Photosynthetic activity,chloroplast ultrastructure,and leaf characteristics of high-light and low-light plants and of sun and shade leaves

The photosynthetic CO₂-fixation rates, chlorophyll content, chloroplast ultrastructure and other leaf characteristics (e.g. variable fluorescence, stomata density, soluble carbohydrate content) were studied in a comparative way in sun and shade leaves of beech (Fagus sylvatica) and in high-light and low-light seedlings.

1. Sun leaves of the beech possess a smaller leaf area, higher dry weight, lower water content, higher stomata density, higher chlorophyll a/b ratios and are thicker than the shade leaves. Sun leaves on the average contain more chlorophyll in a leaf area unit; the shade leaf exhibits more chlorophyll on a dry weight basis. Sun leaves show higher rates for dark respiration and a higher light saturation of photosynthetic CO₂-fixation. Above 2000 lux they are more efficient in photosynthetic quantum conversion than the shade leaves.
2. The development of HL-radish plants proceeds much faster than that of LL-plants. The cotyledons of HL-plants show a higher dry weight, lower water content, a higher ratio of chlorophyll a/b and a higher gross photosynthesis rate than the cotyledons of the LL-plants, which possess a higher chlorophyll content per dry weight basis. The large area of the HL-cotyledon on the one hand, as well as the higher stomata density and the higher respiration rate in the LL-cotyledon on the other hand, are not in agreement with the characteristics of sun and shade leaves respectively.
3. The development, growth and wilting of wheat leaves and the appearance of the following leaves (leaf succession) is much faster at high quanta fluence rates than in weak light. The chlorophyll content is higher in the HL-leaf per unit leaf area and in the LL-leaf per g dry weight. There are no differences in the stomata density and leaf area between the HL- and LL-leaf. There are fewer differences between HL- and LL-leaves than in beech or radish leaves.
4. The chloroplast ultrastructure of shade-type chloroplasts (shade leaves, LL-leaves) is not only characterized by a much higher number of thylakoids per granum and a higher stacking degree of thylakoids, but also by broader grana than in sun-type chloroplasts (sun leaves, HL-leaves). The chloroplasts of sun leaves and of HL-leaves exhibit large starch grains.
5. Shade leaves and LL-leaves exhibit a higher maximum chlorophyll fluorescence and it takes more time for the fluorescence to decline to the steady state than in sun and HL-leaves. The variable fluorescence VF (ratio of fluorescence decrease to steady state fluorescence) is always higher in the sun and HL-leaf of the same physiological stage (maximum chlorophyll content of the leaf) than in the shade and LL-leaf. The fluorescence emission spectra of sun and HL-leaves show a higher proportion of chlorophyli fluorescence in the second emission maximum F₂ than shade and LL-leaves.
6. The level of soluble carbohydrates (reducing sugars) is significantly higher in sun and HL-leaves than in shade and LL-leaves and even reflects changes in the amounts of the daily incident light.
7. Some but not all characteristics of mature sun and shade leaves are found in HL- and LL-leaves of seedlings. Leaf thickness, dry weight, chlorophyll content, soluble carbohydrate level, photosynthetic CO₂-fixation, height and width of grana stacks and starch content, are good parameters to describe the differences between LL- and HL-leaves; with some reservations concerning age and physiological stage of leaf, a/b ratios, chlorophyll content per leaf area unit and the variable fluorescence are also suitable.

     

Fluorescence induction kinetics of green and etiolated leaves by recording the complete in-vivo emission spectra

Complete room temperature fluorescence emission spectra of green and etiolated leaves (Raphanus sativus L., cv. Saxa Treib, Hordeum vulgare L., cv. Villa) are continuously recorded up to 4 min after onset of excitation. In green leaves two emission bands appear, whereas in etiolated leaves only one band is observed. In both cases the emission intensity increases with time, the high-energy band of green leaves decreasing more rapidly than the low-energy band. This phenomenon can be interpreted in terms of energy transfer. During the observation time of the fluorescence induction kinetic no shift of the emission peaks is found within the accuracy of the apparatus (±2nm).     

Heat-induced transfer of protons from chitosan to glycerol phosphate produces chitosan precipitation and gelation

Recently, chitosan dissolved in solutions containing glycerol phosphate (GP) were found to undergo a sol-gel transition when heated and the proposed gelling mechanism was based on increasing hydrophobic interactions with temperature. Subsequently, an investigation of ionization and precipitation behavior of chitosan, including dependencies on temperature, added salt, and fraction of deacetylated monomers (fD) was performed. This latter study revealed important differences in the temperature dependence of pKa of chitosan versus GP and led us to propose an alternative hypothesis for the mechanism of gelation in chitosan-GP systems whereby heat induces transfer of protons from chitosan to glycerol phosphate thereby neutralizing chitosan and allowing attractive interchain forces to form a physical gel. To investigate this specific molecular thermogelling mechanism, temperature ramp experiments on dilute chitosan-GP solutions were performed. Chitosans with fD of 0.72 and 0.98 were used to prepare solutions with a range of molar ratios of GP to chitosan glucosamine monomer of 1.25 to 10 and with 0 or 150 mM added monovalent salt. Light transmittance measurements were performed simultaneously to indicate precipitation in these dilute systems as a surrogate for gelation in concentrated systems. Measured temperatures of precipitation ranged from 15 to 85 degrees C, where solutions with less GP (used in a disodium salt form) had lower precipitation temperatures. A theoretical model using acid-base equilibria with temperature dependent pKa's, including the electrostatic contribution from the polyelectrolyte nature of chitosan, was used to calculate the degree ionization of chitosan (alpha, the fraction of protonated glucosamine monomer) as a function of temperature and showed a significant decrease in alpha with increased temperature due to proton transfer from chitosan to GP. This heat-induced proton transfer from chitosan to GP was experimentally confirmed by 31P NMR measurements during temperature ramp experiments since the chemical shift of 31P of GP is an indicator of its level of protonation. By assuming average temperature independent values of alpha p that were calculated from measured T(p), the model was able to accurately predict measured temperatures of precipitation (T(p)) of all chitosan-GP mixtures. The resulting alpha(p) were temperature independent but increased with increased chitosan fD and with increased salt. Measurements and theory revealed that T(p) can be adjusted in a predictable manner by changing the chitosan-GP molar ratio and thereby systematically tailored to obtain a large range of precipitation temperatures. Finally, similar temperature ramp experiments using inorganic phosphate and MES in place of GP demonstrated that the temperature-induced precipitation of chitosan also occurs with these buffers, confirming that the key feature of the buffer used with chitosan is its ability to absorb heat-stimulated release of chitosan protons and facilitate chitosan neutralization. A theoretical expression for the variation of chitosan ionization degree with temperature in a system composed of two titratable species (chitosan and buffer) was derived and allowed us to establish the required characteristics of the buffer for efficient heat-stimulated proton transfer between a chitosan and the buffer. These results provide a useful explanation for the mechanism of heat-induced gelation of chitosan-based systems that could be exploited for numerous practical applications.     

Variation in Herbivore Damage to Invasive and Native Woody Plant Species in Open Forest Vegetation on Mahé, Seychelles

Enemy release of introduced plants and variation in herbivore pressure in relation to community diversity are presently discussed as factors that affect plant species invasiveness or habitat invasibility. So far few data are available on this topic and the results are inconclusive. We compared leaf herbivory between native and invasive woody plants on Mahé, the main island of the tropical Seychelles. We further investigated variation in leaf herbivory on three abundant invasive species along an altitudinal gradient (50–550 m a.s.l.). The median percentage of leaves affected by herbivores was significantly higher in native species (50%) than in invasive species (27%). In addition, the species suffering from the highest leaf area loss were native to the Seychelles. These results are consistent with the enemy release hypothesis (ERH). While the invasive species showed significant and mostly consistent variation in the amount of leaf damage between sites, this variation was not related to general altitudinal trends in diversity but rather to local variation in habitat structure and diversity. Our results indicate that in the Seychelles invasive woody plants profit from herbivore release relative to the native species and that the amount of herbivory, and therefore its effect on species invasiveness or habitat invasibility, may be dependent on local community structure and composition.     

Anti-tumor necrosis factor-{alpha} therapies attenuate adaptive arteriogenesis in the rabbit

The specific antagonists of tumor necrosis factor-alpha (TNF-alpha), infliximab and etanercept, are established therapeutic agents for inflammatory diseases such as rheumatoid arthritis and Crohn's disease. Although the importance of TNF-alpha in chronic inflammatory diseases is well established, little is known about its implications in the cardiovascular system. Because proliferation of arteriolar connections toward functional collateral arteries (arteriogenesis) is an inflammatory-like process, we tested in vivo the hypothesis that infliximab and etanercept have antiarteriogenic actions. Sixty-three New Zealand White rabbits underwent femoral artery occlusion and received infliximab, etanercept, or vehicle according to clinical dosage regimes. After 1 wk, collateral conductance, assessed with fluorescent microspheres, revealed significant inhibition of arteriogenesis (collateral conductance): 52.4 (SD 8.1), 35.2 (SD 7.7), and 33.3 (SD 10.1) ml x min(-1) x 100 mmHg(-1) with PBS, infliximab, and etanercept, respectively (P < 0.001). High-resolution angiography showed no significant differences in number of collateral arteries, but immunohistochemical analysis demonstrated a decrease in mean collateral diameter, proliferation of vascular smooth muscle cells, and reduction of leukocyte accumulation around collateral arteries in treated groups. Infliximab and etanercept bound to infiltrating leukocytes, which are important mediators of arteriogenesis. Infliximab induced monocyte apoptosis, and neither substance affected monocyte expression of the adhesion molecule Mac-1. We demonstrated that TNF-alpha serves as a pivotal modulator of arteriogenesis, which is attenuated by treatment with TNF-alpha inhibitors. Reduction of collateral conductance is most likely due to inhibition of perivascular leukocyte infiltration and subsequent lower vascular smooth muscle cell proliferation. This is the first report showing a negative influence of TNF-alpha inhibitors on collateral artery growth.     

Fibroblast growth factor (FGF) 18 signals through FGF receptor 3 to promote chondrogenesis

Signaling by fibroblast growth factor (FGF) 18 and FGF receptor 3 (FGFR3) have been shown to regulate proliferation, differentiation, and matrix production of articular and growth plate chondrocytes in vivo and in vitro. Notably, the congenital absence of either FGF18 or FGFR3 resulted in similar expansion of the growth plates of fetal mice and the addition of FGF18 to human articular chondrocytes in culture enhanced proliferation and matrix production. Based on these and other experiments it has been proposed that FGF18 signals through FGFR3 to promote cartilage production by chondrocytes. Its role in chondrogenesis remains to be defined. In the current work we used the limb buds of FGFR3(+/+) and FGFR3(-/-) embryonic mice as a source of mesenchymal cells to determine how FGF18 signaling affects chondrogenesis. Confocal laser-scanning microscopy demonstrated impaired cartilage nodule formation in the FGFR3(-/-) cultures. Potential contributing factors to the phenotype were identified as impaired mitogenic response to FGF18, decreased production of type II collagen and proteoglycan in response to FGF18 stimulation, impaired interactions with the extracellular matrix resulting from altered integrin receptor expression, and altered expression of FGFR1 and FGFR2. The data identified FGF18 as a selective ligand for FGFR3 in limb bud mesenchymal cells, which suppressed proliferation and promoted their differentiation and production of cartilage matrix. This work, thus, identifies FGF18 and FGFR3 as potential molecular targets for intervention in tissue engineering aimed at cartilage repair and regeneration of damaged cartilage.     

Oxidative stress responses during cassava post-harvest physiological deterioration

A major constraint to the development of cassava (Manihot esculenta Crantz) as a crop to both farmers and processors is its starchy storage roots' rapid post-harvest deterioration, which can render it unpalatable and unmarketable within 24–72 h. An oxidative burst occurs within 15 min of the root being injured, that is followed by the altered regulation of genes, notably for catalase and peroxidase, related to the modulation of reactive oxygen species, and the accumulation of secondary metabolites, some of which show antioxidant properties. The interactions between these enzymes and compounds, in particular peroxidase and the coumarin, scopoletin, are largely confined to the vascular tissues where the visible symptoms of deterioration are observed. These, together with other data, are used to develop a tentative model of some of the principal events involved in the deterioration process. Abbreviations: ACMV, African cassava mosaic virus; AFLP, amplified fragment length polymorphism; CAT, catalase; cDNA, complementary deoxyribonucleic acid; CIAT, International Centre for Tropical Agriculture; Cu/ZnSOD, copper/zinc superoxide dismutase; DAB, 3,3-diaminobenzidine tetrahydrochloride; DPPH, 1,1-diphenyl-2-picrylhydrazyl; FeSOD, iron superoxide dismutase; FW, fresh weight; GUS, -glucuronidase; HPTLC, high-performance thin-layer chromatography; HR, hypersensitive response; IEF-PAGE, isoelectric focusing polyacrylamide gel electrophoresis; MAS, marker-assisted selection; MeJa, methyl jasmonate; MnSOD, manganese superoxide dismutase; NADPH, nicotinamide adenine dinucleotide phosphate (reduced form); NBT, nitroblue tetrazolium; PAL, phenylalanine ammonia-lyase; PCD, programmed cell death; PCR, polymerase chain reaction; POX, peroxidase; PPD, post-harvest physiological deterioration; QTL, quantitative trait loci; ROS, reactive oxygen species; RT, room temperature; SAR, systemic acquired resistance; SDS, sodium dodecyl sulfate; SOD, superoxide dismutase     

The role of fibril reinforcement in the mechanical behavior of cartilage

Collagen fibril reinforcement was incorporated into a nonlinear poroelastic model for articular cartilage in unconfined compression. It was found that the radial fibrils play a predominant role in the transient mechanical behavior but a less important role in the equilibrium response of cartilage. The radial fibrils are in tension and can be highly stressed during compression, in contrast to low compressive stresses in all directions for the proteoglycan matrix after a small initial compression. The strain dependent fibril stiffening produces strong nonlinear transient response; the fibrils provide extra stiffness to balance a rising fluid pressure and to restrain stress increase in the proteoglycans. The fibril reinforcement, induced by the fluid pressure and flow, also accounts for a complex pattern of strain-magnitude and strain-rate dependence of cartilage stiffness.