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DOE OSTI2023

Cell‐ and development‐specific degradation controls the levels of mixed‐linkage glucan in sorghum leaves

SUMMARY Mixed‐linkage glucan (MLG) is a component of the cell wall (CW) of grasses and is composed of glucose monomers linked by β‐1,3 and β‐1,4 bonds. MLG is believed to have several biological functions, such as the mobilizable storage of carbohydrates and structural support of the CW. The extracellular levels of MLG are largely controlled by rates of synthesis mediated by cellulose synthase‐like (CSL) enzymes, and turnover by lichenases. Economically important crops like sorghum accumulate MLG to variable levels during development. While in sorghum, like other grasses, there is one major MLG synthase (CSLF6), the identity of lichenases is yet unknown. To fill this gap, we identified three sorghum lichenases (SbLCH1‐3) and characterized them in leaves in relation to the expression of SbCSLF6 , and the abundance of MLG and starch. We established that SbLCH1‐3 are secreted to the apoplast, consistent with a role of degrading MLG extracellularly. Furthermore, while SbCSLF6 expression was associated with cell development, the SbLCH genes exhibited distinct development‐, cell‐type‐specific and diel‐regulated expression. Therefore, our study identifies three functional sorghum MLG lichenases and highlights that MLG accumulation in sorghum leaves is likely controlled by the activity of lichenases that tune MLG levels, possibly to suit distinct cell and developmental needs in planta . These findings have important implications for improving the growth, yield, and composition of sorghum as a feedstock.

Plant Sciences↗

DOE OSTI2014

SU-E-J-85: Leave-One-Out Perturbation (LOOP) Fitting Algorithm for Absolute Dose Film Calibration

Purpose: To introduce an outliers-recognition fitting routine for film dosimetry. It cannot only be flexible with any linear and non-linear regression but also can provide information for the minimal number of sampling points, critical sampling distributions and evaluating analytical functions for absolute film-dose calibration. Methods: The technique, leave-one-out (LOO) cross validation, is often used for statistical analyses on model performance. We used LOO analyses with perturbed bootstrap fitting called leave-one-out perturbation (LOOP) for film-dose calibration . Given a threshold, the LOO process detects unfit points (“outliers”) compared to other cohorts, and a bootstrap fitting process follows to seek any possibilities of using perturbations for further improvement. After that outliers were reconfirmed by a traditional t-test statistics and eliminated, then another LOOP feedback resulted in the final. An over-sampled film-dose- calibration dataset was collected as a reference (dose range: 0-800cGy), and various simulated conditions for outliers and sampling distributions were derived from the reference. Comparisons over the various conditions were made, and the performance of fitting functions, polynomial and rational functions, were evaluated. Results: (1) LOOP can prove its sensitive outlier-recognition by its statistical correlation to an exceptional better goodness-of-fit as outliers being left-out. (2) With sufficient statistical information, the LOOP can correct outliers under some low-sampling conditions that other “robust fits”, e.g. Least Absolute Residuals, cannot. (3) Complete cross-validated analyses of LOOP indicate that the function of rational type demonstrates a much superior performance compared to the polynomial. Even with 5 data points including one outlier, using LOOP with rational function can restore more than a 95% value back to its reference values, while the polynomial fitting completely failed under the same conditions. Conclusion: LOOP can cooperate with any fitting routine functioning as a “robust fit”. In addition, it can be set as a benchmark for film-dose calibration fitting performance.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

DOE OSTI2021

Using 14 C-acetate Pulse-chase Labeling to Study Fatty Acid and Glycerolipid Metabolism in Plant Leaves

Lipids metabolism is comprised of networks of reactions occurred in different subcellular compartments. Isotopic labeling is a good way to track the transformations and movements of metabolites without perturbing overall cellular metabolism. Fatty acids, the building blocks of membrane lipids and storage triacylglycerols, are synthesized in plastids. The immediate precursor for fatty acid synthesis is acetyl-CoA. Exogenous acetate is rapidly incorporated into fatty acids in leaves and isolated plastids because it can diffuse freely through cellular membranes, enter the plastid where it is rapidly metabolized to acetyl-CoA. Therefore, isotope-labeled acetate is often used as a tracer for the investigation of fatty acid synthesis and complex lipid metabolism in plants and other organisms. The basic principle of isotope labeling and its recent technical advances have been reviewed (Allen et al., 2015). The present protocol describes the use of 14C-labeled acetate to determine rates of fatty acid synthesis and degradation and to track the metabolism of glycerolipids in leaves. This method, which is often referred to as acetate pulse-chase labeling, has been widely used to probe various aspects of lipid metabolism (Allen et al., 2015), including the role of autophagy in membrane lipid turnover (Fan et al., 2019) and the interplay between lipid and starch metabolism pathways (Yu et al., 2018).

14C-acetate pulse-chase↗

DOE OSTI2023

Characterization of polysaccharide from Lonicera japonica Thunb leaves and its application in nano-emulsion

The polysaccharides in honeysuckle leaves (PHL) were separated and characterized for the first time. The nano-emulsion stabilized by PHL and whey protein isolate (WPI) were also fabricated based on the ultrasonic method. The results indicated that PHL was mainly composed of glucose (47.40 mol%), galactose (19.21 mol%) and arabinose (20.21 mol%) with the weight-average molecular weight of 137.97 ± 4.31 kDa. The emulsifier concentration, WPI-to-PHL ratio, ultrasound power and ultrasound time had significant influence on the droplet size of PHL-WPI nano-emulsion. The optimal preparation conditions were determined as following: emulsifier concentration, 1.7%; WPI/PHL ratio, 3:1; ultrasonic power, 700 W; ultrasonic time, 7 min. Under the above conditions, the median diameter of the obtained nano-emulsion was 317.70 ± 5.26 nm, close to the predicted value of 320.20 nm. The protective effect of PHL-WPI emulsion on β-carotene against UV irradiation was superior to that of WPI emulsion. Our results can provide reference for the development of honeysuckle leaves.

Li, Yongchao↗

DOE OSTI2020

Characterization of Local and Systemic Impact of Whitefly (Bemisia tabaci) Feeding and Whitefly-Transmitted Tomato Mottle Virus Infection on Tomato Leaves by Comprehensive Proteomics

Tomato mottle virus (ToMoV) is a ssDNA plant begomovirus transmitted to solanaceous crops by the whitefly species complex (Bemisia tabaci), causing stunted growth, leaf mottling, and reduced fruit yield. Using a genetic repertoire of 7 genes, ToMoV pathogenesis includes manipulation of multiple plant biological processes to circumvent antiviral defenses. To further understand the effects of whitefly feeding and whitefly-transmitted ToMoV infection on tomato plants (Solanum lycopersicum ‘Florida Lanai’), we generated comprehensive protein profiles of leaves subjected to feeding by either (1) viruliferous whiteflies harboring ToMoV, or (2) non-viruliferous whiteflies, or (3) a no-feeding control. In each condition (1-3), whiteflies were confined to clip cages on a mature leaf for 3 days. The effects of whitefly feeding and ToMoV infection were measured both locally and systemically by sampling either a mature leaf directly from the site of whitefly feeding, or from a newly formed leaf 10 days post feeding (dpf). At 3 dpf, relatively few biological processes were affected specifically by ToMoV at the site of feeding, and include proteins associated with translation initiation and elongation, and plasmodesmata dynamics. In contrast, the systemic impacts of ToMoV on younger leaves at 10 dpf were more pronounced and included a virus-specific change in plant proteins associated with mRNA maturation and export, RNA-dependent DNA methylation, and other plant anti-viral transcriptional gene silencing systems. Our analysis supports previous findings and provides novel insight into biological processes in tomato altered locally and systemically by whitefly feeding and ToMoV infection.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

Image-Based Methods to Score Fungal Pathogen Symptom Progression and Severity in Excised Arabidopsis Leaves

Image-based symptom scoring of plant diseases is a powerful tool for associating disease resistance with plant genotypes. Advancements in technology have enabled new imaging and image processing strategies for statistical analysis of time-course experiments. There are several tools available for analyzing symptoms on leaves and fruits of crop plants, but only a few are available for the model plant Arabidopsis thaliana (Arabidopsis). Arabidopsis and the model fungus Botrytis cinerea (Botrytis) comprise a potent model pathosystem for the identification of signaling pathways conferring immunity against this broad host-range necrotrophic fungus. Here, we present two strategies to assess severity and symptom progression of Botrytis infection over time in Arabidopsis leaves. Thus, a pixel classification strategy using color hue values from red-green-blue (RGB) images and a random forest algorithm was used to establish necrotic, chlorotic, and healthy leaf areas. Secondly, using chlorophyll fluorescence (ChlFl) imaging, the maximum quantum yield of photosystem II (Fv/Fm) was determined to define diseased areas and their proportion per total leaf area. Both RGB and ChlFl imaging strategies were employed to track disease progression over time. This has provided a robust and sensitive method for detecting sensitive or resistant genetic backgrounds. A full methodological workflow, from plant culture to data analysis, is described.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Coupled Gas-Exchange Model for C4 Leaves Comparing Stomatal Conductance Models

Plant simulation models are abstractions of plant physiological processes that are useful for investigating the responses of plants to changes in the environment. Because photosynthesis and transpiration are fundamental processes that drive plant growth and water relations, a leaf gas-exchange model that couples their interdependent relationship through stomatal control is a prerequisite for explanatory plant simulation models. Here, we present a coupled gas-exchange model for C4 leaves incorporating two widely used stomatal conductance submodels: Ball–Berry and Medlyn models. The output variables of the model includes steady-state values of CO2 assimilation rate, transpiration rate, stomatal conductance, leaf temperature, internal CO2 concentrations, and other leaf gas-exchange attributes in response to light, temperature, CO2, humidity, leaf nitrogen, and leaf water status. We test the model behavior and sensitivity, and discuss its applications and limitations. The model was implemented in Julia programming language using a novel modeling framework. Our testing and analyses indicate that the model behavior is reasonably sensitive and reliable in a wide range of environmental conditions. The behavior of the two model variants differing in stomatal conductance submodels deviated substantially from each other in low humidity conditions. The model was capable of replicating the behavior of transgenic C4 leaves under moderate temperatures as found in the literature. The coupled model, however, underestimated stomatal conductance in very high temperatures. This is likely an inherent limitation of the coupling approaches using Ball–Berry type models in which photosynthesis and stomatal conductance are recursively linked as an input of the other.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2016

XA21-specific induction of stress-related genes following Xanthomonas infection of detached rice leaves

The rice XA21 receptor kinase confers robust resistance to the bacterial pathogen Xanthomonas oryzaepv. oryzae (Xoo). We developed a detached leaf infection assay to quickly and reliably measure activation of the XA21-mediated immune response using genetic markers. We used RNA sequencing of elf18 treated EFR:XA21:GFP plants to identify candidate genes that could serve as markers for XA21 activation. From this analysis, we identified eight genes that are up-regulated in both in elf18 treated EFR:XA21:GFP rice leaves and Xoo infected XA21 rice leaves. These results provide a rapid and reliable method to assess bacterial-rice interactions.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1985

Contrasts among bidirectional reflectance of leaves, canopies, and soils

Simple models are presented for predicting the bidirectional reflectance distribution functions (BRDFs) for soils and plant canopies viewed from various directions. BRDFs are predicted for bare soil, individual leaves, and plant canopies, and the results are compared with measurements and a three coefficient empirical equation. BRDF measurements for corn and soybean leaves are presented to contrast with canopy and soil distributions. Estimates of the soil, canopy, and leaf BRDFs are combined into a model called Cupid to predict BRDFs for complex natural surfaces.

Norman, J. M.↗

NASA NTRS1986

Polarization photometer to measure bidirectional reflectance factor R(55 deg, 0 deg, 55 deg, 180 deg) of leaves

A polarization photometer has been developed for rapidly determining the bidirectional, polarized, and diffuse light-scattering properties of individual leavs illuminated and measured 'in vivo' at an angle of 55 deg (approximately Brewster's angle) in six wavelength bands in the visible and near-IR wavelength regions. The optical performance and data quality of the system are evaluated to estimate the magnitude of the variation in the data attributable to the instrument system and to measurement procedures. The potential to discriminate between three species of oak is demonstrated using data acquired by the polarization photometer. The instrument is being used to increase understanding of the radiation transfer process in plant canopies and specifically to determine how agronomic information of the physical and chemical processes in leaves, plants, and plant canopies is expressed in these light-scattering properties.

Vanderbilt, V. C.↗

NASA NTRS1988

Goniometric observations of light scattered from soils and leaves

The laboratory established at NASA-Goddard to measure and model the light-scattering properties of soil samples and individual plant leaves employs two goniometers: one for the measurement of directional reflectance and transmittance from vertically-mounted leaf samples, and the other for measurement of directional reflectance from such horizontal, semiinfinite particulate surfaces as soil samples. Sample observations of various soil minerals and plant leaves are presented; these goniometric data are compared to the results of a reflectance model from particulate surfaces and those of a ray-tracing model of leaf reflectance and transmittance.

Kestner, Joann M.↗

NASA NTRS1990

Effects of atmospheric CO2 on photosynthetic characteristics of soybean leaves

Soybean (Glycine max. cv. McCall) plants were grown at 500, 1000, and 2000 umol mol (exp -1) CO2 for 35 days with a photosynthetic photon flux of 300 umol m (exp -2) s (-1). Individual leaves were exposed to step changes of photosynthetic photon flux to study CO2 assimilation rates (CAR), i.e., leaf net photosynthesis. In general, CAR increased when CO2 increased from 500 to 1000 umol mol (exp -1), but not from 1000 to 2000 umol mol (exp -1). Regardless of the CO2 level, all leaves showed similar CAR at similar CO2 and PPF. This observation contrasts with reports that plants tend to become 'lazy' at elevated CO2 levels over time. Although leaf stomatal conductance (to water vapor) showed diurnal rhythms entrained to the photoperiod, leaf CAR did not show these rhythms and remained constant across the light period, indicating that stomatal conductance had little effect on CAR. Such measurements suggest that short-term changes in CO2 exchange dynamics for a controlled ecological life support system can be closely predicted for an actively growing soybean crop.

Wheeler, R. M.↗

NASA NTRS1992

Surface roughness considerations for atmospheric correction of ocean color sensors. I - The Rayleigh-scattering component. II - Error in the retrieved water-leaving radiance

The first step in the Coastal Zone Color Scanner (CZCS) atmospheric-correction algorithm is the computation of the Rayleigh-scattering (RS) contribution, L sub r, to the radiance leaving the top of the atmosphere over the ocean. In the present algorithm, L sub r is computed by assuming that the ocean surface is flat. Calculations of the radiance leaving an RS atmosphere overlying a rough Fresnel-reflecting ocean are presented to evaluate the radiance error caused by the flat-ocean assumption. Simulations are carried out to evaluate the error incurred when the CZCS-type algorithm is applied to a realistic ocean in which the surface is roughened by the wind. In situations where there is no direct sun glitter, it is concluded that the error induced by ignoring the Rayleigh-aerosol interaction is usually larger than that caused by ignoring the surface roughness. This suggests that, in refining algorithms for future sensors, more effort should be focused on dealing with the Rayleigh-aerosol interaction than on the roughness of the sea surface.

Gordon, Howard R.↗

NASA NTRS1993

Red edge spectral measurements from sugar maple leaves

Many sugar maple stands in the northeastern United States experienced extensive insect damage during the 1988 growing season. Chlorophyll data and high spectral resolution spectrometer laboratory reflectance data were acquired for multiple collections of single detached sugar maple leaves variously affected by the insect over the 1988 growing season. Reflectance data indicated consistent and diagnostic differences in the red edge portion (680-750 nm) of the spectrum among the various samples and populations of leaves. These included differences in the red edge inflection point (REIP), a ratio of reflectance at 740-720 nm (RE3/RE2), and a ratio of first derivative values at 715-705 nm (D715/D705). All three red edge parameters were highly correlated with variation in total chlorophyll content. Other spectral measures, including the Normalized Difference Vegetation Index (NDVI) and the Simple Vegetation Index Ratio (VI), also varied among populations and over the growing season, but did not correlate well with total chlorophyll content. Leaf stacking studies on light and dark backgrounds indicated REIP, RE3/RE2 and D715/D705 to be much less influenced by differences in green leaf biomass and background condition than either NDVI or VI.

Vogelmann, J. E.↗

NASA NTRS1995

Effects of Raman scattering on the water-leaving radiance

The contribution of Raman scattering to the water-leaving radiance is examined using Monte Carlo simulations. Exit angle information is retained, allowing a comparison of different satellite viewing directions. Chlorophyll values of 0.0, 0.01, 0.1, and 1.0 mg Chl/cu m are simulated. Little directional variability is found, with the exception of the direct solar backscatter direction. The wavelength variability is greatest for low chlorophyll concentrations and is negligible for 1.0 mg Chl/cu m. At 550 nm the Raman contribution ranges from approximately 18% of the total water-leaving radiance for pure water to 3% for 1.0 mg Chl/cu m. At 440 nm the range is from 6% to 2%, indicating that Raman scattering will impact radiance ratios for ocean color satellite algorithms.

Waters, Kirk J.↗

NASA NTRS1999

Effective Elastic Modulus as a Function of Angular Leaf Span for Curved Leaves of Pyrolytic Boron Nitride

A theoretical equation was derived to predict the spring constant (load/deflection) for a simply supported cylindrical section with a line force applied at the center. Curved leaves of PBN were mechanically deformed and the force versus deflection data was recorded and compared to the derived theoretical equation to yield an effective modulus for each leaf. The effective modulus was found to vary from the pure shear modulus for a flat plate to a mixed mode for a half cylinder as a function of the sine of one half the angular leaf span. The spring constants of individual PBN leaves were usually predicted to within 30%.

Kaforey, M. L.↗

NASA NTRS1999

Spring Constants for Stacks of Curved Leaves of Pyrolytic Boron Nitride

Stacks of curved leaves of pyrolytic boron nitride (PBN) were deflected and the force versus deflection data was recorded. From this data, the spring constant for a given spring geometry (radius of curvature of a leaf, width of a leaf, thickness of a leaf, and number of leaves in the stack) was determined. These experiments were performed at room temperature, 500 C and 1000 C. However, temperature was not found to affect the spring constant. The measured values were generally within one order of magnitude of predictions made using a previously derived equation for a simply supported cylindrical section with a line force at the center.

Kaforey, M. L.↗

NASA NTRS2002

Sucrose regulation of ADP-glucose pyrophosphorylase subunit genes transcript levels in leaves and fruits

ADP-glucose pyrophosphorylase (AGPase, EC2.7.7.27) is a key regulatory enzyme in starch biosynthesis. The enzyme is a heterotetramer with two S and two B subunits. In tomato, there are three multiple forms of the S subunit gene. Agp S1, S2 and B are highly expressed in fruit from 10 to 25 days after anthesis. Agp S3 is only weakly expressed in fruit. Sucrose significantly elevates expression of Agp S1, S2 and B in both leaves and fruits. Agp S1 exhibits the highest degree of regulation by sucrose. In fact, sucrose may be required for Agp S1 expression. For excised leaves incubated in water, no transcripts for Agp S1 could be detected in the absence of sucrose, whereas it took up to 16 h in water before transcripts were no longer detectable for Agp S2 and B. Neither Agp S3 nor the tubulin gene is affected by sucrose, demonstrating that this response is specifically regulated by a carbohydrate metabolic signal, and is not due to a general increase in metabolism caused by sucrose treatment. Truncated versions of the promoter for Agp S1 indicate that a specific region 1.3-3.0 kb upstream from the transcription site is responsible for sucrose sensitivity. This region of the S1 promoter contains several cis-acting elements present in the promoters of other genes that are also regulated by sucrose. c2002 Elsevier Science Ireland Ltd. All rights reserved.

Non-NASA Center↗