Seasonal and diurnal leaf orientation,bifacial sunlight incidence,and leaf structure in the sand dune herb Hydrocotyle bonariensis

A conceptual model has been proposed whereby leaf orientation and resulting sunlight exposure dictate functional leaf structure. Specifically, the model states that this relationship is driven by the absolute amount and ratio of incident sunlight on adaxial and abaxial leaf surfaces. To test this model, the relationships between corresponding values of leaf orientation and incident sunlight on both leaf surfaces were measured for the sand dune herb Hydrocotyle bonariensis over a growth season, along with examination of leaf structure. For mature leaves, leaf angle from horizontal and azimuth angle significantly increased over the growing season, indicating diurnal midday avoidance and seasonal maximization of incident sunlight. Consequently, seasonal changes in leaf orientation resulted in an overall decrease in midday sunlight incidence on the adaxial surface and a slight shift in the daily occurrence of peak abaxial incidence. Adaxial surfaces received three to four times more sunlight than abaxial surfaces, and leaf cross-sections revealed relatively thick (564 μm) leaves with multiple adaxial palisade layers and stomata on both leaf surfaces, as predicted by the conceptual model and measured ratio of incident sunlight on adaxial and abaxial leaf surfaces. These data provide further evidence of the relationship between leaf orientation and resulting absolute levels of sunlight incidence on both leaf surfaces, as well as their ratio, and corresponding differences in internal and external leaf structure.     

Investigating CH4 and N2O emissions from eco-engineering wastewater treatment processes using constructed wetland microcosms

Methane (CH₄) and nitrous oxide (N₂O) are important greenhouse gases, because of their contribution to the global greenhouse effect. The present study assessed emissions of N₂O and CH₄ from constructed wetland microcosms, planted with Phragmites australis and Zizania latifolia, when treating wastewater under different biological oxygen demand (BOD) concentration conditions. The removal rate was 95% for BOD and more than 80% for COD in all three pollutant concentrations, both plants’ removal rates of pollutants were at almost the same level, and both were found to resist BOD concentrations as high as 200 mg L⁻¹. When BOD concentrations fell below 200 mg L⁻¹, the soil plant units reached an average of 80–92% T-N and T-P removal rates; however, as the concentrations increased to 200 mg mg L⁻¹ or when during the initial phases of winter, the removal rates for T-N and T-P decreased to less than 70%. With NH₃-N removal, the influences of BOD concentrations and air temperature were more obvious. When BOD concentrations increased to 100 mg L⁻¹ after October, an obvious decrease in NH₃-N removal was detected; almost no nitrification occurred beginning in December at BOD concentrations of 200 mg mg L⁻¹. N₂O and CH₄ emissions showed obvious seasonal changes; higher emissions were observed with higher BOD concentrations, especially among Z. latifolia units. The enumeration of methane-oxidizing bacteria and methane-producing bacteria was also conducted to investigate their roles in impacting methane emissions and their relationships with plant species. The pollutant purification potentials of P. australis and Z. latifolia plant units during wastewater treatment of different pollutant concentrations occurred at almost the same levels. The nutrient outflow and methane flux were consistently higher with Z. latifolia units and higher concentrations of BOD. The more reductive status and higher biomass of methanogens may be the reason for the lower nitrification and higher CH₄ emissions observed with Z. latifolia units and higher concentration systems. The Z. latifolia root system is shallow, and the activity of methanotrophs is primarily confined to the upper portion of the soil. However, the root system of P. australis is deeper and can oxidize methane to a greater depth. This latter structure is more favorable as it is better for reducing methane emissions from P. australis soil plant systems.     

Stomatal Diffusion Resistance of Snap Beans. II. Effect of Light

The effect of light on the stomatal resistance of abaxial and adaxial leaf surfaces of snap beans (Phaseolus vulgaris L.) was studied in the growth chamber and in the field. The adaxial stomata required more light to open than the abaxial stomata; the abaxial stomatal apertures were still about 50% open at 1% full sunlight and light-induced closure was never observed under daytime field conditions. A given value of abaxial stomatal resistance was obtained at a given illumination of the abaxial guard cells whether illumination was adaxial or abaxial.     

Difference in light-induced increase in ploidy level and cell size between adaxial and abaxial epidermal pavement cells of Phaseolus vulgaris primary leaves

Changes in nuclear DNA content and cell size of adaxial andabaxial epidermal pavement cells were investigated using brightlight-induced leaf expansion of Phaseolus vulgaris plants. Inprimary leaves of bean plants grown under high (sunlight) ormoderate (ML; photon flux density, 163 µmol m^–2s^–1) light, most adaxial epidermal pavement cells hada nucleus with the 4C amount of DNA, whereas most abaxial pavementcells had a 2C nucleus. In contrast, plants grown under lowintensity white light (LL; 15 µmol m^–2 s^–1)for 13 d, when cell proliferation of epidermal pavement cellshad already finished, had a 2C nuclear DNA content in most adaxialpavement cells. When these LL-grown plants were transferredto ML, the increase in irradiance raised the frequency of 4Cnuclei in adaxial but not in abaxial pavement cells within 4d. On the other hand, the size of abaxial pavement cells increasedby 53% within 4 d of transfer to ML and remained unchanged thereafter,whereas adaxial pavement cells continuously enlarged for 12d. This suggests that the increase in adaxial cell size after4 d is supported by the nuclear DNA doubling. The differentresponses between adaxial and abaxial epidermal cells were notinduced by the different light intensity at both surfaces. Itwas shown that adaxial epidermal cells have a different propertythan abaxial ones. Key words: Cell enlargement, endopolyploidization, epidermal pavement cells, incident light intensity, leaf expansion, nuclear DNA content, Phaseolus vulgaris     

Efficacy of selected adjuvants for protecting Spodoptera litura nucleopolyhedrovirus from sunlight inactivation

Efficacy of selected adjuvants for protecting Spodoptera litura nucleopolyhedrovirus (SpltNPV) from sunlight inactivation was tested in direct sunlight exposure bioassays and in the field on virus-treated Brassica chinensis var parachinensis plants. All adjuvants, except palm oil, gave significant protection to the virus after 16 h of sunlight exposure, as shown by the larval mortalities. Cumulative mortalities after 14 days show that the virus + Tinopal treatment and the virus not to sunlight treatment attained 100% mortality, while the virus in riboflavin treatment and the crude brown sugar treatment caused mortality of less than 30%. However, the LT₅₀ value of virus + Tinopal was 1.2 times lower than the unexposed virus. All virus-treated plants in the field trials had higher yields and lower damage indices than the control plants. Unlike the results from direct sunlight exposure bioassays, adjuvants, including virus + Tinopal LPW, did not significantly enhance efficacy of the virus against early instars of S. litura in the field. The unformulated virus was as effective as virus + Tinopal LPW and Nurelle-D in protecting the plants from infestation. Thus, in a well-planned pest management program, SpltNPV, without any adjuvant, can be used effectively for controlling S. litura on B. chinensis var parachinensis.     

Effect of different macrophytes in abating nitrogen from a synthetic wastewater

An experiment conducted in an unheated glasshouse from October 2006 to March 2008 studied the efficiency of different macrophytes in reducing NO₃-N and NH₄-N concentrations and loads in synthetic wastewaters. The experimental setup consisted of plastic tanks, filled with gravel and vegetated with Carex elata All., Juncus effusus L., Phragmites australis (Cav.) Trin., Typhoides arundinacea L. Moench (syn Phalaris arundinacea L.) var. picta and Typha latifolia L. There was also a control without vegetation. From January to July, a solution of 50–60 ppm of NH₄-N and NO₃-N was applied monthly; then the input concentration was doubled. The total load at the end of the experimental period was 70.4 g/m² of NO₃-N and 67.3 of NH₄-N. At the end of each month, water was discharged from the tanks and analysed to determine the two nitrogen forms. At the end of the experiment, 33 g/m² of total N (almost 24% of applied N) had disappeared in the control. Among species, the highest abatement was detected in T. latifolia (72 g/m², almost 52% of applied N) and the lowest in J. effusus (35%).A weekly chemical analysis in July showed that a large amount of NH₄-N quickly disappeared in all treatments, while NO₃-N only decreased in the vegetated tanks. In December, NH₄-N had similar dynamics, while NO₃-N increased.All water volumes entering and exiting the tanks were measured in order to evaluate evapotranspiration. T. latifolia showed the highest water consumption, reaching a cumulative value of above 1000 mm.At the end of the experiment, J. effusus presented the highest amount of nitrogen stored in the aerial parts (5.63 g/m²) and T. latifolia the lowest (1.92 g/m²).     

Contribution of hydroxycinnamates and flavonoids to epidermal shielding of UV‐A and UV‐B radiation in developing rye primary leaves as assessed by ultraviolet‐induced chlorophyll fluorescence measurements

Epidermally located ultraviolet (UV)‐absorbing phenolic compounds, flavonoids and hydroxycinnamic acid esters (HCAs), can shield the underlying tissues in plants against harmful UV‐radiation. The relative importance of the two different classes of phenolic compounds for UV‐screening was a matter of recent debate. Using a non‐invasive method based on chlorophyll fluorescence measurements to estimate epidermal UV transmittance, the relationship between epidermal UV shielding and the content of the two different groups of secondary phenolic compounds in the epidermal layers and the underlying photosynthetic mesophyll of developing rye primary leaves grown under supplementary UV‐B radiation was investigated. From the fourth to the tenth day after sowing, epidermally located flavonoids increased in an age‐ and irradiation‐dependent manner, whereas mesophyll flavonoids and epidermal HCAs, mainly ferulic acid and p‐coumaric acid esters, were constitutively present and did not vary in their contents over the observed time period. There was an excellent correlation between epidermal UV‐A and UV‐B absorbances as assessed by chlorophyll fluorescence measurements and contents of epidermal flavonoids. However, HCAs showed an additional contribution to UV‐B shielding. In contrast, mesophyll flavonoids did not seem to play a respective role. When absorbances of the abaxial and adaxial epidermal layers were compared, it became apparent that in fully expanded primary leaves epidermal tissues from both sides were equally effective in absorption of UV‐radiation. However, the earlier and more UV‐exposed abaxial epidermis of young unrolling leaves showed a significantly higher absorption. It is shown that in early stages of development the epidermal HCAs are the dominant UV‐B protective compounds of the primary leaf. This function is increasingly replaced by the epidermal flavonoids during leaf development and acclimation. The application of chlorophyll fluorescence measurements has been proven to be a useful tool for estimating relative contents of these compounds in epidermal tissue.     

An efficient autofluorescence method for screening Limonium bicolor mutants for abnormal salt gland density and salt secretion

With the spread of saline soils worldwide, it has become increasingly important to understand salt-tolerant mechanisms and to develop halophytes with increased salt tolerance. Limonium bicolor is a typical recretohalophyte and has a typical salt excretory structure in the epidermis called the salt gland. A method that can be used to screen a large population of L. bicolor mutants for altered salt gland density and altered salt secretion is needed but is currently unavailable. Leaves of 1-month-old L. bicolor seedlings were processed by three traditional methods [epidermal peel, nail impression, and clearing/differential interference contrast microscope (clearing/DIC) method] and a fluorescence method (fluorescence microscopic examination of cleared leaves). With the fluorescence method, the autofluorescence of salt glands under UV excitation (330–380 nm) was easily distinguished with the least labor and time. The fluorescence method was used to screen ~ 10,000 seedlings (which grew from gamma-irradiated seeds). Four mutants with reduced salt gland density and 15 with increased salt gland density were obtained. Both kinds of mutants will be useful for the isolation of genes involved in salt gland development and salt secretion in L. bicolor and other halophytes. The fluorescence method was also successfully used to observe the salt glands of Aegialitis rotundifolia and the stomata and trichomes of Arabidopsis. The fluorescence method described here will be useful for examining plant epidermal structures that have autofluorescence under UV or other wavelengths.     

Effects of shade treatments on the photosynthetic capacity,chlorophyll fluorescence,and chlorophyll content of Tetrastigma hemsleyanum Diels et Gilg

Tetrastigma hemsleyanum Diels et Gilg was grown under full sunlight and moderate and high levels of shade for one month to evaluate its photosynthetic and chlorophyll fluorescence response to different light conditions. The results showed that T. hemsleyanum attained greatest leaf size and P_n when cultivated with 67% shade. Leaves of seedlings grown with 90% shade were the smallest. Leaf color of plants grown under full sunlight and 50% shade was yellowish-green. The P_n value increased rapidly as PPFD increased to 200 μmol m^?2 s^?1 and then increased slowly to a maximum, followed by a slow decrease as PPFD was increased to 1000 μmol m^?2 s^?1. P_n was highest for the 67% shade treatment and the LSP for this shade treatment was 600 μmol m^?2 s^?1. Full sunlight and 50% shade treatments resulted in significant reduction of ETR and qP and increased NPQ. Chl a, Chl b and total chlorophyll content increased and Chl a/b values decreased with increased shading. Results showed that light intensity greater than that of 50% shade depressed photosynthetic activity and T. hemsleyanum growth. Irradiance less than that of 75% shade limited carbon assimilation and led to decreased plant growth. Approximately 67% shade is suggested to be the optimum light irradiance condition for T. hemsleyanum cultivation.     

Characterization of two peptides isolated from the venom of social wasp Chartergellus communis (Hymenoptera: Vespidae): Influence of multiple alanine residues and C-terminal amidation on biological effects

Chatergellus communis is a wasp species endemic to the neotropical region and its venom constituents have never been described. In this study, two peptides from C. communis venom, denominated Communis and Communis-AAAA, were chemically and biologically characterized. In respect to the chemical characterization, the following amino acid sequences and molecular masses were identified:Communis: Ile-Asn-Trp-Lys-Ala-Ile-Leu-Gly-Lys-Ile-Gly-Lys-COOH (1340.9 Da)Communis-AAAA: Ile-Asn-Trp-Lys-Ala-Ile-Leu-Gly-Lys-Ile-Gly-Lys-Ala-Ala-Ala-Ala-Val-Xle-NH₂ (1836.3 Da).Furthermore, their biological effects were compared, accounting for the differences in structural characteristics between the two peptides. To this end, three biological assays were performed in order to evaluate the hyperalgesic, edematogenic and hemolytic effects of these molecules. Communis-AAAA, unlike Communis, showed a potent hemolytic activity with EC₅₀ = 142.6 μM. Moreover, the highest dose of Communis-AAAA (2nmol/animal) induced hyperalgesia in mice. On the other hand, Communis (10nmol/animal) was able to induce edema but did not present hemolytic or hyperalgesic activity. Although both peptides have similarities in linear structures, we demonstrated the distinct biological effects of Communis and Communis-AAAA. This is the first study with Chartegellus communis venom, and both Communis and Communis-AAAA are unpublished peptides.     

Influence of plant species and wastewater strength on constructed wetland methane emissions and associated microbial populations

Wastewater and treatment processes have been regarded as large contributions to sources of methane (CH₄). The flux rate of CH₄ in constructed wetlands (CWs) was evaluated to test the influence of plant species. Methane emission data showed large temporal and spatial variation ranging from 0 to 16.76 g CH₄ m⁻² day⁻¹. The highest CH₄ flux rate was obtained in the Zizania latifolia systems and higher emission was found with higher influent load. The methanogenic and methanotrophic microbial populations were studied to clarify the mechanisms of CH₄ emission. FISH analysis showed highest amounts of methanogens and methanotrophs in the Z. latifolia and Tytha latifolia systems. Linear regression between CH₄ emission and environmental parameters showed that the regression lines were not forced to pass through the origin, and the slopes of the lines of different systems were allowed to vary between vegetation cover.     

Effects of rabbit gut passage on seed retrieval and germination of three shrub species

It has been known for a few decades that European rabbits consume seeds and fleshy fruits of native woody plants, but relevant factors in the endozoochory processes such as seed predation (chewing and digestion), sexual differentiation, or the temporal pattern of seed recovery have been little evaluated until now. In this study, we examined seed dispersal of three Mediterranean shrub species by wild rabbits through monitoring of seed retrieval and germination after gut passage. Twelve adult wild rabbits (Oryctolagus cuniculus; six males and six females) of similar size and age were fed seeds of three shrub species with fleshy fruits (Crataegus monogyna, Myrtus communis and Pistacia lentiscus). After ingestion of fruits, seeds were retrieved from dung every 12 h for a day and a half. The viability and germination of retrieved seeds were tested along with that of uneaten seeds. Between 5% and 76% of ingested seeds were retrieved from dung, with significant differences between species and sex. Most M. communis seeds were retrieved with 12–24 h after ingestion; almost all C. monogyna seeds were recovered with 0–12 h after ingestion; no seeds of P. lentiscus were recovered. Only in the case of M. communis seed was the recovery rate greater in female than in male. Passage through the rabbit gut significantly increased seed germination in M. communis, and decreased it in C. monogyna. In conclusion, wild rabbits acted in this study as predators of C. monogyna and P. lentiscus seeds and potential dispersers of M. communis seeds.     

Photosynthetic costs and benefits of abaxial versus adaxial anthocyanins in Colocasia esculenta ‘Mojito’

Main conclusion

Anthocyanins in upper (adaxial) leaf tissues provide greater photoprotection than in lower (abaxial) tissues, but also predispose tissues to increased shade acclimation and, consequently, reduced photosynthetic capacity. Abaxial anthocyanins may be a compromise between these costs/benefits. Plants adapted to shaded understory environments often exhibit red/purple anthocyanin pigmentation in lower (abaxial) leaf surfaces, but rarely in upper (adaxial) surfaces. The functional significance of this color pattern in leaves is poorly understood. Here, we test the hypothesis that abaxial anthocyanins protect leaves of understory plants from photo-oxidative stress via light attenuation during periodic exposure to high incident sunlight in the forest understory, without interfering with sunlight capture and photosynthesis during shade conditions. We utilize a cultivar of Colocasia esculenta exhibiting adaxial and abaxial anthocyanin variegation within individual leaves to compare tissues with the following color patterns: green adaxial, green abaxial (GG), green adaxial, red abaxial (GR), red adaxial, green abaxial (RG), and red adaxial, red abaxial (RR). Consistent with a photoprotective function of anthocyanins, tissues exhibited symptoms of increasing photoinhibition in the order (from least to greatest): RR, RG, GR, GG. Anthocyanic tissues also showed symptoms of shade acclimation (higher total chl, lower chl a/b) in the same relative order. Inconsistent with our hypothesis, we did not observe any differences in photosynthetic CO₂ uptake under shade conditions between the tissue types. However, GG and GR had significantly (39 %) higher photosynthesis at saturating irradiance (A _sat) than RG and RR. Because tissue types did not differ in nitrogen content, these patterns likely reflect differences in resource allocation at the tissue level, with greater nitrogen allocated toward energy processing in GG and GR, and energy capture in RG and RR (consistent with relative sun/shade acclimation). We conclude that abaxial anthocyanins are likely advantageous in understory environments because they provide some photoprotection during high-light exposure, but without the cost of decreased A _sat associated with adaxial anthocyanin-induced shade syndrome.     

Action spectra of photosystems II and I and quantum yield of photosynthesis in leaves in State 1

The spectral global quantum yield (Y_II, electrons/photons absorbed) of photosystem II (PSII) was measured in sunflower leaves in State 1 using monochromatic light. The global quantum yield of PSI (Y_I) was measured using low-intensity monochromatic light flashes and the associated transmittance change at 810 nm. The 810-nm signal change was calibrated based on the number of electrons generated by PSII during the flash (4 · O₂ evolution) which arrived at the PSI donor side after a delay of 2 ms. The intrinsic quantum yield of PSI (y_I, electrons per photon absorbed by PSI) was measured at 712 nm, where photon absorption by PSII was small. The results were used to resolve the individual spectra of the excitation partitioning coefficients between PSI (a_I) and PSII (a_II) in leaves. For comparison, pigment–protein complexes for PSII and PSI were isolated, separated by sucrose density ultracentrifugation, and their optical density was measured. A good correlation was obtained for the spectral excitation partitioning coefficients measured by these different methods. The intrinsic yield of PSI was high (y_I = 0.88), but it absorbed only about 1/3 of quanta; consequently, about 2/3 of quanta were absorbed by PSII, but processed with the low intrinsic yield y_II = 0.63. In PSII, the quantum yield of charge separation was 0.89 as detected by variable fluorescence F_v/F_m, but 29% of separated charges recombined (Laisk A, Eichelmann H and Oja V, Photosynth. Res. 113, 145–155). At wavelengths less than 580 nm about 30% of excitation is absorbed by pigments poorly connected to either photosystem, most likely carotenoids bound in pigment–protein complexes.     

Experiment of emergent macrophytes growing in contaminated sludge: Implication for sediment purification and lake restoration

Three emergent macrophytes (Zizania latifolia (Turcz) Hand.-Mazz., Phragmites australis (Cav.) Trin. ex Steud. and Typha angustifolia Linn.) and three different sediments from Lake Dianchi of Yunnan province, China, were studied through orthogonal pot-planting experiment in order to compare the ability of the three emergent macrophytes in dealing with the contaminated sludge and to evaluate the possibility of purifying the sediment through aquatic plant rehabilitation. The results show that the number of sprouts and biomass of all the species growing in the sediment of site 3 were higher than those growing in the sediment of sites 1 and 2; the plants growing in the sediment of site 3 also exhibited the highest root activities; in each sediment, the sequence of root activity of the species was: Z. latifolia > P. australi > T. angustifolia; TP content in the sediments grown with different plants reduced significantly than those of control. These results indicated that these emergent plants were able to grow well in the contaminated sediment though it is black with a strong odor. Z. latifolia shows the highest root activity in the sediment of site 3, from which we can deduce that this plant should be the preferred pioneer species for purifying the sediment. According to their biomass and TN content and TP content, Z. latifolia, P. australi and T. angustifolia retained TN 16.6, 29.8, 12.8, and TP 2.2, 3.6, 3.9 g m^?2, respectively. Based on the climate of Dianchi valley, these plants can be harvested twice in a year. Thus, the amount of nutrient can be removed almost doubly. Therefore, the sludge can be purified through macrophytes restoration and not through sediment dredging, which was proved to be expensive and invalid in the large shallow lakes.     

Origins of non-linear and dissimilar relationships between epidermal UV absorbance and UV absorbance of extracted phenolics in leaves of grapevine and barley

A recent review of climate patterns in Southern Germany has suggested significant increases in ultraviolet (UV) radiation due to decreases in cloud coverage and in cloud frequency which compound the effects of stratospheric ozone depletion. Whether such UV radiation increases result in UV damage of higher plant leaves depends partly on the capacity of UV-absorbing hydroxycinnamic acids and flavonoids located in the plant epidermis to screen out UV radiation. Epidermal UV screening is most often assessed from UV absorbance of whole-leaf extracts but in the present work, this method is critically examined. In grapevine (Vitis vinifera L.), hydroxycinnamic acid as well as mono-hydroxylated and ortho-dihydroxylated flavonoid concentrations increased in parallel with fluorometrically detected adaxial epidermal UV absorbance but only the latter class of flavonoids was associated with epidermal UV absorbance in barley (Hordeum vulgare L). For both species, curvilinear relationships between epidermal and total phenolic UV absorbance were established: initial slopes of the curves differed markedly between species. Modelling suggested that curvilinearity arises from UV-transparent epidermal areas located between vacuoles which are particularly UV-absorbing due to high levels of phenolics. The species-dependent differences were related to allocation of high amounts of phenolics in the mesophyll and abaxial epidermis in barley but not in grapevine. Both factors, optical heterogeneity and variable distribution of phenolics, severely restrict the use of phenolic absorbance to estimate true epidermal screening.     

Structured near-infrared Magnetic Circular Dichroism spectra of the Mn4CaO5 cluster of PSII in T. vulcanus are dominated by Mn(IV) d-d ‘spin-flip’ transitions

Photosystem II passes through four metastable S-states in catalysing light-driven water oxidation. Variable temperature variable field (VTVH) Magnetic Circular Dichroism (MCD) spectra in PSII of Thermosynochococcus (T.) vulcanus for each S-state are reported. These spectra, along with assignments, provide a new window into the electronic and magnetic structure of Mn₄CaO₅. VTVH MCD spectra taken in the S₂ state provide a clear g = 2, S = 1/2 paramagnetic characteristic, which is entirely consistent with that known by EPR. The three features, seen as positive (+) at 749 nm, negative (?) at 773 nm and (+) at 808 nm are assigned as ⁴A → ²E spin-flips within the d³ configuration of the Mn(IV) centres present. This assignment is supported by comparison(s) to spin-flips seen in a range of Mn(IV) materials. S₃ exhibits a more intense (?) MCD peak at 764 nm and has a stronger MCD saturation characteristic. This S₃ MCD saturation behaviour can be accurately modelled using parameters taken directly from analyses of EPR spectra. We see no evidence for Mn(III) d-d absorption in the near-IR of any S-state. We suggest that Mn(IV)-based absorption may be responsible for the well-known near-IR induced changes induced in S₂ EPR spectra of T. vulcanus and not Mn(III)-based, as has been commonly assumed. Through an analysis of the nephelauxetic effect, the excitation energy of S-state dependent spin-flips seen may help identify coordination characteristics and changes at each Mn(IV). A prospectus as to what more detailed S-state dependent MCD studies promise to achieve is outlined.     

Mesophyll distribution of ��antioxidant�� flavonoid glycosides in Ligustrum vulgare leaves under contrasting sunlight irradiance

Background and Aims

Flavonoids have the potential to serve as antioxidants in addition to their function of UV screening in photoprotective mechanisms. However, flavonoids have long been reported to accumulate mostly in epidermal cells and surface organs in response to high sunlight. Therefore, how leaf flavonoids actually carry out their antioxidant functions is still a matter of debate. Here, the distribution of flavonoids with effective antioxidant properties, i.e. the orthodihydroxy B-ring-substituted quercetin and luteolin glycosides, was investigated in the mesophyll of Ligustrum vulgare leaves acclimated to contrasting sunlight irradiance.

Methods

In the first experiment, plants were grown at 20 % (shade) or 100% (sun) natural sunlight. Plants were exposed to 100 % sunlight irradiance in the presence or absence of UV wavelengths, in a second experiment. Fluorescence microspectroscopy and multispectral fluorescence microimaging were used in both cross sections and intact leaf pieces to visualize orthodihydroxy B-ring-substituted flavonoids at inter- and intracellular levels. Identification and quantification of individual hydroxycinnamates and flavonoid glycosides were performed via HPLC-DAD.

Key Results

Quercetin and luteolin derivatives accumulated to a great extent in both the epidermal and mesophyll cells in response to high sunlight. Tissue fluorescence signatures and leaf flavonoid concentrations were strongly related. Monohydroxyflavone glycosides, namely luteolin 4′-O-glucoside and two apigenin 7-O-glycosides were unresponsive to changes in sunlight irradiance. Quercetin and luteolin derivatives accumulated in the vacuoles of mesophyll cells in leaves growing under 100 % natural sunlight in the absence of UV wavelengths.

Conclusions

The above findings lead to the hypothesis that flavonoids play a key role in countering light-induced oxidative stress, and not only in avoiding the penetration of short solar wavelengths in the leaf.     

Characterization and kinetic properties of the purified Trematosphaeria mangrovei laccase enzyme

The properties of Trematosphaeria mangrovei laccase enzyme purified on Sephadex G-100 column were investigated. SDS–PAGE of the purified laccase enzyme showed a single band at 48 kDa. The pure laccase reached its maximal activity at temperature 65 °C, pH 4.0 with K_m equal 1.4 mM and V_max equal 184.84 U/mg protein. The substrate specificity of the purified laccase was greatly influenced by the nature and position of the substituted groups in the phenolic ring. The pure laccase was tested with some metal ions and inhibitors, FeSO₄ completely inhibited laccase enzyme and also highly affected by (NaN₃) at a concentration of 1 mM. Amino acid composition of the pure enzyme was also determined. Carbohydrate content of purified laccase enzyme was 23% of the enzyme sample. The UV absorption spectra of the purified laccase enzyme showed a single peak at 260–280 nm.