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At least 775 records · Page 43

Metabolome and RNA-seq Analysis of Responses to Nitrogen Deprivation and Resupply in Tea Plant (Camellia sinensis) Roots

Nitrogen (N) is an important contributor in regulating plant growth and development as well as secondary metabolites synthesis, so as to promote the formation of tea quality and flavor. Theanine, polyphenols, and caffeine are important secondary metabolites in tea plant. In this study, the responses of Camellia sinensis roots to N deprivation and resupply were investigated by metabolome and RNA-seq analysis. N deficiency induced content increase for most amino acids (AAs) and reduction for the remaining AAs, polyphenols, and caffeine. After N recovery, the decreased AAs and polyphenols showed a varying degree of recovery in content, but caffeine did not. Meanwhile, theanine increased in content, but its related synthetic genes were down-regulated, probably due to coordination of the whole N starvation regulatory network. Flavonoids-related pathways were relatively active following N stress according to KEGG enrichment analysis. Gene co-expression analysis revealed TCS2 , AMT1;1 , TAT2 , TS , and GOGAT as key genes, and TFs like MYB, bHLH, and NAC were also actively involved in N stress responses in C. sinensis roots. These findings facilitate the understanding of the molecular mechanism of N regulation in tea roots and provide genetic reference for improving N use efficiency in tea plant.

Xu, Wenluan↗

Plant root associated chitinases: structures and functions

Chitinases degrade chitin, a linear homopolymer of β-1,4-linked N-acetyl-D-glucosamine (GlcNAc) residues found in the cell walls of fungi and the exoskeletons of arthropods. They are secreted by the roots into the rhizosphere, a complex and dynamic environment where intense nutrient exchange occurs between plants and microbes. Here we modeled, expressed, purified, and characterized Zea mays and Oryza sativa root chitinases, and the chitinase of a symbiotic bacterium, Chitinophaga oryzae 1303 for their activities with chitin, di-, tri-, and tetra-saccharides and Aspergillus niger, with the goal of determining their role(s) in the rhizosphere and better understanding the molecular mechanisms underlying plant-microbe interactions. We show that Zea mays basic endochitinase (ZmChi19A) and Oryza sativa chitinase (OsChi19A) are from the GH19 chitinase family. The Chitinophaga oryzae 1303 chitinase (CspCh18A) belongs to the GH18 family. The three enzymes have similar apparent K M values of (20-40 µM) for the substrate 4-MU-GlcNAc 3 . They vary in their pH and temperature optima with OsChi19A activity optimal between pH 5–7 and 30–40°C while ZmChi19A and CspCh18A activities were optimal at pH 7-9 and 50–60°C. Modeling and site-directed mutation of ZmChi19A identified the catalytic cleft and the active residues E147 and E169 strategically positioned at ~8.6Å from each other in the folded protein. Cleavage of 4-MU-GlcNAc 3 was unaffected by the absence of the CBD but diminished in the absence of the flexible C-terminal domain. However, unlike for the soluble substrate, the CBD and the newly identified flexible C-terminal domain were vital for inhibiting Aspergillus niger growth. The results are consistent with the involvement of the plant chitinases in defense against pathogens like fungi that have chitin exoskeletons. In summary, we have characterized the functional features and structural domains necessary for the activity of two plant root chitinases that are believed to be involved in plant defense and a bacterial chitinase that, along with the plant chitinases, may participate in nutrient recycling in the rhizosphere.

60 APPLIED LIFE SCIENCES↗

Discovery top-down proteomics in symbiotic soybean root nodules

Proteomic methods have been widely used to study proteins in complex biological samples to understand biological molecular mechanisms. Most well-established methods (known as bottom-up proteomics, BUP) employ an enzymatic digestion step to cleave intact proteins into smaller peptides for liquid chromatography (LC) mass spectrometry (MS) detection. In contrast, top-down proteomics (TDP) directly characterizes intact proteins including all possible post-translational modifications (PTMs), thus offering unique insights into proteoform biology where combinations of individual PTMs may play important roles. We performed TDP on soybean root nodules infected by the symbiotic Bradyrhizobium japonicum in both the wildtype bacterium and a nifH- mutant, which lacks the ability to fix nitrogen in the soybean root nodule. TDP captured 1648 proteoforms derived from 313 bacterial genes and 178 soybean genes. Leghemoglobin, the most abundant protein in the sample, existed in many truncated proteoforms. Interestingly, these truncated proteoforms were considerably more abundant in the wildtype relative to the nifH- mutant, implicating protease activity as an important factor in nitrogen fixation. Proteoforms with various PTMs and combinations thereof were identified using an unrestricted open modification search. This included less common PTMs such as myristoylation, palmitoylation, cyanylation, and sulfation. In parallel, we collected high resolution MS imaging (MSI) data of intact proteins and biopolymers (<20 kDa due to current technical limitations) from sections of the soybean root nodules using matrix-assisted laser desorption/ionization (MALDI) coupled to high resolution Orbitrap. Several detected proteoforms exhibited unique spatial distributions inside the infection zone and cortex, suggesting functional compartmentalization in these regions. A subset of peaks from the MALDI-MSI were assigned to proteoforms detected in TDP LCMS data based on matching accurate masses. Many of the proteins detected in both LCMS and MALDI-MSI are currently uncharacterized in UniProt: the PTM and spatial information presented here will be valuable in understanding their biological functions. Taken together, our study demonstrates how untargeted TDP approach can provide unique insights into plant proteoform biology. On-going technology developments are expected to further improve TDP coverage for more comprehensive high-throughput analysis of proteoforms.

59 BASIC BIOLOGICAL SCIENCES↗

Blumenols as shoot markers of root symbiosis with arbuscular mycorrhizal fungi

High-through-put (HTP) screening for functional arbuscular mycorrhizal fungi (AMF)-associations is challenging because roots must be excavated and colonization evaluated by transcript analysis or microscopy. Here we show that specific leaf-metabolites provide broadly applicable accurate proxies of these associations, suitable for HTP-screens. With a combination of untargeted and targeted metabolomics, we show that shoot accumulations of hydroxy- and carboxyblumenol C-glucosides mirror root AMF-colonization in Nicotiana attenuata plants. Genetic/pharmacologic manipulations indicate that these AMF-indicative foliar blumenols are synthesized and transported from roots to shoots. These blumenol-derived foliar markers, found in many di- and monocotyledonous crop and model plants (Solanum lycopersicum, Solanum tuberosum, Hordeum vulgare, Triticum aestivum, Medicago truncatula and Brachypodium distachyon), are not restricted to particular plant-AMF interactions, and are shown to be applicable for field-based QTL mapping of AMF-related genes.

59 BASIC BIOLOGICAL SCIENCES↗

Delayed ripple counter simplifies square-root computation

Ripple subtract technique simplifies the logic circuitry required in a binary computing device to derive the square root of a number. Successively higher numbers are subtracted from a register containing the number out of which the square root is to be extracted. The last number subtracted will be the closest integer to the square root of the number.

Cliff, R.↗

The development of spectro-signature indicators of root disease impacts on forest stands

A field research program was begun in 1969 and intensified in 1970 on the physiology and biophysical responses of second-growth Douglas fir infected with root rot fungus. A double tramway system was suspended between three 100-foot instrument towers to carry sensors for measuring the energy response from above both healthy and infected trees. Processing and analysis was completed of airborne multispectral scanner imagery collected over the Wind River research area in 1969. Likelihood ratio processing of three-channel infrared data and Euclidean distance analysis of ten-channel spectrometer data did not identify incipient root rot infection outside the training sets. In all cases infected fir was misclassified as healthy fir. It was concluded from careful examination of physiological data that Poria root rot infection has little effect on water metabolism and energy exchange. What was identified was a low-grade stress that affects respiration and metabolism over long periods of time. This led to minor changes in the external physical symptoms of Poria-infected trees which was revealed only in the shortwave reflectance data.

Weber, F. P.↗

Computer program to determine roots of polynomials by ratio of successive derivatives

High speed computing finds roots of polynomials with real number coefficients. Ratios of successive polynomial derivatives approach provides accurate roots-of-polynomial computer programs with very high reliability. With derivative ratio method, root analysis can still be done even though the polynomial and its lower order derivatives cannot be evaluated with sufficient accuracy.

Crouse, J. E.↗

New method for propagating the square root covariance matrix in triangular form

A method proposed by Potter of applying the Kalman-Bucy filter to the problem of estimating the state of a dynamic system is described, in which the square root of the state error covariance matrix is used to process the observations. A new technique which propagates the covariance square root matrix in lower triangular form is given for the discrete observation case. The technique is faster than previously proposed algorithms and is well-adapted for use with the Carlson square root measurement algorithm.

Choe, C. Y.↗

Stress analysis, thermomechanical fatique evaluation, and root subcomponent testing of gamma/gamma prime-delta eutectic alloy

Thermomechanical fatigue (TMF) and root subcomponent tensile, creep, and low cycle fatigue (LCF) tests were conducted to determine the capability of a fully lamellar directionally solidified eutectic alloy to sustain the airfoil thermal fatigue and root attachment loads anticipated in advanced, hollow, high work turbine blades. A three dimensional finite element elastic stress analysis was performed on typical advanced hollow eutectic airfoil and root-platform designs to determine appropriate conditions for these tests. Results of TMF tests conducted on longitudinal specimens (stress axis parallel to the solidification direction) containing a simulated leading edge cooling hole pattern indicated the longitudinal TMF properties to be more than adequate for the particular advanced hollow blade analyzed, with the strain range for a 10,000 cycle life being more than 50% above the maximum strain range calculated for the advanced hollow blade.

Sheffler, K. D.↗

"Foreign material" to verify root fusion in welded joints

Foil or thin wire at weld root is used to verify weld penetration. When weld is adequate, material mixes with weld and traces of it diffuse to weld crown. Spectroscopic analysis of samples identifies foreign material and verifies root has fused. Weld roots are usually inaccessible to visual inspection, and X-ray and ultrasonic inspection techniques are not always reliable. Good results are obtained with use of gold/nickel alloy.

Kleint, R. E.↗

Analysis techniques for multivariate root loci

Analysis and techniques are developed for the multivariable root locus and the multivariable optimal root locus. The generalized eigenvalue problem is used to compute angles and sensitivities for both types of loci, and an algorithm is presented that determines the asymptotic properties of the optimal root locus.

Thompson, P. M.↗

The McMillan and Newton polygons of a feedback system and the construction of root loci

The local behaviour of root loci around zeros and poles is investigated. This is done by relating the Newton diagrams which arise in the local analysis to the McMillan structure of the open-loop system, by means of what we shall call the McMillan polygon. This geometric construct serves to clarify the precise relationship between the McMillan structure, the principal structure, and the branching patterns of the root loci. In addition, several rules are obtained which are useful in the construction of the root loci of multivariable control systems.

Byrnes, C. I.↗

Differential wall growth in gravistimulated corn roots: Its timing and regulation

The experiments designed to document cell-wall level changes which occur as a result of their gravistimulation are described. The goal of this research is to elucidate the mechanism and time frame of differential growth following a controlled gravistimulation. To achieve this, rates of wall deposition will be determined by following the incorporation of radioactive monosaccharides into the wall. Complementing this experiment will be a freeze-etch study directed at revealing the spatial arrangment of both newly-deposited microfibrils and microfibrils that were present in the growing root prior to stimulation. The second phase of the proposed research will examine the roles ethylene and Ca(2+) have in the modulation of differential wall changes during gravitropism. Ethylene and Ca(2+) have both been implicated as regulators of the gravitropic response in roots and they have also been implicated as regulators of the gravitropic response in roots and they have also been reported to exert some control on the orientation of microfibrils. Both of these agents will be manipulated in such a way as to reveal whether they have a direct influence on cell wall deposition and microfibrillar alignment during the geotropic response.

Serlin, B. S.↗

Distribution of calmodulin in corn seedlings - Immunocytochemical localization in coleoptiles and root apices

Immunofluorescence techniques have been used to study the distribution of calmodulin in several tissues in etiolated corn (Zea mays, var. Bear Hybrid) seedlings. Uniform staining was seen in the background cytoplasm of most cell types. Cell walls and vacuoles were not stained. In coleoptile mesophyll cells the nucleoplasm of most nuclei was stained as was the stroma of most amyloplasts. The lumen border of mature tracheary elements in coleoptiles also stained. In the rootcap the most intensely stained regions were the cytoplasms of columella cells and of the outermost cells enmeshed in the layer of secreted slime. Nuclei in the rootcap cells did not stain distinctly, but those in all cell types of the root meristem did. Also in the root meristem, the cytoplasm of metaxylem elements stained brightly. These results are compared and contrasted with previous data on the localization of calmodulin in pea root apices and epicotyls and discussed in relation to current hypotheses on mechanisms of gravitropism.

Dauwalder, M.↗

Root damage analysis of aircraft engine blade subject to ice impact

The blade root response due to ice impact on an engine blade is simulated using the NASA in-house code BLASIM. The ice piece is modeled as an equivalent spherical object impacting on the leading edge of the blade and has the velocity opposite to that of the aircraft with direction parallel to the engine axis. The effect of ice impact is considered to be an impulse load on the blade with its amplitude computed based on the momentum transfer principle. The blade response due to the impact is carried out by modal superposition using the first three modes. The maximum dynamic stresses at the blade root are computed at the quarter cycle of the first natural frequency. A combined stress failure function based on modified distortion energy is used to study the spanwise bending damage response at the blade root. That damage function reaches maximum value for very low ice speeds and increases steeply with increases in engine speed.

Reddy, E. S.↗

An investigation of the use of discharge valves and an intake control for improving the performance of N.A.C.A. Roots type supercharger

This report presents the results of an analytical investigation on the practicability of using mechanically operated discharge valves in conjunction with a manually operated intake control for improving the performance of N. A. C. A. Roots type superchargers. These valves, which may be either of the oscillating or rotating type, are placed in the discharge opening of the supercharger and are so shaped and synchronized with the supercharger impellers that they do not open until the air has been compressed to the delivery pressure. The intake control limits the quantity of air compressed to engine requirements by permitting the excess air to escape from the compression chamber before compression begins. The percentage power saving and the actual horsepower saved were computed for altitudes from 0 to 20,000 feet. These computations are based on the pressure-volume cards for the conventional and the modified roots type superchargers and on the results of laboratory tests of the conventional type. The use of discharge valves shows a power saving of approximately 26 per cent at a critical altitude of 20,000 feet. In addition, these valves reduce the amplitude of the discharge pulsations and increase the volumetric efficiency. With slow-speed roots blowers operating at high-pressure differences even better results would be expected. For aircraft engine superchargers operating at high speeds these discharge valves increase the performance as above, but have the disadvantages of increasing the weight and of adding a high-speed mechanism to a simple machine. (author)

Schey, Oscar W↗

Comparative Flight Performance with an NACA Roots Supercharger and a Turbocentrifugal Supercharger

This report presents the comparative flight results of a roots supercharger and a turbocentrifugal supercharger. The tests were conducted using a modified DH-4M2 airplane. The rate of climb and the high speed in level flight of the airplane were obtained for each supercharger from sea level to the ceiling. The unsupercharged performance with each supercharger mounted in place was also determined. The results of these tests show that the ceiling and rate of climb obtained were nearly the same for each supercharger, but that the high speed obtained with the turbocentrifugal was better than that obtained with the roots. The high-speed performance at 21,000 feet was 122 and 142 miles per hour for the roots and turbocentrifugal, respectively.

Schey, Oscar W↗

Chamber free fusion welding root side purging method and apparatus

A method and apparati are presented for non-chamber root side purging in fusion welding of oxygen reactive metals which require that the molten weld zone and local solid areas of the weld seam remaining at high temperatures be shielded from normal atmosphere to prevent degradation of the welded area. The apparati provide an inert atmosphere to the root side of a weld joint through a porous medium whereby the jet-like thrust of the plasma arc actually draws the continuously supplied inert atmosphere into the path of the molten or high temperature solid weld zone. The porous medium is configured so it can be placed at the borders of the weld seam and substantially parallel to the seam without restricting the view of the root side of the seam. The inert gas is dispersed evenly through the porous media and across the weld seam, at the point of arc penetration and in front of and behind the arc. The apparati can be constructed so as to limit the amount of inert gas flow and can be mobile and travel synchronously with the welding arc.

Mcgee, William F.↗