Pinus Muricata Research Articles

Ectomycorrhizal fungi alter soil food webs and the functional potential of bacterial communities.

Most of Earth's trees rely on critical soil nutrients that ectomycorrhizal fungi (EcMF) liberate and provide, and all of Earth's land plants associate with bacteria that help them survive in nature. Yet, our understanding of how the presence of EcMF modifies soil bacterial communities, soil food webs, and root chemistry requires direct experimental evidence to comprehend the effects that EcMF may generate in the belowground plant microbiome. To this end, we grew Pinus muricata plants in soils that were either inoculated with EcMF and native forest bacterial communities or only native bacterial communities. We then profiled the soil bacterial communities, applied metabolomics and lipidomics, and linked omics data sets to understand how the presence of EcMF modifies belowground biogeochemistry, bacterial community structure, and their functional potential. We found that the presence of EcMF (i) enriches soil bacteria linked to enhanced plant growth in nature, (ii) alters the quantity and composition of lipid and non-lipid soil metabolites, and (iii) modifies plant root chemistry toward pathogen suppression, enzymatic conservation, and reactive oxygen species scavenging. Using this multi-omic approach, we therefore show that this widespread fungal symbiosis may be a common factor for structuring soil food webs.IMPORTANCEUnderstanding how soil microbes interact with one another and their host plant will help us combat the negative effects that climate change has on terrestrial ecosystems. Unfortunately, we lack a clear understanding of how the presence of ectomycorrhizal fungi (EcMF)-one of the most dominant soil microbial groups on Earth-shapes belowground organic resources and the composition of bacterial communities. To address this knowledge gap, we profiled lipid and non-lipid metabolites in soils and plant roots, characterized soil bacterial communities, and compared soils amended either with or without EcMF. Our results show that the presence of EcMF changes soil organic resource availability, impacts the proliferation of different bacterial communities (in terms of both type and potential function), and primes plant root chemistry for pathogen suppression and energy conservation. Our findings therefore provide much-needed insight into how two of the most dominant soil microbial groups interact with one another and with their host plant.

Association of Onnia subtriquetra with living and dead bishop pine (Pinus muricata) and shore pine (Pinus contorta var. contorta) in California, USA

AbstractWe report observations of Onnia subtriquetra on bishop pine (Pinus muricata) and shore pine (Pinus contorta var. contorta) from north coastal California. Our identification of this fungus is supported by molecular information, morphological characteristics, and and a description of the observed range of the fungus on the West Coast. These observations represent an expansion of the geographic and host ranges of Onnia subtriquetra, which on the observed sites is generally associated with declining tree condition and the presence of other native forest pathogens and insects.

Positive interactions between mycorrhizal fungi and bacteria are widespread and benefit plant growth.

Bacteria, ectomycorrhizal (EcM) fungi, and land plants have been coevolving for nearly 200 million years, and their interactions presumably contribute to the function of terrestrial ecosystems. The direction, stability, and strength of bacteria-EcM fungi interactions across landscapes and across a single plant host, however, remains unclear. Moreover, the genetic mechanisms that govern them have not been addressed. To these ends, we collected soil samples from Bishop pine forests across a climate-latitude gradient spanning coastal California, fractionated the soil samples based on their proximity to EcM-colonized roots, characterized the microbial communities using amplicon sequencing, and generated linear regression models showing the impact that select bacterial taxa have on EcM fungal abundance. In addition, we paired greenhouse experiments with transcriptomic analyses to determine the directionality of these relationships and identify which genes EcM-synergist bacteria express during tripartite symbioses. Our data reveal that ectomycorrhizas (i.e., EcM-colonized roots) enrich conserved bacterial taxa across climatically heterogeneous regions. We also show that phylogenetically diverse EcM synergists are positively associated with plant and fungal growth and have unique gene expression profiles compared with EcM-antagonist bacteria. In sum, we identify common mechanisms that facilitate widespread and diverse multipartite symbioses, which inform our understanding of how plants develop in complex environments.

Persistent, viable seedbank buffers serotinous bishop pine over a broad fire return interval

BackgroundIn ecosystems where fire has been excluded, pyrosilviculture can restore some processes historically maintained by fire while mitigating risk where fire is inevitable. Pyrosilviculture in crown fire-adapted forests is, however, limited by insight into the temporal window of fire return matching canopy seedbank development. Here, we characterized demographic responses to fire and non-native pine pitch canker infection in a chronosequence of serotinous bishop pine stands burned at high-severity to quantify (1) temporal patterns of seedbank development given seed viability and density, cone production, and tree density, and (2) pine pitch canker incidence across tree ages and sizes to assess how infection might impact stand and seedbank development. We use our findings to elucidate pyrosilvicuture as a means of restoring fire given practical challenges of reintroducing high-severity fire in crown-fire adapted forests embedded within wildland urban interfaces (WUI).ResultsBishop pine produces an abundant, viable seedbank within eight years that persists across developmental stages and age classes. Seed abundance and viability are exceptionally high at even the earliest age (median > 600,000 seeds ha− 1 and 97% viability at 6 years) and remain high, with the oldest stands (36 years) maintaining median densities of > 500,000 seeds ha− 1 and viability of 95%. We additionally learned that pine pitch canker infection is most severe during the sapling stage (8–10 years post-fire), likely altering stand development trajectories as well as potentially limiting recruitment, and thus the aerial seedbank, into the canopy.ConclusionsIn bishop pine and equally-fecund serotinous species, pyrosilviculture appears a viable management tool across a broad fire return window given the early development and persistence of a robust, viable seedbank, allowing managers flexibility in restoring fire to promote forest persistence while simultaneously mitigating wildfire risk. Moreover, pyrosilviculture in pine pitch canker infected stands may also provide disease mitigation. Although the long-term effects of pine pitch canker infection remain unknown, bishop pines’ viable, persistent seedbank suggests that managers can ignite prescribed fire across a broad return interval — as short as eight years and as long as several decades — to promote bishop pine persistence, mitigate disease infection rates, and reduce wildfire risk in WUI-adjacent ecosystems.

A 55,000-Year Reconstruction of Vegetation in the Los Angeles Basin: Insights into Future Change and a Better Guide for the Present

The California Floristic Province is a biodiversity hotspot. Endemic flora is threatened by climate change, habitat fragmentation and destruction. It is also host to the La Brea Tar Pits (LBTP), which in addition to its famous megafauna, preserves a rare long-term plant macrofossil record with taxonomic resolution to genus and species. The LBTP flora has the potential to provide a comprehensive vegetational history for the Los Angeles Basin and a dynamic baseline for modern conservation efforts. We used accelerator mass spectrometry (AMS) radiocarbon dating to establish a 55 ka timeline of plant presence in Los Angeles, California. We identified and radiocarbon dated 188 plant macrofossils from the La Brea Tar Pits collections. Eight distinct phases of vegetation were identified based on the loss or appearance of key taxa in the timeline. Pairwise similarity was calculated between each identified phase to compare changes to phase species makeup through time. The transition periods between phases were compared to existing climate proxy records to better understand forcings behind vegetation shifts. Two transition periods in the record are of interest to modern conservation efforts. First, fog-dependent closed cone pine species, Pinus muricata and Pinus radiata, become extirpated from the Los Angeles Basin at 48 ka. This disappearance corresponds with a period of extended drought. Second, repeated patterns of juniper species replacement during periods of megadrought, and their eventual extirpation at 12.8 ka, indicate the importance of land management choices in the ability of juniper populations to rebound after drought-induced die-off. Understanding the long-term dynamics of plant community structure in a region is crucial to managing landscapes in the context of global change, and these data are now being leveraged to inform local conservation efforts in the city of Los Angeles.

Status of Lompoc Yerba Santa Eriodictyon capitatum (Namaceae): an Endangered Plant in Santa Barbara County, Southern California

Lompoc yerba santa Eriodictyon capitatum (Namaceae) is an evergreen shrub endemic to west Santa Barbara County in coastal southern Calif. The species was listed as rare under the Calif. Endangered Species Act in 1979 and as endangered under the U.S. Endangered Species Act in 2000. At Federal listing in 2000, Lompoc yerba santa was known from four locations (five occurrences) in three geographic areas. In 2022, the species is known from 11 occurrences in four geographic populations of which 10 occurrences are considered extant and 1 occurrence likely extirpated: Orcutt Oil Field in the Solomon Hills, 2 occurrences; Burton Mesa on Vandenberg Space Force Base North, 6 occurrences; La Salle Canyon on Vandenberg Space Force Base South, 1 occurrence; and Santa Ynez Mountains, 2 occurrences. Lompoc yerba santa occurs mostly in chaparral and bishop pine (Pinus muricata) forest, and most occurrences are in sandy soil. The species stands up to 5 m tall and with vertical stems up to 10 cm diameter. Much of its reproduction appears to be vegetative by rhizome production, suggesting low dispersal ability, and it is a habitat specialist with apparent dependence on substrate disturbance and fire. Using all available information and international standards with a precautionary but realistic attitude to uncertainty, Lompoc yerba santa in 2022 meets IUCN criteria for endangered status: the small extent of occurrence is 433 km2 (<5,000 km2) and the small area of occupancy is 2.30 km2 (<10 km2), along with the geographic range being severely fragmented, and the quality of habitat is observed/inferred/projected to be in continuing decline (altered fire regime/fire management practices, climate change with severe drought and increased temperatures).

Does resource exchange in ectomycorrhizal symbiosis vary with competitive context and nitrogen addition?

Ectomycorrhizal symbiosis is essential for the nutrition of most temperate forest trees and helps regulate the movement of carbon (C) and nitrogen (N) through forested ecosystems. The factors governing the exchange of plant C for fungal N, however, remain obscure. Because competition and soil resources may influence ectomycorrhizal resource movement, we performed a 10-month split-root microcosm study using Pinus muricata seedlings with Thelephora terrestris, Suillus pungens, or no ectomycorrhizal fungus, under two N concentrations in artificial soil. Fungi competed directly with roots and indirectly with each other. We used stable isotope enrichment to track plant photosynthate and fungal N. For T. terrestris, plants received N commensurate with the C given to their fungal partners. Thelephora terrestris was a superior mutualist under high-N conditions. For S. pungens, plant C and fungal N exchange were not coupled. However, in low-N conditions, plants preferentially allocated C to S. pungens rather than T. terrestris. Our results suggest that ectomycorrhizal resource transfer depends on competitive and nutritional context. Plants can exchange C for fungal N, but coupling of these resources can depend on the fungal species and soil N. Understanding the diversity of fungal strategies, and how they change with environmental context, reveals mechanisms driving this important symbiosis.

Species diversity of fungal endophytes across a stress gradient for plants.

Foliar fungal endophytes are one of the most diverse guilds of symbiotic fungi found in the photosynthetic tissues of every plant lineage, but it is unclear how plant environments and leaf resource availability shape their diversity. We explored correlations between leaf nutrient availability and endophyte diversity among Pinus muricata and Vaccinium ovatum plants growing across a soil nutrient gradient spanning a series of coastal terraces in Mendocino, California. Endophyte richness decreased in plants with higher leaf nitrogen-to-phosphorus ratios for both host species, but increased with sodium, which may be toxic to fungi at high concentrations. Isolation frequency, a proxy of fungal biomass, was not significantly predicted by any of the same leaf constituents in the two plant species. We propose that stressed plants can exhibit both low foliar nutrients or high levels of toxic compounds, and that both of these stress responses predict endophyte species richness. Stressful conditions that limit growth of fungi may increase their diversity due to the suppression of otherwise dominating species. Differences between the host species in their endophyte communities may be explained by host specificity, leaf phenology, or microclimates.

Disturbance macroecology: a comparative study of community structure metrics in a high‐severity disturbance regime

AbstractMacroecological studies have established widespread patterns of species diversity and abundance in ecosystems but have generally restricted their scope to relatively steady‐state systems. As a result, how macroecological metrics are expected to scale in ecosystems that experience natural disturbance regimes is unknown. We examine macroecological patterns in a fire‐dependent forest of Bishop pine (Pinus muricata). We target two different‐aged stands in a stand‐replacing fire regime: a mature stand with a diverse understory and with no history of major disturbance for at least 40 yr, and one disturbed by a stand‐replacing fire 17 yr prior to measurement. We compare properties of these stands with macroecological predictions from the Maximum Entropy Theory of Ecology (METE), an information entropy‐based theory that has proven highly successful in predicting macroecological metrics in multiple ecosystems and taxa. Ecological patterns in the mature stand more closely match METE predictions than do data from the more recently disturbed, mid‐seral stage stand. This suggests METE's predictions are more robust in late‐successional, slowly changing, or steady‐state systems than those in rapid flux with respect to species composition, abundances, and organisms’ sizes. Our findings highlight the need for a macroecological theory that incorporates natural disturbance, perturbations, and ecological dynamics into its predictive capabilities, because most natural systems are not in a steady state.

Infection of the Native California Grass, Bromus carinatus, by Fusarium circinatum, the Cause of Pitch Canker in Pines.

At Point Reyes National Seashore in California, Fusarium circinatum, the causal agent of pitch canker in pines, was isolated from Pinus muricata, the California native grass, Bromus carinatus, and the introduced grass, Holcus lanatus. All grass plants from which F. circinatum was isolated were symptomless. Pathogenicity of grass isolates was confirmed by inoculation of P. radiata trees, which developed symptoms similar to trees inoculated with a pine isolate of F. circinatum. Isolates from grasses were somatically compatible with isolates recovered from symptomatic pines. B. carinatus grown in a growth chamber was inoculated with a green fluorescent protein-expressing strain of F. circinatum. Segments of inoculated leaves were incubated in moist chambers; after 1 to 2 days, sporulating hyphae were observed growing from leaf tissue. Spores of F. circinatum removed from B. carinatus leaves were confirmed to be fluorescent when illuminated with ultraviolet light. These results raise the possibility that B. carinatus cryptically infected by F. circinatum may be a source of propagules capable of infecting pines.

Parched pines: a quantitative comparison of two multi‐year droughts and associated mass mortalities of bishop pine (Pinus muricata) on Santa Cruz Island, California

AbstractExtreme weather events such as droughts are expected to increase in severity and frequency as the climate changes; it is imperative that land managers be able to monitor associated changes in vegetation health efficiently and across large scales in order to mitigate or prepare for these events. This need motivated deeper study of the die‐off of bishop pine (Pinus muricata) on California's Santa Cruz Island during the 2012–2016 drought. These pines play a keystone role within the island's ecosystem and have experienced two severe droughts and associated mass die‐offs in the past 40 years. In an effort to compare these events, we used meteorological data to track changes in drought severity from 1985 to 2018 coupled with novel methods for forest monitoring to reveal dynamics not detectable by shorter‐duration studies. Leveraging 34 years of 30 m resolution Landsat imagery, we compared vegetation mortality between the two most severe droughts of that time period: 1987–1991 and 2012–2016. We used the slope of decline in the annual median value of three different Vegetation Indices (VIs; NBR, NDMI and NDVI) to compare mortality between the two drought events and to reveal spatial patterns of mortality within the bishop pine forests. Our results indicated that the 2012–2016 drought was the island's harshest in over a century and that it resulted in greater and more widespread mortality of vegetation within bishop pine stands than the 1987–1991 drought. The average VI decline was significantly greater during the 2012–2016 drought than the 1987–1991 drought by a factor of 1.89, 2.09 and 1.84 for NBR, NDMI and NDVI, respectively. Our results aligned with projections of increasing drought severity and associated tree mortality across the region. The temporal monitoring methods developed here can be adapted to study similar landscape scale changes over multiple decades in other forest ecosystems facing similar threats.

Plant-mediated partner discrimination in ectomycorrhizal mutualisms.

Although ectomycorrhizal fungi have well-recognized effects on ecological processes ranging from plant community dynamics to carbon cycling rates, it is unclear if plants are able to actively influence the structure of these fungal communities. To address this knowledge gap, we performed two complementary experiments to determine (1) whether ectomycorrhizal plants can discriminate among potential fungal partners, and (2) to what extent the plants might reward better mutualists. In experiment 1, split-root Larix occidentalis seedlings were inoculated with spores from three Suillus species (S. clintonianus, S. grisellus, and S. spectabilis). In experiment 2, we manipulated the symbiotic quality of Suillus brevipes isolates on split-root Pinus muricata seedlings by changing the nitrogen resources available, and used carbon-13 labeling to track host investment in fungi. In experiment 1, we found that hosts can discriminate in multi-species settings. The split-root seedlings inhibited colonization by S. spectabilis whenever another fungus was available, despite similar benefits from all three fungi. In experiment 2, we found that roots and fungi with greater nitrogen supplies received more plant carbon. Our results suggest that plants may be able to regulate this symbiosis at a relatively fine scale, and that this regulation can be integrated across spatially separated portions of a root system.

Impact of soil pedogenesis on the diversity and composition of fungal communities across the California soil chronosequence of Mendocino.

Understanding how soil pedogenesis affects microbial communities and their in situ activities according to ecosystem functioning is a central issue in soil microbial ecology, as soils represent essential nutrient reservoirs and habitats for the biosphere. To address this question, soil chronosequences developed from a single, shared mineralogical parent material and having the same climate conditions are particularly useful, as they isolate the factor of time from other factors controlling the character of soils. In our study, we considered a natural succession of uplifted marine terraces in Mendocino, CA, ranging from highly fertile in the younger terrace (about 100,000years old) to infertile in the older terraces (about 300,000years old). Using ITS amplicon pyrosequencing, we analysed and compared the diversity and composition of the soil fungal communities across the first terraces (T1 to T3), with a specific focus in the forested terraces (T2 and T3) on soil samples collected below trees of the same species (Pinus muricata) and of the same age. While diversity and richness indices were highest in the grassland (youngest) terrace (T1), they were higher in the older forested terrace (T3) compared to the younger forested terrace (T2). Interestingly, the most abundant ectomycorrhizal (ECM) taxa that we found within these fungal communities showed high homology with ITS Sanger sequences obtained previously directly from ECM root tips from trees in the same study site, revealing a relative conservation of ECM diversity over time. Altogether, our results provide new information about the diversity and composition of the fungal communities as well as on the dominant ECM species in the soil chronosequence of Mendocino in relation to soil age and ecosystem development.

Timing of mutualist arrival has a greater effect on Pinus muricata seedling growth than interspecific competition

Abstract Interactions with symbiotic microbes, such as mycorrhizal fungi, have the potential to greatly influence plant growth, but it is unclear whether field variation in symbiont availability is common and, if so, sufficient to influence interspecific plant competition. In a greenhouse experiment using natural field soils, I varied the timing of ectomycorrhizal inoculation and the presence of an arbuscular mycorrhizal plant competitor, and measured their effects on pine seedling growth. I found that ectomycorrhizal colonization was absent in some field soils, and that in soils without mycorrhizal inoculum, delayed arrival of ectomycorrhizal spores progressively reduced pine seedling growth and favoured growth of the competitor. Competition had significant negative effects on pine seedling growth, but the competition effect was much smaller than the effect of delayed mutualist arrival. Synthesis. The importance of mycorrhizal spore arrival time on pine growth suggests that plants may experience mutualist limitation more frequently than previously expected, and the relative magnitude of seedling responses to mycorrhizal fungi and competing plants show that in some systems mutualism is likely of equal or greater importance compared with interspecific competition in affecting plant community assembly.

Impact of fog drip versus fog immersion on the physiology of Bishop pine saplings.

Fog-drip to the soil is the most obvious contribution of fog to the water budget of an ecosystem, but several studies provide convincing evidence that foliar absorption of fog water through leaf wetting events is also possible. The focus of our research was to assess the relative importance of fog drip and fog immersion (foliar wetting) on leaf gas-exchange rates and photosynthetic capacity of a coastal pine species, Bishop pine (Pinus muricata D.Don), a drought-sensitive species restricted to the fog belt of coastal California and offshore islands. In a controlled experiment, we manipulated fog water inputs to potted Bishop pine saplings during a 3 week dry-down period. Ten saplings were randomly assigned one of two fog treatments: (1) fog drip to the soil and canopy fog immersion, or (2) fog immersion alone. Five saplings were assigned the 'control' group and received no fog water inputs. We found that fog immersion alone significantly increased carbon assimilation rates and photosynthetic capacity of saplings as soil moisture declined compared with those that received no fog at all. The highest carbon assimilation rates were observed in saplings that also received fog drip. Soil moisture was 40% higher in the fog immersion compared with the control group during the dry-down, indicating a reduced demand for soil water in saplings that had only leaves wetted by canopy interception of fog. Leaf-level physiology is more strongly enhanced by fog drip compared with fog immersion, although the results of this study provide evidence that foliar absorption is a viable mechanism by which Bishop pines use fog water and that it can enhance instantaneous plant carbon gain and potentially whole plant productivity.

Competition-function tradeoffs in ectomycorrhizal fungi.

Background. The extent to which ectomycorrhizal fungi mediate primary production, carbon storage, and nutrient remineralization in terrestrial ecosystems depends upon fungal community composition. However, the factors that govern community composition at the root system scale are not well understood. Here, we explore a potential tradeoff between ectomycorrhizal fungal competitive ability and enzymatic function.Methods. We grew Pinus muricata (Bishop Pine) seedlings in association with ectomycorrhizal fungi from three different genera in a fully factorial experimental design. We measured seedling growth responses, ectomycorrhizal abundance, and the root tip activity of five different extracellular enzymes involved in the mobilization of carbon and phosphorus.Results. We found an inverse relationship between competitiveness, quantified based on relative colonization levels, and enzymatic activity. Specifically, Thelephora terrestris, the dominant fungus, had the lowest enzyme activity levels, while Suillus pungens, the least dominant fungus, had the highest.Discussion. Our results identify a tradeoff between competition and function in ectomycorrhizal fungi, perhaps mediated by the competing energetic demands associated with competitive interactions and enzymatic production. These data suggest that mechanisms such as active partner maintenance by host trees may be important to maintaining “high-quality” ectomycorrhizal fungal partners in natural systems.

Fog drip maintains dry season ecological function in a California coastal pine forest

AbstractFog drip is recognized as an important source of water for many ecosystems that often harbor a disproportionate fraction of endemic species. Characterizing and quantifying the ecological importance of fog drip in these ecosystems requires a range of approaches. We report on a multi‐faceted study of Bishop pine (Pinus muricata D. Don) along a coastal‐inland transect on an island off Southern California. Hourly sampling included micrometeorology, sap flux, and soil moisture. Monthly measurements included changes in tree girth, plant water stress, and isotopic values of fogwater, rainwater, and xylem water. These data show that summertime fog drip clearly affected soil moisture and maintained aspects of tree function, including leaf water relations, sap flux dynamics, and growth rates. Although water from fog drip to the soil surface was occasionally taken up by pine trees, as quantified with isotopic measurements and a Bayesian mixing model, this utilization of fog drip was highly variable in space and time. The proportion of fogwater inferred to have been used is also much less than has been demonstrated in more mesic coastal forest ecosystems using isotopic methods. These results thus suggest high ecosystem sensitivity to even moderate amounts of fog drip, a finding with important implications as climate change differentially affects fog and rain patterns.

Coastal fog during summer drought improves the water status of sapling trees more than adult trees in a California pine forest

Fog water inputs can offset seasonal drought in the Mediterranean climate of coastal California and may be critical to the persistence of many endemic plant species. The ability to predict plant species response to potential changes in the fog regime hinges on understanding the ways that fog can impact plant physiological function across life stages. Our study uses a direct metric of water status, namely plant water potential, to understand differential responses of adult versus sapling trees to seasonal drought and fog water inputs. We place these measurements within a water balance framework that incorporates the varying climatic and soil property impacts on water budgets and deficit. We conducted our study at a coastal and an inland site within the largest stand of the regionally endemic bishop pine (Pinus muricata D. Don) on Santa Cruz Island. Our results show conclusively that summer drought negatively affects the water status of sapling more than adult trees and that sapling trees are also more responsive to changes in shallow soil moisture inputs from fog water deposition. Moreover, between the beginning and end of a large, late-season fog drip event, water status increased more for saplings than for adults. Relative to non-foggy conditions, we found that fog water reduces modeled peak water deficit by 80 and 70 % at the inland and coastal sites, respectively. Results from our study inform mechanistically based predictions of how population dynamics of this and other coastal species may be affected by a warmer, drier, and potentially less foggy future.

Soil contamination with silver nanoparticles reduces Bishop pine growth and ectomycorrhizal diversity on pine roots.

Soil contamination by silver nanoparticles (AgNP) is of potential environmental concern but little work has been carried out on the effect of such contamination on ectomycorrhizal fungi (EMF). EMF are essential to forest ecosystem functions as they are known to enhance growth of trees by nutrient transfer. In this study, soil was experimentally contaminated with AgNP (0, 350 and 790 mg Ag/kg) and planted with Bishop pine seedlings. The effect of AgNP was subsequently measured, assessing variation in pine growth and ectomycorrhizal diversity associated with the root system. After only 1 month, the highest AgNP level had significantly reduced the root length of pine seedlings, which in turn had a small effect on above ground plant biomass. However, after 4 months growth, both AgNP levels utilised had significantly reduced both pine root and shoot biomass. For example, even the lower levels of AgNP (350 mg Ag/kg) soil, reduced fresh root biomass by approximately 57 %. The root systems of the plants grown in AgNP-contaminated soils lacked the lateral and fine root development seen in the control plants (no AgNP). Although, only five different genera of EMF were found on roots of the control plants, only one genus Laccaria was found on roots of plants grown in soil containing 350 mg AgNP/kg. At the higher levels of AgNP contamination, no EMF were observed. Furthermore, extractable silver was found in soils containing AgNP, indicating potential dissolution of silver ions (Ag+) from the solid AgNP.Electronic supplementary materialThe online version of this article (doi:10.1007/s11051-015-3246-4) contains supplementary material, which is available to authorised users.

Inter‐ and intraspecific variation in susceptibility to dothistroma needle blight in Britain. How susceptible are Pinus sylvestris and Pinus contorta?

SummaryDothistroma needle blight (DNB), caused by Dothistroma septosporum, is a serious disease in British pine plantations. Natural infection (NI) and artificial inoculation (AI) experiments were conducted to evaluate the susceptibility of Pinus sylvestris and Pinus contorta to DNB, relative to Pinus mugo, Pinus muricata, Pinus nigra ssp. laricio, Pinus nigra ssp. nigra and Pinus ponderosa. A range of P. sylvestris and P. contorta provenances were included in experiments to simultaneously investigate intraspecific variation within these host species. In NI experiments, 20–30 12‐month‐old seedlings of each species/provenance were planted adjacent to infected mature trees at two Scottish sites. In the AI experiment, eighteen 16‐month‐old seedlings of each species/provenance were inoculated with a D. septosporum conidial suspension and maintained under optimum conditions for D. septosporum infection and development. In the NI experiments, the most DNB‐susceptible species were P. muricata and P. ponderosa, while the least susceptible species were P. contorta and P. sylvestris. Pinus nigra spp. nigra and P. nigra spp. laricio had intermediate DNB susceptibility. In the AI experiment, there was no evidence for interspecific variation in DNB susceptibility. Intraspecific variation in DNB susceptibility within P. sylvestris was seen in the NI experiment, but not the AI experiment. Intraspecific variation in DNB susceptibility within P. contorta was not observed in either experiment. In a further AI experiment with 24‐month‐old P. sylvestris and P. contorta seedlings, the virulence of three Scottish D. septosporum isolates (haplotypes) was examined. There was no significant interaction between isolate and relative susceptibility of pine species/provenances. Furthermore, no between‐isolate variation in virulence was observed.