ABSTRACT
The genetic diversity of soil-borne populations of Fusarium oxysporum was assessed using 350 isolates collected from six different French soils. All isolates were characterised by restriction fragment analysis of the PCR-amplified ribosomal intergenic spacer (IGS). Twenty-six IGS types were identified among the 350 isolates analysed. Five to nine different IGS types were detected in each soil. None of the IGS types was common to all of the soils. An analysis of the molecular variance based on IGS type relationships and frequency revealed that the genetic structure of the populations of F. oxysporum varied widely among the soils. Some populations were both highly diverse within the soils and differentiated between the soils. A possible relationship between the intrapopulation or interpopulation level of diversity and some external factors such as the soil type or the crop history was evaluated. A subsample representative of the diversity of the six populations was further characterised by analysing the genomic distribution of two transposable elements, impala and Fot1. One to 10 copies of the impala element were present in most of the isolates, irrespective of their soil of origin. The Fot1 element was only detected in 40% of the isolates originating from the three populations less diverse in terms of IGS types, but in 82.6% of the isolates originating from the three more diverse populations.
ABSTRACT
ABSTRACT The ability of transposon impala to inactivate genes involved in pathogenicity was tested in Fusarium oxysporum f. sp. melonis. Somatic excision of an impala copy inserted in the nitrate reductase-encoding niaD gene was positively selected through a phenotypic assay based on the restoration of nitrate reductase activity. Independent excision events were analyzed molecularly and shown to carry reinsertedimpala in more than 70% of the cases. Mapping of reinserted impala elements on large NotI-restriction fragments showed that impala transposes randomly. By screening 746 revertants on plants, a high proportion (3.5%) of mutants impaired in their pathogenic potential was recovered. According to the kinetics of wilt symptom development, the strains that were impaired in pathogenicity were clustered in three classes: class 1 grouped two strains that never induced Fusarium wilt symptoms on the host plant; class 2 and class 3 grouped 15 and 9 revertants which caused symptoms more than 50 and 30 days after inoculation, respectively. The first results demonstrate the efficiency of transposition in generating mutants affected in pathogenicity, which are usually difficult to obtain by classical mutagenesis, and open the possibility to clone the altered genes with impala as a tag.
ABSTRACT
ABSTRACT The effect of the plant on the diversity of soilborne populations of Fusarium oxysporum was assessed after successive cultures of flax, melon, tomato, and wheat in separate samples of the same soil. Forty soil-borne isolates of F. oxysporum and forty root-colonizing isolates of each plant species were sampled during the first (T0) and fourth (T1) cultures. The population structures were assessed by a genotypic method based on restriction fragment analysis of polymerase chain reaction-amplified ri-bosomal intergenic spacer (IGS) DNA. Sixteen IGS types were defined among the four hundred isolates analyzed. The distributions of soil isolates among IGS types were similar at both sampling times. The structure of F. oxysporum populations associated with the roots of flax or melon did not differ from the structure of soilborne populations. In contrast, the structure of F. oxysporum populations associated with roots of wheat or tomato differed from the structure of soilborne populations. High frequencies were found for IGS type 4 among wheat isolates at both T0 and T1 and for IGS type 11 among tomato isolates at T1. Moreover, a high level of genetic divergence was obtained between IGS types 4 and 11. Our results suggest that tomato and wheat have a selective effect on soilborne populations of F. oxysporum and that this effect seems to be plant specific.
