Spruce Decline and Diaporthe: Incidence, Taxonomy, Virulence, and Tree Susceptibility in Michigan.

In the early 2000s, spruce trees in Michigan began displaying basal needle drop and branch death that slowly progressed upward, symptoms of what we call spruce decline. A survey in 2013 revealed that spruce decline was common throughout Michigan's Lower Peninsula, and Diaporthe was the most likely pathogen causing the cankers associated with these symptoms. Greenhouse inoculation studies completed Koch's postulates, confirming that Diaporthe could cause cankers that cause needle loss and branch death. The five different Diaporthe haplotypes isolated from symptomatic branches during the survey differed in virulence. Haplotypes 2, 4, and 5 were more virulent, and differed from each other by only one or two base pairs using the internal transcribed spacer (ITS) region and did not differ using the ß-tubulin (TUB) gene. These haplotypes were unresolved phylogenetically. Haplotypes 1 and 3 were weakly virulent to avirulent on multiple spruce taxa, and fell into a resolved Diaporthe eres clade. Spruce taxa varied in susceptibility, with Colorado blue spruce (Picea pungens) the most susceptible, followed by Norway (P. abies), then white spruce (P. glauca). Spruce taxa that were much less susceptible were Black Hills (P. glauca var. densata), Serbian (P. omorika), and Meyer spruce (P. meyeri). We demonstrate that one or more Diaporthe species is causing cankers on declining spruce in Michigan, and these cankers elicit symptoms similar to the branch death expressed by declining spruce in the landscape. Future work will focus on further characterizing Diaporthe to species, and determining biotic and abiotic stressors that may predispose spruce trees to express decline symptoms.

Influence of Dew Period and Temperature on Foliar Infection of Greenhouse-Grown Tomato by Colletotrichum coccodes.

Effects of temperature, duration of leaf wetness, and leaf position on foliar infection of greenhouse-grown tomato (cv. Bonnie Best) by Colletotrichum coccodes were determined by inoculating plants with C. coccodes (5.0 × 105 conidia per ml) and keeping them in a dew chamber for 0, 4, 8, 12, 16, 20, or 24 h of wetting at 15, 20, or 25°C. One week after inoculation, leaf disks were placed on the surface of an amended medium on which colonies of C. coccodes were compact and easily identified, and severity of infection was quantified after 4, 6, and 8 days. There was no infection of plants incubated at 15°C, while plants kept at 20 or 25°C had increasing numbers of colonies when leaf wetness duration was extended beyond 12 and 8 h, respectively. Leaf position had a significant effect, with leaves tending to increase in susceptibility as they age. After 24 h of leaf wetness at 25°C, the mean number of colonies per leaf disk from top, middle, and bottom leaves was 23.8, 29.0, and 34.0, respectively.

The acquisition of hypovirulence in host-pathogen systems with three trophic levels.

A major focus of research on the dynamics of host-pathogen interactions has been the evolution of pathogen virulence, which is defined as the loss in host fitness due to infection. It is usually assumed that changes in pathogen virulence are the result of selection to increase pathogen fitness. However, in some cases, pathogens have acquired hypovirulence by themselves becoming infected with hyperparasites. For example, the chestnut blight fungus Cryphonectria parasitica has become hypovirulent in some areas by acquiring a double-stranded RNA hyperparasite that debilitates the pathogen, thereby reducing its virulence to the host. In this article, we develop and analyze a mathematical model of the dynamics of host-pathogen interactions with three trophic levels. The system may be dominated by either uninfected (virulent) or hyperparasitized (hypovirulent) pathogens, or by a mixture of the two. Hypovirulence may allow some recovery of the host population, but it can also harm the host population if the hyperparasite moves the transmission rate of the pathogen closer to its evolutionarily stable strategy. In the latter case, the hyperparasite is effectively a mutualist of the pathogen. Selection among hyperparasites will often minimize the deleterious effects, or maximize the beneficial effects, of the hyperparasite on the pathogen. Increasing the frequency of multiple infections of the same host individual promotes the acquisition of hypovirulence by increasing the opportunity for horizontal transmission of the hyperparasite. This effect opposes the usual theoretical expectation that multiple infections promote the evolution of more virulent pathogens via selection for rapid growth within hosts.

Virulence and Molecular Diversity in Colletotrichum lindemuthianum from South, Central, and North America.

ABSTRACT Isolates of Colletotrichum lindemuthianum (138 total) from Argentina, Brazil, the Dominican Republic, Honduras, Mexico, and the United States were characterized into 41 races based on virulence to 12 differential cultivars of Phaseolus vulgaris. These 41 races were categorized into two groups: those found over a wide geographic area and those restricted to a single country. Races 7, 65, and 73 were widespread. Race 73 was the most common (28%). Race 7 was found once in Argentina and Mexico but at a higher frequency in the United States. Race 65 was found repeatedly in Brazil and the United States. Although 39% of the races were detected repeatedly and three races were widespread, no race was isolated from both P. vulgaris gene pools. Phenetic analyses showed no obvious patterns correlated with virulence clusters. No geographic pattern was evident. Molecular polymorphism generated by random amplified polymorphic DNA confirmed the extensive variability in virulence of C. lindemuthianum. Virulence phenotypes were grouped into 15 clusters. The two largest clusters contained isolates from all the geographic regions sampled. Molecular polymorphism was observed among isolates from races 65 and 73 within and among countries, except among Bra-zilian isolates of race 65. The genetic diversity of C. lindemuthianum was greatest in Mexico and Honduras. Our data suggest that C. lindemuthianum may not be highly structured to specific Phaseolus gene pools.

Host-pathogen interactions in natural populations of Linum marginale and Melampsora lini : III. Influence of pathogen epidemics on host survivorship and flower production.

The epidemiology of rust caused by the fungus Melampsora lini and the effects of infection by this pathogen on its host, the herbaceous perennial Linum marginale, were determined in the field and in garden experiments. There was considerable natural variability in disease levels over the four years (1986-1989) of the study. In two years (1986, 1989) major rust epidemics occurred. In the field, the main effect of disease was to reduce survivorship during the winter following infection. Plants which were heavily infected during the 1986 or 1989 growing seasons had reduced survivorship relative to more lightly infected plants. Melampsora lini infections did not appear to affect survivorship in either 1987 or 1988. Flowering was correlated with environmental factors and the number of stems a plant possessed. A severe drought in 1987 completely inhibited flowering. In the other three years the number of flowers produced by a plant was strongly positively correlated with the number of stems it possessed. Disease levels had no consistent effect on flowering. Controlled garden experiments were also used to examine the response of seedlings and adult plants to infection. These showed that both the timing and severity of disease appears to determine the effect of M. lini infections on L. marginale. Early, severe infection reduced growing season and overwintering survivorship as well as capsule production. However, plants in the field were most often infected only after flowering had begun, and the predominant effect of infection was a reduction in overwintering survivorship. The high variability in disease levels from year to year and the deferred nature of the effect of the rust on its host have significant implications for the design of experiments aimed at assessing the role of diseases in plant communities.

HOST-PATHOGEN INTERACTIONS IN NATURAL POPULATIONS OF LINUM MARGINALE AND MELAMPSORA LINI: I. PATTERNS OF RESISTANCE AND RACIAL VARIATION IN A LARGE HOST POPULATION.

Populations of wild flax, Linum marginale and its associated rust fungus Melampsora lini growing at Kiandra, New South Wales, Australia, were sampled during the 1986-1987 growing season. Thirteen different races of M. lini were detected in a sample of 96 isolates. The distribution of isolates was uneven: race A comprised 73% of the samples; race N, 8%; and race H, 5%; while the remaining races were represented by only one or two samples. The dominance of race A increased over the course of the growing season, comprising 67% of the early season samples and increasing to 78% for those collected late in the season. The overall diversity of the pathogen population decreased late in the growing season, but this trend was not statistically significant. The average virulence of individual isolates of the pathogen population increased during the growing season. This trend was most pronounced among the minor races, where the mean number of differential hosts infected increased from 4.58 for early season samples to 5.12 and 5.08 for mid and late season samples, respectively. In contrast to the virulence pattern in the pathogen, the L. marginale population displayed a more even distribution of resistance. In a sample of 67 plants 10 resistance phenotypes were detected from their pattern of resistance/susceptibility to seven pathogen isolates. No phenotype had a frequency that exceeded 30%. Resistance phenotypes were randomly distributed on both a population level and on a fine scale.

HOST-PATHOGEN INTERACTIONS IN NATURAL POPULATIONS OF LINUM MARGINALE AND MELAMPSORA LINI: II. LOCAL AND REGIONAL VARIATION IN PATTERNS OF RESISTANCE AND RACIAL STRUCTURE.

Spatial variation in the resistance structure of Linum marginale (wild flax) populations to the rust fungus Melampsora lini, and in the racial structure of this pathogen, was investigated by sampling 10 populations distributed throughout the Kosciusko National Park, New South Wales, Australia. Considerable differences were found among populations in the structure of both host and pathogen. Host populations were divided into three broad categories: (1) populations susceptible to all testing races; (2) populations containing a strictly limited number of resistant phenotypes; and (3) populations with a considerable diversity of resistant phenotypes. The pathogen populations also showed a range of diversity. The major differences between these populations were determined by the occurrence and frequency of four common races of pathogen (races A, E, K, and N). These differences were apparent both at a regional spatial scale (over the 100 km separation of the most distant populations) and at a local scale where major differences were detected between two populations only 300 m apart. The distribution of the four common races of M. lini was consistent with the hypothesis that a fitness cost was associated with unnecessary virulence. In general, however, differences in the structure of pathogen populations from genetically very similar host populations implied that, in addition to host resistance genes, other evolutionary forces are also important in determining the genetic structure of individual pathogen populations.

The effect of small-scale environmental changes on disease incidence and severity in a natural plant-pathogen interaction.

The incidence and severity of Rhynchosporium secalis infections were assessed in a large population of Hordeum leporinum. Transects were set out in four directions from five trees to determine the effect of shading. Under the tree canopy 60.3% of H. leporinum plants were infected while only 11.2% were infected away from the canopy. Disease severity, on those plants which were infected, was higher under the canopy (mean 12.4% and 13.0% leaf area diseased for the flag and first leaves, respectively) than away from the canopy (means of 7.8% and 5.0% for the flag and first leaves respectively). Plants under the tree canopy contained on average 23% more nitrogen, raising the possibility that the susceptibility of the host changed in response to nitrogen levels. However, the observed pattern is also consistent with the hypothesis that shade-associated changes in the environment enhanced the ability of the pathogen to infect and develop on the host. The data clearly demonstrate the importance of small-scale environmental factors on natural host-pathogen interactions. These environmental factors may cause differential selection for disease resistance within a host population, which may ultimately lead to the formation of sub-populations with differing levels of resistance.