Genetic Structure of Heterobasidion annosum in White Fir Mortality Centers in California

ABSTRACT The structure of Heterobasidion annosum populations was studied in 15 mixed-conifer sites in central and northern California. Study sites displayed mortality of white fir trees in enlarging discrete patches (mortality centers). At each site, fungal genotypes were defined by somatic compatibility tests. In two sites, further genetic and molecular analyses were performed on field genotypes and on homokaryons obtained by dedikaryotization of field heterokaryons. Isolates were found to be colonizing mostly the roots and the bole sapwood of white fir trees, and no significant infections of other tree species were observed. Each mortality center was characterized by the presence of several fungal genotypes, all belonging to the S intersterility group. Both homokaryotic and heterokaryotic strains were present in all sites. Multiple genotypes were retrieved in individual trees or stumps. Out of 228 fungal genotypes, 86% were found only within a single tree or stump, while 14% had spread to adjacent trees. The two largest genotypes had diameters of 9 and 10 m, and had colonized five and nine trees, stumps, or both, respectively. The maximum distance between two adjacent trees colonized by the same genotype was 6 m, and a highly significant correlation was found between tree diameter and distance of fungal "vegetative" spread. The largest clones were found in areas characterized by high tree and stump densities, and secondary spread of the fungus was more significant in denser stands. In most cases, original infection courts of existing genotypes could be traced to standing trees and not to stumps. The genetic analysis performed in two mortality centers revealed that most local genotypes had different mating alleles, and thus originated from unrelated basidiospores. In a few cases, the same mating allele was shared by two heterokaryons (n+n genome) or by a homokaryon (n genome) and a heterokaryon. Molecular analysis showed that nuclei bearing the same mating allele were identical, providing evidence that the two nuclei forming heterokaryons can act independently in the field and can be shared among isolates, presumably via di-mon mating or by separate matings of different portions of widespread homokaryons.     

Importance of secondary inoculum of Plasmopara viticola to epidemics of grapevine downy mildew

To quantify the magnitude and the spatial spread of grapevine downy mildew secondary sporangia, 4685 Plasmopara viticola single lesion samples were collected from 18 plots spread across central Europe. Disease symptoms were collected on two to 22 sampling dates per plot between 2000 and 2002. Four multiallelic microsatellite markers were used for genotypic identification of pathogen samples. Genetic analysis showed more than 2300 site-specific P. viticola genotypes, indicating that populations are genetically rich demographic units. Approximately 70% of the genotypes were sampled once and 14% were sampled twice throughout the various epidemics. In the 18 populations only seven genotypes (0.3%) were identified more than 50 times. Three genotypes particularly successful in causing disease through secondary cycles showed mainly a clustered distribution. The distance of sporangial migration per secondary cycle was less than 20 m and their plot colonization rate was calculated at around 1–2 m² day⁻¹. Downy mildew epidemics of grapevine are therefore the result of the interaction of a multitude of genotypes, each causing limited (or a few) lesions, and of a dominant genotype able to spread stepwise at plot-scale. These findings contrast with current theories about grapevine downy mildew epidemiology, which postulate that there is massive vineyard colonization by one genotype and long-distance migration of sporangia.     

Phytophthora austrocedrae emerges as a serious threat to juniper (Juniperus communis) in Britain

From 2011 to 2013, Phytophthora austrocedrae was isolated from diseased Juniperus communis exhibiting dieback and mortality at eight geographically separate sites in Scotland and northern England. The pathogen was also confirmed present either by standard PCR of the ITS locus and sequencing or by real‐time PCR on J. communis with symptoms at a further 11 sites in northern Britain. Out of 167 J. communis sampled across the 19 sites, 154 had foliage dieback over all or part of the crown as a result of basal lesions, which extended up the stem. Thirteen sampled trees had aerial branch lesions or discrete stem lesions with no apparent connection to the base of the tree. At 13 sites, dieback was concentrated in areas of poor drainage and/or alongside streams and other watercourses. In artificial inoculation experiments, P. austrocedrae caused rapidly extending stem and root lesions on J. communis and was reisolated from these lesions. Lesions also developed on Chamaecyparis lawsoniana and Chamaecyparis nootkatensis but the pathogen was not reisolated. All P. austrocedrae isolates obtained from J. communis in Britain shared 100% identity across the ITS locus but were distinct at one sequence position from P. austrocedrae isolates collected in Argentina from diseased Austrocedrus chilensis. This study provides clear evidence that P. austrocedrae is a primary pathogen of J. communis and now presents a significant threat to this species in Britain. Pathways for the emergence of P. austrocedrae in Britain, and possible ways in which the pathogen may have spread within the country, are discussed.     

Infection of roots of Dieffenbachia maculata by the foliar blight and soft rot pathogen, Erwinia chrysanthemi

Erwinia chrysanthemi, the organism causing foliage blight, leaf spotting, basal stem rot and root rot of Dieffenbachia maculata, was shown to be capable of infecting via roots. Inoculated plants were sectioned and internal spread of the pathogen determined by 'sandwich' plating between two layers of Miller's selective medium. Light microscopy and SEM confirmed that the pathogen was present throughout the plant in the xylem. Resin ducts, previously claimed as being the route for systemic infection, were not found. Initially confined to the xylem, the pathogen spread intercellularly to the neighbouring parenchyma and, after about 8 days, pockets of infection, often surrounded by periderm–like tissues, were seen.     

Variation in cultural characteristics, pathogenicity, vegetative compatibility and electrophoretic karyotype within field populations of Stagonospora nodorum

Isolates of Stagonospora nodorum , the cause of septoria leaf and glume blotch of wheat, were sampled intensively from single fields in two consecutive years and compared for growth rate, colony morphology, pathogenicity, vegetative compatibility and electrophoretic karyotype. The two populations were highly variable for all characters, even though the isolates in each population originated from a small geographical area. Studies of vegetative compatibility within each population indicated the presence of many genotypes and suggested that clonal spread was very limited. Individual lesions contained a single genotype and therefore, presumably, originated from a single unit of inoculum. However, adjacent lesions, even on the same leaf, frequently contained different genotypes. Ten isolates from population II possessed six different karyotypes, demonstrating that polymorphism for this fundamental feature of the genome occurs within field populations. The population structure revealed by these studies supports the hypothesis that ascospores play a major role in the epidemiology of the disease. On this basis, the observed variation in all the characters, including karyotype, can be explained by recombination during sexual reproduction.     

Long-term development of Heterobasidion annosum in basidiospore-infected Sitka spruce stumps

The extent of colonization by Heterobasidion annosum was measured in 1989 in Sitka spruce stumps that had been inoculated with basidiospores in 1981 and assessed for infection in 1983. Only stumps that had infection in the sapwood in 1983 contained the fungus in 1989. There was a significant positive correlation between the sapwood area colonized in 1983 and the total area colonized in 1989. Between 1983 and 1989, H. annosum spread within the body of stumps and into most primary roots. There were significant correlations between the stump area colonized in 1983 and 1989 and the percentage of stump and root volume colonized in 1989. Roots of 26 excavated stumps were in contact with an average of two roots on each of two living trees. Fourteen roots on eight living trees became infected, and H. annosum spread proximally an average of 126 cm into trees. We believe that this is the first report of the transfer of H. annosum from spruce stumps inoculated with basidiospores to adjacent live trees.     

Host switching between native and non‐native trees in a population of the canker pathogen Chrysoporthe cubensis from Colombia

The purpose of this study was to test the hypothesis that Chrysoporthe cubensis on native trees in South America could be the source of the pathogen that causes severe stem cankers and often mortality in commercially propagated Eucalyptus trees. This was done by investigating populations originating from two adjacent Eucalyptus (Myrtaceae) plantations in Colombia, and wild Miconia rubiginosa trees (Melastomataceae) growing alongside these stands. Polymorphic microsatellite markers were used to quantify allele sizes in 20 and 39 isolates from the two Eucalyptus stands and 32 isolates from adjacent M. rubiginosa trees. Gene and genotypic diversities were calculated from these data, and population differentiation and assignment tests were performed to ascertain whether the populations were genetically different. Results showed that there were no differences between any of the populations using these techniques, and that they can be treated as a single population. Therefore, the results support the hypothesis that host switching has occurred in C. cubensis in Colombia.     

Tracking Phytophthora infestans with SSR markers within and between seasons – a field study in Sweden

The dynamics of a late blight epidemic and sexual reproduction in Phytophthora infestans were studied in an experimental field in mid‐Sweden. The field was inoculated with six isolates of P. infestans taken from another potato field where sexual reproduction of the pathogen was suspected. Three weeks after inoculation single‐lesion leaflets were sampled and the resulting isolates characterized using microsatellites (SSRs) and mating type as markers. Among the 151 isolates analysed, the inoculum genotypes constituted more than 80% of the genotypes found, with three other genotypes making up the remainder. The following year, P. infestans obtained from soil samples taken from this field were analysed, and six novel genotypes were identified. Genotypes from the previous summer’s population were not detected. Analysis of the genotypes recovered was consistent with them being recombinants, with the previous summer’s population acting as parents. These findings are consistent with the hypothesis that oospores produced during a summer epidemic in Sweden can overwinter and cause infection the next year.     

Movement of genotypes of Ceratocystis fimbriata within and among Eucalyptus plantations in Brazil

Ceratocystis wilt on eucalyptus, caused by Ceratocystis fimbriata, was first recognized in 1997 in the state of Bahia, Brazil, but is now known in five other states and in four other countries. C. fimbriata is a native, soilborne pathogen in some parts of Brazil but we hypothesized that genotypes of the pathogen have been moved among plantations in rooted cuttings collected from diseased trees and within plantations on cutting tools. We used six microsatellite markers to identify 78 genotypes of C. fimbriata among 177 isolates from individual trees in 20 eucalyptus plantations. The highest gene and genotypic diversity values were found in plantations on formerly wild Cerrado forest in Minas Gerais, suggesting that the fungus was in the soil prior to planting eucalyptus. In contrast, one or only a few genotypes were found in plantations on previous pastureland (with no woody hosts) in Bahia and S?o Paulo, and most of these genotypes were found in a Bahian nursery or in one of two Bahian plantations that were sources for rooted cuttings. Sources of cuttings tended to be dominated by one or a few genotypes that may have been spread within the plantation on cutting tools.     

Population Dynamics of Fusarium Oxysporum f. Sp. Radicis-lycopersici in Relation to the Onset of Fusarium Crown and Root Rot of Tomato

Fusarium oxysporum f. sp. radicis-lycopersici the causal agent of crown and root rot in tomato comprises two overlapping separate phases: monocyclic and polycyclic. Oversummering inoculum is the source of primary infection (the monocyclic phase) and the spread from plant to plant via root-to-root contact is the source of the secondary infection (the polycyclic phase). In the present work, relationships between initial inoculum density, population dynamics of the pathogen in the root zone of diseased plants, and disease onset were studied. For the monocyclic phase, 55.1% of the variance of disease onset was attributed to the rate of pathogen proliferation in the root zone of plants, and only 12.8% of the variance was attributed to the amount of initial inoculum density. For the polycyclic phase, disease onset was not related to either initial inoculum density or the rate of pathogen proliferation in the root zone. At disease onset, the inoculum density of the pathogen in the root zone of plants infected from oversummering inoculum reached an average of 4.08 log cfu g soil^–1. The inoculum density of the pathogen in the root zone of plants infected by their diseased neighbors was 3.23 log cfu g soil^–1. A large variation in pathogen proliferation rate in the root zone was found among individual plants, suggesting that differences in the level of soil suppressiveness may occur not only between fields, but even in the same field over short distances.     

Characterization of Components of Partial Resistance, Rps2, and Root Resistance to Phytophthora sojae in Soybean

ABSTRACT Phytophthora root and stem rot of soybeans caused by Phytophthora sojae is a serious limitation to soybean production in the United States. Partial resistance to P. sojae in soybeans is effective against all the races of the pathogen and is a form of incomplete resistance in which the level of colonization of the root is reduced following inoculation. Other forms of incomplete resistance include the single dominant gene Rps2 and Ripley's root resistance, which are both race-specific. To differentiate partial resistance from the other types of incomplete resistance, the components lesion length, numbers of oospores, and infection frequency were measured in eight soybean genotypes inoculated with two P. sojae isolates. The Rps2 and root-resistant genotypes had significantly lower oospore production and infection frequency compared with the partially resistant genotype Conrad, while the root-resistant genotype also had significantly smaller lesion lengths. However, the high levels of partial resistance in Jack were indistinguishable from Rps2 in L76-1988, based on the evaluation of these components. Root resistance in Ripley and Rps2 in L76-1988 had similar responses for all components measured in this study. Partial resistance expressed in Conrad, Williams, Jack, and General was comprised of various components that interact for defense against P. sojae in the roots, and different levels of each component were found in each of the genotypes. However, forms of incomplete resistance expressed via single genes in Ripley and Rps2 in L76-1988, could not be distinguished from high levels of partial resistance based on lesion length, oospore production, and infection frequency.     

Disease Progression by Active Mycelial Growth and Biocontrol of Pythium ultimum var. ultimum Studied Using a Rhizobox System

ABSTRACT This study demonstrates that outward growth of mycelium from primary foci through bulk potting mix to roots of adjoining plants can be an important means of spread of damping-off and root rot caused by Pythium ultimum. The use of a rhizobox system, which confines plant roots, enabled us to study the spread of actively growing mycelium between root systems placed at precise distances from each other. In steamed potting mix, hyphae of P. ultimum on average grew 9.6 cm from diseased root tissue compared to 5.3 cm in raw potting mix. The density of mycelium was highest within the first 2 cm from the infected root tissue, decreasing with increasing distances from the roots. Accordingly, the disease on adjacent plants decreased as the distance from infected roots increased. The time required for damping-off of adjacent plants was 3 days slower in raw as compared to steamed potting mix and increased by 2 days for each additional centimeter between the rhizoboxes. The presence of Trichoderma harzianum diminished the production of secondary inoculum and reduced the ability of P. ultimum hyphae to extend through bulk potting mix. In conclusion, the concentration of the primary inoculum, the plant density, the distance separating diseased from healthy roots, the resident microflora, and the presence of an antagonist were shown to be important factors affecting disease spread by mycelial growth.     

Plane decline in European and Mediterranean countries: associated pests and their interactions

Surveys on the spread of the main pests of plane ( Platanus spp.) and on their influence on the decline of this genus in recent years have been carried out in various European countries. Of the primary pests which have been detected, Ceratocystis fimbriata f.sp. platani , agent of canker stain, has spread through several of the northern Mediterranean countries, causing death of trees; Gnomonia platani , agent of anthracnose, has spread mainly in the north-central Mediterranean areas; Microsphaera platani , agent of powdery mildew, has spread throughout almost all Mediterranean countries; so also has the insect Corythucha ciliata , which causes yellowing of the foliage. On trees declining under the interaction of these pests, secondary pests, agents of bark necrosis and, above all, of wood decay, are often detected. Ganoderma spp. are the main wood-decaying pathogen and can cause lethal root rots, especially if favoured by various kinds of stress. The trends and interaction of these pests in northern Italy are considered and control measures are discussed.     

Acacia plantations in Indonesia facilitate clonal spread of the root pathogen Ganoderma philippii

Ganoderma philippii is a root pathogen of many woody plants in tropical regions and is particularly aggressive to Acacia mangium, which is grown on a 6-year rotation for pulpwood in Indonesia. The disease becomes progressively worse over each rotation and control measures have met with limited success. We studied the population genetics of G. philippii to evaluate the role of sexual and asexual reproduction in its mode of spread. Populations were genetically distinct with high levels of inbreeding, and clonal spread to adjacent trees increased after the first rotation. Despite the high levels of genetic diversity seen at all sampling scales, migration rates appear low. Measures to reduce the underground spread of the pathogen as well as methods to prevent the initiation of new infections from basidiospores will be needed to reduce the incidence of root rot in A. mangium plantations.     

Infection and colonization of strawberry by <Emphasis Type="Italic">Gnomonia fragariae</Emphasis> strain expressing green fluorescent protein

Gnomonia fragariae is a poorly studied ascomycete, which was recently demonstrated to be a cause of severe root rot and petiole blight of strawberry. The pathogen was genetically transformed with the GFP as a vital marker and hygromycin resistance gene. Several stable transformants were obtained, which did not differ in their phenotype from the wild type isolate. Using one of the GFP-tagged isolates the infection process and colonization of roots and petioles of host plant by the pathogen were studied. Fluorescence microscopy examinations of the inoculated plants at different time points showed that plant infection occurs 24 h after inoculation and intensively continues during first 3 days. The specific penetration sites on epidermal cells and preferences in colonization for certain root and petiole tissues were observed. The pathogen intensively colonized and destroyed cortex of roots and petioles and spread rapidly longitudinally within intercellular spaces. The petioles were colonized by the hyphae, which grew mostly in the intracellular spaces of the cortical cells while in the roots the intracellular growth of hyphae occurred only in the later stages of infection. The fungus was also capable to infect the vascular tissues of petioles although these were not the primary tissues colonized by the pathogen. The mature ascomata were formed on the infected petiole bases several weeks after the inoculation. This study presents a genetic transformation method for Gnomonia fragariae and it demonstrates details on infection process and colonization of root, crown and petiole tissues of strawberry by the pathogen.     

Temporal and Spatial Spread of Soybean mosaic virus (SMV) in Soybeans Transformed with the Coat Protein Gene of SMV

ABSTRACT Soybean lines transformed with the coat protein (CP) gene of Soybean mosaic virus (SMV) were evaluated for SMV resistance by quantifying the temporal and spatial spread of SMV strain AL-5 released from a point source in the field. The temporal spread of SMV within field plots during 1999 and 2000 was quantified by enzyme-linked immunosorbent assay. The Gompertz model most appropriately described temporal spread. Two SMV CP transformed lines (genotypes) had significantly lower infection rates and significantly lower final SMV incidence values (P 相似文献   

Quantification of within-field spread of soybean mosaic virus in soybean using strain-specific monoclonal antibodies

ABSTRACT Strain-specific monoclonal antibodies were used to follow the temporal increase and spatial spread of soybean mosaic virus (SMV) strain G-5 released from a point source. The use of strain-specific monoclonal antibodies allowed discrimination of within-field temporal and spatial spread of SMV strain G-5 from non-G-5 SMV isolates that originated from exogenous field sources. SMV isolates originating from exogenous sources have potential to alter the temporal and spatial pattern of within-field virus spread, which could potentially affect the choice of models used to quantify within-field pathogen spread. Analysis of SMV epidemics in field-plot experiments indicated that the logistic model was the most appropriate model to describe and compare the temporal spread of SMV among years. On the basis of ordinary runs analyses, within-field spread of SMV strain G-5 was random in 1991 and 1994, but was mostly aggregated in 1992 and 1993. Non-G-5 SMV isolates arising from exogenous sources displayed a random spatial pattern over time. This is the first study in which pathogen incidence (detection of SMV using strain-specific monoclonal antibodies) as opposed to disease incidence (based on symptoms) was employed to monitor and model SMV spread in time and space.     

The role of aggressiveness and competition in the selection of Phytophthora infestans populations

Selection within populations of Phytophthora infestans was investigated by comparing the aggressiveness of single‐lesion isolates on detached leaflets of four potato cultivars with differing levels of race‐nonspecific resistance to P. infestans. The isolates included 23 representative of Northern Ireland genotypes from the early 2000s, used to inoculate previously reported field trials on competitive selection (2003–2005), plus 12 isolates recovered from the 2003 trial. The cultivars were those planted in the previous trials: Atlantic (blight‐susceptible) and Santé, Milagro and Stirling (partially resistant). Very highly significant variation for latent period, infection frequency and lesion area was found between genotypes and cultivars; differences between genotypes were more marked on the more resistant cultivars, but no one genotype was the most aggressive across all. Detached leaflets were also inoculated with mixtures of isolates from each genotype group at three sporangial concentrations: differences in aggressiveness between genotypes were more apparent at lower concentrations and on the more resistant cultivars. Genotype groups that were the most aggressive on the more resistant cultivars tended to be those selected by the same cultivars in the field. A mixture of all isolates of all genotypes was used to inoculate detached leaflets of the same cultivars. With one exception, single spore isolates recovered from any one leaflet belonged to a single genotype, but different genotypes were recovered from different cultivars. Phytophthora infestans isolates from Northern Ireland showed significant variation for foliar aggressiveness, and pathogen genotypes exhibited differential aggressiveness to partially resistant cultivars and interacted competitively in genotype selection.     

Spatial distribution and temporal development of fusarium crown and root rot of tomato and pathogen dissemination in field soil

ABSTRACT The spatial distribution and temporal development of tomato crown and root rot, caused by Fusarium oxysporum f. sp. radicis-lycopersici, were studied in naturally infested fields in 1996 and 1997. Disease progression fit a logistic model better than a monomolecular one. Geostatistical analyses and semivariogram calculations revealed that the disease spreads from infected plants to a distance of 1.1 to 4.4 m during the growing season. By using a chlorate-resistant nitrate nonutilizing (nit) mutant of F. oxysporum f. sp. radicis-lycopersici as a "tagged" inoculum, the pathogen was found to spread from one plant to the next via infection of the roots. The pathogen spread to up to four plants (2.0 m) on either side of the inoculated focus plant. Root colonization by the nit mutant showed a decreasing gradient from the site of inoculation to both sides of the inoculated plant. Simulation experiments in the greenhouse further established that this soilborne pathogen can spread from root to root during the growing season. These findings suggest a polycyclic nature of F. oxysporum f. sp. radicis-lycopersici, a deviation from the monocyclic nature of many nonzoosporic soilborne pathogens.     

Foci of stagonospora nodorum blotch in winter wheat before canopy development

ABSTRACT Stagonospora nodorum blotch (SNB) often develops explosively on upper leaves and glumes of wheat. Inoculum for late season infections may arise from early disease foci in the lower canopy or from recent immigration of wind-dispersed ascospores. Research was conducted to determine if foci of SNB are present and secondary spread has occurred in fields before tiller elongation. We determined the incidence of infection by Stagonospora nodorum for plants sampled at the mid-tillering stage in 96 1-m(2) quadrats in each of two fields. Isolates of S. nodorum were recovered from 32 quadrats, one per infected plant where possible. Multilocus restriction fragment length polymorphism haplotypes were determined for each isolate. Of 55 isolates collected from one field, there were 22 distinct haplotypes. Diseased plants were aggregated in both fields; aggregates sometimes extended to adjacent quadrats. Plants within aggregates were often infected by the same haplotype, suggesting that secondary spread had occurred. Foci overlapped because some aggregates were infected by more than one haplotype. Our results show that genetically diverse populations of S. nodorum were already established in fields before canopy development and were comprised of sometimes overlapping foci undergoing clonal expansion.