Swiss needle cast (SNC) is a foliar disease of Douglas-fir (Pseudotsuga menziesii) caused by Phaeocryptopus gaeumannii (Rohde) Petrak. This fungus is endemic to western North America, where it has historically had little impact in native forests. However, increasing disease severity in western Oregon since the 1990s has prompted renewed interest in P. gaeumannii and SNC. For this study, we analyze multilocus microsatellite genotypes from 482 single-spore isolates from 68 trees across 14 sites in the western Coast Range of Oregon and southwestern Washington. This study assesses genotypic variation and genetic structure at several levels of population hierarchy. Despite the observation that most of the genetic variation occurred within subpopulations, our analyses detected significant differentiation at all hierarchical levels. Clustering among the 482 isolates based on genetic distance clearly supports the existence of two previously described cryptic lineages of P. gaeumannii in the western United States. The two lineages occur in varying proportions along latitudinal and longitudinal gradients in western Oregon and Washington, suggesting a relationship between climate and phylogeography. Sites near Tillamook, Oregon, where SNC is most severe, consist of sympatric subpopulations in which the two lineages comprise roughly equal proportions.
The environment has a strong influence on the abundance and distribution of plant pathogenic organisms and plays a major role in plant disease. Climatological factors may also alter the dynamics of the interactions between plant pathogens and their hosts. Nothophaeocryptopus (=Phaeocryptopus) gaeumannii, the causal agent of Swiss needle cast (SNC) of Douglas‐fir, is endemic to western North America where it exists as two sympatric, reproductively isolated lineages. The abundance of this fungus and the severity of SNC are strongly influenced by climate. We used statistical and population genetic analyses to examine relationships between environment, pathogen population structure, and SNC severity. Although N. gaeumannii Lineage 2 in western Oregon and Washington was most abundant where SNC symptoms were most severe, we did not detect a significant relationship between Lineage 2 and disease severity. Warmer winter temperatures were inversely correlated with foliage retention (AFR) and positively correlated with the relative abundance of Lineage 2 (PL2). However when distance inland, which was strongly correlated with both AFR and PL2, was included in the model, there was no significant relationship between Lineage 2 and AFR. Spring/early summer dew point temperatures also were positively associated with total N. gaeumannii abundance (colonization index (CI)) and inversely correlated with AFR. Warmer summer mean temperatures were associated with lower CI and higher AFR. Our results suggest that the two lineages have overlapping environmental optima, but slightly different tolerance ranges. Lineage 2 was absent from more inland sites where winters are colder and summers are warm and dry, while Lineage 1 occurred at most sites across an environmental gradient suggesting broader environmental tolerance. These relationships suggest that climate influences the abundance and distribution of this ecologically important plant pathogen and may have played a role in the evolutionary divergence of these two cryptic fungal lineages.
Many fungi in the Ophiostomatales are vectored by bark beetles that introduce these fungi directly into their tree hosts. Most of these fungal associates have little effect on their hosts, but some can cause serious diseases. One such fungus, Leptographium wageneri, causes an economically and ecologically important tree disease known as black stain root disease (BSRD). For this study, 159 full genome sequences of L. wageneri were analyzed using a population genomics approach to investigate the epidemiology, dispersal capabilities, and reproductive biology of this fungus. Analyses were performed with SNP haplotypes from 155 isolates of L. wageneri var. pseudotsugae collected in 16 Douglas-fir stands in Oregon and 4 isolates of L. wageneri var. wageneri collected in pinyon pine stands in southern California. These two host-specific varieties appear to be evolutionarily divergent, likely due a combination of factors such as host differentiation and geographic isolation. We analyzed gene flow and population structure within and among Douglas-fir plantations in western Oregon to infer the relative importance of local vs. long distance dispersal in structuring populations of L. wageneri var. pseudotsugae. Long-distance gene flow has occurred between Douglas-fir plantations, contributing to diversity and population structure within stands, and likely reflecting the behavior of an important insect vector. Genetic clustering analyses revealed the presence of unique local clusters within stands and plantations in addition to those common among multiple stands or plantations. Although populations of L. wageneri var. pseudotsugae are primarily asexual, two mating types were present in many stands, suggesting that recombination is at least possible and may contribute to genetic diversity.
Swiss needle cast is a foliar disease of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) that results in premature foliage loss and reduced growth. The causal fungus, Nothophaeocryptopus gaeumannii, was first detected in New Zealand in 1959 and spread throughout the North and South Islands over the following decades. The contemporary genetic structure of the N. gaeumannii population in New Zealand was assessed by analyzing 468 multilocus SSR genotypes (MLGs) from 2,085 N. gaeumannii isolates collected from 32 sites in in the North and South Islands. Overall diversity was lower than that reported from native N. gaeumannii populations in the northwestern U.S., which was expected given that N. gaeumannii is introduced in New Zealand. Linkage disequilibrium was significantly higher than expected under random mating, suggesting that population structure is clonal. Populations of N. gaeumannii in the North and South Island were weakly differentiated, and the isolates collected from sites within the islands were moderately differentiated. This suggests that gene flow has occurred between the N. gaeumannii populations in the North and South Islands, and between the local N. gaeumannii populations within each island. Eighteen isolates of N. gaeumannii Lineage 2, which has previously been reported only from western Oregon, were recovered from two sites in the North Island and four sites in the South Island. The most likely explanation for the contemporary distribution of N. gaeumannii in New Zealand is that it was introduced on infected live seedlings through the forestry or ornamental nursery trade, as the fungus is neither seed borne nor saprobic, and the observed population structure is not consistent with a stochastic intercontinental dispersal event.
Boiling Springs Lake (BSL) is an oligotrophic, acidic geothermal feature where even very low levels of microbial heterotrophic production still exceed autotrophy. To test whether allochthonous leaf litter (LL) inputs fuel this excess, we quantified leaf litterfall, leaching and decomposition kinetics, and measured the impact of organic amendments on production, germination and cell growth, using pyrosequencing to track changes in microbial community composition. Coniferous leaves in BSL exhibited high mass loss rates during leaching and decomposition, likely due to a combination of chemical hydrolysis and contributions of both introduced and endemic microbes. We measured very low in situ (3)H-thymidine incorporation over hours by the dominant chemolithotroph Acidimicrobium (13-65 μg C L(-1) day(-1)), which was inhibited by simple C sources (acetate, glucose). Longer term incubations with additions of 0.01-0.02% complex C/N sources induced germination of the Firmicute Alicyclobacillus within 1-2 days, as well as growth of Acetobacteraceae after 3-4 days. LL additions yielded the opposite successional patterns of these r-selected heterotrophs, boosting production to 30-150 μg C L(-1) day(-1). Growth and germination studies suggest both prokaryotes and fungi likely consume allochthonous organics, and might be novel sources of lignocellulose-degrading enzymes. A model of BSL's C budget supports the hypothesis that allochthonous inputs fuel seasonal microbial heterotrophy, but that dissolved organic C sources greatly exceed direct LL inputs.
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