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DOE OSTI2026

Coupling of high-resolution mass spectrometer and photosynthesis system for comprehensive leaf volatile metabolite profiling

Background Leaf-level biogenic volatile organic compounds (BVOCs) emissions represent a major source of organic gases in the atmosphere, influencing both climate and air quality. These emissions are strongly driven by environmental perturbations, which affect individual plant- to ecosystem-level processes. Uncovering all the BVOCs and understanding how their emissions respond to altered environmental conditions provide critical insights into vegetation-driven changes in atmospheric chemistry. We developed a tandem instrumentation setup that integrates a proton transfer reaction time-of-flight mass spectrometer (PTR-ToF-MS) with parts-per-trillion detection limits and a photosynthetic infrared gas exchange system for the untargeted survey of all the BVOCs. This novel system enables simultaneous, real-time monitoring of BVOC emissions and photosynthetic parameters at the leaf level, offering new opportunities to disentangle the physiological and environmental drivers of VOC release. Furthermore, we established the VOC Analysis and Processing Optimization Resource (VAPOR), an open-access software tool designed for rapid data post-processing and the analysis of the variability of hundreds of BVOCs. We assessed the performance of the tandem system under varying background conditions, using standard gas mixtures and a range of environmental factors. Results Blank emissions were substantially lower for major BVOCs (e.g., isoprene) compared to those observed in plant emissions. Despite this, the observation of background-level VOCs highlights the importance of routinely acquiring and accounting for blank measurements in analyses using the coupled instrumentation. Introduction of known VOC concentrations to the system demonstrated a linear response across different compounds with varying molecular compositions, indicating minimal gas loss regardless of chemical moieties within the coupled instrumentation. We applied the optimized system to investigate the physiological mechanisms driving BVOC emissions across different genotypes of poplar and pennycress. The high mass resolution capabilities of the PTR-ToF-MS, coupled with comprehensive VAPOR-driven data analysis, enabled the identification of several important BVOCs, including methanol and methanethiol; these BVOCs displayed substantial variation across pennycress genotypes and showed concentrations ~ 100–350% higher than the blank. Moreover, isoprene emissions varied significantly among poplar genotypes grown in different potting media. Conclusions Tandem instrumentation offers a powerful tool for profiling volatile molecular markers and elucidating their genetic and environmental underpinnings. This approach enhances our ability to predict BVOC emissions in response to genotype by environmental interactions and contributes to a deeper understanding of vegetation responses to environmental changes.

Biogenic volatile organic compounds↗

DOE OSTI2021

Low soil phosphorus availability triggers maize growth stage specific rhizosphere processes leading to mineralization of organic P

Phosphorus (P) is one of the essential nutrients for all living organisms. High-quality mineral P fertilizer is a finite resource with only an estimated 80-100 years of reserves remaining globally. Both plants and microbes have developed several mechanisms such as secretion of organic acids, acidification of the rhizosphere, and production of extracellular enzymes to enhance the acquisition of P from soils. Increased activity of phosphatase occurs in response to P deficiency as part of P starvation responses which catalyze the hydrolysis of P o from soil organic matter (SOM) and release inorganic P (P i ). However, the specific interactions between plants and microbial communities in the rhizosphere soil triggered by P deficiency are largely unknown. In a greenhouse study we grew maize plants in low and high P containing soils and examined the role of rhizosphere processes on mineralization of organic P (P o ) in soils with low and high P availability and different P speciation 23 (Po dominated versus P i dominated) and determine its effect on plant growth. Plant biomass was determined and rhizosphere soils, and soil samples from pots with and without plants were collected at key vegetative growth stages (VGS). A variety of biogeochemical parameters (e.g. microbial biomass C and P, potential phosphatase activities, plant biomass P, soil P species etc.) were determined using both conventional (sequential chemical extraction) and advanced (e.g. high resolution mass spectrometry (FTICR-MS) and 3129 P solution NMR) techniques. Here, we used phospholipid fatty acid (PLFA) and DNA based microbial community analysis to track changes in microbial community structure and diversity. Low P availability induced changes in biogeochemical processes and microbial community composition in the rhizosphere soils of plants grown in low P containing soil. At early VGS, the plants in low P soil were visually struggling which correlated with greater rhizosphere potential acid phosphatase activity and a reduction in P o fractions. At late VGS, the plants appeared to recover which correlated with a decrease in Meh (III) extractable P, an increase in microbial biomass C and P and greater total P in the plant biomass (roots, shoots and leaves). In high P containing soil, on the other hand, greater P availability masked these rhizosphere processes. Our results confirmed the degradation of SOM and Po mineralization in rhizosphere soils driven by microbe and plant need for P. P deficiency may have favored the abundance of fungi which utilized easily degradable root exudates for the production of extracellular enzymes responsible for the mineralize of SOM P o . This study highlights the intertwined mechanisms involved, and emphasizes the importance of, P o sources for plant nutrition and survival in marginal soils.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1985

Biogeochemical factors which regulate the formation and fate of sulfide in wetlands

Coastal wetland areas occupy a small percentage of the terrestrial environment yet are extremely productive regions which support rapid rates of below ground bacterial activity. Wetlands appear to be significant as biogenic sources of gaseous sulfur, carbon, and nitrogen. These gases are important as tracers of man's activities, and they influence atmospheric chemistry. The interactions among wetland biogeochemical processes regulate the anaerobic production of reduced gases and influence the fate of these volatiles. Therefore, spatial and temporal variations in hydrology, salinity, temperature, and speciation and growth of vegetation affect the type and magnitude of gas emissions thus hindering predictive estimates of gas flux. The research is divided into two major parts, the first is the biogeochemical characterization of a selected tidal wetland area in terms of factors likely to regulate sulfide flux; the second is a direct measurement of gaseous sulfur flux as related to changes in these biogeochemical conditions. Variant factors affecting sulfide flux include the wetlands' tidal range, seasonal salinity, and other hydrological conditions, grass species and plant growth, soil composition, and microbial activity.

Hines, Mark E.↗

DOE OSTI2023

Seasonality regulates the structure and biogeochemical impact of ectomycorrhizal fungal communities across environmentally divergent neotropical dry forests

Abstract Ectomycorrhizal (ECM) symbioses support forest functioning globally, yet both the structure and function of ECM fungal communities in seasonally dry neotropical forests (SDTFs), known for extreme heterogeneity in vegetation and edaphic properties, remain under characterized. Here, we evaluated the relative influences of seasonal versus spatial variation in ECM fungal community structure in soils from four environmentally divergent SDTFs. We also assessed the importance of biotic and abiotic drivers of SDTF ECM fungal community structure at regional scales, as well as ECM impacts on soil carbon (C) and nitrogen (N) cycling. ECM fungal frequency, relative abundance and richness all increased in the wet season, but spatial rather than seasonal effects explained more variation in community composition. Across the four SDTFs investigated, differences in tree communities drove ECM fungal community turnover more than geographic distances, site abiotic conditions or soil chemistry. Although soil moisture and ECM tree basal area were stronger predictors of soil biogeochemistry, incorporating ECM fungal community composition and relative abundance added explanatory power to models of soil C and N cycling in the wet season. Synthesis : Our results highlight the importance of seasonality and plant community composition in shaping different aspects of SDTF ECM fungal community structure and diversity as well as the potential for both the plant and fungal components of ECM symbioses to impact soil functioning across heterogenous SDTFs. Furthermore, our findings suggest that alterations in SDTF plant community composition due to climate or land‐use change will have important consequences for ECM fungal diversity and associated effects on soil biogeochemical cycling.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2021

Vegetation Type and Decomposition Priming Mediate Brackish Marsh Carbon Accumulation Under Interacting Facets of Global Change

Coastal wetland carbon pools are globally important, but their response to interacting facets of global change remain unclear. Numerical models neglect species-specific vegetation responses to sea level rise (SLR) and elevated CO 2 (eCO 2 ) that are observed in field experiments, while field experiments cannot address the long-term feedbacks between flooding and soil growth that models show are important. Here, we present a novel numerical model of marsh carbon accumulation parameterized with empirical observations from a long-running eCO 2 experiment in an organic rich, brackish marsh. Model results indicate that eCO 2 and SLR interact synergistically to increase soil carbon burial, driven by shifts in plant community composition and soil volume expansion. Furthermore, newly parameterized interactions between plant biomass and decomposition (i.e. soil priming) reduce the impact of eCO 2 on marsh survival, and by inference, the impact of eCO 2 on soil carbon accumulation.

54 ENVIRONMENTAL SCIENCES↗

NASA NTRS2013

United States Forest Disturbance Trends Observed Using Landsat Time Series

Disturbance events strongly affect the composition, structure, and function of forest ecosystems; however, existing U.S. land management inventories were not designed to monitor disturbance. To begin addressing this gap, the North American Forest Dynamics (NAFD) project has examined a geographic sample of 50 Landsat satellite image time series to assess trends in forest disturbance across the conterminous United States for 1985-2005. The geographic sample design used a probability-based scheme to encompass major forest types and maximize geographic dispersion. For each sample location disturbance was identified in the Landsat series using the Vegetation Change Tracker (VCT) algorithm. The NAFD analysis indicates that, on average, 2.77 Mha/yr of forests were disturbed annually, representing 1.09%/yr of US forestland. These satellite-based national disturbance rates estimates tend to be lower than those derived from land management inventories, reflecting both methodological and definitional differences. In particular the VCT approach used with a biennial time step has limited sensitivity to low-intensity disturbances. Unlike prior satellite studies, our biennial forest disturbance rates vary by nearly a factor of two between high and low years. High western US disturbance rates were associated with active fire years and insect activity, while variability in the east is more strongly related to harvest rates in managed forests. We note that generating a geographic sample based on representing forest type and variability may be problematic since the spatial pattern of disturbance does not necessarily correlate with forest type. We also find that the prevalence of diffuse, non-stand clearing disturbance in US forests makes the application of a biennial geographic sample problematic. Future satellite-based studies of disturbance at regional and national scales should focus on wall-to-wall analyses with annual time step for improved accuracy.

Masek, Jeffrey G.↗

NASA NTRS1985

Vegetation canopy discrimination and biomass assessment using multipolarized airborne SAR

Multipolarized airborne Synthetic Aperture Radar (SAR) data were acquired over a largely agricultural test site near Macomb, Illinois, in conjunction with the Shuttle Imaging Radar (SIR-B) experiment in October 1984. The NASA/JPL L-band SAR operating at 1.225 GHz made a series of daily overflights with azimuth view angles both parallel and orthogonal to those of SIR-B. The SAR data was digitally recorded in the quadpolarization configuration. An extensive set of ground measurements were obtained throughout the test site and include biophysical and soil measurements of approximately 400 agricultural fields. Preliminary evaluation of some of the airborne SAR imagery indicates a great potential for crop discrimination and assessment of canopy condition. False color composites constructed from the combination of three linear polarizations (HH, VV, and HV) were found to be clearly superior to any single polarization for purposes of crop classification. In addition, an image constructed using the HH return to modulate intensity and the phase difference between HH and VV returns to modulate chroma indicates a clear capability for assessment of canopy height and/or biomass. In particular, corn fields heavily damaged by infestations of corn borer are readily distinguished from noninfested fields.

Ulaby, F. T.↗

DOE OSTI2021

A micrometeorological flux perspective on brush management in a shrub-encroached Sonoran Desert grassland

Woody plant encroachment typically limits the forage productivity of managed rangelands and alters a panoply of semiarid ecosystem processes and services. Intervention strategies to reduce woody plant abundance, collectively termed “brush management”, often lack observations to quantify and interpret changes in ecosystem processes. Furthermore, comparative studies between treated and untreated areas should account for heterogeneity since plant composition, microclimate, topoedaphic factors, and historical land use can substantially vary over short distances in drylands. Here, in this work, we quantify ecosystem responses to brush management after a single aerial herbicide application on an 18 hectare shrub-encroached grassland (savanna) in southern Arizona, USA. We conducted a pre- and post-treatment comparison of a flux tower site in the treated area with that of a tower in a nearby control site. The comparison, spanning a seven year period, included: (1) ground, airborne, and satellite-based measurements of vegetation structure, and (2) eddy covariance measurements. The herbicide treatment defoliated the dominant shrub (velvet mesquite, Prosopis velutina) and led to a temporary reduction in summer greening, but full foliar recovery occurred within two years. Contrary to expectations, perennial grass cover decreased and bare soil cover increased on the treated site. Relative amounts of evapotranspiration were reduced, while carbon uptake increased during the 2 year post-treatment period at the treated site due to a higher water use efficiency in the following spring. During mesquite recovery, carbon uptake was enhanced by higher gross primary productivity and accompanied by a decrease in ecosystem respiration relative to the untreated site. Mesquite recovery was facilitated by access to deep soil water, carbohydrate reserves in rooting systems, and a lower competition from reduced perennial grass cover.

54 ENVIRONMENTAL SCIENCES↗

NASA NTRS1973

Land use mapping and modelling for the Phoenix Quadrangle

The author has identified the following significant results. The land use of the Phoenix Quadrangle in Arizona had been mapped previously from aerial photographs and recorded in a computer data bank. During the ERTS experiment, changes in land use were detected, first with the ERTS-simulation photographs, then with the ERTS-1 images when they became available. In each case, the I2S color additive viewer was used as the primary image enhancement tool, operated in a multispectral mode. A search was made for a method of creating hard copy color composite images of the best combinations of multiband composites from ERTS-1, mostly by photographic and diazo processes. The I2S viewer was also used to enhance changes between successive images by quick flip techniques or by registering with different color filters. Improved interpretation of land use change resulted, and a map of changes in the Phoenix Quadrangle was compiled using magnified ERTS-1 images alone. The first level of a standard land use classification system was successfully used. Between the ERTS-1 images for August and November, some differences were detected that could be caused by seasonal characteristics of vegetation or by change in use.

Place, J. L.↗

NASA NTRS2004

Comparison of Landsat-7 Enhanced Thematic Mapper Plus (ETM+) and Earth Observing One (EO-1) Advanced Land Imager

We compare images from the Enhanced Thematic Mapper Plus (ETM+) sensor on Landsat-7 and the Advanced Land Imager (ALI) instrument on Earth Observing One (EO-1) over a test site in Rochester, New York. The site contains a variety of features, ranging from water of varying depths, deciduous/coniferous forest, and grass fields, to urban areas. Nearly coincident cloud-free images were collected one minute apart on 25 August 2001. We also compare images of a forest site near Howland, Maine, that were collected on 7 September, 2001. We atmospherically corrected each pair of images with the Second Simulation of the Satellite Signal in the Solar Spectrum (6S) atmosphere model, using aerosol optical thickness and water vapor column density measured by in situ Cimel sun photometers within the Aerosol Robotic Network (AERONET), along with ozone density derived from the Total Ozone Mapping Spectrometer (TOMS) on the Earth Probe satellite. We present true-color composites from each instrument that show excellent qualitative agreement between the multispectral sensors, along with grey-scale images that demonstrate a significantly improved ALI panchromatic band. We quantitatively compare ALI and ETM+ reflectance spectra of a grassy field in Rochester and find < or equal to 6% differences in the visible/near infrared and approx. 2% differences in the short wave infrared. Spectral comparisons of forest sites in Rochester and Howland yield similar percentage agreement except for band 1, which has very low reflectance. Principal component analyses and comparison of normalized difference vegetation index histograms for each sensor indicate that the ALI is able to reproduce the information content in the ETM+ but with superior signal-to-noise performance due to its increased 12-bit quantization.

Pedelty, Jeffrey A.↗

NASA NTRS1973

Growth and decline of vegetation on mine dumps

The author has identified the following significant results. The main objective is to determine the extent to which can be used to differentiate between mine dumps having varying degrees of vegetative cover. At this stage it is clear that the various mine dumps can be located and identified. Differences in vegetative cover can be seen and measured. Patterns of vegetative growth, some characteristic to particular dumps, can also be seen. It is therefore tentatively concluded that mine dumps can be differentiated with respect to their vegetative cover on the imagery received to date, and this is reported as a significant result. Subsequent imagery showing seasonal variations should facilitate this program. In addition to work on the mine dumps, a photogeological project has been initiated on geotectonics of South Africa. Studies of ERTS-1 images has indicated that major structures (faults, folds, and linear features) associated with the three geotectonic environments can be identified. In addition, major as well as relatively minor stratigraphic subdivisions can be recognized by their color tones. Results obtained warrant continuation of this study using color composite prints enlarged to a scale of 1:500,000.

Gilbertson, B. P.↗

NASA NTRS1973

Remote sensing of physiographic soil units of Bennett County, South Dakota

A study was conducted in Bennett County, South Dakota, to establish a rangeland test site for evaluating the usefulness of ERTS data for mapping soil resources in rangeland areas. Photographic imagery obtained in October, 1970, was analyzed to determine which type of imagery is best for mapping drainage and land use patterns. Imagery of scales ranging from 1:1,000,000 to 1.20,000 was used to delineate soil-vegetative physiographic units. The photo characteristics used to define physiographic units were texture, drainage pattern, tone pattern, land use pattern and tone. These units will be used as test data for evaluating ERTS data. The physiographic units were categorized into a land classification system. The various categories which were delineated at the different scales of imagery were designed to be useful for different levels of land use planning. The land systems are adequate only for planning of large areas for general uses. The lowest category separated was the facet. The facets have a definite soil composition and represent different soil landscapes. These units are thought to be useful for providing natural resource information needed for local planning.

Frazee, C. J.↗

NASA NTRS1993

Use of spectral analogy to evaluate canopy reflectance sensitivity to leaf optical property

The spectral variation of canopy reflectance is mostly governed by the absorption properties of the elements, hence the leaves, since their intrinsic scattering properties show very little spectral variation. The relationship between canopy reflectance and leaf reflectance measured at the red edge over sugar beet canopies was used to simulate canopy reflectance from leaf reflectance spectra measured over the whole spectral domain. The results show that the spectral analogies found allows accurate reconstruction of canopy reflectance spectra. Explicit assumptions about the very low spectral variation of leaf intrinsic scattering properties are thus indirectly justified. The sensitivity of canopy reflectance (rho(sub c)) to leaf optical properties can then be investigated from concurrent spectral variations of canopy (delta rho(sub c)/delta lambda) and leaf reflectance (delta rho(sub l)/delta lambda): (delta rho(sub c))/(delta rho(sub l)) = ((delta rho(sub c))/(delta lambda) ((delta rho( sub l))/(delta lambda))(sup -1)). This expression is strictly valid only when the optical properties of the soil background or the other vegetation elements such as bark are either spectrally flat or do not contribute significantly to canopy reflectance. Simulations using the SAIL and PROSPECT models demonstrate that the sensitivity of canopy reflectance to leaf reflectance is significant for large vegetation cover fractions in spectral domains where absorption is low. In these conditions, multiple, scattering enhances the leaf absorption features by a factor that can be greater than 2.0. To override the limitations of the SAIL model for the description of the canopy architecture, we tested the previous findings on experimental data. Concurrent canopy and leaf reflectance spectra were measured for a range of sugar beet canopies. The results show good agreement with the theoretical findings. Conclusions are drawn about the applicability of these findings, with particular attention to the potential detectability of leaf biochemical composition from canopy reflectance sensed from space.

Baret, Frederic↗

NASA NTRS1978

The meaning of desert color in earth orbital photographs

The color of desert surfaces as seen in earth orbital photographs is indicative of soil composition. Apollo-Soyuz photographs of the Sturt and Simpson Deserts of Australia confirm that sand grains become redder as the distance from the source increases. Reddening is caused by a thin iron-oxide coating on individual sand grains and can be used, in some cases, to map relative-age zones. Photographs of the Western (Libyan) Desert of Egypt indicate three distinct and nearly parallel color zones that have been correlated in the field with: (1) arable soil composed of quartz, clay, and calcium carbonate particles; (2) relatively active sand with or without sparse vegetation; and (3) relatively inactive sand mixed with dark (desert-varnished) pebbles. The youngest sands are in the form of longitudinal dunes, which are migrating to the south-southeast along the prevailing wind direction. Some of the young dune fields are encroaching on the western boundary of the fertile Nile Valley.

El-Baz, F.↗

DOE OSTI2022

Age and chemistry of dissolved organic carbon reveal enhanced leaching of ancient labile carbon at the permafrost thaw zone

Abstract. Climate change will alter the balance between frozen and thawed conditions in Arctic systems. Increased temperatures will make the extensive northern permafrost carbon stock vulnerable to decomposition and translocation. Production, cycling, and transport of dissolved organic carbon (DOC) are crucial processes for high-latitude ecosystem carbon loss that result in considerable export off the Arctic landscape. To identify where and under what conditions permafrost DOC is mobilized in an Arctic headwater catchment, we measured radiocarbon (14C) of DOC and assessed DOC composition with ultraviolet–visible spectroscopy (UV–Vis) of surface waters and shallow and deep subsurface porewaters from 17 drainages in the Barrow Environmental Observatory in Alaska. Samples were collected in July and September 2013 to assess changes in age and chemistry of DOC over time. DOC age was highly variable ranging from modern (19 ‰ Δ14C) to approximately 7000 BP (−583 ‰ Δ14C). DOC age increased with depth, over the summer as the active layer deepened, and with increasing drainage size. DOC quality indicators reflected a DOC source rich in high molecular-weight and aromatic compounds, characteristics consistent with vegetation-derived organic matter that had undergone little microbial processing, throughout the summer and a weak relationship with DOC age. In deep porewaters, DOC age was also correlated with several biogeochemical indicators (including dissolved methane concentration, δ13C, and the apparent fractionation factor), suggesting a coupling between carbon and redox biogeochemistry influencing methane production. In the drained thawed lake basins included in this study, DOC concentrations and contributions of vegetation-derived organic matter declined with increasing basin age. The weak relationship between DOC age and chemistry and consistency in DOC chemical indicators over the summer suggest a high lability of old DOC released by thawing permafrost.

54 ENVIRONMENTAL SCIENCES↗

NASA NTRS1995

Urban remote sensing applications: TIMS observations of the City of Scottsdale

A research program has been initiated between Arizona State University and the City of Scottsdale, Arizona to study the potential applications of TIMS (Thermal Infrared Multispectral Scanner) data for urban scene classification, desert environmental assessment, and change detection. This program is part of a long-term effort to integrate remote sensing observations into state and local planning activities to improve decision making and future planning. Specific test sites include a section of the downtown Scottsdale region that has been mapped in very high detail as part of a pilot program to develop an extensive GIS database. This area thus provides excellent time history of the evolution of the city infrastructure, such as the timing and composition of street repavement. A second area of study includes the McDowell intensive study by state and local agencies to assess potential sites for urban development as well as preservation. These activities are of particular relevance as the Phoenix metropolitan area undergoes major expansion into the surrounding desert areas. The objectives of this study in urban areas are aimed at determining potential applications of TIMS data for classifying and assessing land use and surface temperatures. Land use centers on surface impermeability studies for storm runoff assessment and pollution control. These studies focus on determining the areal abundance of urban vegetation and undeveloped soil. Highly experimental applications include assessment and monitoring of pavement condition. Temperature studies focus on determining swimming pool area and temperature for use in monitoring evaporating and urban water consumption. These activities are of particular relevance as the Phoenix metropolitan area undergoes major expansion into the surrounding desert area.

Christensen, Philip R.↗

DOE OSTI2020

Modifying flowering time in maize

The present invention provides compositions and methods for modulating the reproductive transition in plants, such as grasses (e.g., maize). In particular, the invention provides methods for enhancing agronomic properties in plants by modulating expression of GRMZM2G171650 (zmm22) or homologs thereof. Modulation of expression of one or more additional genes which affect reproductive transition such as zagl1, in conjunction with such modulation of expression is also contemplated. Nucleic acid constructs for down-regulation of GRMZM2G171650 are also contemplated, as are transgenic plants, and products produced therefrom, that demonstrate altered development such as extended flowering time and display associated phenotypes such as enhanced yield of vegetative biomass, improved digestibility, and increased disease resistance. Plants described herein may be used, for example, as improved forage or feed crops or in biofuel production.

Kaeppler, Shawn↗

DOE OSTI2023

Disruption of p -coumaroyl-CoA:monolignol transferases in rice drastically alters lignin composition

Grasses are abundant feedstocks that can supply lignocellulosic biomass for production of cell-wall-derived chemicals. In grass cell walls, lignin is acylated with p-coumarate. These p-coumarate decorations arise from the incorporation of monolignol p-coumarate conjugates during lignification. A previous biochemical study identified a rice (Oryza sativa) BAHD acyltransferase (AT) with p-coumaroyl-CoA:monolignol transferase (PMT) activity in vitro. In this study, we determined that that enzyme, which we name OsPMT1 (also known as OsAT4), and the closely related OsPMT2 (OsAT3) harbor similar catalytic activity toward monolignols. We generated rice mutants deficient in either or both OsPMT1 and OsPMT2 by CRISPR/Cas9-mediated mutagenesis and subjected the mutants’ cell walls to analysis using chemical and nuclear magnetic resonance methods. Our results demonstrated that OsPMT1 and OsPMT2 both function in lignin p-coumaroylation in the major vegetative tissues of rice. Notably, lignin-bound p-coumarate units were undetectable in the ospmt1 ospmt2-2 double-knockout mutant. Further, in-depth structural analysis of purified lignins from the ospmt1 ospmt2-2 mutant compared with control lignins from wild-type rice revealed stark changes in polymer structures, including alterations in syringyl/guaiacyl aromatic unit ratios and inter-monomeric linkage patterns, and increased molecular weights. Furthermore, our results provide insights into lignin polymerization in grasses that will be useful for the optimization of bioengineering approaches for the effective use of biomass in biorefineries.

59 BASIC BIOLOGICAL SCIENCES↗