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.     

Epidemiology of sharka disease in Spain

PPV was first detected in Spain in 1984 in Japanese plum ( Prunus salicina Lindl) cv. Red Beaut and spread very quickly to other Japanese and European plums and apricot cultivars but left peach cultivars unaffected. In the years following the detection of PPV, the predominant aphid species visiting Prunus orchards in Mediterranean areas were Aphis gossypii followed by Aphis spiraecola , the latter being the main aphid species found at present. Both species are considered to be the main vectors of PPV in Spanish early Prunus growing areas. Spatial analysis of the spread of PPV-D in Japanese plum and apricot trees confirmed the lack of significant association between immediately adjacent trees. The observed spatial pattern of sharka suggests a lack of movement of PPV-viruliferous aphid vectors to immediately adjacent trees and indicates their preferential movement to trees several tree spaces away. PPV-D is the only type currently present in Spain, with the exception of a PPV-M outbreak that was detected in and successfully eradicated from Aragón in 2002. The short-distance spread of PPV-M infection occurred as far as 12 m along the rows of peach trees. However, PPV-D has not been observed to spread through peach cultivars, despite being grown in the vicinity of heavily infected plots of apricot or Japanese plum trees.     

Spatial pattern analysis of citrus canker-infected plantings in são paulo, Brazil, and augmentation of infection elicited by the asian leafminer

ABSTRACT Eradication of Asiatic citrus canker (ACC) has become increasingly difficult over the last decade, following the introduction of the Asian leafminer into Brazil and Florida, which has led to changes in the eradication protocols. The present study, undertaken in Brazil, was aimed at characterizing the spatial patterns of ACC in commercial citrus plantings to gain better understanding of the dynamics of the disease subsequent to introduction of the leafminer. The spatial patterns of ACC were mapped in 326 commercial citrus plantings and statistically assessed at various spatial dimensions. The presence of "within-group" aggregation in each plot was examined via beta-binomial analysis for groups of trees parsed into three-by-three-tree quadrats. The relative intensity of aggregation was expressed as a binomial index of dispersion (D) and heterogeneity among plots expressed as the intracluster correlation coefficient, rho. The population of data sets was found to fall into three D categories, D < 1.3, 1.3 3.5. These categories then were related to other spatial characteristics. The binary form of Taylor's power law was used to assess the overdispersion of disease across plots and was highly significant. When the overall population of plots was parsed into D categories, the Taylor's R (2) improved in all cases. Although these methods assessed aggregation well, they do not give information on the number of foci or aggregations within each plot. Therefore, the number of foci per 1,000 trees was quantified and found to relate directly to the D categories. The lowest D category could be explained by a linear relationship of number of foci versus disease incidence, whereas the higher two categories were most easily explained by a generalized beta function for the same relationship. Spatial autocorrelation then was used to examine the spatial relationships "among groups" composed of three-by-three-tree quadrats and determine common distances between these groups of ACC-infected trees. Aggregation was found in >84% of cases at this spatial level and there was a direct relationship between increasing D category and increasing core cluster size, and aggregation at the among-group spatial hierarchy was generally stronger for the within-row than for the across-row orientation. Clusters of disease were estimated to average between 18 and 33 tree spaces apart, and the presence of multiple foci of infection was commonplace. The effectiveness of the eradication protocol of removing all "exposed" trees within 30 m surrounding each "ACC-infected tree" was examined, and the distance of subsequent infected trees beyond this 30-m zone from the original focal infected tree was measured for each plot. A frequency distribution was compiled over all plots to describe the distance that would have been needed to circumscribe all of these outliers as a theoretical alternative protocol to the 30-m eradication protocol. The frequency distribution was well described by a monomolecular model (R(2) = 0.98) and used to determine that 90, 95, and 99% of all newly infected trees occurred within 296, 396, and 623 m of prior-infected trees in commercial citrus plantings, respectively. These distances are very similar to previously reported distances determined for ACC in residential settings in Florida.     

Spatial patterns of the Citrus leprosis virus and its associated mite vector in systems without intervention

Leprosis is caused by the Citrus leprosis virus cytoplasmic type and is vectored by the mite Brevipalpus yothersi. Miticide applications, which cost $54 million annually, are based on inspection for the presence of mites. The aim of the present study was to characterize the spatial patterns of B. yothersi-infested trees and trees with leprosis symptoms for further improvement in sampling and disease control. The presence of mites and the occurrence of leprosis were assessed over two years in 1160 Valencia trees and 720 Natal trees in a commercial sweet orange grove in Sao Paulo State, Brazil. To assess the natural growth and dispersal of mites and leprosis, mite populations were not controlled during the experimental period. Maps of mite-infested trees and trees with leprosis symptoms were analysed at three different levels of spatial hierarchy using complementary methods, i.e. among adjacent trees within and across rows, within quadrats, and the strength and orientation of aggregation among quadrats. The study showed that the spatial patterns of virus-infected and mite-infested trees were different, with a strong aggregation pattern of trees with leprosis symptoms that increased over time. Conversely, the spatial pattern of B. yothersi showed randomness or weak aggregation at all three spatial hierarchical levels. Disease incidence increased steadily in plots of both cultivars, unlike in mite-infested trees where incidence fluctuated over time. These results have important implications for the development of better management strategies for leprosis. Sampling methods and action thresholds for mite control should consider primary disease inoculum in addition to the incidence of mites.     

Spatial and temporal analyses of citrus sudden death as a tool to generate hypotheses concerning its etiology

ABSTRACT Citrus sudden death (CSD), a new disease of unknown etiology that affects sweet orange grafted on Rangpur lime, was visually monitored for 14 months in 41 groves in Brazil. Ordinary runs analysis of CSD-symptomatic trees indicated a departure from randomness of symptomatic trees status among immediately adjacent trees mainly within rows. The binomial index of dispersion (D) and the intraclass correlation (k) for various quadrat sizes suggested aggregation of CSD-symptomatic trees for almost all plots within the quadrat sizes tested. Estimated parameters of the binary form of Taylor's power law provided an overall measure of aggregation of CSD-symptomatic trees for all quadrat sizes tested. Aggregation in each plot was dependent on disease incidence. Spatial autocorrelation analysis of proximity patterns suggested that aggregation often existed among quadrats of various sizes up to three lag distances; however, significant lag positions discontinuous from main proximity patterns were rare, indicating a lack of spatial association among discrete foci. Some asymmetry was also detected for some spatial autocorrelation proximity patterns, indicating that within-row versus across-row distributions are not necessarily equivalent. These results were interpreted to mean that the cause of the disease was most likely biotic and its dissemination was common within a local area of influence that extended to approximately six trees in all directions, including adjacent trees. Where asymmetry was indicated, this area of influence was somewhat elliptical. Longer-distance patterns were not detected within the confines of the plot sizes tested. Annual rates of CSD progress based on the Gompertz model ranged from 0.37 to 2.02. Numerous similarities were found between the spatial patterns of CSD and Citrus tristeza virus (CTV) described in the literature, both in the presence of the aphid vector, Toxoptera citricida. CSD differs from CTV in that symptoms occur in sweet orange grafted on Rangpur lime. Based on the symptoms of CSD and on its spatial and temporal patterns, our hypothesis is that CSD may be caused by a similar but undescribed pathogen such as a virus and probably vectored by insects such as aphids by similar spatial processes to those affecting CTV.     

Increase and Patterns of Spread of Citrus Tristeza Virus Infections in Costa Rica and the Dominican Republic in the Presence of the Brown Citrus Aphid, Toxoptera citricida

ABSTRACT Citrus tristeza virus (CTV) was monitored for 4 years by monoclonal antibody probes via enzyme-linked immunosorbent assay in four citrus orchards in northern Costa Rica and four orchards in the Dominican Republic following the introduction of the brown citrus aphid, Toxoptera citricida. The Gompertz nonlinear model was selected as the most appropriate in most cases to describe temporal increase of CTV. Ordinary runs analysis for association of CTV-positive trees failed to show a spatial relationship of virus status among immediately adjacent trees within or across rows. The beta-binomial index of dispersion for various quadrat sizes suggested aggregations of CTV-positive trees for all plots within the quadrat sizes tested. Spatial autocorrelation analysis of proximity patterns suggested that aggregation often existed among quadrats of various sizes up to four lag distances; however, significant lag positions discontinuous from the main proximity pattern were rare. Some asymmetry was also detected for some spatial autocorrelation proximity patterns. These results were interpreted to mean that, although CTV-positive trees did not often influence immediately adjacent trees, virus transmission was common within a local area of influence that extended two to eight trees in all directions. Where asymmetry was indicated, this area of influence was somewhat elliptical. The spatial and temporal analyses gave some insight into possible underlying processes of CTV spread in the presence of T. citricida and suggested CTV spread was predominantly to trees within a local area. Patterns of longer-distance spread were not detected within the confines of the plot sizes tested. Longer-distance spread probably exists, but may well be of a complexity beyond the detection ability of the spatial analysis methods employed, or perhaps is on a scale larger than the dimensions of the plots studied.     

Epidemiology of sharka disease in France

Plum pox virus was first detected in France in the 1960s. Both PPV-D and PPV-M strains are present but epidemics related to the PPV-M strain detected in the late 1980s are the most problematic. The two PPV strains have unequal distributions in peach and apricot orchards and different prevalences. More than 20 different aphid species have been identified as vectors of PPV but most of them do not colonize Prunus species. Thus, aphids involved in the spread of PPV in orchards are essentially visiting aphids. The main sources of inoculum for the vectors are leaves and fruits of infected stone-fruit trees. Spontaneous, wild and ornamental Prunus species such as Prunus dulcis , P. spinosa or P. pissardii are susceptible to PPV isolates found in France but their role as a reservoir in sharka epidemics is probably negligible. The disease spreads rapidly in orchards but the rate of progression may vary according to the identity of the PPV strain and the Prunus species. Analysis of spatial patterns of disease has shown that secondary spread by aphids frequently occurs over short distances in the orchards (aggregated patterns) but also that dissemination at longer distances (of several hundred metres) is a common event.     

Factors Affecting the Spread of Plum pox virus Strain M in Peach Orchards Subjected to Roguing in France

ABSTRACT We evaluated the impact of roguing on the spread and persistence of the aggressive Plum pox virus strain M (PPV-M) in 19 peach orchard blocks in Southern France. During a 7- to 10-year period, orchards were visually inspected for PPV symptoms, and symptomatic trees were removed every year. Disease incidence was low in all orchards at disease discovery and was <1% in 16 of the 19 orchard blocks. The spread of Sharka disease was limited in all 19 blocks, with an annual disease incidence between 2 and 6%. However, new symptomatic trees were continuously detected, even after 7 to 10 years of uninterrupted control measures. An extended Cox model was developed to evaluate to what extent tree location, orchard characteristics, environment, and disease status within the vicinity influenced the risk of infection through time. Eleven variables with potential effect on tree survival (i.e., maintenance of a tree in a disease- free status through time) were selected from survey data and databases created using a geographical information system. Area of the orchard, density of planting, distance of a tree from the edge of the orchard block sharing a boundary with another diseased orchard, and distance to the nearest previously detected symptomatic tree had a significant effect on the risk for a tree to become infected through time. The combined results of this study suggest that new PPV-M infections within orchards subjected to roguing resulted from exogenous sources of inoculum, disease development of latent infected trees, as well as infected trees overlooked within the orchards during visual surveys. A revision of the survey and the roguing procedures used for more effective removal of potential sources of inoculum within the orchards and in the vicinity of the orchards would improve disease control suppression of PPV.     

Resistance to plum pox potyvirus in apricots1

The susceptibility of 115 apricot cultivars to plum pox potyvirus (PPV) has been examined, since 1981, in the experimental plots of the Pomology Institute at Naoussa and Skydra, Makedonia (GR). Inoculation was assured by aphids, transmitting strain PPV-M (Marcus) from naturally infected trees in adjacent peach orchards. For each cultivar, four to six trees were examined for at least 4 years. Observations on symptoms were made on leaves early in May and on fruits at maturity. Most cultivars expressed severe disease symptoms. Those without symptoms were inoculated by grafting onto heavily infected old apricot trees. The grafted shoots were tested for PPV in the following year by ELISA and on the woody indicator GF305. The cultivars which were rated as resistant after artificial inoculation and ELISA came from North America: Early Orange, Stella, NJA2, Sunglo, Veecot, Harlayne, Goldrich and Henderson. Most of these have been crossed with quality cultivars for the creation of resistant hybrids. The PPV resistance of large numbers of these apricot hybrids is now under investigation.     

Characterizing the spatial pattern of Abutilon theophrasti seedling patches

Spatial biology of weed populations is the study of weed patches and their relevant patch-level processes. In this context, a patch was defined as an area in which individuals are aggregated into discrete subdivided populations. Four Abutilon theophrasti seedling patches in two continuous maize production fields were surveyed using a contiguous grid of quadrats between 1995 and 1997. Surveyed area was dependent on patch size and ranged from 96 m² to 1134 m². Within each area, all seedlings were counted in each 1 m × 0.75 m quadrat in June, just before post-emergence weed control, and in mid-July after all weed control practices were completed. The spatial pattern observed in the seedling distribution maps was single or multiple focal points of high seedling density that decreased with distance from the focal point. Two-directional correlograms corroborated this visual observation, such that A. theophrasti seedling density in neighbouring quadrats was spatially autocorrelated, and correlation strength decreased with distance separating quadrats. Autocorrelation coefficients decreased at a greater rate across crop rows than parallel to crop rows. Visually, patch shape was elliptical and oriented in the direction of field traffic. Factors affecting patch-level processes of spatial aggregation, stability and edge dynamics were considered.     

Spatial patterns of leprosis and its mite vector in commercial citrus groves in Brazil

In order to describe the dissemination of Citrus leprosis virus (CiLV) and Brevipalpus phoenicis , the Tenuipalpidae mite which transmits it, as well as to generate data for future development of better sampling and disease control procedures, analyses were conducted at three levels of spatial hierarchy. Over 100 distribution maps of mite-infested plants and plants with CiLV symptoms were constructed after evaluation of 174 sweet orange commercial grove blocks from four citrus regions of São Paulo State, Brazil. Spatial correlation between maps of mite-infested plants and those with CiLV symptoms was very low and mostly not significant. Spatial dependency between adjacent plants was incipient as ordinary runs analysis showed that percentages of aggregated sequences within or across rows were very low for both mite-infested and diseased plants. Index of dispersion ( D ) values for all quadrat sizes suggested aggregation of plants with symptoms within quadrats, but much less aggregation for mite-infested plants. Values of log( A ) and b were higher than 0 and 1, respectively, indicating a general and significant aggregation of infested/diseased plants inside quadrats. However, the degree of aggregation of plants with symptoms was higher than that of infested plants. Aggregation in each grove was positively correlated to the incidence of infested/diseased plants. Spatial autocorrelation also showed a higher strength of aggregation for plants with symptoms than for infested ones. This is the first time that spatial patterns of leprosis and B. phoenicis have been described.     

Spatial Patterns of Microsclerotia of Verticillium dahliae in Soil and Verticillium Wilt of Cauliflower

ABSTRACT The spatial patterns of microsclerotia of Verticillium dahliae in soil and wilt symptoms on cauliflower were determined at three sites in each of two fields in 1994 and 1995. Each site was an 8 x 8 grid divided into 64 contiguous quadrats (2 by 2 m each). Soil samples were collected to a depth of 15 cm with a probe (2.5 cm in diameter), and samples from four sites in each quadrat were bulked. Plants in each quadrat were cut transversely, and the number of plants with vascular discoloration and the number without discoloration were recorded. The soil was assayed for microsclerotia by the modified Anderson sampler technique. Lloyd's index of patchiness (LIP) was used as an indicator to evaluate the aggregation of microsclerotia in the field. Spatial autocorrelation and geostatistical analyses were also used to assess the autocorrelation of microsclerotia among quadrats. The LIP for microsclerotia was greater than 1, indicating aggregation of propagules; however, the degree of aggregation at most sites was not high. Significant autocorrelation within or across rows was detected in some spatial autocorrelograms of propagules, and anisotropic patterns were also detected in some oriented semivariograms from geostatistical analyses for microsclerotia, indicating the influence of bed preparation in the fields on pathogen distribution. The parameter estimates p and theta in the beta-binomial distribution and the index of dispersion (D) associated with the distribution were used to assess the aggregation of diseased plants at each site. A random pattern of wilt incidence was detected at 7 of 12 sites, and an aggregated pattern was detected at 5 of 12 sites. The degree of aggregation was not high. A regular pattern of wilt severity was detected at all sites. The high disease incidence (77 to 98%) observed at 11 of the 12 sites could be explained by high inoculum density.     

Analysis of the Spatial Patterns of Pierce's Disease Incidence in the Lower San Joaquin Valley in California

ABSTRACT The incidence of Pierce's disease (PD), caused by Xylella fastidiosa, was monitored in 11 naturally infested commercial vineyards to determine the presence of an X. fastidiosa vector, Homalodisca coagulata (glassy-winged sharpshooter [GWSS]), to examine the spatial patterns of the disease and elucidate possible influences of surrounding environments. Disease incidence ranged from <1 to 65% among vineyards in 2001. Our efforts to trap or capture potential disease vectors have indicated that the GWSS is the most likely vector. Disease incidence doubled in most vineyards during the 2002 production season. Spatial patterns of symptomatic vines in 2001 and 2002, as determined by ordinary runs analysis, showed strong evidence for within- and across-row aggregation of infected vines. In most fields, they were no disease gradients observed relative to GWSS source (e.g., citrus). Within fields, however, disease incidence displayed strong spatial dependence and a high degree of anisotropy, indicating strongly aggregated patterns of disease with distinct directional orientation. The within-row (0 degrees ) and across-row (90 degrees ) orientations generally were the predominant directions of increased disease incidence, consistent with vine-to-vine spread of X. fastidiosa. We concluded that the distribution of PD in vineyards reflected the feeding pattern of vectors carrying X. fastidiosa. Based on these results, effective PD management is likely to be based on practices that reduce significant insect vector populations and remove infected vines as soon as identified and on the use of resistant cultivars.     

Development of an on-site plum pox virus detection kit based on immunochromatography

Sharka disease, caused by plum pox virus (PPV), is the most serious viral disease of stone fruit trees. Among the eight known strains of the virus, PPV-D is the most important due to its recent global spread. Although enzyme-linked immunosorbent assay (ELISA) is the most common approach for diagnosing sharka, it involves time-consuming steps and requires expensive equipment and trained technicians. In this study, an on-site PPV detection kit based on immunochromatography was developed using polyclonal antibodies against the coat protein (CP) of a PPV-D isolate. The immunochromatographic (IC) assay kit was as sensitive as a commercial ELISA system for detecting Japanese PPV-D isolates. Moreover, it was easy to use (a one-step procedure), and results could be obtained on-site within 15 min without special laboratory equipment. The IC assay kit detected the virus from every aerial part of symptomatic Japanese apricot trees. In a detailed study of viral localization in leaves, the most suitable plant parts for use in the IC assay were symptomatic mesophyll tissues and the region from the petiole to the main vein. A positive reaction was also observed using the CP of other major (PPV-M and PPV-Rec) and minor (PPV-EA, PPV-W, and PPV-T) strains.     

Spatio-temporal patterns of pre-harvest brown rot epidemics within individual peach tree canopies

Tree canopies are architecturally complex and pose several challenges for measuring and characterizing spatial patterns of disease. Recently developed methods for fine-scale canopy mapping and three-dimensional spatial pattern analysis were applied in a 3-year study to characterize spatio-temporal development of pre-harvest brown rot of peach, caused by Monilinia fructicola, in 13 trees of different maturity classes. We observed a negative correlation between an index of disease aggregation and disease incidence in the same tree (r?=??0.653, P?<?0.0001), showing that trees with higher brown rot incidence had lower aggregation of affected fruit in their canopies. Significant (P?≤?0.05) within-canopy aggregation among symptomatic fruit was most pronounced for early-maturing cultivars and/or early in the epidemic. This is consistent with the notion of a greater importance of localized, within-tree sources of inoculum at the beginning of the epidemic. Four of five trees having >10 blossom blight symptoms per tree showed a significant positive spatial association of pre-harvest fruit rot to blossom blight within the same canopy. Spatial association analyses further revealed one of two outcomes for the association of new fruit rot symptoms with previous fruit rot symptoms in the same tree, whereby the relationship was either not significant or exhibited a significant negative association. In the latter scenario, the newly diseased fruit were farther apart from previously symptomatic fruit than expected by random chance. This unexpected result could have been due to uneven fruit ripening in different sectors of the canopy, which could have affected the timing of symptom development and thus led to negative spatial associations among symptoms developing over time in a tree.     

Control and monitoring: monitoring and eradication of sharka in south-east Italy over 15 years

When the first foci of sharka were discovered in Puglia region (south-east Italy) in the late 1980s, the regional agricultural authorities launched a programme for Plum pox virus (PPV) monitoring and disease eradication. The infecting virus strain was identified as PPV-D. From 1989 to 1993, a strong eradication campaign was successfully carried out involving 13 plum and 2 apricot orchards with different levels of infection. During 1994–2000, besides plum, apricot and peach, monitoring was extended to sweet cherry. At that time, surveys and testing did not reveal any new PPV focus, but the eradication of infected trees continued in a couple of orchards. In 2001–05, particular attention was paid to peach, as devastating PPV-M outbreaks had developed in other areas of the country. A new PPV focus was found in apricot, caused by PPV-Rec, which was promptly eradicated. In the following two years, surveys in the once infected orchard and surrounding peach plantings did not detect any virus spread. The endeavour has taken 15 years making this PPV monitoring and eradication programme the longest in Italy. Its overall results indicate that the fruit tree industry in Puglia region can now be regarded as essentially PPV-free.     

Spatial pattern analysis of hop powdery mildew in the pacific northwest: implications for sampling

ABSTRACT The spatial pattern of hop powdery mildew was characterized using 3 years of disease incidence data collected in commercial hop yards in the Pacific Northwest. Yards were selected randomly from yards with a history of powdery mildew, and two to five rows were selected for sampling within each yard. The proportion of symptomatic leaves out of 10 was determined from each of N sampling units in a row. The binomial and the beta-binomial frequency distributions were fit to the N sampling units observed in each row and to SigmaN sampling units observed in each yard. Distributional analyses indicated that disease incidence was better characterized by the beta-binomial than the binomial distribution in 25 and 47% of the data sets at the row and yard scales, respectively, according to a log-likelihood ratio test. Median values of the beta-binomial parameter theta, a measure of small-scale aggregation, were near 0 at both sampling scales, indicating that disease incidence was close to being randomly distributed. The variability in disease incidence among rows sampled in the same yard generally increased with mean incidence at the yard scale. Spatial autocorrelation analysis, used to measure large-scale patterns of aggregation, indicated that disease incidence was not correlated between sampling units over several lag distances. Results of a covariance analysis showed that heterogeneity of disease incidence was not dependent upon cultivar, region, or time of year when sampling was conducted. A hierarchical analysis showed that disease incidence at the sampling unit scale (proportion of sampling units with one or more diseased leaves) increased as a saturation-type curve with respect to incidence at the leaf level and could be described by a binomial function modified to account for the effects of heterogeneity through an effective sample size. Use of these models permits sampling at the sampling unit scale while allowing inferences to be made at the leaf scale. Taken together, hop powdery mildew was nearly randomly distributed with no discernable foci, suggesting epidemics are initiated from a well-distributed or readily dispersible overwintering population. Implications for sampling are discussed.     

Diversity of Plum pox virus isolates in Bosnia and Herzegovina

Sixteen Plum pox virus (PPV) isolates from several stone fruit cultivars, host species, orchards and geographical areas of Bosnia and Herzegovina were selected for typing, using serotype-specific monoclonal antibodies (MAbs) and PCR–RFLP, targeting the 3' terminal region of the coat protein (CP) and P3-6K₁ with restriction enzymes Rsa I and Dde I. Four PPV isolates were identified as PPV-M by serology and PCR; eight isolates were identified as PPV-D based on PCR–RFLP on both genomic regions, but were not recognized by the D-specific MAb4DG5. Four isolates from plum were identified as natural D/M recombinants (PPV-Rec), based on conflicting results of CP and P3-6K₁ typing. To investigate the genetic diversity of Bosnian PPV isolates in more detail, five isolates (three PPV-Rec, one PPV-M and one PPV-D) were partially sequenced in the region spanning the 3' terminal part of the NIb gene and the 5'-terminal part of the CP gene, corresponding to nucleotides 8056–8884. Nucleotide sequence alignment of recombinant isolates showed that they were closely related at the molecular level to previously characterized recombinants from other European countries, and shared the same recombination break point in the 3' terminal part of the NIb gene. This is the first report of naturally infected Prunus trees with PPV-M, PPV-D and PPV-Rec in Bosnia and Herzegovina. The high variability of the Bosnian PPV isolates fits with the presence of this virus in the country over a long period.     

Use of SADIE statistics to study spatial dynamics of plant disease epidemics

Using a previously developed stochastic simulation model for plant disease epidemics, the relationship of the SADIE aggregation statistic I _a with initial epidemic conditions, spore dispersal distance, sampling quadrat size and other spatial statistics was investigated. Most variation in I _a was attributable to the initial spatial pattern of infected plants and sampling quadrat size. The importance of initial spatial pattern on SADIE clustering indices (for patches and gaps) was also demonstrated using a number of selected data sets. Correlation of I _a with clustering indices was close to 1·0. Epidemics arising from the regular and random initial patterns resulted in the smallest and greatest I _a values, respectively, at sampling times after disease spread had occurred. Furthermore, the variability in I _a between simulation runs also varied greatly with initial patterns, being lowest and greatest for the clumped and random initial patterns, respectively. I _a increased initially and then decreased with increasing incidence, especially for the clumped and random initial patterns. Overall, the effect of median spore dispersal distance on I _a was very small, especially for the random initial pattern. The correlation between I _a and intraclass correlation was generally small and varied greatly between initial patterns. However, there was a high positive correlation between I _a and a parameter describing the rate of decline of autocorrelation over spatial lags, indicating that I _a, clustering indices and autocorrelations measure some common properties of patterns.