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Exploring the metabolic differences directly on tissues is essential for the comprehensive understanding of how multicellular organisms function. Mass spectrometry imaging (MSI) is an attractive technique toward this goal; however, MSI in metabolomics scale has been hindered by multiple limitations. This is most notable for single cell level high-spatial resolution imaging because of the limited number of molecules in small sampling size and the low ionization yields of many metabolites. Several on-tissue chemical derivatization approaches have been reported to increase MSI signals of targeted compounds, especially in matrix-assisted laser desorption/ionization (MALDI)-MSI. Herein, we adopt a combination of chemical derivatization reactions, to selectively enhance metabolite signals of a specific functional group for each consecutive tissue section. Three well-known on-tissue derivatization methods were used as a proof of concept experiment: coniferyl aldehyde for primary amines, Girard's reagent T for carbonyl groups, and 2-picolylamine for carboxylic acids. This strategy was applied to the cross-sections of leaves and roots from two different maize genotypes (B73 and Mo17), and enabled the detection of over six hundred new unique metabolite features compared to without modification. Statistical analysis indicated quantitative variation between metabolites in the tissue sections, while MS images revealed differences in localization of these metabolites. Combined, this untargeted approach facilitated the visualization of various classes of compounds, demonstrating the potential for untargeted MSI in the metabolomics scale.

In the White Mountains of California, eight bristlecone pine (Pinus longaeva) tree-ring width chronologies were developed from trees at upper treeline and just below upper treeline along North- and South-facing elevational transects from treeline to ~90 m below. There is evidence for a climate-response threshold between approximately 60-80 vertical m below treeline, above which trees have shown a positive growth-response to temperature and below which they do not. Chronologies from 80 m or more below treeline show a change in climate response and do not correlate strongly with temperature-sensitive chronologies developed from trees growing at upper treeline. Rather, they more closely resemble lower elevation precipitation-sensitive chronologies. At the highest sites, trees on South-facing slopes grow faster than trees on North-facing slopes. High growth rates in the treeline South-facing trees have declined since the mid-1990s. This suggests the possibility that the climate-response of the highest South-facing trees may have changed and that temperature may no longer be the main limiting factor for growth on the South aspect. These results indicate that increasing warmth may lead to a divergence between tree growth and temperature at previously temperature-limited sites.

Aim: To understand how the biophysical environment influences patterns of infection by non-native blister rust (caused by Cronartium ribicola) and mortality caused by native mountain pine beetles (Dendroctonus ponderosae) in whitebark pine (Pinus albicaulis) communities, to determine how these disturbances interact, and to gain insight into how climate change may influence these patterns in the future. Location: High-elevation forests in south-west Montana, central Idaho, eastern and western Oregon, USA. Methods: Stand inventory and dendroecological methods were used to assess stand structure and composition and to reconstruct forest history at sixty 0.1-ha plots. Patterns of blister rust infection and mountain pine beetle-caused mortality in whitebark pine trees were examined using nonparametric Kruskal-Wallis ANOVA, Mann-Whitney 17-tests, and Kolmogorov-Smirnov two-sample tests. Stepwise regression was used to build models of blister rust infection and mountain pine beetle-related mortality rates based on a suite of biophysical site variables. Results: Occurrence of blister rust infections was significantly different among the mountain ranges, with a general gradient of decreasing blister rust occurrence from east to west. Evidence of mountain pine beetle-caused mortality was identified on 83% of all dead whitebark pine trees and was relatively homogenous across the study area. Blister rust infected trees of all ages and sizes uniformly, while mountain pine beetles infested older, larger trees at all sites. Stepwise regressions explained 64% and 58% of the variance in blister rust infection and beetle-caused mortality, respectively, indicating that these processes are strongly influenced by the biophysical environment. More open stand structures produced by beetle outbreaks may increase the exposure of surviving whitebark pine trees to blister rust infection. Main conclusions: Variability in the patterns of blister rust infection and mountain pine beetle-caused mortality elucidated the fundamental dynamics of these disturbance agents and suggests that the effects of climate change will be complex in whitebark pine communities and vary across the species' range. Interactions between blister rust and beetle outbreaks may accelerate declines or facilitate the rise of rust resistance in whitebark pine depending on forest conditions at the time of the outbreak.

The Border Lakes Region of Minnesota and Ontario has long been viewed as a fire‐dependent ecosystem. High‐severity fire in the region's near‐boreal forests has been a focus of ecological research and public fascination. However, the surface fire history within this transnational wilderness landscape has received more limited attention. We used an interdisciplinary, dendroecological approach to characterize the surface fire history of the region, assess potential drivers of historical surface fires, and document the ecological legacies of frequent fires within the red pine forests of the Boundary Waters Canoe Area Wilderness (BWCAW) in northern Minnesota. We used tree‐ring and fire atlas data to reconstruct multi‐century surface fire records for 101 sites and document age structure and composition at 32 sites across the BWCAW. Stratification of these sites relative to their proximity to a primary travel and trade corridor used first by Indigenous groups and later by Euro‐American fur traders through the late 1800s provided strong evidence of human augmentation of fires. The patterns of fire activity, fire–climate relationships, and forest development indicate that traditional landuse by Anishinaabeg (Ojibwe) increased rates of local surface fire and played an important role in shaping the landscape. The decline of traditional subsistence practices by the Border Lakes Anishinaabeg coincided with a sharp decline in surface fires and a period of abundant tree establishment. In the absence of repeat surface fires, many red pine sites have shifted in composition, increased in stem density, and grown vulnerable to forest‐type conversion through future high‐severity fire. These results highlight the need for active fire reintroduction to red pine forests of the Great Lakes Region and underscore the importance of collaboration and guidance from Indigenous Knowledge Keepers in this process. A blended knowledge approach to fire restoration that directly engages with Indigenous perspectives and cultural practices can perpetuate the distinctive character of the largest remaining tracts of long‐lived pine forest in the Great Lakes Region. Carefully developed fire restoration practices would enhance the visitor use experience within one of the most frequently visited wilderness areas in the United States while re‐engaging directly with Indigenous knowledge and traditional cultural practices.

Dopant-assisted atmospheric pressure chemical ionization (dAPCI) is a soft ionization method rarely used for gas chromatography-mass spectrometry (GC-MS). The current study combines GC-dAPCI with tandem mass spectrometry (MS/MS) for analysis of a complex mixture such as lignin pyrolysis analysis. To identify the structures of volatile lignin pyrolysis products, collision-induced dissociation (CID) MS/MS using a quadrupole time-of-flight mass spectrometer (QTOFMS) and pseudo MS/MS through in-source collision-induced dissociation (ISCID) using a single stage TOFMS are utilized. To overcome the lack of MS/MS database, Compound Structure Identification (CSI):FingerID is used to interpret CID spectra and predict best matched structures from PubChem library. With this approach, a total of 59 compounds were positively identified in comparison to only 22 in NIST database search of GC-EI-MS dataset. This study demonstrates the effectiveness of GC-dAPCI-MS/MS to overcome the limitations of traditional GC-EI-MS analysis when EI-MS database is not sufficient. Graphical Abstract ᅟ

[This corrects the article DOI: 10.3389/fpls.2019.00860.].[This corrects the article DOI: 10.3389/fpls.2019.00860.].

Non-native, invasive earthworms are altering soils throughout the world. Ecological cascades emanating from these invasions stem from rapid consumption of leaf litter by earthworms. This occurs at a midpoint in the trophic pyramid, unlike the more familiar bottom-up or top-down cascades. These cascades cause fundamental changes (“microcascade effects”) in soil morphology, bulk density, and nutrient leaching, and a shift to warmer, drier soil surfaces with a loss of leaf litter. In North American temperate and boreal forests, microcascade effects can affect carbon sequestration, disturbance regimes, soil and water quality, forest productivity, plant communities, and wildlife habitat, and can facilitate other invasive species. These broader-scale changes (“macrocascade effects”) are of greater concern to society. Interactions among these fundamental changes and broader-scale effects create “cascade complexes” that interact with climate change and other environmental processes. The diversity of cascade effects, combined with the vast area invaded by earthworms, leads to regionally important changes in ecological functioning.

We examined a set of five proxy reconstructions of the Pacific Decadal Oscillation (PDO) to test whether the choice of reconstruction affected the association between the PDO and widespread forest fires in the western United States. Exact binomial tests suggest the PDO has little direct impact on wildfires, with a statistically‐significant association between the phase of the PDO and regional fire activity observed with only one reconstruction. Region‐wide fires were not consistently associated with specific phase combinations of ENSO and the PDO. Any conclusion that extensive wildfires are more or less common when the PDO is in one phase or the other depends entirely on the choice of PDO reconstruction. Without a better understanding of low‐frequency behavior in the north Pacific prior to 1900, efforts using proxy data to determine whether or not the PDO affects wildfire activity in the western United States will produce ambiguous results. Key Points The PDO is believed to influence wildfire activity over one to several decades The choice of reconstruction determines the inferred PDO/fire relationship Without guidance from paleoclimatology, PDO‐fire tests will remain uncertain

New dates from culturally modified red pine rediscovered in the Boundary Waters Canoe Area Wilderness in northern Minnesota provide an opportunity to merge tree-ring records of human land use with archaeological records, historical travel accounts, and traditional knowledge to enhance understanding of Anishinaabeg land tenure in the Wilderness. Records from 244 culturally modified trees (CMTs) demonstrate varying intensities of human use along historical water routes, notably the Border Route that connected Grand Portage to Rainy Lake and Lake of the Woods during the North American fur trade. Crossdated modification years from 119 CMTs provide direct evidence of human-landscape interaction along historical travel routes utilized by Anishinaabeg and Euro-American traders from the mid-1700s to the early 1900s. This CMT network preserves a fading biological record of fur-trade-era cultural history that contributes to a growing cross-cultural conversation on the storied traditional use of a cultural landscape that is now the most visited federal wilderness area in the United States.

Drought and mountain pine beetle (Dendroctonus ponderosae Hopkins) outbreaks have affected millions of hectares of high-elevation conifer forests in the Northern Rocky Mountains during the past century. Little research has examined the distinction between mountain pine beetle outbreaks and climatic influence on radial growth in endangered whitebark pine (Pinus albicaulis Engelm.) ecosystems. We used a new method to explore divergent periods in whitebark pine radial growth after mountain pine beetle outbreaks across six sites in western Montana. We examined a 100-year history of mountain pine beetle outbreaks and climate relationships in whitebark pine radial growth to distinguish whether monthly climate variables or mountain pine outbreaks were the dominant influence on whitebark pine growth during the 20th century. High mortality of whitebark pines was caused by the overlapping effects of previous and current mountain pine beetle outbreaks and white pine blister rust infection. Wet conditions from precipitation and snowpack melt in the previous summer, current spring, and current summer benefit whitebark pine radial growth during the following growing season. Whitebark pine radial growth and climate relationships were strongest in sites less affected by the mountain pine beetle outbreaks or anthropogenic disturbances. Whitebark pine population resiliency should continue to be monitored as more common periods of drought will make whitebark pines more susceptible to mountain pine beetle attack and to white pine blister rust infection.