haplotypes, which coincided with VCGs, among 60 isolates of F. oxysporum from tomato. Five race 3 isolates from California were of the same genomic DNA RFLP haplotype as a race 2 isolate from the same location, and all 13 race 3 isolates clustered together into a subgroup in the neighbor joining tree. Collective evidence suggests that race 3 in California originated from the local race 2 population.

Potential for Dispersal of Fusarium oxysporum f. sp. erythroxyli by Infested Seed.

Fusarium oxysporum f. sp. erythroxyli causes a vascular wilt of the narcotic plant coca (Erythroxylum coca var. coca). To determine whether this pathogen can be transmitted by infested seed, fruit from symptomatic and asymptomatic plants was collected from different coca-growing areas in Peru and from an experimental field site in Hawaii. A total of 202 fruit from Peru and 69 fruit from Hawaii were surface-disinfested and separated into five parts: pedicel, pericarp, seed coat, endosperm, and cotyledons. After the pedicel and pericarp were removed from the seed coat, the seed was surface disinfested again. Each fruit part was plated separately. Both F. oxysporum and F. moniliforme were recovered from fruit collected in Peru. Both species were isolated from all parts of some fruit. F. oxysporum was isolated from 33% of the fruit plated and most (35%) of these isolates were obtained from the seed coat. Slightly greater numbers of isolates (57%) were recovered from asymptomatic plants than from symptomatic plants (43%). Only F. oxysporum was isolated from fruit collected in Hawaii. Most of these isolates (59%) were from the pedicels of fruit collected from symptomatic plants. Out of 91 isolates of F. oxysporum, 21 were pathogenic to coca seedlings in a bioassay. Six of these pathogenic isolates were originally from the pedicel of the fruit, eight from the pericarp, four from the seed coat, and three from the endosperm. No isolates from the cotyledons were pathogenic. Most of the pathogenic isolates (76%) were from symptomatic plants. The pathogenic isolates were characterized using random amplified polymorphic DNA analysis and vegetative compatibility groups. Based on these analyses, two different subpopulations of the forma specialis erythroxyli were found in Peru, whereas only one was present in Hawaii. These data indicate that infested seed may contribute significantly to dissemination of this pathogen because seed is collected by growers and planted fresh or fermented briefly before planting.

Genetic Characterization by RAPD Analysis of Isolates of Fusarium oxysporum f. sp. erythroxyli Associated with an Emerging Epidemic in Peru.

ABSTRACT An epidemic of vascular wilt caused by Fusarium oxysporum f. sp. erythroxyli is currently occurring on Erythroxylum coca var. coca in the coca-growing regions of the Huallaga Valley in Peru. Random amplified polymorphic DNA (RAPD) analysis of isolates of the pathogen was undertaken to elucidate its genetic complexity, as well as to identify a specific DNA fingerprint for the pathogen. Two hundred isolates of Fusarium were collected from 10 coca-growing regions in Peru. Of these, 187 were confirmed to be F. oxysporum, and 143 of the F. oxysporum were shown to be pathogens of coca by a root-dip pathogenicity test. The pathogens could be grouped into two subpopulations based on RAPD analysis, and no polymorphism in RAPD pattern was observed among isolates of either subpopulation. Both subpopulations were present in the central Huallaga Valley, where earliest reports of the epidemic occurred. RAPD analysis could easily distinguish the isolates of F. oxysporum f. sp. erythroxyli from the nonpathogenic isolates of F. oxysporum from E. coca var. coca, indicating its utility in DNA fingerprinting.

Hyphal Elongation of Glomus fasciculatus in Response to Root Exudates.

The spore germination rates on water agar of the vesicular-arbuscular mycorrhizal fungus Glomus fasciculatus were highest at water potentials of -4 to -6 bars. Root exudates from plants grown in a sterile nutrient solution, with or without phosphorus, did not affect germination. Root exudates collected from 2-, 4-, and 6-week-old Trifolium repens cv. ;Ladino' seedlings that were deprived of P enabled hyphal growth from germinated Glomus fasciculatus spores of 21.4, 14.7, and 7.6 mm, respectively. Hyphal elongation in the presence of exudates from plants grown with P, or in the absence of exudates, was negligible (<1 mm). Root P at 2 weeks was not significantly different between plants grown with and without P. There were no significant differences between the quantities of exudates from plants grown with or without P at 2, 4, and 6 weeks. The data suggest that it is the quality of exudates from plants experiencing P deprivation that is important in stimulating vesicular-arbuscular mycorrhizal hyphal elongation.

Effect of clindamycin on the in vitro activity of amikacin and gentamicin against gram-negative bacilli.

The in vitro effect of clindamycin on the inhibitory and bactericidal activity of amikacin (BB-K8) and gentamicin against Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa was examined by the checkerboard technique in microtiter plates. Clindamycin (15 mug/ml) produced statistically significant increases in the minimal bactericidal concentrations of amikacin and gentamicin against E. coli and Klebsiellae at 2 and 4 h of incubation. The minimal bactericidal concentration against P. aeruginosa was not affected. Higher concentrations of clindamycin (20 to 25 mug/ml) reduced the minimal inhibitory and bactericidal concentrations of amikacin and gentamicin for E. coli at 18 h of incubation. The synergistic bactericidal activity of amikacin and carbenicillin against E. coli, but not P. aeruginosa, was also inhibited slightly by clindamycin (15 mug/ml). The clinical implications of this inhibition of the early bactericidal in vitro activity of aminoglycosides by clindamycin remain to be determined. Although these in vitro results have not been studied in clinical infections, it is conceivable that slight interference in early bacterial killing could alter the outcome of infection in the immunosuppressed patient.