The xylem of anisohydric Quercus alba L. is more vulnerable to embolism than isohydric codominants.

The coordination of plant leaf water potential (ΨL ) regulation and xylem vulnerability to embolism is fundamental for understanding the tradeoffs between carbon uptake and risk of hydraulic damage. There is a general consensus that trees with vulnerable xylem more conservatively regulate ΨL than plants with resistant xylem. We evaluated if this paradigm applied to three important eastern US temperate tree species, Quercus alba L., Acer saccharum Marsh. and Liriodendron tulipifera L., by synthesizing 1600 ΨL observations, 122 xylem embolism curves and xylem anatomical measurements across 10 forests spanning pronounced hydroclimatological gradients and ages. We found that, unexpectedly, the species with the most vulnerable xylem (Q. alba) regulated ΨL less strictly than the other species. This relationship was found across all sites, such that coordination among traits was largely unaffected by climate and stand age. Quercus species are perceived to be among the most drought tolerant temperate US forest species; however, our results suggest their relatively loose ΨL regulation in response to hydrologic stress occurs with a substantial hydraulic cost that may expose them to novel risks in a more drought-prone future.

Is xylem of angiosperm leaves less resistant to embolism than branches? Insights from microCT, hydraulics, and anatomy.

According to the hydraulic vulnerability segmentation hypothesis, leaves are more vulnerable to decline of hydraulic conductivity than branches, but whether stem xylem is more embolism resistant than leaves remains unclear. Drought-induced embolism resistance of leaf xylem was investigated based on X-ray microcomputed tomography (microCT) for Betula pendula, Laurus nobilis, and Liriodendron tulipifera, excluding outside-xylem, and compared with hydraulic vulnerability curves for branch xylem. Moreover, bordered pit characters related to embolism resistance were investigated for both organs. Theoretical P50 values (i.e. the xylem pressure corresponding to 50% loss of hydraulic conductance) of leaves were generally within the same range as hydraulic P50 values of branches. P50 values of leaves were similar to branches for L. tulipifera (-2.01 versus -2.10 MPa, respectively), more negative for B. pendula (-2.87 versus -1.80 MPa), and less negative for L. nobilis (-6.4 versus -9.2 MPa). Despite more narrow conduits in leaves than branches, mean interconduit pit membrane thickness was similar in both organs, but significantly higher in leaves of B. pendula than in branches. This case study indicates that xylem shows a largely similar embolism resistance across leaves and branches, although differences both within and across organs may occur, suggesting interspecific variation with regard to the hydraulic vulnerability segmentation hypothesis.

Coarse roots prevent declines in whole-tree non-structural carbohydrate pools during drought in an isohydric and an anisohydric species.

Predicted increases in the frequency and severity of droughts have led to a renewed focus on how plants physiologically adjust to low water availability. A popular framework for understanding plant responses to drought characterizes species along a spectrum from isohydry to anisohydry based on their regulation of gas exchange and leaf water potential under drying conditions. One prediction that arises from this theory is that plant drought responses may hinge, in part, on their usage of non-structural carbohydrate (NSC) pools. For example, trees that respond to drought by closing stomates (i.e., isohydric) are predicted to deplete NSC reserves to maintain metabolism, whereas plants that keep stomata open during water stress (i.e., anisohydric), may show little change or even increases in NSC concentration. However, empirical tests of this theory largely rely on aboveground measurements of NSC, ignoring the potentially conflicting responses of root NSC pools. We sought to test these predictions by subjecting potted saplings of Quercus alba L. (an anisohydric species) and Liriodendron tulipifera L. (an isohydric species) to a 6 week experimental drought. We found that stem NSC concentrations were depleted in the isohydric L. tulipifera but maintained in the anisohydric Q. alba-as predicted. However, when scaled to whole-plant NSC content, the drought-induced decreases in stem NSCs in L. tulipifera were offset by increases in root NSCs (especially soluble sugars), resulting in no net change to whole-plant NSC content. Similarly, root sugars increased in Q. alba in response to drought. This increase was concurrent with declines in growth, suggesting a potential trade-off between allocation of photoassimilates to root sugars vs biomass during drought. Collectively, our results suggest that the responses of NSC in coarse roots can differ from stems, and indicate a prominent role of coarse roots in mitigating drought-induced declines in whole-tree NSC pools.

Canopy gradients in leaf functional traits for species that differ in growth strategies and shade tolerance.

In temperate deciduous forests, vertical gradients in leaf mass per area (LMA) and area-based leaf nitrogen (Narea) are strongly controlled by gradients in light availability. While there is evidence that hydrostatic constraints on leaf development may diminish LMA and Narea responses to light, inherent differences among tree species may also influence leaf developmental and morphological response to light. We investigated vertical gradients in LMA, Narea and leaf carbon isotope composition (δ13C) for three temperate deciduous species (Carpinus caroliniana Walter, Fagus grandifolia Ehrh., Liriodendron tulipifera L.) that differed in growth strategy (e.g., indeterminate and determinate growth), shade tolerance and leaf area to sapwood ratio (Al:As). Leaves were sampled across a broad range of light conditions within three vertical layers of tree crowns to maximize variation in light availability at each height and to minimize collinearity between light and height. All species displayed similar responses to light with respect to Narea and δ13C, but not for LMA. Light was more important for gradients in LMA for the shade-tolerant (C. caroliniana) and -intolerant (L. tulipifera) species with indeterminate growth, and height (e.g., hydrostatic gradients) and light were equally important for the shade-tolerant (F. grandifolia) species with determinate growth. Fagus grandifolia had a higher morphological plasticity in response to light, which may offer a competitive advantage in occupying a broader range of light conditions throughout the canopy. Differences in responses to light and height for the taller tree species, L. tulipifera and F. grandifolia, may be attributed to differences in growth strategy or Al:As, which may alter morphological and functional responses to light availability. While height was important in F. grandifolia, height was no more robust in predicting LMA than light in any of the species, confirming the strong role of light availability in determining LMA for temperate deciduous species.

Dynamics of stem water uptake among isohydric and anisohydric species experiencing a severe drought.

Predicting the impact of drought on forest ecosystem processes requires an understanding of trees' species-specific responses to drought, especially in the Eastern USA, where species composition is highly dynamic due to historical changes in land use and fire regime. Here, we adapted a framework that classifies trees' water-use strategy along the spectrum of isohydric to anisohydric behavior to determine the responses of three canopy-dominant species to drought. We used a collection of leaf-level gas exchange, tree-level sap flux and stand-level eddy covariance data collected in south-central Indiana from 2011 to 2013, which included an unusually severe drought in the summer of 2012. Our goal was to assess how patterns in the radial profile of sap flux and reliance on hydraulic capacitance differed among species of contrasting water-use strategies. In isohydric species, which included sugar maple (Acer saccharum Marsh.) and tulip poplar (Liriodendron tulipifera L.), we found that the sap flux in the outer xylem experienced dramatic declines during drought, but sap flux at inner xylem was buffered from reductions in water availability. In contrast, for anisohydric oak species (Quercus alba L. and Quercus rubra L.), we observed relatively smaller variations in sap flux during drought in both inner and outer xylem, and higher nighttime refilling when compared with isohydric species. This reliance on nocturnal refilling, which occurred coincident with a decoupling between leaf- and tree-level water-use dynamics, suggests that anisohydric species may benefit from a reliance on hydraulic capacitance to mitigate the risk of hydraulic failure associated with maintaining high transpiration rates during drought. In the case of both isohydric and anisohydric species, our work demonstrates that failure to account for shifts in the radial profile of sap flux during drought could introduce substantial bias in estimates of tree water use during both drought and non-drought periods.

Calcium and aluminum cycling in a temperate broadleaved deciduous forest of the eastern USA: relative impacts of tree species, canopy state, and flux type.

Ca/Al molar ratios are commonly used to assess the extent of aluminum stress in forests. This is among the first studies to quantify Ca/Al molar ratios for stemflow. Ca/Al molar ratios in bulk precipitation, throughfall, stemflow, litter leachate, near-trunk soil solution, and soil water were quantified for a deciduous forest in northeastern MD, USA. Data were collected over a 3-year period. The Ca/Al molar ratios in this study were above the threshold for aluminum stress (<1). Fagus grandifolia Ehrh. (American beech) had a median annual stemflow Ca/Al molar ratio of 15.7, with the leafed and leafless values of 12.4 and 19.2, respectively. The corresponding Ca/Al molar ratios for Liriodendron tulipifera L. (yellow poplar) were 11.9 at the annual time scale and 11.9 and 13.6 for leafed and leafless periods, respectively. Bayesian statistical analysis showed no significant effect of canopy state (leafed, leafless) on Ca/Al molar ratios. DOC was consistently an important predictor of calcium, aluminum, and Ca/Al ratios. pH was occasionally an important predictor of calcium and aluminum concentrations, but was not a good predictor of Ca/Al ratio in any of the best-fit models (of >500 examined). This study supplies new data on Ca/Al molar ratios for stemflow from two common deciduous tree species. Future work should examine Ca/Al molar ratios in stemflow of other species and examine both inorganic and organic aluminum species to better gauge the potential for, and understand the dynamics of, aluminum toxicity in the proximal area around tree boles.

Meteorological influences on stemflow generation across diameter size classes of two morphologically distinct deciduous species.

Many tree species have been shown to funnel substantial rainfall to their stem base as stemflow flux, given a favorable stand structure and storm conditions. As stemflow is a spatially concentrated flux, prior studies have shown its impact on ecohydrological and biogeochemical processes can be significant. Less work has been performed examining stemflow variability from meteorological conditions compared to canopy structural traits. As such, this study performs multiple regressions: (1) to examine stemflow variability due to event-based rainfall amount, intensity, mean wind speeds, and vapor pressure deficit; (2) across three diameter size classes (10-20, 21-40, and >41 cm DBH); and (3) for two common tree species in the northeastern USA of contrasting canopy morphology--Liriodendron tulipifera L. (yellow poplar) versus Fagus grandifolia Ehrh. (American beech). On the whole, multiple regression results yielded significant positive correlations with stemflow for rainfall amount, intensity, and mean wind speed and a significant negative correlation for vapor pressure deficit (VPD). Tree size altered stemflow-meteorological condition relationships, where larger trees strengthened indirect stemflow-VPD and direct stemflow-rainfall and stemflow-intensity associations. Canopies of rougher bark and lower branch angle (represented by L. tulipifera) enhanced correlations for nearly all meteorological conditions via greater stemflow residence time (and longer exposure to meteorological conditions). Multiple regressions performed on leafless canopy stemflow resulted in an inverse relationship with wind speeds, likely decoupling stemflow sheltered solely on bark surfaces from VPD influences. Leaf presence generally increased direct stemflow associations with rainfall intensity, yet diminished stemflow-rainfall relationships. F. grandifolia canopies (exemplifying structures of smoother bark and greater branch angle) strengthened differences in stemflow associations with rainfall/mean wind speed between leaf states. These findings are placed in a conceptual interception loss path analysis, which shows the potential to alter common interception loss estimates in high stemflow stands.

Foraging strategies in trees of different root morphology: the role of root lifespan.

Resource exploitation of patches is influenced not simply by the rate of root production in the patches but also by the lifespan of the roots inhabiting the patches. We examined the effect of sustained localized nitrogen (N) fertilization on root lifespan in four tree species that varied widely in root morphology and presumed foraging strategy. The study was conducted in a 12-year-old common garden in central Pennsylvania using a combination of data from minirhizotron and root in-growth cores. The two fine-root tree species, Acer negundo L. and Populus tremuloides Michx., exhibited significant increases in root lifespan with local N fertilization; no significant responses were observed in the two coarse-root tree species, Sassafras albidum Nutt. and Liriodendron tulipifera L. Across species, coarse-root tree species had longer median root lifespan than fine-root tree species. Localized N fertilization did not significantly increase the N concentration or the respiration of the roots growing in the N-rich patch. Our results suggest that some plant species appear to regulate the lifespan of different portions of their root system to improve resource acquisition while other species do not. Our results are discussed in the context of different strategies of foraging of nutrient patches in species of different root morphology.

Highly efficient uptake of ultrafine mesoporous silica nanoparticles with excellent biocompatibility by Liriodendron hybrid suspension cells.

The characteristics of the interactions co-cultures of ultrafine mesoporous silica nanoparticles (MSNs) and the Liriodendron hybrid suspension cells were systematically investigated using laser scanning confocal microscope (LSCM) and scanning electron microscopy (SEM). Using fluorescein isothiocyanate (FITC) labeling, the LSCM observations demonstrated that MSNs (size, 5-15 nm) with attached FITC molecules efficiently penetrated walled plant cells through endocytic pathways, but free FITC could not enter the intact plant cells. The SEM measurements indicated that MSNs readily aggregated on the surface of intact plant cells, and also directly confirmed that MSNs could enter intact plant cells; this was achieved by determining the amount of silicon present. After 24 h of incubation with 1.0 mg mL(-1) of MSNs, the viability of the plant cells was analyzed using fluorescein diacetate staining; the results showed that these cells retained high viability, and no cell death was observed. Interestingly, after the incubation with MSNs, the Liriodendron hybrid suspension cells retained the capability for plant regeneration via somatic embryogenesis. Our results indicate that ultrafine MSNs hold considerable potential as nano-carriers of extracellular molecules, and can be used to investigate in vitro gene-delivery in plant cells.

Identification of conserved and novel microRNAs from Liriodendron chinense floral tissues.

BACKGROUND: Liriodendron chinense (L. chinense) is an endangered basal angiosperm plant in China because of its low reproductive efficiency. Recently, miRNAs have obtained great attention because they can play important roles. Through high throughput sequencing technique, large amount of miRNAs were identified from different plant species. But there were few studies about the miRNAs in the basal angiosperms especially in the sexual reproduction process. RESULTS: Deep sequencing technology was applied to discover miRNAs in L. chinense flowers at different stages. After bioinformatic analysis, 496 putative conserved miRNAs representing 97 families and 2 novel miRNAs were found. Among them, one is previously regarded as gymnosperm specific. Their expressions were further validated by Real-time PCR for 13 selected miRNAs. Putative targeting genes were predicted and categorized with gene ontology (GO) analysis. About ten percents of the targets are involved in the reproduction process. Further expressional analysis showed that many of these miRNAs were highly related to the reproductive growth. CONCLUSIONS: This is the first comprehensive identification of conserved and novel miRNAs in L. chinense. The data presented here might not only help to fill the gap of miRNA registered about basal angiosperm plants but also contribute to understanding the evolution of miRNAs. The differential expression of some of the miRNAs and the prediction of their target genes are also helpful in understanding the regulation of L. chinense sexual reproduction.