Integration of metabolite with transcript and enzyme activity profiling during diurnal cycles in Arabidopsis rosettes

ABSTRACT: BACKGROUND: Genome-wide transcript profiling and analyses of enzyme activities from central carbon and nitrogen metabolism has shown that transcript levels undergo marked and rapid changes during diurnal cycles and after transfer to darkness, whereas changes of enzyme activities are smaller and delayed. In the starchless pgm mutant, where sugars are depleted every night, there are accentuated diurnal changes of transcript levels. Enzyme activities do not show larger diurnal changes; instead they shift towards the levels found in wild-type after several days of darkness. These results indicate that enzyme activities change slowly, integrating the changes of transcript levels over several diurnal cycles. RESULTS: To generalize this conclusion, 137 metabolites were profiled using GC-MS and LC-MS. Amplitudes of the diurnal changes of metabolites in pgm were (with the exception of sugars) similar or smaller than in wild-type. The average levels shifted towards those found after several days of darkness in wild-type. Examples include increased levels of many amino acids due to protein degradation, decreased levels of many fatty acids, increased tocopherol and decreased myo-inositol. Many metabolite-transcript correlations were found and the proportion of transcripts correlated with sugars increased dramatically in the starchless pgm mutant. CONCLUSION: Rapid diurnal changes of transcripts are integrated over time to generate quasi-stable changes across large sectors of metabolism. The slow response of enzyme activities and metabolites implies that correlations between metabolites and transcripts are due to regulation of gene expression by metabolites, rather than metabolites being changed as a consequence of a change in gene expression

Delamination growth in composite materials

The Double Cantilever Beam (DCB) and the End Notched Flexure (ENF) specimens are employed to characterize MODE I and MODE II interlaminar fracture resistance of graphite/epoxy (CYCOM 982) and graphite/PEEK (APC2) composites. Sizing of test specimen geometries to achieve crack growth in the linear elastic regime is presented. Data reduction schemes based upon beam theory are derived for the ENF specimen and include the effects of shear deformation and friction between crack surfaces on compliance, C, and strain energy release rate, G sub II. Finite element (FE) analyses of the ENF geometry including the contact problem with friction are presented to assess the accuracy of beam theory expressions for C and G sub II. Virtual crack closure techniques verify that the ENF specimen is a pure Mode II test. Beam theory expressions are shown to be conservative by 20 to 40 percent for typical unidirectional test specimen geometries. A FE parametric study investigating the influence of delamination length and depth, span, thickness and material properties on G sub II is presented. Mode I and II interlaminar fracture test results are presented. Important experimental parameters are isolated, such as precracking techniques, rate effects, and nonlinear load-deflection response. It is found that subcritical crack growth and inelastic materials behavior, responsible for the observed nonlinearities, are highly rate-dependent phenomena with high rates generally leading to linear elastic response

Delamination growth in composite materials

Research related to growth of an imbedded through-width delamination (ITWD) in a compression loaded composite structural element is presented. Composites with widely different interlaminar fracture resistance were examined, viz., graphite/epoxy (CYCOM 982) and graphite/PEEK (APC-2). The initial part of the program consisted of characterizing the material in tension, compression and shear mainly to obtain consistent material properties for analysis, but also as a check of the processing method developed for the thermoplastic APC-2 material. The characterization of the delamination growth in the ITWD specimen, which for the unidirectional case is essentially a mixed Mode 1 and 2 geometry, requires verified mixed-mode growth criteria for the two materials involved. For this purpose the main emphasis during this part of the investigation was on Mode 1 and 2 fracture specimens, namely the Double Cantilever Beam (DCB) and End Notched Flexure (ENF) specimens

Urine patch detection using LiDAR and RPAS/UAV produced photogrammetry

In grazed dairy pastures, the largest N source for both nitrate (NO3-) leaching and nitrous oxide (N2O) emissions is urine-N excreted by the animals. Additional application of N on urine patches as fertilizer may increase these losses so adapting N-fertilisation in these areas is necessary. The objective of this study was to examine the use of a tractor mounted LiDAR (Light Detection and Ranging) system to accurately identify and quantify areas affect by excess N, such as urine and dung. To do so, a controlled experiment was designed in a paddock with no recent exposure to animals or N fertilisation. Synthetic urine was randomly applied within two 20m x 20m blocks and weekly LiDAR scans were taken for 5 weeks. LiDAR based contour maps of the pasture canopy were shown to accurately detect the asymmetric urine patches as well as calculate a percent area of urine based high N as early as one week after a simulated grazing event. Further, weekly flights were taken with a remotely piloted aircraft system (RPAS/UAV) to have aerial footage of the trial. Resulting mosaic of RGB and NIR images were used to create photogrammetric based contour maps. Both approaches (LiDAR and photogrammetry) show no significant difference in the identification and sizing of urine patch cluster