Rob Broekman

Stratospheric ozone reduction and ecosystem processes: enhanced UV-B radiation affects chemical quality and decomposition of leaves of the dune grassland species Calamagrostis epigeios

This study reports changes in the plant’s chemical composition and the decomposition of this plant material under enhanced solar UV-B radiation. Calamagrostis epigeios, a dominant grass species in the dune grassland in The Netherlands, was grown outdoor on an experimental field under ambient and enhanced solar UV-B (5 and 7.5 kJ m-2 day-1 UV-BBE, respectively), corresponding to about 15% stratospheric ozone depletion. After one growing season aerial plant parts were harvested. The decomposition of this harvested leaf material was studied in a dune grassland and on the above mentioned experimental field under ambient (5 kJ m-2 day-1 UV-BBE) and enhanced (7.5 kJ m-2 day-1 UV-BBE) radiation, using litter bags. The chemical quality of the leaves grown under enhanced solar UV-B changed. There was an increase in the leaf content of lignin, while no significant changes occurred for the content of α-cellulose, hemicellulose and tannins under enhanced UV-B. In the field, the rate of decomposition of leaf material grown under enhanced UV-B (with an increased content of lignin) was reduced. The content of lignin of the decomposing leaf material increased, but less under exposure to enhanced UV-B. The latter may be explained by photodegradation of the lignin. The consequences of enhanced UV-B radiation for carbon fluxes in the dune grassland ecosystem are discussed.

UV-B Absorbance and UV-B absorbing compounds (para-coumaric acid) in pollen and sporopollenin: the perspective to track historic UV-B levels.

UV-B absorbance and UV-B absorbing compounds (UACs) of the pollen of Vicia faba, Betula pendula, Helleborus foetidus and Pinus sylvestris were studied. Sequential extraction demonstrated considerable UV-B absorbance both in the soluble (acid methanol) and insoluble sporopollenin (acetolysis resistant residue) fractions of UACs, while the wall-bound fraction of UACs was small. The UV-B absorbance of the soluble and sporopollenin fraction of pollen of Vicia faba plants exposed to enhanced UV-B (10 kJ m(-2) day(-1) UV-B(BE)) was higher than that of plants that received 0 kJ m(-2) day(-1) UV-B(BB). Pyrolysis gas chromatography-mass spectrometry (py-GC-MS) analysis of pollen demonstrated that p-coumaric acid and ferulic acid formed part of the sporopollenin fraction of the pollen. The amount of these aromatic monomers in the sporopollenin of Vicia faba appeared to increase in response to enhanced UV-B (10 kJ m(-2) day(-1) UV-B(BE)). The detection limit of pyGC-MS was sufficiently low to quantify these phenolic acids in ten pollen grains of Betula and Pinus. The experimental data presented provide evidence for the possibility that polyphenolic compounds in pollen of plants are indicators of solar UV-B and may be applied as a new proxy for the reconstruction of historic variation in solar UV-B levels.

Relation between heavy metal concentrations in salt marsh plants and soil

The aim of the research reported here was to investigate the relation between heavy metal concentrations in salt marsh plants, extractability of the metals from soil and some soil characteristics. In April 1987, Spartina anglica and Aster tripolium plants and soil were collected from four salt marshes along the Dutch coast. The redox potential of the soil between the roots of the plants and at bare sites was measured. Soil samples were oven-dried and analyzed for chloride concentration, pH, fraction of soil particles smaller than 63 microm (f < 63 microm), loss on ignition (LOI) and ammonium acetate and hydrochloric acid extractable Cd, Cu and Zn concentrations. The roots and shoots of the plants were analyzed for Cd, Cu and Zn. Because drying of the soil prior to chemical analysis might have changed the chemical speciation of the metals, and therefore the outcome of the ammonium acetate extraction, a second survey was performed in October 1990. In this survey A. tripolium plants and soil were collected from two salt marshes. Fresh and matched oven-dried soil samples were analyzed for water, ammonium acetate and diethylene triaminepentaacetic acid (DTPA) extractable Cd, Cu and Zn concentrations. The soil samples were also analyzed for f < 63 microm, LOI and total (HNO(3)/HCl digestion) metal concentrations. Soil metal concentrations were correlated with LOI. Drying prior to analysis of the soil had a significant effect on the extractability of the metals with water, ammonium acetate or DTPA. Plant metal concentrations significantly correlated only with some extractable metal concentrations determined in dried soil samples. However, these correlations were not consistently better than with total metal concentrations in the soil. It was concluded that extractions of metals from soil with water, ammonium acetate or DTPA are not better predictors for metal concentrations in salt marsh plants than total metal concentrations, and that a major part of the variation in metal concentrations in the plants cannot be explained by variation in soil composition.

Effect of elevated atmospheric CO2 on growth, photosynthesis and water relations of salt marsh grass species

Abstract The C3 grass species Scirpus maritimus L. and Puccinellia maritima (Huds.) Parl., and the C4 grass species Spartina anglica C.E. Hubbard and Spartina patens (Ait.) Muhl. were grown at ambient (340 p.p.m. CO2) and elevated (580 p.p.m. CO2) atmospheric CO2 concentration, at low (10 mM NaCl) and high salinity (250 mM NaCl) under aerated and anaerobic conditions in the culture solution. The relative growth rate of both the C3 grass species was enhanced with atmospheric CO2 enrichment, no such increase was found in the C4 grass species. High salinity reduced growth of the C3 species tested, but this relative growth reduction was not prevented by elevated CO2 concentration. The growth increase at elevated CO2 of Scirpus maritimus and Puccinellia maritima is greater under aerated than under anaerobic solution conditions. Water-use efficiency of all species was increased by elevated CO2. In the case of Scirpus (C3), this increase was caused by increased net photosynthesis, for Spartina patens (C4) photosynthesis was not increased, but transpiration was reduced. The water potential of the shoot was less negative under conditions of CO2 enrichment, in particular at increased salinity (250 mM NaCl).

Flavonoid concentrations in three grass species and a sedge grown in the field and under controlled environment conditions in response to enhanced UV-B radiation.

An investigation was carried out to find whether enhanced levels of UV-B radiation induce increased concentrations of flavonoids in the leaves of the grass species Deschampsia antarctica, Deschampsia borealis and Calamagrostis epigeios and the sedge Carex arenaria. Whether the enhanced levels of UV-B influenced the proportions of the various flavonoids in the leaves was also studied. Increased flavonoid concentrations would improve the UV-B shielding of UV-B susceptible tissues. Using HPLC analysis the flavonoids orientin and luteolin were identified in D. antarctica, orientin in D. borealis and tricin in C. arenaria. Neither flavonoid concentrations nor the proportion of the various flavonoids in climate room grown D. antarctica and D. borealis plants differed between individuals grown under 0, ambient or elevated UV-B levels. After 12 weeks of growth biomass production and shoot-to-root ratios of D. antarctica were not affected by elevated UV-B radiation. Greenhouse grown C. epigeios plants contained higher concentrations and different proportions of flavonoids grown under elevated levels of UV-B than when grown under ambient or 0 UV-B. In C. epigeios plants grown in their natural habitat in the field under ambient or elevated levels of UV-B, flavonoid concentrations and proportions were the same in plants from both treatments. In the leaves of the sedge C. arenaria grown in a greenhouse flavonoid concentrations and proportions were not affected by UV-B radiation. Leaves were harvested four times during the growing season from C. arenaria plants grown in their natural habitat in the field under ambient or elevated levels of UV-B. Leaves harvested in January contained higher concentrations of flavonoids when grown under elevated UV-B than when grown under ambient UV-B radiation. In leaves harvested in May, September and December flavonoid concentrations were the same in plants grown under ambient or elevated UV-B. The proportion of the different flavonoids was the same for both treatments in all months. These results indicate that constitutive levels of flavonoids in these grass and sedge species are adequately high to protect them against ambient and elevated levels of solar UV-B radiation.

Stratospheric ozone depletion: High arctic tundra plant growth on Svalbard is not affected by enhanced UV-B after 7 years of UV-B supplementation in the field

The response of tundra plants to enhanced UV-B radiation simulating 15 and 30% ozone depletion was studied at two high arctic sites (Isdammen and Adventdalen, 78° N, Svalbard).The set-up of the UV-B supplementation systems is described, consisting of large and small UV lamp arrays, installed in 1996 and 2002. After 7 years of exposure to enhanced UV-B radiation, plant cover, density, morphological (leaf fresh and dry weight, leaf thickness, leaf area, reproductive and ecophysiological parameters leaf UV-B absorbance, leaf phenolic content, leaf water content) were not affected by enhanced UV-B radiation. DNA damage in the leaves was not increased with enhanced UV-B in Salix polaris and Cassiope tetragona. DNA damage in Salix polaris leaves was higher than in leaves of C. tetragona. The length of male gametophyte moss plants of Polytrichum hyperboreum was reduced with elevated UV-B as well as the number of Pedicularis hirsuta plants per plot, but the inflorescence length of Bistorta vivipara was not significantly affected. We discuss the possible causes of tolerance of tundra plants to UV-B (absence of response to enhanced UV-B) in terms of methodology (supplementation versus exclusion), ecophysiological adaptations to UV-B and the biogeographical history of polar plants

The occurrence of p-coumaric acid and ferulic acid in fossil plant materials and their use as UV-proxy.

The applicability of p-coumaric acid and ferulic acid concentrations or ratios in (sub)fossil plant remnant as UV-B proxies relies on various aspects, which are discussed in this paper and will be illustrated with some experimental data. A newly developed THM-micropyrolysis–gas chromatography–mass spectrometry method was tested on various spores, pollen and other plant remains, which were analysed for the presence of the UV-absorbing compounds p-coumaric acid and ferulic acid. This revealed that these supposed building-blocks of sporopollenin appear to be present in pollen of many plant species but also in moss spores. The development of this micropyrolysis method paved the way for the quantitative analysis of UV-absorbing compounds in case only a small amount of analyte is available, for example for fossil pollen and spores but also other small palynomorphs and plant fossils. The use of this technique will provide a better insight in the plant responses to UV-radiation, the chemistry of pollen and spores, their fossil counterparts and furthermore the means for a further development of a proxy for the reconstruction of past UV-B radiation.

No divergence in Cassiope tetragona: persistence of growth response along a latitudinal temperature gradient and under multi-year experimental warming.

BACKGROUND AND AIMS The dwarf shrub Cassiope tetragona (Arctic bell-heather) is increasingly used for arctic climate reconstructions, the reliability of which depends on the existence of a linear climate-growth relationship. This relationship was examined over a high-arctic to sub-arctic temperature gradient and under multi-year artificial warming at a high-arctic site. METHODS Growth chronologies of annual shoot length, as well as total leaf length, number of leaves and average leaf length per year, were constructed for three sites. Cassiope tetragona was sampled near its cold tolerance limit at Ny-Ålesund, Svalbard, at its assumed climatic optimum in Endalen, Svalbard, and near its European southern limit at Abisko, Sweden. Together these sites represent the entire temperature gradient of this species. Leaf life span was also determined. Each growing season from 2004 to 2010, 17 open top chambers (OTCs) were placed near Ny-Ålesund, thus increasing the daily mean temperatures by 1·23°C. At the end of the 2010 growing season, shoots were harvested from OTCs and control plots, and growth parameters were measured. KEY RESULTS All growth parameters, except average leaf length, exhibited a linear positive response (R(2) between 0·63 and 0·91) to mean July temperature over the temperature gradient. Average leaf life span was 1·4 years shorter in sub-arctic Sweden compared with arctic Svalbard. All growth parameters increased in response to the experimental warming; the leaf life span was, however, not significantly affected by OTC warming. CONCLUSIONS The linear July temperature-growth relationships, as well as the 7 year effect of experimental warming, confirm that the growth parameters annual shoot length, total leaf length and number of leaves per year can reliably be used for monitoring and reconstructing temperature changes. Furthermore, reconstructing July temperature from these parameters is not hampered by divergence.

Quantification of UV-B flux through time using UV-B-absorbing compounds contained in fossil Pinus sporopollenin.

UV-B radiation currently represents c. 1.5% of incoming solar radiation. However, significant changes are known to have occurred in the amount of incoming radiation both on recent and on geological timescales. Until now it has not been possible to reconstruct a detailed measure of UV-B radiation beyond c. 150 yr ago. Here, we studied the suitability of fossil Pinus spp. pollen to record variations in UV-B flux through time. In view of the large size of the grain and its long fossil history, we hypothesized that this grain could provide a good proxy for recording past variations in UV-B flux. Two key objectives were addressed: to determine whether there was, similar to other studied species, a clear relationship between UV-B-absorbing compounds in the sporopollenin of extant pollen and the magnitude of UV-B radiation to which it had been exposed; and to determine whether these compounds could be extracted from a small enough sample size of fossil pollen to make reconstruction of a continuous record through time a realistic prospect. Preliminary results indicate the excellent potential of this species for providing a quantitative record of UV-B through time. Using this technique, we present the first record of UV-B flux during the last 9500 yr from a site near Bergen, Norway.

Ecophysiological response of Crambe maritima to airborne and soil-borne salinity

BACKGROUND AND AIMS There is a need to evaluate the salt tolerance of plant species that can be cultivated as crops under saline conditions. Crambe maritima is a coastal plant, usually occurring on the driftline, with potential use as a vegetable crop. The aim of this experiment was to determine the growth response of Crambe maritima to various levels of airborne and soil-borne salinity and the ecophysiological mechanisms underlying these responses. METHODS In the greenhouse, plants were exposed to salt spray (400 mM NaCl) as well as to various levels of root-zone salinity (RZS) of 0, 50, 100, 200 and 300 mM NaCl during 40 d. The salt tolerance of Crambe maritima was assessed by the relative growth rate (RGR) and its components. To study possible salinity effects on the tissue and cellular level, the leaf succulence, tissue Na(+) concentrations, Na(+) : K(+) ratio, net K(+)/Na(+) selectivity, N, P, K(+), Ca(2+), Mg(2+), proline, soluble sugar concentrations, osmotic potential, total phenolics and antioxidant capacity were measured. KEY RESULTS Salt spray did not affect the RGR of Crambe maritima. However, leaf thickness and leaf succulence increased with salt spray. Root zone salinities up to 100 mM NaCl did not affect growth. However, at 200 mM NaCl RZS the RGR was reduced by 41 % compared with the control and by 56 % at 300 mM NaCl RZS. The reduced RGR with increasing RZS was largely due to the reduced specific leaf area, which was caused by increased leaf succulence as well as by increased leaf dry matter content. No changes in unit leaf rate were observed but increased RZS resulted in increased Na(+) and proline concentrations, reduced K(+), Ca(2+) and Mg(2+) concentrations, lower osmotic potential and increased antioxidant capacity. Proline concentrations of the leaves correlated strongly (r = 0.95) with RZS concentrations and not with plant growth. CONCLUSIONS Based on its growth response, Crambe maritima can be classified as a salt spray tolerant plant that is sensitive to root zone salinities exceeding 100 mM NaCl.