Verticillium Wilt Evaluation of Olive Breeding Selections Under Semi-Controlled Conditions.

Genetic resistance is the most recommended measure to control verticillium wilt in olive (VWO), a vascular disease caused by the soil-borne fungus Verticillium dahliae, which has promoted the development of olive breeding programs aimed at obtaining new resistant and highly yielding cultivars in recent years. Screening has been commonly performed under controlled conditions in grow chamber after artificial inoculation during the early stage of breeding programs, but additional evaluation is necessary to confirm previous results as well as to test for additional agronomic traits. During this study, 20 breeding selections initially classified as resistant to the disease have been re-evaluated in artificially infested soils under natural environmental conditions. The maximum disease incidence (52.6%) was reached at 26 months after planting, and the disease intensity index reached the maximum value of 38.5% at 29 months after planting. Nine breeding selections consistently confirmed the previous results regarding resistance to V. dahliae infection; however, contradictory results, compared with those of previous evaluations under controlled conditions in grow chambers, were obtained for the rest of selections tested, thereby underlining the need for long-term experimentation under natural environmental conditions. Additional positive agronomic traits, such as early bearing, were also observed for some of the resistant selections, but plant vigor varied. Some seem highly promising for release as new cultivars when characterization of other important agronomic traits is completed in the future.

Spatiotemporal analysis of spread of infections by Verticillium dahliae pathotypes within a high tree density olive orchard in southern Spain.

The development of Verticillium wilt epidemics in olive cv. Arbequina was studied from November 1999 to May 2003 in a drip-irrigated, nontillage orchard established in a soil without a history of the disease at Córdoba, southern Spain. Disease incidence measured at 1-month-intervals increased from 0.2 to 7.8% during this period. Verticillium dahliae infecting the trees was characterized as defoliating (D) or nondefoliating (ND) pathotypes by a specific, multiplex-polymerase chain reaction (PCR) assay. Of the symptomatic trees, 87.2 and 12.8% were infected by the D or ND pathotypes, respectively. Dynamics of disease incidence were described by a generalized logistic model with a multiple sigmoid pattern. In the fitted model, the infection rate was highest in the winter to spring period and decreased to minimum values in the summer to fall period. Binary data of disease incidence was analyzed for point pattern and spatial correlation, either directly or after parsing them in contiguous quadrats. Overall, ordinary runs analysis indicated a departure from randomness of disease within rows. The binomial index of dispersion, interclass correlation, and Taylor's power law for various quadrat sizes suggested aggregation of diseased trees within the quadrat sizes tested. Spatial analysis by distance indices showed a nonrandom arrangement of quadrats containing infected trees. Spatial pattern was characterized by the occurrence of several clusters of infected trees. Increasing clustering over time was generally suggested by stronger values of clustering index over time and by the increase in the size of patch clusters. Significant spatial association was found in the clustering of diseased trees over time across cropping seasons; however, clustering was significant only for infections by D V. dahliae, indicating that infections by the D pathotype were aggregated around initial infections. The number and size of clusters of D V. dahliae-infected trees increased over time. Microsatellite-primed PCR assays of a representative number of V. dahliae isolates from diseased trees indicated that the majority of infecting D isolates shared the fingerprinting profile with D V. dahliae isolated from soil of a naturally infested cotton field in close proximity to the orchard, suggesting that short distance dispersal of the pathogen from this soil to the olive orchard may have occurred.

Simultaneous Detection of the Defoliating and Nondefoliating Verticillium dahliae Pathotypes in Infected Olive Plants by Duplex, Nested Polymerase Chain Reaction.

Pathogen-free certified planting material and accurate detection of Verticillium dahliae pathotypes infecting the plant are key components of successful management of Verticillium wilt of olive. Use of a nested-polymerase chain reaction (PCR) procedure developed in earlier studies for in planta detection of the defoliating (D) and nondefoliating (ND) V. dahliae pathotypes resulted in ambiguous detection of the pathogen in some cases, due to heterologous amplification of the D-associated marker in ND-infected olive plants. In the present study, an improved procedure was developed that eliminates ambiguity and reduces time and cost for detection of D and ND V. dahliae in olive. The improved procedure is based on the simultaneous amplification of both an ND- and a new D-specific marker by means of duplex, nested PCR. The procedure was effective in the rapid and unequivocal detection of the D and ND V. dahliae in both artificially inoculated, own-rooted olive plants and naturally infected adult olive trees of different cultivar, age, and growing conditions. Furthermore, the duplex, nested-PCR procedure detected simultaneously the D and ND pathotypes in adult olive trees naturally infected by both pathotypes and in young olive plants that were double-inoculated with D and ND isolates under controlled conditions.