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At least 163 records · Page 9

A best-practice guide to predicting plant traits from leaf-level hyperspectral data using partial least squares regression

Partial least squares regression (PLSR) modelling is a statistical technique for correlating datasets, and involves the fitting of a linear regression between two matrices. One application of PLSR enables leaf traits to be estimated from hyperspectral optical reflectance data, facilitating rapid, high-throughput, non-destructive plant phenotyping. This technique is of interest and importance in a wide range of contexts including crop breeding and ecosystem monitoring. The lack of a consensus in the literature on how to perform PLSR means that interpreting model results can be challenging, applying existing models to novel datasets can be impossible, and unknown or undisclosed assumptions can lead to incorrect or spurious predictions. We address this lack of consensus by proposing best practices for using PLSR to predict plant traits from leaf-level hyperspectral data, including a discussion of when PLSR is applicable, and recommendations for data collection. Further, we provide a tutorial to demonstrate how to develop a PLSR model, in the form of an R script accompanying this manuscript. This practical guide will assist all those interpreting and using PLSR models to predict leaf traits from spectral data, and advocates for a unified approach to using PLSR for predicting traits from spectra in the plant sciences.

54 ENVIRONMENTAL SCIENCES↗

Contrasting coordination of non‐structural carbohydrates with leaf and root economic strategies of alpine coniferous forests

Non-structural carbohydrates (NSCs), as the labile fraction and dominant carbon currency, are essential mediators of plant adaptation to environments. However, whether and how NSC coordinates with plant economic strategy frameworks, particularly the well-recognized leaf economics spectrums (LES) and root economics space (RES), remains unclear. We examined the relationships between NSC and key plant economics traits in leaves and fine roots across 90 alpine coniferous populations on the Tibetan Plateau, China. We observed contrasting coordination of NSC with economics traits in leaves and roots. Leaf total NSC and soluble sugar aligned with the leaf economic spectrum, conveying a trade-off between growth and storage in leaves. However, NSC in roots was independent of the root economic spectrum, but highly coordinated with root foraging, with more starch and less sugar in forage-efficient, thinner roots. Further, NSC-trait coordination in leaves and roots was, respectively, driven by local temperature and precipitation. These findings highlight distinct roles of NSC in shaping the above- and belowground multidimensional economics trait space, and NSC-based carbon economics provides a mechanistic understanding of how plants adapt to heterogeneous habitats and respond to environmental changes.

54 ENVIRONMENTAL SCIENCES↗

Wood‐density has no effect on stomatal control of leaf‐level water use efficiency in an Amazonian forest

Forest disturbances increase the proportion of fast-growing tree species compared to slow-growing ones. To understand their relative capacity for carbon uptake and their vulnerability to climate change, and to represent those differences in Earth system models, it is necessary to characterise the physiological differences in their leaf-level control of water use efficiency and carbon assimilation. We used wood density as a proxy for the fast-slow growth spectrum and tested the assumption that trees with a low wood density (LWD) have a lower water-use efficiency than trees with a high wood density (HWD). We selected 5 LWD tree species and 5 HWD tree species growing in the same location in an Amazonian tropical forest and measured in situ steady-state gas exchange on top-of-canopy leaves with parallel sampling and measurement of leaf mass area and leaf nitrogen content. We found that LWD species invested more nitrogen in photosynthetic capacity than HWD species, had higher photosynthetic rates and higher stomatal conductance. Furthermore, contrary to expectations, we showed that the stomatal control of the balance between transpiration and carbon assimilation was similar in LWD and HWD species and that they had the same dark respiration rates.

54 ENVIRONMENTAL SCIENCES↗

Tree architectural characteristics and stem and leaf functional traits for 17 individuals in the Central Amazon

Given recent increases in tree mortality rates in the Amazon forest following extreme drought and wind events, we tested if lower wood density and acquisitive plant functional traits were associated with increased growth and mortality for common co-occurring trees in the Central Amazon. Research was conducted at the ZF2 Research Station located north or Manaus, Brazil, managed by the Instituto Nacional de Pesquisas da Amazônia (INPA). Seventeen trees of different species with similar sizes but a range in wood density (WD) and wood traits were felled, then assessed for 27 different individual functional parameters, including whole tree architecture, stem xylem anatomical and hydraulic traits and leaf traits. Wood logs were collected at DBH, 50% stem length and at 100% stem length (at the base of the canopy). For wood anatomy samples, n=3-6 subsamples from each height. For leaf samples, 30 leaves were collected from the upper sunlit canopy. The methodology is detailed in the accompanying manuscript. The trait data are summarized in this file: "Trait_Summary.CSV". Summary Trait code abbreviations and units are described in this file: "Sample_Info_Traits_Summary.CSV". Stem traits measured along the bole from the base of the tree (DBH, diameter breast height), mid-stem, and base of the canopy are described in these files: "Sapwood_Area_height.CSV"; "Species_Info_height.CSV"; "Sample_Info_height.CSV"

54 ENVIRONMENTAL SCIENCES↗

Celestial leaf amplitudes

Celestial amplitudes may be decomposed as weighted integrals of AdS 3 -Witten diagrams associated to each leaf of a hyperbolic foliation of spacetime. We show, for the Kleinian three-point MHV amplitude, that each leaf subamplitude is smooth except for the expected light-cone singularities. Moreover, we find that the full translationally-invariant celestial amplitude is simply the residue of the pole in the leaf amplitude at the point where the total conformal weights of the gluons equals three. This full celestial amplitude vanishes up to light-cone contact terms, as required by spacetime translation invariance, and reduces to the expression previously derived by Mellin transformation of the Parke-Taylor formula.

79 ASTRONOMY AND ASTROPHYSICS↗

Quantitative trait locus mapping combined with variant and transcriptome analyses identifies a cluster of gene candidates underlying the variation in leaf wax between upland and lowland switchgrass ecotypes

Switchgrass (Panicum virgatum L.) is a promising warm-season candidate energy crop. It occurs in two ecotypes, upland and lowland, which vary in a number of phenotypic traits, including leaf glaucousness. To initiate trait mapping, two F 2 mapping populations were developed by crossing two different F 1 sibs derived from a cross between the tetraploid lowland genotype AP13 and the tetraploid upland genotype VS16, and high-density linkage maps were generated. Quantitative trait locus (QTL) analyses of visually scored leaf glaucousness and of hydrophobicity of the abaxial leaf surface measured using a drop shape analyzer identified highly significant colocalizing QTL on chromosome 7K (Chr07K). Using a multipronged approach, we identified a cluster of genes including Pavir.7KG077009, which encodes a Type III polyketide synthase-like protein, and Pavir.7KG013754 and Pavir.7KG030500, two highly similar genes that encode putative acyl-acyl carrier protein (ACP) thioesterases, as strong candidates underlying the QTL. The lack of homoeologs for any of the three genes on Chr07N, the relatively low level of identity with other switchgrass KCS proteins and thioesterases, as well as the organization of the surrounding region suggest that Pavir.7KG077009 and Pavir.7KG013754/Pavir.7KG030500 were duplicated into a fast-evolving chromosome region, which led to their neofunctionalization. Furthermore, sequence analyses showed all three genes to be absent in the two upland compared to the two lowland accessions analyzed. This study provides an example of and practical guide for trait mapping and candidate gene identification in a complex genetic system by combining QTL mapping, transcriptomics and variant analysis.

59 BASIC BIOLOGICAL SCIENCES↗

Lattice light-sheet microscopy allows for super-resolution imaging of receptors in leaf tissue

Plant leaf tissues are difficult to image via fluorescent microscopy, largely due to the presence of chlorophyll and other pigments that provide large background fluorescence. An advantage of Lattice Light-Sheet microscopy is its use of Bessel beams that illuminate a thin focal region of interest for microscopy, allowing for the excitation of fluorescent molecules within this region without surrounding chlorophyll-like objects outside of the region of interest. Here, we apply STORM Super-resolution techniques to observe Receptor-Like Kinases in Arabidopsis thaliana leaf cells. By applying this technique with the Lattice Light-Sheet, we can localize immune response proteins in sub-100 nm length scales and reconstruct three-dimensional locations of proteins within individual leaf cells. Using this technique, we observed the effect of the elicitors ATP and flg22, where we observed a significant degree of internalization of cognate receptors P2K1 and FLS2. We were also able to similarly observe differences in colocalization due to stimulation with these elicitors, where we observe proteins on the membrane becoming less colocalized as a result of stimulation, suggesting an immune response mechanism involving receptors internalizing via pathways distinct to the receptor. Further, these data show the Lattice Light-Sheet’s capabilities for imaging tissue with problematic background fluorescence that otherwise makes super-resolution fluorescence microscopy difficult.

59 BASIC BIOLOGICAL SCIENCES↗

DUET - Distribution of Understory using Elliptical Transport: A mechanistic model of leaf litter and herbaceous spatial distribution based on tree canopy structure

Heterogeneity in surface fuels produced by overstory trees and understory vegetation is a major driver of fire behavior and ecosystem dynamics. Previous attempts at predicting tree leaf and needle litter accumulation over time have been constrained in scope to probabilistic models that consider a limited number of key factors influencing tree litter dispersal patterns and decomposition processes. Here we present a mechanistic model for estimating variation in surface fuels called the Distribution of Understory using Elliptical Transport (DUET). DUET uses a pre-generated voxelated canopy array and distributes the leaf litter from the trees in an elliptical shape based on tree species characteristics, wind data, and location-specific features. DUET then calculates grass growth between trees and around the litter patterns and sets moisture levels based on relative humidity. In this study, we use DUET to simulate spatial distributions of leaf litter and illustrate model sensitivity to several parameters, providing inputs for testing fuel spatial heterogeneity impacts on fire behavior using process-based models.

54 ENVIRONMENTAL SCIENCES↗

Leaf-cutting ant (Atta cephalotes) nests may be hotspots of methane and carbon dioxide emissions in tropical forests

Leaf-cutting ants of the genus Atta are widely distributed throughout the American tropics and subtropics and rival other herbivores in the consumption of surrounding foliage. Although numerous studies have been conducted on the role these insects play in herbivory and organic matter dynamics, only a handful of studies have examined their impacts on soil greenhouse gas emissions. Our study investigated fluxes of carbon dioxide (CO 2 ) and methane (CH 4 ) from three nests of Atta cephalotes using a portable greenhouse gas analyzer, and measured CO 2 and CH 4 emissions from soils containing nest holes that ranged 5.2–152.1 g CO 2 -C and –1.1 to 15,264.7 mg CH 4 -C m -2 day -1 , respectively. Fluxes of CO 2 and CH 4 were positively correlated above nest holes, but not in patches of soil away from leaf-cutting ant nests. Nearby non-nest soil emissions were significantly lower, ranging from 0.6 to 6.0 g CO 2 -C and –1.3 to 0.77 mg CH 4 -C m -2 day -1 . Fluxes of both gases among nests and among holes within a single nest were highly variable. This preliminary dataset is small in scale both temporarily and geographically, but the discovery of substantial greenhouse gas fluxes from Atta cephalotes nests may have important implications for carbon budgets of tropical and subtropical American forests. Further work will be necessary to determine the mechanisms behind enhanced greenhouse gas emissions from leaf-cutting ant nests, and how this may alter ecosystem-scale CO 2 emissions and CH 4 sink strength in tropical forest soils.

54 ENVIRONMENTAL SCIENCES↗

Grass Evolutionary Lineages Can Be Identified Using Hyperspectral Leaf Reflectance

Hyperspectral remote sensing has the potential to map numerous attributes of the Earth’s surface, including spatial patterns of biological diversity. Grasslands are one of the largest biomes on Earth. Accurate mapping of grassland biodiversity relies on spectral discrimination of endmembers of species or plant functional types. We focused on spectral separation of grass lineages that dominate global grassy biomes: Andropogoneae (C 4 ), Chloridoideae (C 4 ), and Pooideae (C 3 ). We examined leaf reflectance spectra (350–2,500 nm) from 43 grass species representing these grass lineages from four representative grassland sites in the Great Plains region of North America. Here, we assessed the utility of leaf reflectance data for classification of grass species into three major lineages and by collection site. Classifications had very high accuracy (94%) that were robust to site differences in species and environment. We also show an information loss using multispectral sensors, that is, classification accuracy of grass lineages using spectral bands provided by current multispectral satellites is much lower (accuracy of 85.2% and 61.3% using Sentinel 2 and Landsat 8 bands, respectively). Our results suggest that hyperspectral data have an exciting potential for mapping grass functional types as informed by phylogeny. Leaf-level hyperspectral separability of grass lineages is consistent with the potential increase in biodiversity and functional information content from the next generation of satellite-based spectrometers.

59 BASIC BIOLOGICAL SCIENCES↗

Incorporating the Acclimation of Photosynthesis and Leaf Respiration in the Noah‐MP Land Surface Model: Model Development and Evaluation

Abstract Realistic simulation of leaf photosynthetic and respiratory processes is needed for accurate prediction of the global carbon cycle. These two processes systematically acclimate to long‐term environmental changes by adjusting photosynthetic and respiratory traits (e.g., the maximum photosynthetic capacity at 25°C ( V cmax,25 ) and the leaf respiration rate at 25°C ( R 25 )) following increasingly well‐understood principles. While some land surface models (LSMs) now account for thermal acclimation, they do so by assigning empirical parameterizations for individual plant functional types (PFTs). Here, we have implemented an Eco‐Evolutionary Optimality (EEO)‐based scheme to represent the universal acclimation of photosynthesis and leaf respiration to multiple environmental effects, and that therefore requires no PFT‐specific parameterizations, in a standard version of the widely used LSM, Noah MP. We evaluated model performance with plant trait data from a 5‐year experiment and extensive global field measurements, and carbon flux measurements from FLUXNET2015. We show that observed R 25 and V cmax,25 vary substantially both temporally and spatially within the same PFT ( C.V. >20%). Our EEO‐based scheme captures 62% of the temporal and 70% of the spatial variations in V cmax,25 (73% and 54% of the variations in R 25 ). The standard scheme underestimates gross primary production by 10% versus 2% for the EEO‐based scheme and generates a larger spread in r (correlation coefficient) across flux sites (0.79 ± 0.16 vs. 0.84 ± 0.1, mean ± S.D.). The standard scheme greatly overestimates canopy respiration (bias: ∼200% vs. 8% for the EEO scheme), resulting in less CO 2 uptake by terrestrial ecosystems. Our approach thus simulates climate‐carbon coupling more realistically, with fewer parameters.

Ren, Yanghang [Department of Earth System Science ↗

Arabidopsis thaliana zinc accumulation in leaf trichomes is correlated with zinc concentration in leaves

Zinc (Zn) is a key micronutrient for plants and animals, and understanding Zn homeostasis in plants can improve both agriculture and human health. While root Zn transporters in plant model species have been characterized in detail, comparatively little is known about shoot processes controlling Zn concentrations and spatial distribution. Previous work showed that Zn hyperaccumulator species such as Arabidopsis halleri accumulate Zn and other metals in leaf trichomes. To date there is no systematic study regarding Zn accumulation in the trichomes of the non-accumulating, genetic model species A. thaliana. Here, we used Synchrotron X-Ray Fluorescence mapping to show that Zn accumulates at the base of trichomes of A. thaliana. Using transgenic and natural accessions of A thaliana that vary in bulk leaf Zn concentration, we demonstrate that higher leaf Zn increases total Zn found at the base of trichome cells. Our data indicates that Zn accumulation in trichomes is a function of the Zn status of the plant, and provides the basis for future studies on a genetically tractable plant species to understand the molecular steps involved in Zn spatial distribution in leaves.

59 BASIC BIOLOGICAL SCIENCES↗

Boosts in leaf-level photosynthetic capacity aid Pinus ponderosa recovery from wildfire

Forests mitigate climate change by sequestering massive amounts of carbon, but recent increases in wildfire activity are threatening carbon storage. Currently, our understanding of wildfire impacts on forest resilience and the mechanisms controlling post-fire recovery remains unresolved due to a lack of empirical data on mature trees in natural settings. Here, we quantify the physiological mechanisms controlling carbon uptake immediately following wildfire in mature individuals of ponderosa pine (Pinus ponderosa), a wide-spread and canopy-dominant tree species in fire-prone forests. While photosynthetic capacity was lower in burned than unburned trees due to an overall depletion of resources, we show that within the burned trees, photosynthetic capacity increases with the severity of damage. Our data reveal that boosts in the efficiency of carbon uptake at the leaf-level may compensate for whole-tree damage, including the loss of leaf area and roots. We further show that heightened photosynthetic capacity in remaining needles on burned trees may be linked with reduced water stress and leaf nitrogen content, providing pivotal information about post-fire physiological processes. Our results have implications for Earth system modeling efforts because measurements of species-level physiological parameters are used in models to predict ecosystem and landscape-level carbon trajectories. Finally, current land management practices do not account for physiological resilience and recovery of severely burned trees. Our results suggest premature harvest may remove individuals that may otherwise survive, irrevocably altering forest carbon balance.

54 ENVIRONMENTAL SCIENCES↗

Stomatal closure as a driver of minimum leaf conductance declines at high temperature and vapor pressure deficit in Quercus

Abstract Rising global temperatures and vapor pressure deficits (VPDs) are increasing plant water demand and becoming major drivers of large-scale plant mortality. Controlling transient leaf water loss after stomatal closure (minimum stomatal conductance [gmin]) is recognized as a key trait determining how long plants survive during soil drought. Yet, substantial uncertainty remains regarding how gmin responds to elevated temperatures and VPD and the underlying mechanisms. We measured gmin in 24 Quercus species from temperate and Mediterranean climates to determine whether gmin was sensitive to a coupled temperature and VPD increase. We also explored mechanistic links to phenology, climate, evolutionary history, and leaf anatomy. We found that gmin in all species exhibited a nonlinear negative temperature and VPD dependence. At 25 °C (VPD = 2.2 kPa), gmin varied from 1.19 to 8.09 mmol m−2 s−1 across species but converged to 0.57 ± 0.06 mmol m−2 s−1 at 45 °C (VPD = 6.6 kPa). In a subset of species, the effect of temperature and VPD on gmin was reversible and linked to the degree of stomatal closure, which was greater at 45 °C than at 25 °C. Our results show that gmin is dependent on temperature and VPD, is highly conserved in Quercus species, and is linked to leaf anatomy and stomatal behavior.

Zailaa, Joseph (ORCID:000000019103190X)↗

Mechanisms underlying leaf photosynthetic acclimation to warming and elevated CO 2 as inferred from least‐cost optimality theory

Abstract The mechanisms responsible for photosynthetic acclimation are not well understood, effectively limiting predictability under future conditions. Least‐cost optimality theory can be used to predict the acclimation of photosynthetic capacity based on the assumption that plants maximize carbon uptake while minimizing the associated costs. Here, we use this theory as a null model in combination with multiple datasets of C 3 plant photosynthetic traits to elucidate the mechanisms underlying photosynthetic acclimation to elevated temperature and carbon dioxide (CO 2 ). The model‐data comparison showed that leaves decrease the ratio of the maximum rate of electron transport to the maximum rate of Rubisco carboxylation ( J max / V cmax ) under higher temperatures. The comparison also indicated that resources used for Rubisco and electron transport are reduced under both elevated temperature and CO 2 . Finally, our analysis suggested that plants underinvest in electron transport relative to carboxylation under elevated CO 2 , limiting potential leaf‐level photosynthesis under future CO 2 concentrations. Altogether, our results show that acclimation to temperature and CO 2 is primarily related to resource conservation at the leaf level. Under future, warmer, high CO 2 conditions, plants are therefore likely to use less nutrients for leaf‐level photosynthesis, which may impact whole‐plant to ecosystem functioning.

Smith, Nicholas G.↗

PHOTO‐SENSITIVE LEAF ROLLING 1 encodes a polygalacturonase that modifies cell wall structure and drought tolerance in rice

Summary The biosynthesis and modification of cell wall composition and structure are controlled by hundreds of enzymes and have a direct consequence on plant growth and development. However, the majority of these enzymes has not been functionally characterised. Rice mutants with leaf‐rolling phenotypes were screened in a field. Phenotypic analysis under controlled conditions was performed for the selected mutant and the relevant gene was identified by map‐based cloning. Cell wall composition was analysed by glycome profiling assay. We identified a photo‐sensitive leaf rolling 1 ( psl1 ) mutant with ‘napping’ (midday depression of photosynthesis) phenotype and reduced growth. The PSL1 gene encodes a cell wall‐localised polygalacturonase (PG), a pectin‐degrading enzyme. psl1 with a 260‐bp deletion in its gene displayed leaf rolling in response to high light intensity and/or low humidity. Biochemical assays revealed PG activity of recombinant PSL1 protein. Significant modifications to cell wall composition in the psl1 mutant compared with the wild‐type plants were identified. Such modifications enhanced drought tolerance of the mutant plants by reducing water loss under osmotic stress and drought conditions. Taken together, PSL1 functions as a PG that modifies cell wall biosynthesis, plant development and drought tolerance in rice.

Zhang, Guangheng↗

Seasonal trends in leaf-level photosynthetic capacity and water use efficiency in a North American Eastern deciduous forest and their impact on canopy-scale gas exchange

Vegetative transpiration (E) and photosynthetic carbon assimilation (A) are known to be seasonally dynamic, with changes in their ratio determining the marginal water use efficiency (WUE). Despite an understanding that stomata play a mechanistic role in regulating WUE, it is still unclear how stomatal and nonstomatal processes influence change in WUE over the course of the growing season. As a result, limited understanding of the primary physiological drivers of seasonal dynamics of canopy WUE remains one of the largest uncertainties in earth system model projections of carbon and water exchange in temperate deciduous forest ecosystems. Here, we investigated seasonal patterns in leaf-level physiological, hydraulic, and anatomical properties, including the seasonal progress of the stomatal slope parameter (g 1 ; inversely proportional to WUE) and the maximum carboxylation rate (V cmax ). V cmax and g 1 were seasonally variable; however, their patterns were not temporally synchronized. g 1 generally showed an increasing trend until late in the season, while V cmax peaked during the midsummer months. Seasonal progression of V cmax was primarily driven by changes in leaf structural, and anatomical characteristics, while seasonal changes in g 1 were most strongly related to changes in V cmax and leaf hydraulics. Using a seasonally variable V cmax and g 1 to parameterize a canopy-scale gas exchange model increased seasonally aggregated A and E by 3% and 16%, respectively.

54 ENVIRONMENTAL SCIENCES↗

Single-cell proteomics of Arabidopsis leaf mesophyll reveals dynamic protein responses to water-deficit stress

Background The application of single-cell omics tools to biological systems can provide unique insights into diverse cellular populations and their heterogeneous responses to internal and external perturbations. Thus far, most single-cell studies in plant systems have been limited to RNA-sequencing approaches, which only provide indirect readouts of cellular functions. Results Here, we present a single-cell proteomics workflow for plant cells that integrates tape-sandwich protoplasting, piezoelectric cell sorting, nanoPOTS sample preparation, and ion mobility-based MS data acquisition method for label-free single-cell proteomics analysis of Arabidopsis leaf mesophyll cells. From a single leaf protoplast, over 3,000 proteins were quantified with high precision. The workflow is demonstrated to identify stress associated changes in protein abundance by analyzing 117 protoplasts from well-watered and water-deficit stressed plants. Additionally, we describe a new approach for constructing covarying protein networks at the single-cell level and demonstrate how single-cell protein covariation analysis can reveal previously unrecognized protein functions while also capturing stress-induced changes in protein–protein dynamics. Conclusions The label-free scProteomic approach presented here represents a significant advance through the demonstration of a facile protoplast isolation method combined with deep and precise proteomic coverage of Arabidopsis leaf mesophyll cell types. We believe this study will serve as an informative reference to future plant scProteomic investigations.

Arabidopsis↗