DK9700093 DK9700093

Environmental Science and Technology Department Annual Report 1996 Environmental Science and Technology Department Annual Report 1996

Edited by A. Jensen, G. Gissel Nielsen, V. Gundersen, O.J. Nielsen, H. 0stergard and A. Aarkrog

Risp National Laboratory, Roskilde, Denmark February 1997

VOL 281 18 Abstract

The Environmental Science and Technology Department aspires to develop new ideas and methods for industrial and agricultural production through basic and applied research thus exerting less stress and strain on the environment. The Department endeavours to develop a com­ petent scientific basis for future production tech ­ nology and management methods in industrial and agricultural production. The research approach in the Department is mainly experimen ­ tal. Selected departmental research activities during 1996 are introduced and reviewed in seven chapters: 1. Introduction, 2. Atmospheric Che ­ mistry and Air Pollution, 3. Gene Technology and Population Biology, 4. Plant Nutrition and Nu­ trient Cycling, 5. Trace Analysis and Reduction of Pollution in the Geosphere, 6. Ecology and 7. Other Activities. The Department's contribution to national and international collaborative research programmes are presented together with informa ­ tion about the use of its large experimental facilities. Information about the Department's contribution to education and training are in ­ cluded in the report along with lists of publica ­ tions, publications in press, lectures and poster presentations at international meetings. The names of the scientific and technical staff mem­ bers, visiting scientists, Postdoctoral fellows, Ph D. students and M.Sc. students are also listed.

ISBN 87-550-2284-7 ISSN 0106-2840 ISSN 0906-8090

Riso 1997

? Ris0-R-972(EN) Contents

1 Introduction 5 1.1 The Department of Environmental Science and Technology 5 1.2 Research Programmes 5 1.3 Research Projects 8 1.4 New Strategy and Organisation of the Research at Rise National Laboratory 8 2 Atmospheric Chemistry and Air Pollution 10 2.1 The Gas Phase 10 2.2 The Liquid Phase 14 2.3 Atmospheric Environmental Chemistry 17 2.4 Chemistry of Combustion Products 23 2.5 Development of New and Alternative Fuels 26 3 Gene Technology and Population Biology 33 3.1 Epidemiology and Population Biology of the Powdery Mildew Fungus, Erysiphe gram inis f.sp. hordei 33 3.2 Molecular Biology of the Powdery Mildew Fungus, Erysiphe gram inis f.sp. hordei 39 3.3 Seed Borne Diseases of Barley and Wheat 40 3.4 Disease Resistance in Barley and Wheat 40 3.5 Stress-Related Barley Peroxidases 41 3.6 Genetic Analysis of Resistance to Leptospheria maculans (stat. con. Phoma lingam) in Oilseed Rape and Characterization of Fungal Isolates 43 3.7 Gene Mapping in Barley 43 3 8 Mapping of the Brassica Genomes 45 3.9 Quality of the Barley Grain 45 3.10 Risk Assessment of Genetically Modified Plants 47 3.11 Seeds for Educational Purposes 50 4 Plant Nutrition and Nutrient Cycling 51 4.1 Genetical Background for Plant Uptake of Minerals and Organic Compounds 51 4.2 Biogeochemical Cycling of Carbon and Nitrogen in Agro-Ecosystems 52 4.3 Root-Microbe Symbioses 55 4.4 Plant Constituents 63 4.5 Climate Change Experiment 64 5 Trace Analysis and Reduction of Pollution in the Geosphere 67 5.1 Chemical Analysis 67 5.2 Trace Metals in the Geosphere 69 5.3 Organic Matter and Pollution in the Geosphere 71 5.4 Pretreatment and Conversion Processes of Agricultural and Industrial Waste 73 6 Ecology 77 6.1 Biosphere Atmosphere Exchange 77 6.2 Marine Tracers 82 6.3 Radioecological Models 86 6.4 Contamination Physics 100 6.5 Ecophysiology 106 6.6 Radioecology 107

Ris0-R-972(EN) 3 7 Other Activities 111 7.1 Molecular Dynamics and Photochemistry 111 7.2 Dosimetry and Industrial Irradiation 1 12 8 Special Facilities 113 8.1 The RIM1 Field Station 113 8.2 Dyskaergard, the Experimental Farm 113 8.3 Open Top Chamber Facility 113 8.4 Riso Environmental Risk Assessment Facility, RERAF 113 9 Publications 116 9.1 Refereed Journals 116 9.2 Books 1 19 9.3 Printed Abstracts and Proceedings 120 9.4 Research Reports 123 9.5 Other Publications 124 9.6 Presentations (Oral and Posters) 125 10 Papers accepted for Publication 129 11 Education 13 I 111 Titles of Flonour 131 112 Ph D. Theses 131 11.3 M. Sc. Theses 131 114 External Examiners 13 I 11.5 External Teaching and Lectures 132 12 Exchange of Scientists 133 13 Guest Lectures 13 5 14 Committee Membership 136 14.1 National 136 14.2 International 137 15 Seminars and Courses Organized 139 16 Scientific Results and Finances 140 16.1 Scientific Results 140 16.2 Finances 141 17 Personnel 142 17.1 Scientific staff 142 17.2 Technical Staff 143 17.3 Office Staff 144 17.4 Ph D. Students 144 17.5 M. Sc. Students 144 17.6 B Sc Students 144 17.7 Apprentices 144 18 Acronyms 145

4 Ris0-R-972(EN) 1 Introduction

1.1 The Department of Environmental 1.2 Research Programmes Science and Technology 1.2.1 Atmospheric Chemistry and Research Objectives Air Pollution The Department is engaged in research to establish the scientific basis for new methods in The effect of human activity on the global at­ industrial and agricultural production. Through mosphere has become increasingly evident during basic and applied experimental research, the the last decades. Changes in the atmospheric Department aspires to develop methods and composition and chemistry influence both local, technology for industrial and agricultural produc ­ regional and global climate. tion, exerting less stress on the environment. The goals of research, on natural and anthro ­ pogenic influences on the atmosphere, are to Approach understand the role of the different forcing The Department's expertise spans a wide range of functions. Understanding the fundamental pro ­ subjects including atmospheric chemistry, chemi ­ cesses is absolutely essential in building a cal kinetics in the liquid and gas phase, geoche ­ predictive capability of these forcings. The mistry, geochemical modelling, hydrochemistry, complete atmospheric degradation for each analytical chemistry, process chemistry, plant individual compound must be known in order to molecular biology, plant pathogenicity, plant quantify known, and as yet unknown, potential genetics, population biology, plant nutrition, atmospheric environmental problems. nutrient cycling, ecophysiology, marine and The new substitute compounds, the HFCs, terrestrial ecology, radioecology and tracemetal have been shown to be environmentally accept ­ ecology. able. New fire-fighting agents and new solvents The results of research and development are will be investigated for their environmental disseminated internationally to companies, insti ­ acceptability. A study of alternative fuels and tutions, organizations and public authorities additives will be a major part of our future through scientific publications, research reports, research. Global warming potentials, GWPs, are lectures and posters at scientific - and other used to characterize the relative contributions to professional meetings, personal communication radiative forcing. GWPs are provided from model with collaborators and through teaching courses calculations and depend largely on the atmos­ at universities. pheric lifetime and the infrared absorption The research and development activities in the spectrum of a species. Atmospheric research aims Department are planned for three years and at establishing a firm scientific basis for sensible reassessed every year. The research activities are and effective legislative measures, to reduce the mainly funded directly from the government or industrial and agricultural impact on the atmos­ from National Science Research Councils. How­ pheric environment. In order to model and predict ever, national and European research program ­ changes in the atmospheric composition, it is mes, private foundations and commercial con ­ essential to develop a better understanding of the tracts also make substantial contributions to the chemical processes in the atmosphere. Deposition total budget of the Department. Scientific results of airborne pollutants can give rise, either to and finances can be seen in Chapter 16. direct injuries to plants and animals, or to indirect damage through bio-accumulation in ecosystems

Riso-R-972(EN) 5 Research activities include basic atmospheric plants and weeds, and between crop plants and chemistry, gas kinetics, determination of the pathogens. influence of atmospheric processes, determina ­ Studies of plant population biology are neces ­ tion of primary and secondary pollutants, sary to predict the consequences of using new transport and dispersion of air pollutants and the genotypes possessing transgenes. Introgression of effect of air pollution on trees, crop plants and genes from crop plants to their wild relatives is terrestrial ecosystems. being studied to assess the risks of releasing genetically modified plants Further, the evolutio ­ 1.2.2 Gene Technology and Population nary processes imposed by interactions between Biology crop plants and wild plant species are being studied. Oilseed rape and wild Brassica species The Department aims to establish the scientific are used as model systems for these studies. basis for breeding crop plants with new and Experiments in population biology and theore ­ stronger resistance to diseases and with improved tical studies of interactions between populations nutrient efficiency Crop plants, highly resistant are important in the understanding of the ecosy­ to diseases and efficient in nutrient uptake, are a stems. prerequisite for low input plant production, and will reduce a number of environmental problems 1.2.3 Plant Nutrition and Mineral Cycling related to modern high-yield plant production. The research includes modelling of evolutionary The Department is investigating the scientific processes and the detection of gene products, basis for low input in plant production based on which are important for resistance and virulence. biologically fixed nitrogen and enhanced phos ­ The host-pathogen system, barley and barley phorus uptake mediated by VA-mychorrhiza, powdery mildew Erysiphe graminis f.sp. hordei, together with highly efficient nutrient assimila­ is used as a main model system for an obligate tion by crop plants and effective nutrient re­ fungus. Til/clia caries, which causes common cycling in the agro ecosystem burst in barley, and Leptospheria maculans, which The research provides basic scientific informa ­ causes root rot in oilseed rape, are also tion about the mechanisms and processes investigated. involved in six aspects: 1) research on symbiotic Much of the research effort in plant biology nitrogen fixation, which highlights the exchange and genetics, is devoted to the identification of of compounds between the host plant and the genetic markers. The use of genetic variability, microsymbiont across the membrane interface; 2) related to attributes of economical and environ ­ studies of the interaction between VA-mychor- mental importance in crop improvement, can be rhiza, plant roots and the rhizosphere; 3) studies greatly facilitated by the use of genetic markers in on turn-over of nitrogen from crop residues and the selection process. Understanding regulation farmyard manure; 4) biological transfer of nitro ­ and expression of genes, and the characterization gen from legumes to non-legumes; 5) plant of gene products, is important for developing uptake of organic micro pollutants and 6) the plants with improved characteristics, e g. a better effect of air pollution with ammonia on plants. quality of grain proteins or new proteins pro ­ The research emphasizes biological, physiolo ­ duced in the grains. Transformation techniques gical, biochemical and chemical processes in ­ also, are implemented to improve grain quality volved in the transfer of plant nutrients through for low input farming. Barley, wheat and oilseed the Soil-Plant-Air-Continuum. The research also rape are of interest in this context. provides basic information about the processes Identification of changes in genetic variability, involved in the turn-over of carbon, nitrogen and under different environmental conditions, can be phosphorus in agro ecosystems, helping to reduce enhanced by a better understanding of the inter ­ losses of nutrients to the atmosphere, ground action between plant species, as well as the inter ­ water or to streams and lakes. action between plants and pathogens. The A better understanding of the biological and che­ research involves interrelationships between crop mical processes in the soil-plant-atmosphere system will lead to a reduced requirement for energy­

6 Riso-R-972(EN) consuming fertilizers and to a reduced loss of and homogenization, without contamination from nutrients. The microbial biomass in the soil is of metals. Research on trace elements in plant fundamental importance to the mineralization of nu ­ materials grown on different soil types, using trients in plant residues and animal manure. Special different methods of cultivation, is planned in co­ attention is directed towards processes involved in operation with the Riso plant research pro ­ the cycling of nitrogen and turn-over of organic grammes on genetics and plant nutrition. matter. Nitrogen is essential for plants, but also a The research also contributes to the scientific risk to the environment through leaching of nitrate basis for developing the wet oxidation technique and volatilization of some other nitrogen com­ for pre-treatment and conditioning of biomass, pounds. The study of these processes will provide straw and wood chips. Wet oxidation readily the basic information needed for economical, solubilizes lignin in straw and the product is ecological and sustainable plant production. susceptible to enzymatic treatment and fermenta ­ Symbioses between plant roots and microorga­ tion. In the field of upgrading biomass, we aim at nisms are directly involved in the nutrient uptake by converting straw to ethanol by means of wet plants. Nitrogen is provided by Rhizobium bacteria oxidation followed by fermentation. fixing atmospheric nitrogen in symbiosis with legu­ The research in applied geochemistry includes mes. Our research in this area is particularly directed studies on polycyclic organic matter (especially to the exchange of signal components between azaarenes in polluted soils), rare and precious bacteria and host plant across the peribacteroid metals, and ferromanganese complexes in marine membrane. The soil-plant transport of mineral nutri ­ sediments The analytical equipment for organic ents, especially phosphorus, is mediated by analysis was extended with a GC/MS/MS and arbuscular mycorrhizal fungi in symbiosis with analytical procedures for analysis of triazines in most plant species. In our programme, we study the sediments were developed. influence of soil-P status on the mycorrhizal The use of municipal sludge as fertilizers in transport of phosphoms, and the efficiency of farming raises the problem of uptake of com­ different fungi. pounds, such as LAS and DEPH by plants In order to fertilize the different parts of fields in Development of analytical procedures for accordance with the natural variation in topography, analysis of LAS and DEPH in sludge, soil and texture and fertility, we are developing systems to plants has started. continuously measure plant vigour and the position The Danish R&D Center for Decontamination of the machinery in the field. of Soils and Sediment, an umbrella organization between five environmental research institutions, 1.2.4 Trace Analysis and Reduction of initiated a project to develop techniques for soil Pollution in the Geosphere and ground water remediation by bioventing and air sparging. The research programme aims at understanding fundamental processes and developing methods 1.2.5 Ecology for chemical analysis of agricultural products and conversion of biomass to useful products, in a Research within ecology and mineral cycling pertinent and effective way. seeks to improve the scientific understanding of Trace elements in the human food chain and the impacts of environmental pollution by radio­ in agricultural products are investigated within nuclides and heavy metals on marine and terres­ the national research programme of food techno ­ trial ecosystems, with emphasis on the effects on logy, F0TEK The High Resolution Inductively natural and semi-natural ecosystems. The Coupled Argon Plasma Mass Spectrometer is research also aims at establishing a better under ­ indispensable for the analysis of more than 60 standing and knowledge about the transport and elements in products from 200 different farmers. fate of pollutants by the use of stable- and For this purpose, a TIMS and an advanced radioisotopes, and trace-metal techniques. computer programme for data calculation from Forty years experience in the measurements of additional calibrations were developed together environmental radioactivity gives us expertise in with sophisticated tools for sample preparation low-level measurements and radiochemical

Riso-R-972(EN) 7 analysis. After the installation of a HRICPMS, environmental perspective, the study of biogenic new possibilities for low-level analysis exist. A emissions may identify those substances, which major effort is directed towards an application of in particular may contribute to crucial global pro ­ this instrument for the determination of very cesses, such as the greenhouse effect and the long-lived radionuclides, e.g. 99 Tc and 2U240pu jn depletion of the ozone layer environmental samples. The aim is to improve the sensitivity obtained by radiochemical analysis by one to two orders of magnitude. If this effort is 1.3 Research Projects successful, the size of many environmental samples can be reduced substantially, which is of Detailed information about all research projects in great advantage because global sampling creates the Department can be obtained from WWW at difficulties with shipments of larger samples. http://www.risoe.dk/pbk/pbkhome.html The radioecological research focuses on four aspects: 1) Radionuclides, discharged from La Hague and Sellafield, are used to trace the 1. 4 New Strategy and Organisation of advection and dispersion of pollutants with the water in the general circulation of the ocean the Research at Rise National currents. The brown algae P'ucus vesiculosus, Laboratory were used as bioindicators, as these plants accumulate tracers, e g. technetium. 2) Radioeco ­ A new strategy and a new organisation were logical modelling concentrated on marine com­ developed at Rise during 1996. According to the partment models and the assessment of doses to new strategy of July 1996, environmental man from the dumping of radioactive material in research is no longer an independent area, and the Arctic Seas. 3) Contamination physics is research in radioecology is included in the area developing a basic knowledge and practical related to nuclear safety as follows: methods for decontaminating areas after the 1) Environmental aspects will be an integral part Chernobyl contamination. Further, we study of all future research programmes and projects indoor aerosol deposition by using neutron-activ- at Rise. atable tracers as substitutes for airborne conta ­ 2) The research group on radioecology is minants, such as bacteria, fungi and radon transferred to the Nuclear Safety Department progeny, which are attached to ambient aerosols. 3) The Environmental Science and Technology 4) Research in radioecology, comprising studies Department is re-named as the Plant Biology of the assessment of previous releases in the and Biogeochemistry Department. Arctic region and in the Ob river system, as well 4) From 1997, the Plant Biology and Biogeo­ as the assessment of potential releases, especially chemistry Department will be organized in in the Arctic region. five research programmes and three special In 1996, Rise initiated an intercomparison of facility units. measurements of radionuclides in sediments. A 5) The research programmes on Plant Nutrition total of 21 laboratories participated in the inter ­ and Nutrient Cycling and Ecology1 will be comparison: 14 from the Nordic countries, 6 from merged into one programme on Plant the Baltic republics, and one from Japan. Ecosystems and Nutrient ('ycling . Radioactive tracer techniques are used in 6) A new research programme on Plant-Microbe ecophysiological studies on lipid metabolism and Symbiosis will be established. copper toxicity to rainbow trout and European 7) In 1998, a new research programme on Plant eel. Products and Products Quality will be added The biosphere-atmosphere exchange studies to the existing five research programmes and investigate deposition of stable elements in the three special facility units. ecosystems in addition to biogenic emissions. Deposition of airborne pollutants may influence Dr. Ame Jensen is Head of the Plant Biology the plant production, but may also have an impact and Biogeochemistry Department on especially vulnerable ecosystems. In a larger

8 Riso-R-972(EN) Research Programmes A tmospheric ('hem is try. Head: Dr. Ole John Nielsen. Trace Elements and Organic Compounds. Head: Dr. Vagn Gundersen. Resistance Biology and Plant Genetics. Head: Dr. Hanne Ostergard. Plant Microbe-Symbioses. Head: Dr. Henrietta Giese (constituted). Plant Products and Products Quality. Head: position will be opened in 1998. Plant Ecosystems and Nutrient Cycling. Head: Dr. Gunnar Gissel Nielsen.

Special Facility Units Growth chambers. Riso Environmental Risk Assessment Facility (RERAT), green houses, the Open Top Chamber Facility (OTC) and the experimental farm, Dyskcergard. Head. Dr. Vagner Haahr Riso High Dose Reference Laboratory. Head: Dr. Arne Miller. Riso Integrated Environmental Facility (R1MI). Head: Dr Kim Pilegaard

This will be the last report from the Environmental Science and Technology Depart ­ ment but we hope our readers will take interest in the annual report of the Plant Biology and Biogeochemistry Department.

Riso-R-972(EN) 9 2 Atmospheric Chemistry and Air Pollution

Introduction (O.J. Nielsen) Summary of Projects

The goals of all scientific research on natural and 2.1 The Gas Phase anthropogenic influences on the atmosphere are to understand the role of the different forcing 2.1.1 Atmospheric Chemistry of HFCs and functions. Understanding the fundamental pro ­ HCFCs (J Sehested) cesses is absolutely essential in building a predic ­ tive capability on these forcings. The complete Hydrofluorocarbon 134a (1,1,1,2-tetrafluoro- atmospheric degradation scheme must be known ethane) is an important CFG replacement used in for each individual compound in order to quantify automotive air-conditioning and domestic refrige­ known and future unknown potential atmospheric ration systems. The current global production of environmental problems. In the atmospheric FtFC-134a is 1.5 x 105 tonnes yr"’ and is anti ­ chemistry research programme, this issue is cipated to double in the next 25 years. The HFC- addressed by laboratory studies in the gas and 134a concentration in the remote atmosphere is liquid phase and by field investigations. 1.5 ppt and is increasing exponentially at a rate of The research on the atmospheric chemistry of approximately 100% yr"1. alternative CFCs is coming to an end. The new The atmospheric degradation of HFC-134a substitute compounds, the HFCs, have been (CF3CFH2) proceeds via the formation of shown to be environmentally acceptable. New CF3CFHO radicals. Long pathlength FTIR fire-fighting agents and new solvents will still environmental chamber techniques were used to have to be investigated for their environmental study the atmospheric fate of CF3CFHO radicals acceptability. Alternative fuels and additives will The atmospheric oxidation of FTFC-134a is be a major part of our research in the future. initiated by reaction with the OH radical. The Global warming potentials, GWPs, are used to alkyl radical formed in reaction (I) adds characterize the relative contributions to radiative molecular oxygen to give the peroxy radical forcing. GWPs are provided from model calcula­ cf,cfho2 tions and depend largely on the atmospheric lifetime and the infrared absorption spectrum of a (1) cf,cfh2 + oh^cf,cfh + h2o species Many compounds not only contribute (2) cf,cfh + o2 + m -^cf,cfho2 + m directly, but also indirectly in many ways. Methane, a simple greenhouse gas, creates ozone Reaction with NO is an important fate of and water which again are greenhouse gases and CF,CFH02 radicals in the atmosphere influence the lifetime of yet other compounds. Bi- ogenically emitted compounds, such as isoprene (3) CF,CFH02 + NO -» CF3CFHO + N02 and dimethylsulfide, generate atmospheric parti ­ culates during their atmospheric degradation. A Reaction (3) produces N02 and the alkoxy decision on emission reductions will depend on radical CF,CFHO. Two competing reaction the relative contributions and we scientists will pathways were identified for CF,CFHO radicals, have to refine the methods of calculating GWPs. reaction with 02 and decomposition via C-C bond We have modified and used a simple ID model cleavage. and calculated GWPs with great success. This work will continue in the future. (4) CF3CFHO + 02 -* CF3C(0)F + H02 (5) CFjCFHO + M -* CF, + HC(0)F + M

10 RiS0-R-972(EN) Decomposition gives a CF3 radical and self reaction of CF,CFH02 radicals, or via the HC(0)F which are expected to be of no CF3CFH02 + NO reaction. Figure 2.1.1 shows environmental concern. Reaction with 02 pro ­ the yield of CF3C(0)F as a function of 02 partial duces CF3C(0)F which is incorporated into rain- pressure in experiments with and without NO cloud-sea water and hydrolyzed to CF3COOH. added. As seen in Figure 2.1.1, the behaviour of While the concentration of CF3COOH in rain ­ CF3CFHO radicals was dependent on the method water is not expected to be toxic to living by which they were generated Decomposition organisms, concerns have been raised that the was much more important when CF3CFHO mildly phytotoxic CF3COOH could accumulate radicals were produced via the CF3CFH02 + NO to problematic concentrations. reaction than when they were produced via the In the laboratory, CF3CFHO radicals were self reaction of CF3CFH02 radicals. produced by two different reactions: either via the

1.0 Without NO A A tu 0.8 o

u A/ p-T u 0.6 o / 2 /A 0.4 > c5 With NO o s 0.2

0.0 0 200 400 600 800 [02], Torr

T'igure 2.1.1 Yield of ('J-’f '(())}•' observed during the photolysis of HMY]34a/Cl/()fN 2 mixtures at 269K and 800 Torr total pressure as a function of the ()2 partial pressure with (fdled circles) and without (filled triangles) NO present.

The data can be analysed in terms of a “hot is of no atmospheric importance. The fact that alkoxy radical effect”. In this analysis we assume CF3CFHO radicals formed in reaction (3) have an that reaction (3) produces vibrationally excited increased tendency to decompose, limits the CF3CFHO radicals which either decompose formation of CF3C(0)F and hence CF,COOH in rapidly or are thermalized via collisions with the the atmospheric degradation of HFC-134a. We surrounding molecules. The thermalized radicals estimate that the CF3COOH yield from then react with 02, reaction (4), or decompose, atmospheric oxidation of HFC-134a is 7-19% as reaction (5). opposed to the previous estimate of In the atmosphere, reaction (3) controls the approximately 30%. A similar effect has been fate of CF3CFH02 radicals while the self reaction

Ris0-R-972(EN) 11 observed in the degradation of HFC-236cb undergoes quick ignition and is, therefore, an (CF3CF2CFH2) excellent diesel fuel. The reactions of dioxane and trioxane are shown in Table 2.1.2. No effect 2.1.2 EUROVOC (J. Sehested ) on k(R+02) of the total pressure was observed. The atmospheric degradation of dioxane is Rise is a part of a research programme sponsored ethylene glycol di formate. There are two by the EU which aims at making control important atmospheric products from trioxane, strategies for the release of volatile organic methylene diformate and a lactone. Finally, we compounds to the atmosphere to reduce the ozone studied the atmospheric degradation of formation over Europe. The contribution from dimethoxy methane and dimethyl carbonate. We Rise is to measure rate constants of atmospheric studied the first step in the atmospheric importance in the degradation of VOCs (volatile degradation of dimethoxy methane, the reaction organic compounds) as input for the models to with hydroxyl radicals, and obtained 4.9x1 O'12 determine the atmospheric degradation schemes. cm3 molecule" 1 s’1. In addition, the products of In 1996, we studied the degradation of dimethoxy dimethoxy methane in the atmosphere were methane, dimethyl carbonate, dioxane, trioxane, shown to be 25% dimethyl carbonate, 69% dimethyl ether, and CF3CFH2 (HFC-134a, as methoxy methyl formate and 6% unknown described above). products. For dimethyl carbonate, we obtained a Dimethyl ether: The reaction of CH3OCH2 + rate constant for the reaction with hydroxyl 02 was studied as a function of temperature. This radicals of 3. lxlO" 13 cm3 molecule" 1 s'. reaction gives the peroxy radical, CH30CH202, at Additional kinetic results are shown in Table atmospheric pressure and two formaldehyde 2.1.2. Finally, the products of the atmospheric molecules and a hydroxyl radical at low pressure, oxidation were shown to be 56% approximately 30% at 12 mbar. It is the first time CH3OC(0)OCHO and 44% decomposition, that this type of reaction mechanism has been probably to C02, and two formaldehydes observed at room temperature. This reaction is These results are used in the EUROVOC not very important in the atmosphere, where most programme to determine the chemical of the dimethyl ether is oxidized at pressures mechanisms for the degradation of VOCs. The between 200 and 1000 mbar total pressure. obtained chemical mechanism is going to be However, the reaction may be active in diesel checked in a 200m3 smog chamber in Valencia, engines and may well explain why dimethyl ether Spain, in 1997.

Table 2 1.2 Rate constants for reactions for three VOCs studied at Rise

F+RH r+o2 R+R ro2+ro2 ro2+no ro2+no2 1,4-Dioxane 24 0.87 3.3 0.73 0.60 1.3 1,3,5-Trioxane 11 0.77 8.5 0.91 0.56 Dimethyl carbonate 6.4 — 6.5 1.27 1.2 1.2

Units: 10'" cm' molecule" s" Uncertainties in the range of 10-20%

References Peelers J., Pultau V. (1992) Rate constants of Alternative Fluorocarbon Environmental Acceptability CF,CFH02+NO.Proceedings of CEC/EUROTRAC Study. W.M.O. Global Ozone Research and Moni ­ Workshop on Chemical Mechanisms Describing toring Project, Report #20, Scientific assessment of Tropospheric Processes. J. Peelers (cd ). September stratospheric ozone, Vol. 2. 1989. 1992. DeMore II.R.. Sander S.P., Golden DM.. Hampson Tromp T.K., Ko M.K.W.. Rodriguez J.M., Sze A'. I). R.I., Kurylo M.J., Howard ('.J., Ravishankara A.R., (1995) Potential accumulation of a CFC- Kolb C.K, Molina M.J (1994) Chemical kinetics rcplacemcnt degradation product in seasonal and photochemistry data for use in stratospheric wetlands. Nature 376: 327. modelling, evaluation no. 11. JPL Publication 94: 26.

12 Ris0-R-972(EN) Wallington T.J., Nielsen O.J. (1991) Pulse radiolysis of benzene, employing pulse radiolysis combined CF3CFHO2 radicals in the gas phase at 298 K. with time-resolved UV-spectroscopy The tro­ Chem. Phys. Lett. 187: 73. pospheric oxidation of benzene is initiated by the Wallington T.J., Schneider W.F., Worsnop D.R., Nielsen O.J., Sehested ./, DeBruyn W.J., Shorter reaction with hydroxyl radicals, and we have J.A. (1994) The environmental impact of CFC identified two competing reactions where the replacements - MFCs and HCFCs. Environ. Sci. products were identified by the ultraviolet spectra Tcchnol. 28(7): 320-325. and kinetics of the shortlived adduct, HO-C6H6 and the stable product, C6H5OH 2.1.3 Oxidation of Benzene in the Troposphere (P. Pagsberg, E. Bjergbakke, (la) OH + C6H6 -» HO-C6H6 (75%) A. El. Sillesen) (lb) OH + C6H6 ->H + C6H6OH (25%)

The chemical reaction mechanisms involved in UV-spectra of HO-C6H6 and C6H5OH are the tropospheric oxidation of benzene are very shown in Figure 2.1.3. Based on the UV- complex, and detailed experimental investigations spectrum of HO-C6H6 we have been able to study of the various intermediate species are required in the kinetics of subsequent reactions which are of order to identify the most important pathways and importance in the troposphere. products. We have been engaged in a CEC- project, CYTROXA, in collaboration with four (2) HO-C6H6 + 02 ^ H0-C6H6-02 other European research laboratories. In our (3) HO-Cf)Hf, + N02 -> products laboratory, we have studied the oxidation of

H-C„H H0-C„H

C H OH 6 5

Wavelength / nm Figure 2.1.3 UV spectra ofHO-CJA, H-( \J16 and (\.H6 ()H.

13 References used for reduction of nitrogen oxides from power Bjerghakke K., Sillesen A., I’agsberg I’. (1996) UV plants. The complex reaction mechanisms have spectrum and kinetics of hydroxy cyclohexadienyl been studied for many years and detailed radicals. J. Phys. Cheni. 100: 5729-57.16. computer modelling has been employed as a tool Zetzsch Devohler I’., I’agsberg I’., Barnes /., Glavas for optimizing the performance of this DeNOx S. (1996) Cycling of OH and H02 by NOx in the tropospheric oxidation of aromatics. In: CEC R&D technology. In order to establish a realistic Programme in the field of environment 1994-1996. computer model, one has to include about 150 Final Report on CYTROXA, C. Zetzsch (cd.) elementary reactions involving free radicals Contract no. EV5V-CT91-0109 European Commis ­ acting as chain carriers, e g. OH and NH2. sion. Recently, the reaction NH2 + N02 -+ pro ­ ducts, has been the subject of experimental 2.1.4 UV-Absorption Cross Section of Nitrous studies to establish the reaction mechanism and Acid (P. Pagsberg, K Ratajczak, various product channels. However, published A’. Bjerghakke, AH. Sillesen) experimental results on reaction products and estimated branching ratios suggested that this We are now engaged in a new EU-project, reaction deserved further study. In collaboration FORMONA, in collaboration with eight other with a visiting scientist, H. Meunier, Assumption European laboratories. The main tasks are con ­ College, Worcester, MA, USA, we have in ­ centrated on laboratory studies and field measure­ vestigated the reaction employing pulse radiolysis ments of the Formation and Occurance of Nitrous combined with time-resolved infrared diode laser Acid in the Atmosphere. Detection of nitrous acid spectroscopy. Using this method, we have been in ambient air can be carried out using able to study the kinetics of NH2 radicals in the Differential Optical Absorption Spectroscopy, presence of N02 Formation of the main DOAS, based on the characteristic UV-spectrum products, N20 and NO was observed on the same of MONO in the range of 300-400nm. However, time scale in accordance with the following in the case of HONO, this technique suffers from reactions: large uncertainties because of published values of the UV-absorption cross section of HONO which (la) NH2 + N02 -> N20 + H20 (59+3)% differ by as much as a factor of four. The main (lb) NH2 + N02 -» H2NO + NO (40+5)% reason for these large uncertainties is the low stability of nitrous acid due to the reaction Considering the overall rate constant, our HONO + HONO -► H20 + NO + N02 where the value of (1.35+0.15)xl0" n molecule" 1 crnV 1 is in product N02 absorbs in the same spectral region. fair agreement with the results of previous In our laboratory, we have employed pulse investigations. However, our values of the radiolysis of SFr/H20/N0 mixtures to initiate the branching ratios differ significantly from fast and clean source reaction OH + NO + M —> experimental results recently obtained in other HONO + M where we have observed the laboratories. formation of nitrous acid on a time scale of 10- 100 psec without interference from N02 which is Reference formed on a much longer time scale. Thus, we Meunier II., I’agsberg I’., Sillesen A. (1996) Kinetics believe that this method will allow us to and branching ratios of the reactions NH; + NO: -> determine a reliable UV-absorption spectrum and N20 + Hs and NH2 + N02 -> H2NQ + NO studied by absorption cross section required for field pulse radiolysis combined with time-resolved infra ­ measurements of nitrous acid by DOAS. red diode laser spectroscopy. Chem. Phys. Lett. 261: 277-282. 2.1.5 Mechanism and Ranching Ratio of the Reaction NH2 + NO -> Products 2.2 The Liquid Phase (K. Sehcsted, (P. Pagsberg, H. Meunier, A H. Sillesen) ,/. Holcman, F. Jacobsen)

Thermal DeNOx or Selective Non-Catalytic Re­ The influence of Transition Metal Ions, especially duction of nitrogen oxides by ammonia is widely iron and manganese, on the oxidation processes

14 Ris0-R-972(EN) in the atmospheric liquid phase was studied by which proceeds until one of the oxidants is stopped-flow and pulse radiolysis experiments. practically exhausted. The mechanism of the The temperature dependence of the oxidation catalysis consists of oxidation of Mn 2 to Mn 3 by kinetics of Fe2+ by 03, at pH 0-3, was measured ozone with k(Mn 2 + 03) = 1.5 x 103 M 1 s’1. No in the temperature range 5-40°C. Activation para ­ OH radicals are formed during reaction, and Mn 3 meters of the reactions involved in formation and is subsequently reduced to Mn 2 by H202 k(Mn 3 decay of the ferryl ion (iron(IV)), FeO2 were + H202) = (0.03-1.1) x 105 M’1 s’1. Mn 2' determined. The reaction of Fe2' + FeO2 was therefore contributes to reduction of the oxidation found to branch into two channels forming capacity in the aqueous phase of the atmosphere iron(HI)-dimer, Fe(OH)2Fe4+ and Fe3'. The yield by catalytic action. (Jacobsen et al. c, in press). of the dimer, Fe(OH)2Fe4 , increases with tem­ Manganese forms a superoxide complex, perature at the expense of the Fe3 yield. On the Mn0 2 , which was studied in pulse radiolysis by basis of the overall rate constant and relative three distinct routes Mn(I) + 02, Mn(U) ± 02" and yield of Fe(OH)2Fe4', the activation energy was Mn(UI) ± H202. The stability of this complex was determined for both channels. The activation found to be governed by the two equilibria: parameters of the hydrolysis of the ferryl ion and Mn 2’ + 02" <-» Mn0 2 (1,-1) and Mn 2 + H02 its reaction with H202 were also determined. <-> Mn0 2 + H (6, -6). Both forward and reverse (Jacobsen et al. a, in press). rate constants of the reactions involved in these The rate constants for reactions between the equilibria were determined: k, = (1.5 ± 0.2) x 108 M" ferryl ions and some selected compounds of 's’1, k, =(6.5 ±1.0) x 10V; kfi = (l.l ± 0.2) x 10h cloud water were obtained in acid solutions. For NT's"1, k^ = (6.5 ± 1.0) x 106 M’V1 yielding K, ., = inorganic reactants: kHN02 = 1.1 x 104 NT’s krl_ = (2.3 ±0.5) x 104M"' and 1^ = 0.17 ±0.5. 1.0 x 102 NT’s"1, kHS03„= 4.5 x 105 NT's"', kMn(I]| = 1.0 The metal-oxy complex Mn0 2 decays by self­ x 104 M’V were obtained and for the organic reaction with k(Mn0 2 ± Mn0 2 ) = (6.0 ± 1.0) x 106 reactants kHC(X)11 = 160 NT's-1, kaL3COOH = 3.1 NT's"1, IVT's’1 and in acid solutions also, by reaction with kcH2o = 400 NT s , knurorH3 = 16 NT s , knI3rl!20l i H02, k(Mn0 2 ± H02) = (1.0 ± 0.3) x 107mV. In = 2.5 x 103 NT’s"1, kr6H5oii = 4.0 x 103 M's’1 and both cases, stochiometric amounts of H202 are kc6H5coon = 80 Mf's'1 were measured. The reaction formed as the end product. mechanisms, in the case of inorganic compounds, Mn(I) was formed by reduction of Mn" with H- proceed by electron transfer and a good atoms. It has an absorption spectrum with maxima correlation between log(k) and the standard at 290 and 340 nm with e290 = 1300 ± 200 M^cm’1 reduction potential, E°, was obtained. The and e34i) = 1000 ± 150 M'1cm"1. It reacts with reaction mechanisms, in the case of organic oxygen with k(Mn(I) ± 02) = (6.0 ± 1.0) x 106 M"'s" compounds, are very similar to that of the OH '. Mn(UI) reacts with hydrogen peroxide with radical eg. H-abstraction, and a fairly good k(Mn(IU) + H202) = (2.8 ± 0.3) x 103 M’V1 in ­ correlation between log(k) and the bond creasing with pH due to another hydrolytic form of dissociation energy, BDE, was obtained. Mn(IU). (Jacobsen cl al. d, accepted, Jacobsen et al Activation parameters were measured for the 1996). reaction of FeO2' with HN02 (Ea= 34.5 kJ/mol); As part of the EU contract RINOXA 2, DMSO, Mn 2' (Ea= 21.3 kJ/mol); HCOOH (Ea= 22.3 DMS02, methansulfonic acid, methansulfinic acid kJ/mol); CH20 (E,= 44.5 kJ/mol) and C6H5OH and hydroxy methansulfinic acid, which are (Ea= 28.1 kJ/mol). At higher pH, the lifetime of oxidation products from biogenic dimethylsulfide, FeO1" is substantially reduced, which is ratio­ were studied with respect to their oxidation by OH nalized in terms of an acid-base equilibrium radicals and reactions with oxygen. DMSO is between the two hydrolytic forms of these species oxidized to methansulfinic acid and methyl radicals, with pKa = 2.0. (Jacobsen et al. b, submitted, which in turn react with oxygen forming Holcman 1996). methylperoxy radicals. The rate constants of OH An otherwise slow and strongly pH-dependent with DMS02 and methansulfonic acid are 1.5 x 107 reaction (03 + H202 —>202 + H20) is, in the M"1 s"1, whereas the rate constants with the other presence of Mn"', accelerated by a catalytic chain compounds are (6-7) x 109 M~' s’1. All the secondary destruction of ozone and hydrogen peroxide, radicals react with oxygen with a rate of (1 -2) x 10J

15 M~l s'1. The methansulfmic acid was studied the same chemical system to determine the pK previously (Sehested and Holcman 1996) and a when the equilibrium between OH and O’ or OD chain oxidation was explored. It was, therefore, of and O’ is established. The two methods give interest to study hydroxomethansulfinic acid similar results with pK(OH) = 12.0 ± 0.2, in good (HOMSEA) more closely. Upon oxidation by OH agreement with the literature, and pK(OD) = 12.6 radicals, HOMSEA splits into formaldehyde and the ± 0.2. The OD radical is a weaker acid in D20 S02" radical. A slight difference in absorption than the OH radical in H20. The isotope effect on spectra in neutral and acid solution is preliminary the equilibrium constant is mainly due to reverse ascribed to a protonation of S02 to HS02. The reaction 2, which is 3 times slower in D20 than in bimolecular product is dithionite which is unstable H20. (Hickel el cil. 1996). in acid solution. The reaction of S027HS02 with Kinetics of the electron transfer from OH- oxygen is fast k = 2.0 x 10J M"1 s’1 and 027H02 is adducts of various derivatives of DNA-pyri- formed. No chain oxidation as found in methan­ midine bases, uracil and uridine, to mytomycine- sulfmic acid was observed. C (MMC) has been initiated. Both mytomycin As there was no difference in absorption spectra and the base derivatives are used in experimental or extinction in Ar-saturated acidic solutions therapy, and the main purpose of this study is to containing only HOMSEA or HOMSEA and S02 verify the hypothesis of the correlation between (HSOf), it was concluded that the H atom reacts electron transfer kinetics and therapeutic activity. with HOMSEA by formation of methanol and S02". Rate constant and yields were determined for (Sehested 1996, Sehested and Holcman 1996). several hydroxyl-, methyl-, sulphur- and fluorine- The kinetics of the reaction between hydrated substituted bases. electron and perbromate, Br04" + eaq" —> Br03’ + 03 decomposition in acid solutions, pH 0-4, is O', was investigated by combining kinetic propagated by radicals H02 and OH. Formation measurements of the formation of ferricyanide in of H202 is detected during the course of the the reaction between perbromate and ferrocianide decomposition. Addition of H202 to vessels of with measurements of the yield of ferri cyanide various sizes has revealed that both initiation and formed by pulse radiolysis of aqueous N20 and termination reactions are surface catalyzed. The Br04" solutions containing ferro- and ferri- precise nature of the surface reaction, converting cyanide. ozone molecules into OH radicals and/or H202 is The data obtained indicate that whereas the not fully understood. The formation of H202 electron transfer from ferrocyanide to perbromate plays a crucial role in the initiation of the chain takes place adiabatically, yielding Br04‘ in the process by the formation of chain propagating most stable configuration, the reaction of perbro ­ radicals OH and H02. From both y-irradiation mate with the hydrated electron yields excited and the thermal decay of O,, it is concluded that states of Br042, in which a Br-0 bond is broken. the surface reactions account for most of the Owing to the stability of Br04" a subsequent termination which, in the homogeneous phase, is cage-back reaction between O' and Br03" cannot too ineffective to play any significant role due to compete with other relaxation processes by which very low radical concentrations. (Sehested and O’ and BrO, diffuse away from each other. Holcman 1996). (Klaning and Sehested, submitted). The rate constant of the reactions References OH + OH" ^ O + H20 (k,, k2) and Hickel B., Corfitzen If., Sehested K. (1996) OD + OD o 0 + D20 (k3,k4) Measurements of the rale constant of the reactions was measured by pulse radiolysis in the forward 0H+0H»0+H20 and 0D+0D>>0 +D20 in forward and reverse directions. Kinetic determina ­ and reverse directions. At 22°C the bimolecular tions of the pK's of OH and OD radicals. J Phys rate constants (in M"1 s’1) are k, = 6.3 ± 1.3) x 109 Chcm. 100: 17186-90. and k2 = (1.0 ± 0.2) x 106 in H20 and k3 = (5 ± 1) Holcman J. (1996) Fcrrvl ion and manganese(lll) x 109 and k4 = (3.5 ± 0.7) x 105 in D20. The pK s reactions with simple compounds. RINOXA of the OH and OD radicals were calculated in meeting in Essen, Germany 22-23 February 1996. non-equilibrium conditions by the ratio of the k, Jacobsen F., Holcman J., Sehested K. (1996) Oxidation and k2. Experiments were also performed using of transition metal ions by ozone and the role of

16 Ris0-R-972(EN) resulting intermediates. Iron and manganese. Final precursor, acetaldehyde, was poor. The emissions report of the EC R and D project: RINOXA, inventories for traffic VOC, e g. toluene, appear to Removal and Interconversion of Oxidants in the work satisfactorily, while the emissions inventories Atmospheric Aqueous Phase, p. 113-147. Contract EF5V-CT93-0317, European Commission. for other VOC sources, especially biogenic alkene Jacobsen F, llolcman J., Sehested K. (a) Activation sources, are most uncertain. Thus, the measured parameters of ferry! ion reactions in aqueous acid values of ethylene and CrC5 alkenes were 6-23 solutions. Int. J. Chem. Kind. In press. times higher than the standard modelled values. Jacobsen F, Flolcman J., Sehested K. (b) Reactions of Both the emissions of NOx and biogenic alkenes the ferryl ion with some cloud water constituents. appear to affect the ozone formation (Nielsen et al. Env ironmental Science and Technology . Submitted. 1996b). Jacobsen /•.. Holcman J., Sehested K. (c) Effects of manganese on the oxidation capacity of the troposphere. Proceedings of the 7th European References Symposium on Physico-Chemical Behaviour of Nielsen T., Samuelsson U., Grennfelt P , Thomsen E.I. Atmospheric Pollutants. The Oxidizing Capacity of (1981) Peroxyacetvl nitrate in long-range trans­ the Troposphere. B. Larsen (ed ). Zitelle. Venice, ported polluted air. Nature 293: 553-555. Italy 2-4 October 1996. In press. Nielsen T.. Hansen A.M, Thomsen F.L. (1982) A Jacobsen F., Holcman J., Sehested K. (d) Man- convenient method for preparation of pure standards ganese(II)-superoxide complex in aqueous solution. of peroxyacelyl nitrate for atmospheric analyses. J. Phvs. Chem. Accepted. Atmos. Environ. 16: 2447-2450. Kldning fSehested K. On the mechanism of the Nielsen T., Granby K., Egelov A. II, Hummelshoj /’., reaction of the hydrated electron with perbromate. Jensen F.P., Skov H. (1994) Lille Valby, Denmark. Submitted. In: T. Cvitas. D. Kiev (eds ). The TOR Network. A Sehested K. (1996) Oxidation of DMSO, DMS02, Description of TOR Measurements Stations, pp. 36- methansulfonic acid, methansulfinic acid and 45. EUROTRAC. International Scientific Secretary hydroxy methansulfinic acid by the OH radical. ISS. Garmisch-Partenkirchen, Germany. RINOXA meeting in Zitelle, Venice. Italy 1-4 Nielsen T., Egelov All, Granby A', Skov //. (1995) October 1996. Observations on particulate organic nitrates and Sehested K., Holcman J. (1996) A pulse radiolysis study unidentified components of NOv . Atmos. Environ. of the OH radical induced autoxidation of methan- 29: 1757-1769. sulfmic acid. Radiat. Phvs. Chem. 47. 357-60. Nielsen T., Pilegaard K., Egelov A. II., Granby K . Hummelshoj P., Jensen NO., Skov H. (1996a) Atmospheric nitrogen compounds: Occurrence, composition and deposition. Sci. Total Environ. 2.3 Atmospheric Environmental 189/190: 459-465. Chemistry Nielsen T., Hertel 0., Christensen C.S., Egelov A H.. Granby K., Hansen A.B., Platz J., Skov II (1996b) Evaluation of the Danish ACDEP model to simulate 2.3.1 Comparison of Field Measurements formation of tropospheric ozone, other photo ­ and a Trajectory Model (T. Nielsen) chemical oxidants and atmospheric reaction pro ­ ducts. 7th European Symposium on Physico- Chemical Behaviour of Atmospheric Pollutants. The The Danish ACDEP, Atmospheric Chemistry and Oxidizing Capacity of the Troposphere. European Deposition model reasonably describes the levels Commission, Brussels, Belgium, p. 0-47. and diurnal variations of measured concentrations of the photochemical oxidants, ozone and PAN, when the emissions of C2-C5-alkenes were increased by a 2.3.2 Atmospheric Nitrogen Compounds factor of 10. The measurements were performed in (T. Nielsen) June and July, 1995 at Lille Valby (Nielsen et al. 1994), an agricultural site situated 35 km west of The tropospheric ozone in the northern hemisphere Copenhagen. The alkene model also gave a fair has increased in this century by a factor of three description of the concentration levels of NOy and (Volz-Thomas and Kley 1989). The annual increase the different NOv compounds (Nielsen et al. 1995 in the last decade has been about 1-2% (Scheel et al. and 1996a), although it cannot be excluded that a in press). The atmospheric formation of ozone in correct model PAN (Nielsen et al. 1981 and 1982) open land is controlled by the presence of NOx at may give a wrong answer, as the modelling of the least during summertime (Nielsen et al. 1996). Bifunctional organic nitrates have been found both

17 in gas phase and as unidentified bi- and/or multi­ tants between water and suspended solid matter is functional organic nitrates. These are associated determined by the solid matter’s content of organic with particles in smog-chamber experiments and are carbon. Besides, the distribution of polar organic formed by reactions of alkenes with OH radicals compounds is also affected by polar interactions during the day and with N03 radicals at night-time between functional groups in the substrate and the (Nielsen et al. 1995). These compounds are con­ compound itself (Nielsen et al. a, accepted). From sidered to be a possible source of N02 in remote the data of Yamasaki et al. (1982), the following areas and thus may affect the formation of ozone relation between vapour pressure and particle from the photooxidation of CO and CH4 (Ayers el proportion values for non-polar compounds can be al. 1992). The atmospheric levels of the particulate estimated at 20°C and a particle concentration of 30 organic nitrates, PON, i.e. organic nitrates asso­ pg m"3: (vapour pressure mm Hg, particle ciated with particles, have earlier been determined to proportion %), (1*1 O'5, 1), (8*1 O'7, 10), (8* I O'8 , 50), be 14±5 ng(N) m"3 (0.25% of total NOy) in an open (7*1 O'9, 90) and (6*10" ll), 99). Although the land area in Denmark (Nielsen et al. 1995). extrapolation of these data to organic nitrates should Table 2.3.2 shows the estimated vapour be done cautiously, it is estimated that the major pressures of different organic nitrates. The gas to VOC source for PON formation appears to be particle distribution is dependent on the vapour biogenic emissions, terpenes and sesquiterpenes. It pressure, temperature and particle concentration, but can also be assumed that the PON compounds the nature of the particles could also affect the probably are multifunctional organic nitrates distribution. Thus, the distribution of organic pollu­ (Nielsen et al b, submitted).

Table 2.3.2 Estimated vapour pressure (P, mm Hg) of different organic nitrates at 20°C .

Number of Dihydroxy Hydroxy Carbonyl Alkyl carbon nitrates nitrates nitrates nitrates atoms 5 0.001 0.05 0.2 3 10 4*1 0'5 0.002 0.006 0.08 15 2*1 O'6 7*1 0'5 2*1 0‘4 0.003 20 6* 1O'8 3*10' (’ 8*10-6 1 * 10 4 25 2*1 O'9 9*1 O'8 3*1 O'7 4*1 O'6

: P is calculated from the expression log P= -(0.2884±0.0072)*no. of C-atoms -(1,630±0.110)*no. of nitrate groups -(1.662±0.065)*no. of hydroxy groups -(1.115±0.079)*no. of carbonyl groups +(3.458±0.094), n=165, r=0.96, p<0.001 . The applied data for the correlation includes C7-C2<) hydrocarbons, C,-C18 alcohols, C2-C10 diols, C5-C18 carbonyls, Q-C5 alkyl nitrates and C2-C3 hydroxynitrates and dinitrates (CRC 1982, Karnes and Schurath 1992, Hallquist et al. 1996,Saxena el al. 1996). Most of the data were achieved by extrapolating to 20°C. Some of the data (Saxena et al. 1996) were at 25°C.

References viour of Atmospheric Pollutants. The Oxidizing Avers (IP.. Penkelt S.A.. Gillett RAP., Bandy B. Capacity of the Troposphere. European Commis ­ Galbally IP., Meyer C.P., Klsworth CM., Bentley sion. Brussels, Belgium, p. 0-21. SI'.. Forgan BAf. (1992) Evidence for photo ­ Karnes ,/., Schurath (J. (1992) Alkyl nitrates and bi­ chemical control of ozone concentrations in un­ functional nitrates of atmospheric interest: Henry 's polluted marine air. Nature 360: 446-449. law constants and their temperature dependencies. J. CRC (1982) Handbook of Chemistry and Physics. 62nd Atmos. Chem. 15: 79-95. Edition. CRC Press, Inc. Boca Raton. Florida. Nielsen 71, Fgelov A.If., Granby K, Skov //. (1995) Hallquist M, ff'angherg /., Ljungstrom F. (1996) Observations on particulate organic nitrates and Atmospheric chemistry of dicarbonyl products from unidentified components of NOy. Atmos Environ. the atmospheric oxidation of selected terpenes. 7th 29: 1757-1769. European Symposium on Physico-Chemical Beha ­

18 Ris0-R-972(EN) Nielsen T., Pilegaard A., Egelov A.H., Granhv K., (2) 5NQJ-T + OH-* [5NQH-OH]* Hummelshaj P., Jensen N.O., Skov //. (1996) (3) 5NQ-OH* + 02 —> products Atmospheric nitrogen compounds: Occurrence, composition and deposition. Sci. Total Environ. (4) [5NQH-OH]'+ + 02-> products 189/190: 459-465. Nielsen 71, Platz J., Granby A., Hansen A.B., Skov //., The rate constants obtained were: Egelov, A.H. (b) Particulate organic nitrates: k, = (5.4±0.5)xl0 9 dm3cm"'s'1, Sampling and night/day variation. J. Atmos. Chem. Submitted. k2 = (1.4+0.1 )x 109 dm3cm"'s"', Nielsen 71, Siigur A'., Ilelweg ('., Jorgensen ()., Hansen k3 = (1.1 ±0.1 )x 106 dm3cm"ls"1 and P.E., Kirso U. (a) Sorption of polycyclic aromatic k4 = (8.7±0.6)x 105 dm3cm"’s'1 compounds to humic acid as studied by high (Feilberg et al. in press). The results show that performance liquid chromatography. Environ. Sci. the lifetime of 5NQ in water droplets, with Tcchnol. Accepted. Saxena P., Hildemann L.M. (1996) Water-soluble respect to the reaction with OH, is between 15 organics in atmospheric particles: A critical review minutes and 1 hour. Further investigations of the literature and application of thermodynamics include similar experiments with 6- and 8-nitro- to identify candidate compounds. J. Atmos. Chem. quinoline and 4-nitroquinoline-N-oxide. 24: 57-109. Schee! HE., Ancellel G., Areskoug H. Beck J., Muer References D. de, Egelov A.H, Esser P., Etienne A., Ferenczi Feilberg A., Holcman J., Lohse (’., Nielsen 71, Sehested Z, Geis II., Grab be G., Granby A., Gomiscek B., K. Reactions of N-heteroaromatics with OH and 0: Haspra Kesele N., Klasinc L., Laurila 71, in the aqueous phase. In: Proceedings of Lindskog A., Mower J., Nielsen 71, Roemer A/., EUROTRAC Symposium '96. Borrell P. M.. Borrcll Schmitt R., Simmonds P., Sladkovic 77., Solberg S., P., Kelly K., Seiler W. (eds.) Computational I ’arolsos C., Waal L. de. TOR task group 1 report, Mechanics Publications. In press. EUROTRAC, International Scientific Secretariat, Nielsen 71, Ramdahl 71, Bjorseth A. (1983) The fate of Garmisch-Partcnkirchen. Germany. In press. airborne polycyclic organic matter. Environ. Hllh. Volz-Thomas A., Kiev D. (1989) Evaluation of the Perspec. 47. 103-114. Montsouris series of ozone measurements made in Sera N., Eukuhara A'., Tokiwa H. (1994) Detection of the nineteenth century . Nature 332: 240-242. nitro-azabenzo[a]pyrene derivatives in the semi- Yamasaki H., Kuwata A., Miyamoto H. (1982) Effects volatile phase originating from airborne particulate of temperature on aspects of airborne polycyclic matter, diesel and gasoline vehicles. Mutagenesis 9: aromatic hydrocarbons. Environ. Sci. Technol. 16: 47. 189-194.

2.3.3 Water Photooxidation Reactions (A. Feilberg. J. Holcman , 71 Nielsen, K. 2.3.4 Soot-Liquid Water Partitioning Sehested) (T. Nielsen, K. Siigur)

Atmospheric pollution of nitro derivatives of The need to predict the transport of toxic che ­ polycyclic aromatic compounds has evoked con ­ micals in air, water and soil/sediment systems has cern, as some of the compounds are strong generated considerable interest in the develop ­ genotoxics (Nielsen el at. 1983/ The presence of ment of environmental models as convenient nitroazaarenes and a nitroazaarene-N-oxide has tools. Soot is known as a chemically complex recently been demonstrated in diesel exhaust mixture of amorphous polymerized organic gases and ambient air (Sera et al. 1994). A survey material and graphitic elemental carbon (Katrinak of the atmospheric degradation of nitro-azaarenes et al. 1992). Some of the PAC bound to the soot has, therefore, been initiated. The first results we may be released after their deposition in lakes, have obtained concern reactions of 5-nitro- rivers or on the ground and become dissolved, quinoline, 5NQ, in atmospheric water droplets while others remain in the particle phase. using pulse radiolysis of alkaline and acidic Information about partitioning between particle aqueous solutions. Kinetics of the following phase and water can be obtained from the reactions were investigated. partitioning coefficients between organic carbon and water, Koc . Considering the huge number of (1) 5NQ + OH -* 5NQ-OH* polluting PAC in the environment, there is a great

19 need of data to evaluate the partitioning of PAC sorbent and solute characteristics. Koc values between water and soot particles. from literature were correlated with the capacity During the transport of soot particles in the coefficients (k’) on the humic acid HPLC column atmosphere, their surfaces are readily oxidized by (Table 2.3.4.1). From these results, it can be seen common photochemical oxidants, such as 03, and that log Koc increased with the size of the thereby begin to resemble humic acid molecules molecule for both PAH and N-PAC. The Koc (Haumaier and Zech 1995). These were used as values for PAH of the same molecular weight model substrates for soot particles, having the (dibenzanthracenes) vary very slightly. However, same properties as those reported for soot Koe values of dibenzacridines decrease in the particles - alcohol/phenol, carbonyl, carboxylic order of: acid, quinone/hydroquinone (Haumaier and Zech 3.4.5.6- dibenzacridine > 1,2,3,4-dibenzacridine > 1995). Aldrich humic acid was chemically immo­ 1.2.5.6- dibenzacridine > 1,2,7,8-dibenzacridine. bilized to the silanol surface of the column Correspondingly, Koc for 7,8-benzoquinoline was material for use with HPLC (Nielsen et al. larger than those for acridine, phenanthridine and accepted). 5.6- benzoquinoline. The reason for this appears Studies focused on the sorption mechanisms to be the shielding of the nitrogen atom by the have concluded that the degree of sorption of benzene rings which decreases the ability of the PAC, i.e. the particulate water distribution, can be nitrogen atom to form hydrogen bonds. modelled fairly well with the help of a few

Table 2.3.4.1 Logarithms of measured capacity coefficients on the humic acid column, estimated logK ot values and log Koc values from literature.

Compound logk logK„c" logK oc Compoimd logk ’ logK. oc ~ N-PAC (lit) X-PAH quinoline -0.85 2.91 3.05 9-acetylanthracenc 0.04 4.23 isoquinoline -0.71 3.12 9-anthraccnecarboxamidc -0.05 4.11 4-azaflourenc -0.35 3.65 9-antliracenecarboxylic acid 0.21 4.48 methyl ester acridine -0.11 4.02 4.11 9-bromoanthraccne 0.66 5.16 5.6-benzoquinolinc -0.06 4.09 9-chloroanthracene 0.53 4.97 7,8-benzoquinoline -0.02 4.15 9-cyanoanthracenc 0.36 4.72 phenanthridine -0.07 4.08 9-fonnylanthraccne -0.08 4.05 1. 2-bcnzacridine 0.56 5.02 9-mcthoxyanthracene 0.18 4.45 10-azabenzo(a)pyrene 1.14 5.87 9-methylanthraccnc 0.47 4.88 1.2.3,4-dib enzacridinc 1.34 6.17 9-nitroanthracene 0.35 4.70 1,2.5,6-dibenzacridinc 1.26 6.06 anthraquinonc 0 10 4.33 1.2.7.8-dibcnzacridinc 1.14 5.87 1-bromopyrenc 1.01 5.68 3.4.5,6-dibcnzacridinc 1.40 6.26 2-bromopyrcne 1.00 5.67 carb azole 0.39 4.76 4-bromopyrene 1.04 5.72 PAH 1.3-dibromopyrene 1.36 6.21 naphtlialenc -0.28 3.77 3.32 1.6-dibromopyrenc 1.25 6.04 fluorcnc 0.35 4.70 1.8 -dibromopyrcne 1.35 6.19 anthracene 0.33 4.66 4.87 0,S-PAC phenan Irene 0.33 4.67 4.81 dibenzofuran -0.01 4.17 1,2 -benzanthracene 0.98 5.63 5.48 dibcnz.othiophenc 0.29 4.60 benzo(a)pyrene 1.41 6.28 6.30 X-N-PAC 1.2.3.4-dibenzanthraccne 1.59 6.55 2-hydroxy quinoline 2.7" 1.2.5.6-dibenz anthracene 1.52 6.44 quinoline-N-oxidc 2.0* ” 1,2.7,8-dibenzanthraccne 1.62 6.59 N-mcthylquinoliniumiodide 3.1"

JogK oc = (1.49 ± 0.13)xlogk ’ + (4.18 ± 0.10) r" = 0.96 calculated from logK vc=logK oc (naphthalenc)+ 1.50x(logk'(X) 95% wa ,cr-logk ‘(naphthalenc) 9 5.„watcr )

20 Ris0-R-972(EN) In Table 2.3.4.2, there is an example of different small amount, <1%, bound to soot particles, situations in the atmosphere or in a lake where unless the soot sedimentation rate is very fast. soot particles have been deposited. The amount of The second column describes a situation where compounds in the bound phase was calculated there is a water uptake of 10% by the soot particle from the partition coefficients (Table 2.3.4.1) and in the humid environment. This case is the it was supposed that equilibrium had been reverse of the previous one, as most of the achieved. The first column describes a situation compounds, >99.5%, are bound to the soot in a lake with an area of 1 km2 and a depth of 5m. particles. The third column depicts a soot particle The average concentration of the soot in the air in a cloud droplet. Here it is surrounded by a over the lake is 10 pg/m 3 and the deposition water drop with a diameter of 0.1 mm, thus velocity 0.1 cm/s. In this case, there is an input of making the water concentration as high as 99%. 25kg of soot/km 2 to water in one month, giving a In this case, it is possible to notice the differences concentration of soot in the lake of 5xl0" 9 kg/L. between the various compounds. It can be seen Assuming that the fraction of organic carbon that with a lower molecular weight, more polar (OC) in soot is 0.5, the concentration in the lake compounds have a lower fraction in the bound will be 2.5x1 0"9 kg[OC]/L. Most of the phase. Table 2.3.4.2 includes only a few example compounds will be in dissolved form and a very compounds.

Table 2.3.4.2 The amount of bound compounds in different situations.

The percent of the compound bound with soot particles Compound lake’ Soot particle Soot particle (10%water)* (99%water)*** quinoline 2'10"4 99.73 80.41 3,4,5,6-dibenzacridine 0.45 99.9999 99.99 naphthalene 1.510"3 99.96 96.75 benzo(a)pyrene 0.47 99.9999 99.99 1,2,7,8-dibenzanthracene 0.96 99.9999 99.99 anthraquinone 5.1' 10"3 99.9896 99.08 9-nitroanthracene 0.013 99.9956 99.61 dibenzothiophene 9.9' 10"3 99.9944 99.50 quinoline-N-oxide 2.5*10" 5 97.83 33.56

*2.5x 10 9 kg|OC]/L; "0.45 kg[OC]/L; **'5.()5x 10"3 kg[OC]/L

References Haumaier L, Zech If. (1995) Blade carbon - possible source of highly aromatic compounds of soil humic acids. Org. Geochem. 23: 191-196. Katrinak K.A., Rez /'.. Buseck PR. (1992) Structural variation in individual carbonaceous particles from an urban aerosol. Environ. Sci. Technol. 26: 1967- 1976 Nielsen T„ Siigur K., flefweg Jorgensen ()., Hansen P.K., Kirso U. Sorption of polycyclic aromatic com ­ pounds to humic acid as studied by high performance liquid chromatography. Environ. Sci. Technol. Accepted.

21 2.3.5 Water-Octanol Partitioning (C. Helweg, context. However, the Kow of many chemicals is T. Nielsen) unknown. Among these chemicals are the nitrogen-containing polycyclic aromatic com­ pounds (N-PACs) which are present in coal and tar pollutions. Therefore, methods for the estima­ tion of Kow were examined with N-PACs and polycyclic aromatic hydrocarbons (PAHs) as model compounds. The major work was done using the HPLC method. This method is based on the linear relationship between capacity coeffi­ cients in a HPLC system and partition coefficients. This relationship, once established, makes the prediction of Kow from measured HPLC capacity coefficients possible. The results show that compounds, differing in polarity and hydrogen bonding ability, exhibit 2 • different relationships between capacity coeffi­ cients on a given HPLC column and Kows 1 -! I (Helweg et al. a, submitted). Furthermore, it has -10 12 been shown that different columns work best for log k' different groups of compounds. A Diol bonded reversed phase column gave more accurate KI)W Figure 2.3.5.1 Linear relationships between predictions for polar N-PACs than for that of the measured log k' and reference log Kow for the traditionally applied octadecylsilylsilica (ODS) Diol column at 35% MeOH. column (Helweg et al. a, submitted). The Diol column stationary phase material possesses Partition coefficients are key parameters in the hydroxy groups capable of hydrogen bonding, evaluation of the fate and hazard of chemicals in similar to the hydroxy group of octanol. the environment. In particular, the octanol-water partition coefficient (Kow ) is widely used in this

Pooled data □ Nonpolar data ■ Polar data

Chromatographic system

Figure 2.3.5.2 Mean square error. MSF, calculated for different chromatographic systems. Non-polar compounds are mainly PAH while polar compounds are N-PAC. The number after the column type designates the amount of methanol/water (v/v) used.

22 Ris0-R-972(EN) Substituted PAH is another group of environ ­ submitted) from Kow, is not valid for substituted mentally important compounds for which Kows PAH (Helweg et al. b, submitted). are not known. Kows have, therefore, been Parallel with the HPLC method, methods for estimated for a range of substituted anthracenes theoretical prediction of log Kow were using HPLC. Electronic and steric structure was investigated. A simple model based on size found to be important for Kow . A comparison (calculated as graph theoretical indices or with Koc (partition coefficient between humic molecular weight), nitrogen occurrence and steric acid and water) revealed, that different shielding, predicted log Kow of the model mechanisms are responsible for partitioning in the compounds N-PAC and PAH very well (Helweg two systems. Therefore, the frequently made et al. c, in preparation). prediction of Kou (Nielsen et al. a, accepted and b,

1 + HPLC o Rekker O) a Simple O u "O0) o •K3 A T)

A

-1 - 1 4 5 6 7 Reference log Kovv

Figure 2.3.5.3 Deviations between reference log Kow and predicted log K,m. for the best HPLC methods. Iiekkers/-fragment method and a simple predictive model.

References the sorption of polycyclic aromatic compounds to Helweg C., Nielsen 7'., Hansen P.E. (a) Determination humic acid. Talanta. Submitted. of octanol-water partition coefficients of polar polycyclic aromatic compounds (N-PAC) by high 2.4 Chemistry of Combustion performance liquid chromatography. Chemospherc. (E. Larsen , H. Egsgaard. J.F. Submitted. Products Helweg ('.. Nielsen /’., Hansen P.E. (b) QSPRs com ­ Jensen, K. T. Poulsen) pared with HPLC experiments for the estimation of Kow for N-PAC compounds. Proceedings of QSAR 2.4.1 Pressurized Laser Pyrolysis of Coal 96. Submitted. Helweg Nielsen /., Hansen P.E (c) Determination of Kow of substituted polycyclic aromatic compounds. The investigation of coal samples using flash In preparation. pyrolysis has continued. In a high-pressure Nielsen T., Siigur K., Helweg C.. Jorgensen O.. Hansen pyrolysis cell, foils of coal are irradiated with a P.E., Kirso f ’ (a) Sorption of polycyclic aromatic powerful C02 laser. The reactions are the result compounds to humic acid studied by apply ing high of very fast heating rates approaching 105°C/s performance liquid chromatography. Environ. Sci. and final temperatures above 1500°C, hence Tcchnol. Accepted. Nielsen 7., Helweg C., Siigur K., Kirso U. (b) simulating the conditions in power plants Application of capacity coefficients to characterize utilizing pulverized coal combustion. The cell is connected on-line to a GC/MS system facilitating

23 the analysis of the gaseous products. The analysis Experiments with coals, originating from includes the product distribution as well as the mines world-wide, are being studied in the isotopic composition. In Figures 2.4.1.1 and pressure range 1-50 bar and in atmospheres of 2.4.1.2 the experimental set-up is shown. Ar, N2, He, H2 and D2. The analysis has con ­ centrated on the low-molecular-weight hydro ­ carbons methane, ethane, ethylene, acetylene, propane. In particular, ethylene and acetylene may carry information on the underlying reaction mechanisms. This is being studied using the incorporation of deuterium. The experiments will be concluded with studies of the free methylene, CH2, generated by SF6-sensitized laser pyrolysis of diazomethane.

Reference Thomsen MS., Egsgaard II. (1995) Pressurized laser pyrolysis of coal. J. Anal. Appl. Pyrolysis 54: 243- 250.

2.4.2 Tar Cracking/Conversion in Gasifiers for Biomass

The use of biomass in combined power/heat Figure 2.4.1.1 The experimental set-up showing plants is obviously the most direct way of using the CO2 laser and the high-pressure pyrolysis the C02 neutral fuels. However, in using low cell connected to the GC/MS system. temperature pyrolysis/gasification of biomass, large amounts of tar products are often formed When the gas is intended to be used in engines for the production of electricity, the tar is a highly undesirable product, which can result in serious damage to the engines. The concentration of tar products and its chemical composition are strongly dependent on the actual conditions for pyrolysis/gasification. The tar products can be removed by a number of processes including thermal cracking, catalytic transformation, steam refinering and partial oxidation. We have investigated the tar reduction in a two-step experimental oven, where the process can be studied under well defined conditions. The preliminary experiments have focused on the partial oxidation of tar. Figure 2.4.2.1 shows the two-step oven used for the pyrolysis and the transformation of the tar products. The oven consists of two separate sections with independent temperature controls. The first Figure 2.4.1.2 The vacuum system for the section is used for the pyrolysis and the second sampling handling of diazoalkanes. for the transformation of the tar.

24 Ris0-R-972(EN) Figure 2.4.2.1 A photo of the experimental two-step oven used.

The first experiments demonstrated that the tar References products in the pyrolysis gas can be partially Jensen P.A., Larsen K., Jorgensen K.H. Tar reduction oxidized without being accompanied by the by partial oxidation. 9th European Bioenergy Con ­ oxidation of CO. Thus, the gas maintains a ference. Copenhagen 24-27 June 1996. Jorgensen K.LL, Larsen E., Jensen P.A. Tjaerened- relatively high heating value. Figure 2.4.2 2 brydning i lavtemperatur forgasscre til biomasse. shows the reduction of tar as a function of Ease 1. Rise Report. (ENS journal no. 138.3/95- temperature and oxygen. The tar is generated by 0003). pyrolysis of wheat straw at 500°C. 2.4.3 Flash Pyrolysis

0.7 Pyrolysis is a powerful analytical technique which can be applied to complex non-volatile 0.6 - samples. Three projects are briefly discussed: 0.5 -

0.4 - 0% 02 Waste Samples 0.3 - In collaboration with the Systems Analysis 0.2 - 2% O, Department, Riso, waste samples from the Danish chemical company, Prom Kemi, were analysed. The samples were pyrolysed/combusted Temperature, °C at 600-900°C in the DIN 53 436 furnace used to Figure 2.4.2.2 Tar amount as function of simulate chemical fires on a small laboratory temperature and oxygen injection in the scale. The decomposition/combustion products secondary reaction chamber were analysed on-line by FTIR In addition, samples were collected for the analysis for HCN,

25 HC1 and complex organic compounds by only oxygen can be incorporated into the fuel, GC/MS maintaining the overall emission characteristics. The results were evaluated by the Systems Nevertheless, the presence of oxygen may give Analysis Department, Riso, with respect to differ­ rise to very significant changes in the combustion ent scenarios, including the various types of fires processes due to its role as a reaction centre for and the transport and dispersion of the concerted elimination processes. In addition, the combustion products in the atmosphere. elimination of thermodynamically strongly favoured molecules, such as formaldehyde, may Coal Samples be a driving force in certain reactions, as in the A number of coal samples of different origin case of dimethylether. In sum, these properties were investigated by flash pyrolysis to determine may affect the reactions of both the intact fuel the amount of volatiles released at moderate molecule and the intermediates in the combustion temperatures, i.e. in the range 300-500°C. It process. Thus, we gain some possibilities to appears to be a fast and reliable method, which control the reaction paths in the combustion can be used for the calculation of the optimal process and hence, to design new and alternative mixing ratio of different coal types, hence fuels. avoiding the risk of fires in coal mills and flash The design of a fuel system mimicking the back in the burner systems in power plants. Thus, excellent properties of dimethylether as a diesel coal which otherwise could not have been used, fuel, but exhibiting more favourable physical pro ­ can now be burned safely. perties, such as being a low-viscous liquid at ambient temperature, is the ultimate goal. Chalk Samples Nevertheless this may be possible, as the Chalk may contain minor, but nevertheless combustion chemistry of dimethylether is rather significant amounts of organic compounds. Chalk well established. The initial abstraction of a samples were flash pyrolysed to measure and hydrogen atom followed by the thermally induced identify possible organic compounds in the decomposition of the methoxymethyl radical are material in order to complete the energy balance the key features. A strategy for the design of in the production of clinker alternative fuels could be based on the reversal of the initiation and the thermal processes as outlined in Figure 2.5.1. 2.5 Development of New and Alternative Fuels (H Egsgaard, E. Larsen, The Dimethyl Ether Case O.J, Nielsen, J. Platz, J. Sehesled, R. Stuhbe)

The research on new and alternative fuels relies on the interplay between direct studies of premixed flames and reaction kinetics of Initiation Thermal process Combustion chains conceivable elementary reactions. This provides a basis on the molecular level, for the development of new fuel systems. In addition, this research New & Alternative Fuels - A Strategy may contribute to a reduction of emissions from energy production as well as improved efficiency CH3CHO CHXHO + CH-O in after-treatment. 0 CH.O —

In situ fuel synthesis Initiation/combustion 2.5.1 New and Alternative Fuels - How to Figure 2.5.1 A strategy for new fuels. gain Success ? In this context, the 1,3-dioxolanes are particularly All present concepts seem to be based on oxygen- interesting candidates, as they undergo a facile containing compounds. From an environmental thermally-induced ring opening, yielding two point of view, this is logical, in the sense that molecules of oxo-compounds (Figure 2.5.1).

26 Ris0-R-972(EN) Thus, 1,3-dioxolane gives rise to formaldehyde the combustion chemistry and the fuel and acetaldehyde. A major combustion process characteristics of dimethylether. The CH yield of for acetaldehyde is the formation of acetyl 1,3-dioxolane was included in Figure 2.5.2.1. The radicals followed by thermal decomposition to a very low yield suggests a novel combustion methyl radical and CO Thus, the combustion of chemistry and makes it an interesting candidate the parent 1,3-dioxolane may be expected to take for further investigations. the same favourable route as that of dimethylether. Our activities on new and alternative fuels will, in the following, be illustrated with results obtained from the investigations of dimethylether and 1,3-dioxolane, respectively. Flame ionization of alkanes, dimethylether and 1,3-dioxolane

2.5.2 Screening New Fuels: The Soot Potential

The particle (soot) emission from engines is a problem of major concern. The mechanisms for the formation of soot from simple low-molecular- weight fuels have not yet been definitely established. However, it should be noted that 1,3-dioxolane there are essentially two current mechanisms proposed for soot formation in flames. The most DME widely accepted mechanism is based on free radical reactions and the other on an ionic mechanism. The CH radical is the most reactive organic radical known In its ground state, it possesses the Carbon content unique combination of lone pair and a vacant '- Figure 2.5.2.1 The relative yields of CH from orbital. The reaction between CH and acetylene simple alkanes, dimethylether and 1,3-dioxolane, seems to be of key importance for the formation respectively. of higher alkynes/PAH ’s in flames via the C3H2 product. The C3H2 radical, as well as its The blue light is one of the most notable features protonated counterpart, C3H3 , may enter reaction of hydrocarbon flames. It has been known for cycles leading to high molecular compounds and almost a century that the emission at 43 lnm is ultimatively to soot. The ionic mechanism is due to CH*. It is important to note that the initiated by the chemi-ion C3H3 , i.e. the pro- electronic excitation is not the result of the high pargylic cation, which rapidly grows by the temperature within the flame, but is brought addition of alkynes, e g. acetylene. In addition, about by highly energetic chemical reactions The the reaction between the CH radical and atomic reaction between C2H* and O seems to be a major oxygen is responsible for the enhanced ionization source of the CH* chemiluminescence. The C2H* in flames and provides the basis for the flame radical itself is the result of various and rather ionization detector. Thus, the determination of the complex reactions of acetylene and CH. Thus, it relative sensitivities of various compounds pro ­ seems reasonable to assume that the CH* vides a straightforward measurement of the emission correlates to the acetylene/CH relative yields of the CH radicals. The relative concentrations within the flame and hence, that yields from a large number of compounds were the CH* emission may be used as a diagnostic determined. An illustrative example is shown in tool in the prediction of the sooting properties of Figure 2.5.2.1. It is apparent that dimethylether a given compound. on a molecular basis, yields only 50% of the CH In collaboration with the Optics and Fluid radicals that may be expected on a carbon basis. Dynamics Department, Rise, we investigated However, this is in excellent agreement with both premixed flames of dimethylether/- and methane/

27 oxygen using a small diode-array spectrometer. oxygen flame. The CH* emission reveals a pro ­ The set-up was used to investigate the spatial nounced maximum at the reaction zone; the distribution of the chemiluminescence of the emission from OH* (3 12nm), on the other hand, flames. Figure 2.5.2.2 shows a 3D plot of the persists into the burnt gas region. luminescence from a premixed dimethylether/

0 ■ ■ D ■ ■

Figure 2.5.2.2 The spatial resolution of emission spectra from a premixed dimethylether/oxygen flame.

28 Ris0-R-972(EN) A series of experiments, carefully maintaining provides a stable gas flow of the fuel being mixed the flame size, makes the comparison of with preheated oxygen and finally burnt on dimethylether and methane flames possible. stainless steel capillary. Figure 2.5.3 (see end of Figure 2.5.2.3 shows the CH* emission from Chapter 2) shows the mass spectra of premixed premixed flames of dimethylether/- and methane/ methane, dimethylether, ethanol and 1,3- oxygen as a function of the load. It is interesting dioxolane, respectively. to note the significantly lower CH* yield from The spectra of the oxygen-containing mole­ dimethylether on an energy basis. cules show, in comparison with that of methane, a lower content of hydrocarbon!c ions at the high mass end. This is indicative of a lower particle Chemiluminescence from methane and dimethylether flames emission upon combustion. In addition, the intensity of several ions, e g. m/z 31 and m/z 45 directly reflects important routes in the 17500 - Methane underlying combustion process. The ions are due to protonation of formaldehyde and acetaldehyde 12500 " formed within the flame. DME

7500 - 2.5.4 Reaction Kinetics at Ambient to Medium Temperature and Pressure

2500 ' The reaction kinetics related to the oxidation of dimethylether were studied in depth. Thus, -2500 absolute rate constants for the self reaction of methoxymethyl radical, the reaction of methoxy- Fuel, (kW) methyl radicals with oxygen and the thermal Figure 2.5.2.3 C,H * emission as a function of the decomposition of methoxymethyl radical were load in premixed dimethylether/- and methane/ determined. It should be noted that, at ignition oxygen flames. temperatures, the formation of peroxy radicals becomes reversible. However, the intermolecular To make the optical methods available on a hydrogen transfer in the peroxy radical, followed smaller scale, a traditional flame photometric by decomposition to formaldehyde, may remain detector, as used in combination with gas an important ignition process, as in the scheme. chromatography, has been modified to monitor The reaction has been demonstrated to be the specifically the CH* emission at 431nm. This dominant route at low pressure and temperatures. technique appears very promising and enables an This is primarily due to the decomposition of the easy determination of the CH* emission using excited CH30CH200 radicals before collision samples

2.5.3 Premixed Flames of Alternative Fuels

Riso National Laboratory has a long tradition in the development of methods for direct analysis. The novel Riso flame-ion tandem mass spectro ­ meter includes a high temperature supersonic molecular beam sampling system, which gives CH30CH200------[ 'CH2OCH2OOH1 ------2 CH20 + "OH unique possibilities for studies on the global and elementary level of the combustion process. In order to handle liquid fuels, a unit consisting of a The kinetics of the unimolecular high pressure pump and a heated stainless steel decomposition of the methoxy radicals is very evaporation reservoir was constructed. This unit interesting. In Figure 2.5.4, the decay rate as a

Ris0-R-972(EN) 29 function of temperature can be seen. At The fast-flow/MS system has previously been approximately 1000°C, the rate of reaction will used in connection with EPR-detection of exceed the gas kinetic collision frequency, and selected free radical/molecule reactions. Laser then this unimolecular reaction becomes the most pyrolysis has likewise been used in both important loss reaction for methoxymethyl fundamental studies of pyrolytical reactions and radicals in the dimethylether combustion. as applied to characterization fossil fuels. At present, the controlled abstraction of hydrogen from dimethylether by free radicals has been demonstrated, whereas the laser heating process remains a major project for 1997. The low pressure facility enables the study of thermal reactions under conditions minimizing secondary and bi-molecular reactions. At the same time, the reactions are the result of single collisions with an inductively heated non- catalytic surface (Au). The detection principle is based on a true soft ionization mode (field ionization) supplementary with collision activa­ tion of selected ions. In the present context, the capabilities of the set-up are well illustrated by the pyrolysis of 1,3-dioxolane. Figure 2.5.5 shows the field ionization spectra of 1.3- dioxolane without, and following pyrolysis. The formation of significant products at m/z 30 and m/z 44, respectively, is in clear evidence. The products are assigned to formaldehyde (M=30) Figure 2.5.4 Pseudo first order decay rates as a and acetaldehyde (M=44), respectively. Thus, the function of temperature as observed for the facile thermal decomposition of 1,3-dioxolane to methoxymethyl radical. formaldehyde and acetaldehyde was demon ­ strated. 2.5.5 High Temperature Processes Related to the Use of Alternative Fuels b 0 In the case of dimethylether, a unique thermal process has, as discussed above, been unambi ­ guously demonstrated to play a key-role in the novel combustion chemistry. Thus, studies of thermally induced reactions may be expected to JlL play a dominant role in the development of new, alternative fuel additives. Investigations of free a radicals and kinetically stable molecules in the medium to high temperature range, 400-1000°C, seem particularly to be in demand. With this in .j mind, two major efforts were initiated this year, namely 1) the combination of a fast-flow modulated molecular beam mass spectrometer with infrared laser sentisized laser heating and 2) the re-installation of the facility for pyrolysis at very low pressure combined with real-time mass Figure 2.5.5 Field ionization mass spectra of spectrometric analysis. 1,3-dioxolane a) without and b) following pyrolysis at 1404 K.

30 Ris0-R-972(EN) 2.5.6 Characterization of Fuel Properties connection with the combustion of dimethylether. The NOx emission from engines fuelled by The use of a particular compound as fuel for dimethylether is reduced to approximately 50%; either the Otto- or Diesel-engines depends on a this value corresponds to the reduction of CH and rather complex series of physical/chemical consequently, the reduction of prompt NOx . It properties including vapour pressure, viscosity should, however, be noted that NOx emission and ignition temperature. Direct testing of from engines in general, is believed to originate selected fuel systems on single-cylinder engines predominantly from the thermal process. On the is part of the co-operation between the Technical other hand, the relative importance of two routes University of Denmark and Riso on the for NOx formation remains at present development of new and alternative fuel systems. unanswered. Investigations on one of the most promising systems, i.e. the 1,3-dioxolanes are in progress. References Egsgaard H. (1996) Investigation of the initial reactions 2.5.7 NOx-Formation and DeNO, Chemistry of the calcote mechanism for soot formation. J. Am. Soc. Mass Spcctrom. 7: 559-64. Egsgaard II., Larsen E. (1996) Direct sampling tandem Formation of NOx may take place by a number of mass spectrometry of premixed flames. Proceedings different mechanisms. Motor fuel is essentially 44th Conf. Mass Spcctrom. Allied Top. Portland. free of nitrogen and hence the NOx generated in 12-16 May 1996. p. 420a-b. engines must be produced in reactions involving Egsgaard II., Larsen Sehested J., Nielsen O.J. molecular nitrogen present in the air. Two distinct (1996) Intermediates in dimethylether flames. mechanisms are apparently of significance, Proceedings Comb. Inst.. Funchal. Portugal 1-4 April 1996. p. 18.3.1-3.4. namely the thermal formation, formally due to the Sehested J., Sehested A., PlatzJ., Egsgaard II., Nielsen reaction of molecular nitrogen and oxygen, and O.J. Oxidation of dimethylether: Absolute rate the so-called prompt NOx, which is due to the constants for the self reaction of CH3OCH2 radicals, reaction between molecular nitrogen and CH the reaction of CH3OCH: with 02, and the thermal radicals. The latter is particularly interesting in decomposition of CH,OCH2 radicals. Submitted.

31 09/04/96

4

of

1

#

Scan

.le:13oxo8 Fi

;

j

3

of

1

#

Scan

2 f u e l

: F i l e

Figure 2.5.3 Mass spectra of premixed flames of methane/-, dimethylether/-, ethanol/- and 1,3- dioxo lane/oxygen .

32 Ris0-R-972(EN) 3 Gene Technology and Population Biology

Introduction (H. Ostergard) new genotypes, e.g. genotypes possessing trans ­ genes. In this context, we are studying gene An understanding of interactions between plants introgression from crop plants to their wild rela­ and pathogens is important for developing strate­ tives to assess the risks of releasing genetically gies for sustainable agriculture with reduced use modified plants. Further, the evolutionary pro ­ of chemicals. A large part of our research is cesses imposed by interactions between crop devoted to the study of the mechanisms of plant- plants and wild plant species, eg. weeds, are pathogen interactions (Sections 3.1-3.6). This being studied. Oilseed rape and wild Brassica includes modelling evolutionary processes and species are used as model systems for these the detection of gene products, which are import ­ studies (Sections 3.8 and 3.10). ant for resistance and virulence. The establish ­ ment of genetic markers and analysis of genetic diversity are important tools for these studies. Summary of Projects The host-pathogen system, barley and barley powdery mildew Erysiphe graminis f.sp. hordei 3.1 Epidemiology and Population is used as a main model system for an obligate Biology of the Powdery Mildew fungus. However, Tillelia caries, which causes common burst in barley and Leptospheria Fungus, Erysiphe graminis f.sp. maculans, which causes root rot in oilseed rape hordei (H. Ostergard, C, Damgaard, are also considered. R. O’Hara, M.F. Lyngkjcer) Methods for identification, characterization and quantification of genetic variability are essen ­ The fungus Erysiphe graminis f.sp. hordei infects tial to many basic and applied research pro ­ barley leaves and causes reductions in both yield grammes in the agricultural and environmental and quality of grain. For many years plant breeders sciences. Exploitation of genetic variability, have developed varieties with resistance genes, but related to traits of economic and environmental in most cases the resistance breaks down within a importance in crop plant improvement, can be few years because the pathogen population changes greatly facilitated by the use of marker-assisted towards increased virulence (Hovmoller and selection (Sections 3.7 and 3.8). In particular, the Ostergard 1996). However, one group of host new methods for identifying molecular markers resistance alleles, called mlo, has been successfully may help in understanding the genetic basis of used in barley for over fifteen years without the many agronomic traits, which are of a quantita ­ mildew adapting to it (Lyngkjser cl al. 1996). We tive nature and governed by the interaction of are involved in research into the mechanisms by many Quantitative Trait Loci (QTLs) impossible which mlo operates. to assay independently. An understanding of the Another method, used to control mildew on regulation and expression of genes and the char ­ barley, is the application of fungicides (Brown acterization of gene products is important for 1996). Again, the fungus adapts to the disease developing plants with improved characteristics, control strategy and isolates of mildew, which are e.g. a better quality of grain proteins or new resistant to the fungicides used, evolve. This proteins produced in the grains (Sections 3.6 and evolution is, however, slower than the evolution 3.9) Also transformation techniques are imple ­ towards virulence. Optimal disease control relies on mented to improve grain quality for low input a detailed understanding of the epidemic develop ­ farming. Barley is the main crop plant of interest, ment of the disease (e.g. Damgaard and Ostergard but wheat and oilseed rape are of increasing 1996a, Damgaard and Ostergard, in press) and the interest. effect of host resistance and fungicides on the Studies of plant population biology are neces ­ parameters of importance for growth of the fungal sary for predicting the consequences of using population.

Riso-R-972(EN) 33 Our research is in close collaboration with the variation in the number of vimlence genes and European research network established through the resistance genes with pathogens carrying several, COST Action 817, i.e. population studies of but not all, vimlence genes available. Similarly, the airborne pathogens on cereals as a mean of plants have only a few resistance genes. In order to improving strategies for disease control (Ostergard account for what is observed, it has been suggested and Masson, in press). that there is a cost of carrying unnecessary vimlence genes and resistance genes, respectively. However, Co-Evolution of Virulence and of Host Resistance there is little experimental evidence to support this For pathogens to be able to grow on host plants, hypothesis. We have examined an alternative they must have genes that will allow them to explanation for the observed patterns. By using a overcome the host ’s resistance, i.e. vimlence genes. mathematical model, we have investigated the It has been argued that pathogens would evolve into possibility that vimlence genes and resistance genes having all possible vimlence genes enabling growth in a gene-for-gene system could be maintained on all available hosts Similarly hosts would carry purely by the dynamics of host and pathogen all of the resistance genes available. However, this is populations (Damgaard and 0stergard 1996b) not seen in natural populations. Instead, there is

Virulence: Ne = HXXK) ,\V = KXX) Ne = 2(X) Ne = 50

Resistance: Ne = 50 Ne = 2(X) Ne = l(XX) Ne = l(XXX)

Figure 3.1.1 The effect of genetic drift on allele frequencies. The frequency of the virulence allele and the resistance allele after 5,000 generations is a decreasing function of increased genetic drift. Ne is the effective population size, and a reduction in the effective population size increases genetic drift.

We assumed that the host population was made vimlence locus and one resistance locus, can be up of many sub-populations, with little migration maintained in the sub-population, and the amount of between them. Infection by the pathogen reduced variation depends on several factors, such as the size the fitness of the host, and so increased the chances of the populations and the amount of migration that the sub-population would become extinct. We between the sub-populations. Importantly, this have shown that in this model, variation at one model creates variation similar to that seen in the

34 Ris0-R-972(EN) real world, without having to assume that there is a these m/o-virulent isolates, HL3/5, and its near- cost of virulence or resistance. Instead, the isogenic, avimlent progenitor GE3, were examined modelling suggests that we can explain what is in an experimental system using the isogenic occurring in the real world in terms of population barley lines Riso-R (mlo5 resistance allele) and dynamics. Riso-S (Mlo susceptibility allele) (J0rgensen 1975). A time course was conducted in which Powdery Mildew Infection of Barley with the early host responses in relation to fungal mlo Resistance Gene development were observed at one hour intervals Barley with resistance, based on the effect of from 10 to 24 hours after inoculation (Lyngkjasr m/o-alleles, is highly effective in preventing and Ostergard 1996). penetration by fungal hyphae of host epidermal Development and growth of the virulent and cells. The resistance is correlated with rapid avirulent E. graminis isolates, respectively, on deposition of papillae in the epidermal cell wall. both barley isolines were similar up to first In susceptible barley lines with the dominant penetration attempt, 11 to 12 hours after inocula ­ M/r;-allele, papillae are also produced, but many tion. When the m/o-virulent isolate attacked Riso- more of these are penetrated, resulting in R leaves, about 20% of appressoria succeeded haustorium formation and successful infection. penetration and formed haustoria. However, for In Europe, there is no indication of increase in the avirulent isolate, less than 1% of fungal disease severity in barley with mlo-based appressoria succeeded penetration and formed resistance. However, two ‘laboratory ’ isolates of haustoria. On Riso-S about 25% of appressoria E. graminis have been described as m/o-virulent from both E. graminis isolates succeeded penetra ­ (Schwarzbach 1979, Lyngkjasr et al. 1995) One of tion and formed haustoria, see Figure 3.1.2.

%

.2 *c Rise S-HL3/5 © s Riso S-GE3 55

tt=< ---- Riso R-HL3/5 © bD 55 s 4> V ---- Riso R-GE3 hm 0U Hours after inoculation

Figure 3.1.2 Mean percentage of haustoria formed during attempted infection by the virulent HL3/5 and the avirulent GE3 near-isogenic E. graminis isolates on leaf epidermal cells of barley isolines Riso-R (mlo5 resistance allele) and Riso-S (Mlo susceptibility allele) at 10 to 24 hours after inoculation. Means are based on observation of 100 appressoria forming conidia on each of four leaves. Error bars represent 95% confidence intervals.

Riso-R-972(EN) 35 Our investigation further confirmed, that the was included in these tests. We confirmed that effects of the /u/oJ-alleles on resistance, are in ­ there was contamination/ segregation in the first creased frequency of papillae beneath appressoria seed lot, but to a much smaller extent than that at the time of penetration, larger papillae size and found in Zurich. These tests were conducted on larger callose deposition at attempted penetration seedlings. After this, we exchanged isolates and sites. However, our results do not imply earlier new seed samples between all laboratories and formation of papilla in the Riso-R line, as earlier made guidelines for the assessment. A new test suggested. was conducted on seedlings and in Zurich, also The hypothesis that mlo alleles induce deregu­ on detached leaves. The same results were lation of a product involved in basic obtained in all three laboratories with only little physiological processes in the plant (Wolter el at. variation. It was concluded that quantitative 1993) may be used to explain these observations. mildew tests on seedlings are much more reliable The virulent K gram inis isolates may be able to than those on detached leaves and care has to be inhibit or overcome a main mlo resistance factor. taken in the way assessment is performed. This will increase the penetration efficiency of the virulent K graminis isolate on the resistant Development of Fungicide Resistance barley line, but not on the susceptible. In Another method of disease control, successfully addition, a general "back up" resistance is used to control mildew on barley, is the application working in the Riso-R line. This is supported by of fungicides. Although isolates of mildew, which the findings that phenolic compound synthesis are resistant to the fungicides used, have been provides back up to support the main resistance observed for many years, some fungicides still factor in mlo5 barley and that changes in this appear to provide effective control. We are engaged system do not increase the infection efficiency of in a programme of work in collaboration with the the avirulent isolate, as long as the main mlo5 Danish Institute of Plant and Soil Science, Lyngby, resistance factor operates effectively (Carver el and the Royal Veterinary and Agricultural Univer ­ at. 1995). If however, the main mlo5 resistance sity, Copenhagen, to investigate the evolution of factor is suppressed by 2-deoxy-D-glucose, fungicide resistance. We hope this work will avirulent isolates are found to be controlled by provide explanations of the observed patterns of the back up system (Lyngkjasr el al. in press). fungicide resistance and will help us devise effective The //i/ovirulent isolate is also sensitive to the mildew control strategies which use lower doses of phenolic back up system. fungicides, and hence are more environmentally In collaboration with the Department of Plant friendly. Science, Swiss Federal Institute of Technology in Standard methods for measuring resistance of the Zurich, Switzerland, and the Sainsbury Labora ­ powdery mildew to fungicides, involve spraying tory in Norwich, U K., a range of progeny leaves with different doses of fungicide and then isolates, from a cross between an w/o-virulent infecting them with mildew. The amount of mildew and an avirulent isolate, were tested for infection corresponding to each dose is measured, and then efficiency on a mlo5 resistant barley variety and the dose, at which half of the infecting mildew on two mutated m!o5 resistant lines with would be killed, the ED50, or median effective dose, modified resistance (Freialdenhoven el al. 1996). is estimated. Although this method is well In preliminary studies, the Zurich group obtained established and used routinely throughout Europe, inconsistent results in different replications of the the test has to be refined to enable us to use the tests. These tests were conducted on detached results for genetic analyses As an example, we have leaves. The reason for the inconsistent results investigated the effect of inoculum density on was first thought to be contamination of seed estimated ED50 values. Standard fungicide tests obtained from the Norwich group. Because of were carried out using three fungicides, our experience with mlo resistance and propiconazol, triadimenol and fenpropimorph. One quantitative assessment of mildew infection, Riso isolate of mildew was inoculated onto the leaves at

36 Riso-R-972(EN) three different inoculum densities. For two of the visited Rise to analyse his field data describing fungicides, triadimenol and fenpropimorph, we the epidemic development of Erysiphe graminis showed that the estimated ED50 value increased with f.sp. hordei on specific leaf stages measured on the inoculum density. We then constructed a single leaves sampled in a field plot during a mathematical model of the infection process which season. Disease was measured as leaf area suggested that at higher densities, unsuccessful covered, as well as the proportion of green area infections would be masked by the large number of covered (Pons-Kuhnemann 1994). The total area successful infections, and this would increase the of fully developed single leaves at a specific leaf observed EDS0 value. We would have to conclude stage was nearly constant during the sampling that the ED5() value cannot be considered as a period, whereas the green leaf area decreased constant unless the inoculum density is controlled. linearly. The infected leaf area of single leaves One possible reason why the evolution of increased as did the proportion of green leaf area fungicide resistance is slow, when compared to infected. When the average overall leaves were evolution of virulence, is that the resistant mildews considered, these proportions decreased slightly could be less fit. In other words, in the absence of after 14 days and very much again at the end of fungicide they are out-competed by susceptible the period. Even if this effect has often been mildew. To test this, we took a mildew isolate which found, proportions are usually considered in is very resistant to three fungicides, propiconazol, estimating disease increase rates (eg. Kranz triadimenol and fenpropimorph, and grew several 1975). We considered single leaf observations large populations of it for thirty generations. It is (Figure 3.1.3), and logistic growth curves were hoped that natural mutation will occur so that a fitted to estimate disease increase rates (rate of mutant which is susceptible to one, or more, of the increase in infected area when the area is small) fungicides is produced. If this mutant is fitter than and maximal infected leaf area (infected area or the original isolate, then it will increase in frequency number of colonies which can be maintained by in the population. By testing sample isolates from the resources in the leaf, i.e. infected area late in the population for resistance to the fungicides we the epidemic) (0stergard and Pons 1996). This can see whether this has occurred. As well as analysis assumes that the epidemic on a single allowing us to test the hypothesis that fungicide leaf is initiated from aerial spores when the leaf resistance incurs a cost, it will also give us a pair of emerges and that new colonies are made by isolates differing only in their fungicide sensitivity, spores produced on the same leaf. which could be used for further work. The disease increase rates were calculated for leaf stages 5, 6, 7 and 8, respectively. The logistic Influence of Density on Epidemic Development growth curves fitted the observations well, Understanding basic growth processes of plant indicating that one disease increase rate could pathogen populations, epidemic development, account for all factors influencing the growth of under field conditions is important for optimal the pathogen population. The maximal infected disease control. Epidemic development is influ ­ leaf area was estimated to be very low, between enced by numerous factors characteristic of a 2% and 20% of green leaf area, and high values specific host genotype and pathogen genotype of the disease increase rates were obtained. The under specific environmental conditions, e.g. rates were nearly identical among leaves of genetic variation in infection efficiency, latent different stages, but the maximal infected area period, spore production and infectious period, decreased. This indicates that adult resistance with all factors potentially varying during the may influence the population dynamic late in the season in response to varying host resistance, epidemic of a leaf and not, as often suggested, plant growth and senescence. through decreased disease increase rates early in As part of a COST 817 collaboration, J. Pons, the epidemic. Justus-Liebig University of Giessen, Germany,

Ris0-R-972(EN) 37 180 -

Ieaf2 ltaf3 Ieaf4 leaf? Icaf6 lean leafH Ieaf9

0 10 20 30 40 50 60 Days Figure 3.1.3 Observed changes in infected leaf area for leaves at stage 1 to 9 (seedlings being stage 1) with starting date May 22.

References Hovmoller M, Ostergard II. (1996) Prediction of durability Brown J.K.M. (1996) Fungicide resistance in barley of powdery' mildew resistance in autumn sown and powdery mildew: from genetics to crop protection. springh sown barley varieties. In Proceedings of 9th Proceedings of the 9th European and Mediterranean Cereal Rusts and Powdery' Mildews Conference. 2-6 Cereal Rusts and Powdery Mildews Conference, pp. September 1996, Luntcrcn, The Netherlands. G.H.J. 259-267. Kema, RE. Niks and R.A. Damcn (eds.) Cereal Carver T.L.W., /even R.J., Lvngkjter MF. (1995) Plant Rusts and Powdery Mildews Bulletin, Vol. 24. cell defences to powdery mildew of Graminea. D.R. Supplement 1996: 105-108. Walters. J.D Scholes. R.J. Bryson. N.D. Paul, N. Jorgensen J.II. (1975) Identification of powdery McRoberts (cds.) Phy siological responses of plants mildew resistant barley mutants and their allelic to pathogens. Aspects of Applied Biology 42: 257- relationship. Anonymous (ed.) Barley Genetics III. 266. Proceedings of the third international barley Damgaard C., Ostergard 11. (1996a) Density dependent genetics symposium. Ginching 1975. Munchcn: growth of powdery mildew (Krysiphe graminis f.sp. Verlag Karl Thiemig: 446-455. hordei) on a partial resistant barley variety: Kranz J. (1975) Das Abklingcn von Bcfallskurvcn. Z. Infection efficiency and spore production. E. PflKrankh. PfiSchutz 82: 655-664. Limpcrt. M R. Finckh, M S. Wolfe (eds.) Integrated Iyngkjcer M.F., Ostergard II. (1996) Powdery mildew control of cereal mildews ;md rusts: Towards co ­ infection of barley with the mlo resistance gene. In ordination of research across Europe. COST 817. Proceedings of 9th Cereal Rusts and Powdery European Commission, pp. 241-245. Mildews Conference, 2-6 September 1996. Dcnngaard ('.. Ostergard I!. (1996b) Genetic variation Lunteren, The Netherlands. G.H.J. Kcma, R E. Niks. in the gcnc-for-gcnc sy stem. In Proceedings of 9th R.A. Damen (eds.) Cereal Rusts and Powdery Cereal Rusts and Powdery Mildews Conference, 2-6 Mildews Bulletin, Vol. 24. Supplement 1996: 58-60. September 1996. Luntcrcn, The Netherlands. G.H.J. Lvngkjter M.F., Jensen IIP., Ostergard II. (1995) A Kcma. R E. Niks, R.A. Damcn (cds.) Cereal Rusts Japanese powdery mildew isolate with exceptional and Powdery Mildews Bulletin. Vol. 24. large infection efficiency on Mlo-rcsistant barley. Supplement 1996: 284-286. Plant Pathology 44: 786-790. Damgaard C, Ostergard II. Density dependent growth Lvngkjter A/., Carver T.W.L., /even R.J. Suppression of and life history evolution of polycyclic leaf resistance to Krysiphe graminis f.sp. hordei pathogens: A continuous time growth model. conferred by the mlo5 barley powdery mildew Journal of Phytopathology . In press. resistance gene. Physiol. Mol. Plant Pathol. In L'reialdenhoven A., Peterhansel C., Kurth ./., Kreuzaler press. Schulze-Lefert P. (1996) Identification of genes Lvngkjter MF., Jorgensen J.II, Ostergard II. (1996) Er required for the function of non-racc-specific mlo Mlo-resistens i byg holdbar? 13. Danske Plante ­ resistance to powdery mildew in barley. Plant Cell s'

38 Ris0-R-972(EN) Pons-Kuhnemann J. (1994) Struktur und Dynamik dcr YAC clones as a probe was adapted. Due to the Fungizidresistcnz in Populationen von Erysiphe large amounts of repetitive DNA in Egh cold graminis f.sp. hordei bei verschiedenen Applika- SINE, Short Interspersed Elements, and LINE, tionsstratcgien mil Triadimenol und Ethirimol. Ph D.dissertation, Justus-Liebig University Giessen, Long Interspersed Elements, sequences are added Germany. to the YAC probe. In order to block the yeast Schwarzbach E. (1979) Response to selection for ribosomal DNA, which cross hybridizes to Egh virulence against the ml-o based mildew resistance ribosomal sequences cold ribomal, DNA is added in barley, not fitting the gcnc-for-gene hypothesis. as well. Inserts from the positive cDNA clones Barley Genetic Newsletter 9: 85-88. have been isolated and will be mapped to confirm Walter M, ffollrichter K., Salamini Schulze-Lefert linkage to the avirulence gene. P. (1993) The mlo resistance alleles to powdery mildew infection in barley trigger a developmental!} controlled defence mimic phenotype. Molecular and Functional Studies of two Differentially General Genetics 239: 122-128 Expressed Egh denes Ostergard II, Masson JP. COST 817 - Population studies To analyse the function of the two novel Egh of airborne pathogens on cereals as a mean of genes, Egh7 and Eghl6 (Justesen et al. 1996), improving strategies for disease control. Annual report which are differentially expressed in germinating 1995. In press. Ostergard II., Pons./. (1996) Epidemic development of a conidia, we want to express them in vitro to make barley powdery mildew population. In Proceedings polyclonal antibodies and to localize in situ on of 9th Cereal Rusts and Powdery Mildews Egh mycelium. For that purpose, the genes were Conference, 2-6 September 1996, Lunteren, The cloned into the E. coli pET and the yeast pYES Netherlands. G.H.J. Kcrna. R E. Niks. R.A. Damcn expressions vectors. In the pET construction, the (eds.) Cereal Rusts and Powdery Mildews Bulletin, gene is regulated by the strong inducible E coli Vol. 24. Supplement 1996: 279-283. promoter T7 and in the pYES system, the promoters from the genes themselves are used 3.2 Molecular Biology of the Powdery So far, we have not been able to detect any Mildew Fungus, Erysiphe graminis expression in the E. coli system, but polyclonal f.sp. hordei (H. Giese, S.K. Christiansen , antibodies against germinating Egh conidia are M. Nicolaisen) produced which may provide a more sensitive assay. Linkage Map of Erysiphe graminis f.sp. hordei (Egh) and Map Based Cloning of Avirulence Transformation of Egh Genes A significantly better frequency of Egh cells, In the process of making a linkage map based on expressing GUS after biolistic transformation, our two Egh crosses, we are routinely screening has been obtained. This is primarily due to the for, and mapping cDNA clones which display use of a construct containing the promoter region polymorphisms between the parents. Establish ­ from the highly and constitutively expressed Egh ment of colinearity between the two maps glyceraldehyde-3-phosphate dehydrogenase gene, requires cDNAs showing polymorphisms in both gpd. Transformation efficiencies were determined crosses and therefore, a large number of cDNAs in four independent experiments and the best have to be analysed. Clones are sequenced to results, obtained for the constructions containing provide expressed sequence tags for the map. the gpd promoter region and that of the Egh p- Our strategy, in which Egh YACs (Borbye tubulin gene, were 40 blue cells/shot and 20 blue and Giese 1994) potentially containing avirulence cells/shot, respectively. In addition, we have genes are used to identify cDNAs that co­ worked on reducing cell injury which is a major segregate with avirulence genes, has led to the problem in biolistic transformation. Transforma ­ isolation of a number of cDNAs. The selected tion conditions were optimized and shock-attenu ­ YAC clones, containing restriction bands which ating mechanisms tested. As a result, we have are linked to an avirulence gene, were used to now obtained GUS expressing conidia which isolate cDNA clones from a library based on were developed from transformed basal cells, mRNA from germinating conidi a. For this indicating that some of the Egh hyphae are not purpose, a routine screen of cDNA libraries using only alive, but also conidiating after the particle

Ris0-R-972(EN) 39 gun shooting. With a view to obtaining stable obligate biotroph Erysiphe graminis f.sp. hordei. transformants, we have made a construction con ­ Gene 170 :131-135. taining the gpd promoter region in front of the Sherwood J.E., Somerville S.(\ (1990) Sequence of the Erysiphe graminis f.sp. hordei gene encoding 11- Neurospora crassa benomyl resistance gene, tubulin. Nucleic Acids Res. 18: 1052. benL K . 3.3 Seed Borne Diseases of Barley Characterization of the Egh G lyceraldehyde-3 - Phosphate Dehydrogenase Gene, gpd and Wheat (.S'.K. Christiansen. H. Cstergard) Few genes have been isolated from Egh and obligate biotrophis pathogens in general. The Stinking smut, a seed borne disease of wheat, sequence of the gpd gene is conserved across barley stripe and loose smut, similar diseases of species and allows for comparison of primary barley caused by the fungi Tillelia caries , structures, sequence similarity and evolutionary Pyrenophora graminea and IJslilago nuda , re­ studies. The complete gpd gene from Egh was spectively, have previously been controlled by sequenced and characterized (Christiansen el al. pesticides. Crop production without pesticide submitted). It contains typical promoter elements application is an important goal in modem and has 3 introns; one is positioned in the 5’ agriculture and a project on “Resistance Towards untranslated region of the gene. The deduced Seed Borne Diseases” has been initiated in amino acid sequence has 87% similarity to gpd collaboration with, the Danish Institute of Plant genes from other Ascomycete fungi. This is at the and Soil Science, Lyngby, and the Danish same level as previously estimated among these breeders. Riso is responsible for the identification fungi. Comparisons at the DNA level reveal of molecular markers that can be used for similarities of only around 70% which is 10% identification of the stinking smut disease at an lower than previously reported. In an early stage of infection and for differentiating evolutionary tree based on the sequences from 18 isolates of the fungus. Samples of the Tillelia fungal gpd genes, Egh falls into the group of caries fungus from different geographical places Ascomycetes at a basal position. The regulatory in Denmark were obtained. Methods and media to region of the Egh gpd gene has no homology to grow the fungus artificially have been corresponding sequences in other filamentous established. Ascomycetes. In another project included in the Research Codon usage was determined for the four Centre for Organic Agriculture, Cereals and characterized Egh genes tub 2 (Sherwood and Pulses Group, the prospectives for resistant culti- Somerville 1990), Egh~ and Eghl6 (Justesen et vars in organic farming will be considered. At al. 1996) gpd (Christiansen el al. submitted ) and Riso, strategies for the use of resistance genes found to be similar for all four genes. The results towards stinking smut on wheat will be of the codon usage analysis suggest that Egh is developed in relation to modelling the population more flexible in the choice of nucleotides in the dynamics of the corresponding pathogen, Tillelia wobble position than other fungi. caries.

References 3.4 Disease Resistance in Barley and Borbye I... Giese If. (1994) Genome manipulation in Wheat (JH. Jorgensen) recalcitrant species: construction and characteriza ­ tion of a yeast artificial chromosome (YAC) library from Erysiphe graminis f.sp. hordei, an obligate Resistance from Different Sources fungal pathogen of barley. Gene 144:107-11. A collection of barley cultivars from Turkey and Christiansen S., Justesen A.E.. Giese II. Disparate three collections of wild barley (Hordeum sequence characteristics of the Erysiphe graminis spontaneum ), sampled at three locations in f.sp. hordei glyccraldchyde-3-phosphate dehydro ­ Turkey, were tested with selected powdery genase gene. Curr. Gen. Submitted. mildew isolates from Turkey and from the stock Justesen I. Somerville S.. Christiansen S., Giese II. (1996) Isolation and characterization of two novel collection at Riso. This was done by C. Lower, genes expressed in germinating conidia of the Justus-Liebig University of Giessen, Germany, as part of a COST-817 collaboration.

40 Riso-R-972(EN) In collaboration with the Nordic Gene Bank at References Alnarp, Sweden, and cereal scientists in the other Jorgensen J.H. (1996) Effect of three suppressors on the Nordic countries, the spring wheat accessions at expression of powdery mildew resistance genes in the gene bank were sown in the field and barley. Genome 39: 492-498. Thomsen S.B., Jensen II. P.. Jensen ./., Skou J.P.. evaluated for an array of traits, mainly morpholo ­ Jorgensen J.H. Localization of a resistance gene and gical. At Rise, where we got an early, uniform identification of sources of resistance to leaf stripe. attack of wheat powdery mildew, scoring for Plant Breeding. Submitted. powdery mildew reaction was also done. The gene for barley leaf stripe resistance, the 3.5 Stress-Related Barley Peroxidases so-called Vada resistance gene, has been named (S.K Rasmussen, B.K. Kristensen, P S. Nielsen Rdgla. It is located on barley chromosome 2 at a H. Kollist) distance of about 20 cM from powdery mildew resistance gene MILa. Eleven barley cultivars/ Plant peroxidases are monomeric enzymes con ­ lines selected for leaf stripe resistance have taining a heme-group, stabilized by four disulfide previously been tested with different leaf stripe bridges and containing calcium. Most peroxi ­ isolates. The results are now reported and they dases are glycosylated, influencing the mobility indicate the presence of three genetically different in polyacrylamide gels. Hence, molecular weights sources of race-specific resistance in addition to between 30 to 45 kDa have been assigned to gene lidgla (Thomsen el al ., submitted). peroxidases analysed from a wide range of plant During 1996, a country-wide research and species. With regard to their subcellular localiza­ development project, The Cereal Network, was tion, it is possible to divide peroxidases in two initiated. One sub-project, “Resistance to Leaf groups, cell wall peroxidases and vacuolar Diseases of Barley and Wheat ”, investigates eight peroxidases, presumably participating in distinct diseases. Potential sources of primarily partial physiological processes. Cell wall peroxidases resistance are traced and seed samples brought catalyze the last step of lignification, crosslinking home for muliplication. In collaboration with the of structural wall proteins, and crosslinking of Danish Institute for Soil and Plant Research, phenolic compounds coupled to carbohydrate Lyngby, the respective pathogen populations are polymers in the wall. Besides this, cell wall sampled, characterized and multiplied so that peroxidases can oxidize ascorbic acid and thereby representative pathogen populations can be used protect against pollutant oxygen radicals. The to test the plant samples. physiological function of the vacuolar peroxi ­ dases is still obscure. However, they are capable Mutational Dissection of Powdery Mildew of utilizing the same artificial substrates as the Resistance Gene Mia 12 cell wall peroxidases, although with variable In a mutation study, several more or less powdery efficiency. mildew susceptible mutants were identified. Most of them carried mutations in gene M!al2. Three Vacuolar and Extracellular Localization of the mutants had mutated in other genes, presumably twoStress-Related Barley Peroxidases Prx7 and genes that are involved in the plants' defence Prx8 in Transgenic Tobacco reaction. The effect of these three "suppressor" Two barley peroxidases, Prx7 and Prx8, are genes on the phenotypic expression of different expressed in barley leaves infected with the powdery mildew resistance genes was tested powdery mildew fungus. We are currently (Jorgensen 1996). The results showed that some characterizing their physiological function during resistance genes remained unaffected by the the infection, with the aim of exploiting this suppressors, whereas others were changed in knowledge in the breeding of future culture phenotypic expression. This implies that different plants. Their cDNA sequence indicates that Prx8 resistance genes employ different, but over ­ is a cell wall peroxidase, while Prx7 presumably lapping, spectra of defence processes, or signal is localized in the vacuole. Using transgenic transduction pathways. tobacco plants, we showed that Prx8 is sent to the cell wall and that the N-terminal signalpeptide is cleaved before secretion, similar to barley leaving

Ris0-R-972(EN) 41 a pyroglutamate as the amino terminal residue. and the distinct 10 kD polypeptide is in progress The glycosylation level in barley and tobacco is with respect to the induction pattern and their the same, and tobacco proves an attractive system function. for the production of recombiant plant peroxidases (Kristensen et al. in press). Similarity, transgenic tobacco, expressing Prx7, was produced to support studies of subcellular localization and to make tobacco lines expressing Egh sporer 2. germtubc I germtubc Cell wall both peroxidases by crossing. Vacoules were prepared from leaf protoplast from tobacco lines expressing Prx7, and western blot showed that Vesicular transport Prx7 is located in the vacuoles. This presumably Golgiapparatuiggl shows that the C-terminal extension, present in Prx7, serves as a vacuolar targeting signal. Attempts to prepare vacuoles from barley leaves x * failed, but tissue separation in combination with immunodetection showed that Prx7 is expressed Barley empidermal cel in the epidermis, while mRNA studies showed that Prx8 is expressed in the mesophyll. These two peroxidases serve as a valuable tool for studying the different roles of distinct peroxidases in the pathogen defense response of Figure 3.5.1 A naive drawing of the epidermal the plant. The first three crystal structures of plant cell layer. The cellular localization of Prx' and peroxidases have been resolved. Among these PrxH in the tissue 12 hrs after inoculation of a three is the barley seed peroxidase BP1 barley leaf with a fungal spore. (Henriksen et al. 1996) which presumably is a vacuolar peroxidase as is Prx7, because BP1 also has two signalpeptides in the preprotein Effects of Ozoneon Ascorbic Acid in Harley (Rasmussen et al. 1991). The systematic and Leaves sequencing of all expressed peroxidases in the It has been demonstrated that the grain yield in model plant Arabidopsis thaliana has identified spring barley compared to spring wheat, is less two new classes not previously found (Kjaersgard influenced by ozone. As a step in understanding cl at. in press), thus providing new insights into the physiological-biochemical basis of this the structure-function analysis of plant peroxi ­ difference, the influence of ozone on the diurnal dases course of ascorbic acid in cell walls was measured in barley and wheat. This was done in Differential Display of mRNA in Infected the open top facilities as a joint project between Harley Leaves DMU og Riso. The ascorbic acid levels in leaf In a search for genes, expressed following cell walls of ozonated barley were found to be infection of barley with powdery mildew fungus, more stable compared to the levels in wheat. In we did a differential display RT-PCR analysis addition, ozone flux to the plasmalemma in with epidermis RNA isolated from infected barley appeared to be less, due to lower stomatal versus uninfected plants. Twelve PCR fragments conductance and thicker cell wall, while ascorbic were isolated as putatively differential expressed acid had negligible influence (Kollist et al. 1996). genes, cloned, and characterized by sequencing. Whether these differences between species in the Data base search identified a ribosomal protein, a early stage of plant development may explain the 10 kD photosystem II protein and a third with no reduced effect of ozone on grain yield in barley, known function. The analysis of the 10 kD remains to be seen. photosystem II protein showed that at least two forms are present in barley leaves (Nielsen and Rasmussen 1996). The analysis of the third clone,

42 Ris0-R-972(EN) References shown to segregate for resistance. Finally, Ilenriksen A., Welinder K.G., Gajhede M (1996) The integration mapping is being carried out to Structure of a barley grain peroxidase at basic pH. combine results from different mapping experi ­ In: Plant Peroxidases. Biochemistry and Physiology . ments and disease resistance tests. IV international symposium 6-10 July 1996. Univer- sitat fur Bodenkultur, Institut fur Chcmie. Wienna, Two DNA sequences, one specific for virulent Austria. Abstract. isolates and one specific for weakly virulent Kjcersgard I. VII., Jespersen H.M., Rasmussen S.K., isolates of Leptosphaeria maculans were IVelinder K.G. Sequence and RT-PCR expression characterized. Primer pairs were designed for analysis of two peroxidases from Arabidopsis both sequences and used on a selection of thaliana belonging to a novel evolutionary branch of isolates. Disease tests were performed to identify plant peroxidases. Plant Mol. Biol. In press. highly- and weakly-virulent isolates. Analysis for Kollist //., Afoldau II, Afortensen /,., Rasmussen S.K., Bolt-Jorgensen L. (1996) Ascorbate levels and pigment production, typical for avirulent isolates, ozone decay in cell walls of barley and wheat was determined using the Czapek-Dox test. PGR seedling. In: C. Obingcr. U. Burner, R. Ebcrmann. based analysis for the presence of a sequence C. Pcnel. H. Greppin (cds.) Plant Peroxidases. shown to be specific for highly virulent isolates Biochemistry and Phy siology . University of Geneva (Taylor and Borgmann 1993) was also carried Press. Geneva. Switzerland, pp. 358-362. out. There was a good correspondance between Kristensen B.K., Brandt ,/., Bojsen A'., Thordal- Christensen If., Kerbv K.B., Collinge D.B., virulence characteristics and the presence or Mikkelsen J.D., Rasmussen S.K. (1997). Expression absence of the cloned sequences, but a few of a defence-related intercellular barley peroxidase isolates gave variable results and are being re­ in transgenic tobacco. Plant Science 122: 173-182. tested. Kristensen B.K., Rasmussen S.K. (1996) Expression of In conclusion, there appears to be significant the stress-related intercellular barley peroxidase differences in the genomes of weakly- and Prx8 in barley and transgenic tobacco. In: C. Obingcr, U. Burner, R. Ebermann. C. Pencl. H. highly-virulent isolates of L. maculans. The Greppin (cds.) Plant Peroxidases: Biochemistry and cloned sequences may provide a quick tool for Physiology. University of Geneva Press, Geneva. the identification of isolates and may facilitate Switzerland, pp. 317-321. the development of a PGR based method for the Nielsen /US’, Rasmussen S.K. (1996) A barley estimation of infection severity. photosystem II associated 10 kD protein (Accession No. X97771) shows that at least two forms arc present in monocot plants (PGR96-108) Plant Reference Physiol. 112: 1398. Taylor ,/., Borgmann I. (1993) An unusual repetitive Rasmussen S.K., Welinder K.G., Hejgaard J. (1991) element from highly virulent isolates of Leptosphae ­ cDNA cloning, characterization and expression of ria maculans and evidence of its transfer to weakly an endosperm-specific barley peroxidase. Plant Mol. virulent isolates. Molecular Plant-Microbe Interac­ Biol. 16: 317-327. tions 7: 181-188.

3.7 Gene Mapping in Barley (H. Giese, J. 3.6 Genetic Analysis of Resistance to .Jensen, A. Heiberg) Leptospheria maculans (stat. con. Phoma litigant) in Oilseed Rape and Two crosses, one between the two-rowed spring Characterization of Fungal Isolates varieties, Alexis and Regatta, and the other (H. Giese, L. Butterworth) between the six-rowed winter varieties, Sonate and Marinka, have provided the basis for con ­ In collaboration with the John Innes Institute, structing genetic maps of the barley genome. U.K., genetic analysis is being carried out on a About 200 progeny lines in each cross were recombinant inbred population of oilseed rape scored using RFLP and RAPD markers using RFLP markers. In addition, a backcross A large number of traits were analysed for all population was tested for resistance towards spring barley progeny lines in the field at 3 Leptospheria maculans, both in the field and in the different localities by the Danish barley breeders, laboratory, at the John Innes Institute. Three field Sejet Plantbreeding, the Pajbjerg Foundation, and trials were carried out and the population was the Abed Plant Breeding Station. QTL analysis

Ris0-R-972(EN) 43 has identified loci which influence lodging, analysed selected lines for chromosome abnor ­ heading, necrosis, yield, resistance towards rusts malities and found some which show varying and Rhynchosporium, and the malting quality ploidy levels. traits, viscosity, protein, maltextract and RF value (see Figure 3.7.1). Microsatellite Markers for Identification of Barley Varieties and Mapping Microsatellite markers are now under develop ­ ment for barley and a project has been initiated in Barley chromosome 6 collaboration with the Alfred Jorgensen Labora ­ tory, Denmark. Forty commercially available MWG620 microsatellite primers were used to establish the 10.32 technique in the laboratory. A method, using F14-300 denaturing polyacrylamide gels and silver 6.02 IF3-400 staining, was established and the parents of the 14.13 mapping populations were screened for poly ­ 012-475 morphisms. Thirteen primer pairs were found to 12.21 be informative in the tested material. Q10-700 A PCR based cloning strategy, to facilitate 15.75 cloning of sequences bordering microsatellites, L08-1400 QTL for resistance was attempted. Many microsatellite sequences 14.34 towards leaf rust and were cloned, but too many cloning artifacts Rhynchosporium K14-1300 5.24 occurred to make the method reliable. MWG820 16.28 Localization of Barley Resistance Genes bBE54H towards Bipolaris sorokiniana 7.05 B. sorokiniana causes severe damage to barley bBE54B crops in Thailand and in collaboration with the 13.24 008-650 Kasetsart University, Thailand, an EU collabora ­ 19.50 tion project was initiated to identify disease- Psb56 resistant genes. Two barley lines from Thailand 5.10 AKB2 and BRB2R were crossed and 75 double MWG897 haploid, DH, lines from F, kernels were made. 7.20 The AKB2 line is a two-rowed early maturing K06-750 line which is drought and heat tolerant, but not Figure 3. 1 Location on chromosome 6 of a resistant towards spot blotch caused by Bipolaris OTL which controls resistance against leaf rust sorokiniana. The other line, RBR2R, is a late and Rhynchosporium in double haploid progeny maturing six-rowed line which is intolerant of lines from the cross between Alexis' and heat and drought, but resistant towards spot Regatta blotch. Fourty-two isolates of Bipolaris sorokiniana In collaboration with Nordic barley breeding were collected from different barley growing institutes, the Agricultural University, Norway, areas in Thailand. An aggressive and a non- and Svalof/Weibull AB, Sweden, winter hardi ­ aggressive isolate will be selected for disease ness, yield and disease resistance were assessed. resistance tests in the laboratory. Progeny lines Winter hardiness was tested in the Senate have been propagated and are currently in field Marinka cross, but as weather conditions were trials in Thailand. The 75 progeny lines were too severe, all lines died in Norway and the test analysed using 15 RFLP markers and 30 RAPD will be repeated. Rhynchosporium resistance tests markers. The cross is highly polymorphic and were also carried out in Norway and will be mapping will shortly be carried out. repeated to get results from a 2-year study. The Agricultural Research Institute, Iceland, has

44 Ris0-R-972(EN) 3.8 Mapping of the Brassica Genomes During guest visits from INRA, Le Rheu, (R.B. Jorgensen) France and SLU, Svalov, Sweden in January, February and April 1996, the in situ hybridization We have developed PCR-based DNA markers in technique was applied to B. napus, B. carnpestris , oilseed rape, B. napus , and the related species B. B. juncea , B. adpressa, Raphanus raphanistrum oleraeea, B. carnpestris (B. rapa) and B. juncea and some of their interspecific hybrids. Two (e g. Heneen et al. 1995, Mikkelsen et al. 1996, different ribosomal probes were used and made Jorgensen el al. 1995, Chen et al. a, in press and the identification of individual chromosomes b, submitted). The DNA methods applied were possible. RAPD and recently also microsatellites. The studies have revealed the extent of transfer of References genetic material, the introgression from oilseed Chen B.Y., Cheng B.F., Jorgensen R.B., Heneen W.K. (a) Production and cytogenetics of Brassica cam- rape to the related species, and a knowledge of pestris-alhoglabra chromosome addition lines. the importance in the evaluation of transgene Thcor. App. Genet. In press. transfer to wild relatives. Chen B.Y., Jorgensen R.B., Cheng B.F., Heneen W.K. We have produced a map of the nine (b) Identification and chromosomal assignment of chromosomes of the C-genome of oilseed rape RAPD markers linked with seed colour in a Brassica from five different backcrosses between oilseed campestris-alboglabra addition line. Hercditas. In rape and B. carnpestris ((B napus x B cam- press. Heneen W., Chen B Y., Cheng B.F., Simonsen I ., pestris) x B. carnpestris) reported by Mikkelsen Jorgensen R.B., Danik J. (1995) Characterization et al. (1996), Jorgensen et al. (1995), Chen et al. of the A and C genomes of Brassica carnpestris L. (a, in press and b, submitted). The oilseed rape and B. alboglabra Bailey. Hercditas 125: 251-267, involved in these crosses were both the modem Jorgensen R.B., Chen B Y., Cheng II. F., Heneen IV.K., and the old varieties (Drakkar, Westar and Topas, Simonsen I'. (1995) RAPD mapping of the Brassica > 100 plants per crossing combination) as well as alboglabra cromosome of a B. campestris- alboglabra addition line. Chromosome Research 4: a line of resynthesized oilseed rape made for 111-114. breeding purposes. Our marker-based maps for Kresovich S, Szewcmcfadden A.K., Bliek .S’. A/.. the five different crosses, showed that 32 of the McPerson J.R. (1995) Abundance and characteri ­ markers, approximately 1/3 of the applicable zation of simple-sequence repeats (SSRs) isolated markers, were general to all five maps, and from a size-fractionated genomic library of Brassica markers were often found in the same linkage napus L. (rapeseed). Theor. Appl. Gen. 91: 206-211. Mikkelsen T.R., Jensen J., Jorgensen R.B. (1996) combinations. Our results point to a genomic Inheritance of oilseed rape (Brassica napus) RAPD stability within parts of the C-genome, with markers in a backcross progeny with Brassica intergenomic, as well as intragenomic, recom­ carnpestris. Thcor. Appl. Genet. 92: 492-497. bination being frequent in other parts of the genome. Consistent RAPD markers, found in all 3.9 Quality of the Barley Grain five crosses, are being converted into SCARs for (S.K. Rasmussen, K.S. Johansen, FI. Brinch- easy and fast identification of synteny groups. Petersen) Microsatellites delivered by Cornell University, USA (Kresovich et al. 1995) are being integrated Barley grain can be regarded as a bioreactor with into the maps. unique properties, as it contains large amounts of Linkage between agronomic traits and DNA useful compounds and provides an environment markers has been established in collaboration for the stable storage of proteins. Among the with the Swedish Agricultural University, SLU in advantages of using barley, rather than fermen ­ Svalov. Genes encoding contents of erucic acid tation, are that plants are self-supplied with and oil, as well as the flower colour (relevant to energy and the production facilities are available insect damage) were mapped. Rise has been at the farms. Most of the proteins in the barley responsible for the mapping of traits by molecular grains are seed specific storage proteins with no methods and SLU for referring the marker metabolic function and they are deposited as linkage groups to chromosome number by proteinbodies in the cell. Some of this storage karyotype analysis.

Ris0-R-972(EN) 45 protein can be substituted with other proteins tory, Denmark. The amino acid sequence showed without a negative effect on the seed formation. It similarity to clones in the EMBL database from is our goal to investigate the potential for the oilseed rape and maize. No function has been production and storage of new protein in barley assigned to these two sequences. The maize grain by means of genetic transformation. Cereal phytase is a homo dimer of a subunit that starchy endosperm is one of the major food migrates as a 38kD protein in a SDS-gel. The sources in the world for humans and animals. Our amino acid composition of soybean, maize and success in fulfdling the future need for im­ the putative wheat phytase is similar. A titer of provement in food mass and quality, depends on antibodies raised against mung beans at the our insight into the molecular mechanisms University of Calcutta, India, recognizes the controlling grain filling. partially purified phytase, but not the Aspergillus phytase, Phytase Novo. Primers designed from Increasing the Phytase Activity in Barley the peptide sequences are currently used in a The availability of phosphorus in grains from coupled reverse transcription - PCR-based cereals is limited for monogastric animals, cloning of the cDNA or directly on genomic because the phosphate is stored in the molecule DNA phytate, wyn-inositol hexakisphoshate. Phytate is hydrolyzed by the enzyme phytase which Active Plant Serpins liberates phosphate and lower inositolphosphates. The recent discovery that plants express active Phytases have been purified from mung bean, serpins, serine proteinase inhibitors, was further soybean and maize, and a few from wheat, barley exploited by kinetic analyses of recombinant or rice. We hope to purify, sequence and clone a plant serpins produced in E. coli. In particular, phytase from wheat. the barley proteins Zx and BSZx were shown to be strong inhibitors of cathepsin G, chymotrypsin PO i '1 and trypsin, respectively, due to overlapping PO reactive sites (Dahl et al. 1996a). Trypsin from j >' / Fusarium was not inhibited by BSZx (Dahl et al. // / PO 1996b). The wheat chymotrypsin inhibitor WSZl po 7 1/ -1 1/ // (Rosenkrands et al. 1994, Rasmussen et al. PO / / PO 1996a) could also inhibit cathepsin G (Dahl et al. 4 1996). The dominant grain albumin in many barley varieties, protein Z4, shows some myo inositol hexakisphospate inhibitory action towards cathepsin G, but acts as a substrate when tested with chymotrypsin and trypsin, indicating that it is cleaved at the reactive Figure 3.9.1 Structure of phytin. The backbone is site bond. The primary structure of one of the myo-inositol with six phosphate groups and protein Z7 variants was analysed in detail by phytic acid, which can complex with Zn and ( a to sequencing the purified protein and by cDNA form the salt phytin. cloning (Rasmussen et al. 1996b). From this, it was shown that BSZ7 has an acetylated Ala as Extracts from wheat bran have a very high the N-terminus, is non glycolsylated, in contrast phytase activity and we have partially purified to mammalian serpins and is the only serpin from phytase from this source. The enzyme is very barley grains which can be isolated in an stable at 4°C and is inactivated at a pH >7.0. A uncleaved form. Future experiments will be number of chromatographic procedures have been directed towards elucidating if the cellular tried out with limited success in order to obtain a function is regulatory or protective. pure phytase. The partially purified phytase was subjected to Improving Grain Quality SDS gel-electrophoresis and selected bands were Transformation of the cereals barley and wheat cut out. The protein in this gel slice was digested will be used to improve the quality of the grain and peptides sequenced at the Carlsberg Labora ­ for fodder or baking and to support low imput

46 Ris0-R-972(EN) farming. Promotors for seed specific production Relation to Release of Genetically Modified have been isolated and characterized and are now Plants, which was brought to a successful ending included in vector constructs. Since this autumn, in May. 1996. The programme was a collabora ­ barley immature embryos are being bombarded tion with the University of Aarhus and the Royal with gold particles coated with the DNA Veterinary and Agricultural University, Copen ­ constructs and the regeneration of plantlets from hagen, and was part of the Danish Biotechnology callus-forming tissue is in progress. Research Programme. Riso participates in the evaluation of applications for EU marketing and References in the national releases of transgenic plants. Dahl .S’IF., Rasmussen S.K., Hejgaard J. (1996a) Heterologous expression of three plant serpins with distinct inhibitory specificities. J. Biol. Chem. 271: 2508.1-25085. Dahl .S'. If ., Rasmussen S.K., Petersen L.C., Hejgaard J. (1996b) Inhibition of coagulation factors by recombinant barley serpin BSZx. FEES Lett. 394: 165-168. Rasmussen S.K., Dahl S. It'., Hejgaard J. (1996a) A recombinant wheat serpin with inhibitory activity. Plant Mol. Biol. 30: 673-677. Rasmussen S.K., Klausen ./, Hejgaard J., Svensson B., Svendsen 1. (1996b) Primary structure of the plant serpin BSZ7 having the capacity of chymotrypsin inhibition. Biochcm. Biophys. Acta 1297: 127-130. Rosenkrands /., Hejgaard J., Rasmussen S.K., Bjorn S.K. (1994) Serpins from wheat grain. FEES Lett. 343: 75-80.

3.10 Risk Assessment of Genetically Modified Plants (1IB. Jorgensen, IP. Hauser, L. Landbo, T.R. Mikkelsen, B. Andersen. Figure 3.10.1 B. campestris (the small plant in E. Ibsen, H. 0stergard) the centre) is a rather common weed in winter oilseed rape fields. A transgenic crop may transfer the transgene to other plant species in natural or cultivated Analysis of Spontaneous Hybridization and environments through hybridization with wild Backcrossingbetween Oilseed Rape and B. relatives. The impact of a transgene introgressed campestris to a wild species is dependent on the transgenic Our previous data indicate that spontaneous character, as well as on the biology of the wild hybrids between oilseed rape and weedy Danish species. If a transgene provides an increased B. campestris are easily obtained. The hybrids fitness to the plant, it may be able to invade also backcrossed to the wild species in the field. natural ecosystems and affect their composition This has been reported in Jorgensen (1996), of species. The purpose of the research is to Jorgensen et al. (1996), Mikkelsen et al. (1996a) elucidate interactions between transgenes, the and Landbo et al. (1996). Seeds harvested on recipient plants and the ecosystem. This is individual B. campestris plants, from an obtained by studying fundamental mechanisms in additional number of natural populations, were plant population biology using molecular germinated and checked for the presence of genetics. Oilseed rape and its related wild species interspecific hybrids. Hybrids were not found in Brassica campestris and B. juncea are used as seed lots from autumn-flowering B. campestris. model systems. Part of the research has been In summer-flowering B campestris, as a weed in carried out in the programme, Risk Assessment in fields of oilseed rape, we found hybrids, once again confirming the spontaneous hybridization (Landbo et al. 1996).

Riso-R-972(EN) 47 B. campestris and interspecific hybrids, Spontaneous Hybridization between Crop produced by controlled crosses with two trans ­ Plants and Wild Relatives in Denmark genic lines of B. napus. tolerant to the herbicide In collaboration with the Danish Nature and Basta, were grown together to assess the extent of Forest Agency, a report has been produced from backcrossing. A large number of transgenic the available literature regarding the hybridization plants developed from seeds harvested on the between crop plants and wild relatives in hybrids, and some transgenic plants with a B. Denmark (Mikkelsen and Jorgensen, in press). campestris-Y\ke morphology were selected for This background information is needed in the further analysis. Most of the selected plants were evaluation of risks of transgene transfer from clearly backcross plants of which a few were genetically modified crops to related plants. almost identical to B campestris (chromosome number 2n=20, high pollen fertility) and had a Fitness of Hybrids between Weedy Brassica high seed set in crosses with genuine B. campestris and Cultivated Oilseed Rape, B. campestris (Mikkelsen et al. 1996a). We napus conclude that rapid gene flow from oilseed rape We study the fitness of hybrids between oilseed to B. campestris is possible. The distribution of rape and B. campestris with respect to environ ­ oilseed rape genetic markers in a backcross mental and genetic variation, in order to evaluate progeny (B. napus x B campestris) x B the possibilities for spontaneous introgression of campestris has been reported by Mikkelsen et al. transgenes and other genetic material from the (1996b). crop to the weed. To investigate at what life stages a possible Analysis of Spontaneous Hybridization between selection against hybridization takes place, and to Brassica napus and B. juncea study whether individuals, varieties and popula ­ Mixed populations of B. juncea and the oilseed tions differ in their ability to hybridize and rape variety, Drakkar, were sown. Interspecific backcross, two generations of hybridizations (F, hybrids were identified by isozyme analysis of and Fj/BCO were performed between different B plants from the seeds harvested from both campestris populations and B. napus varieties. species. In a 3:1 proportion between B. juncea In the first crossing generation, plants from and B. napus. we found 2.3% and 1.1% spontane ­ each of three weedy Danish B. campestris ous interspecific hybrids respectively with B. populations and three B. napus varieties were juncea and B. napus as the female. In a 10:1 intercrossed, and the fruit and seed set estimated. proportion, the percentage of hybrids was 1.1% The seeds/offspring plants were grown in the and 1.0%, and in a 13:1 proportion, the frequency field, and monitored for survival and repro ­ of hybrids was 0.3% and 1.3%. Previously, we duction. Analysis of the fitness data was reported 3% of interspecific hybrids harvested performed in co-operation with the University of from a small population of B. juncea growing in a Minnesota, St. Paul, USA The results show that B. napus field (Frello et al. 1995). Apparently, at some lifestages, hybrids have a lower fitness there was no effect from the proportion of B. than pure B. campestris , but at other lifestages a juncea plants on the frequency of hybrids with significantly higher fitness. Combining the fitness oilseed rape as the mother. In contrast, the components into an estimate of lifetime fitness, frequency of hybrids with B. juncea as the mother survival components multiplied by reproductive was dependent on the proportion between the components, indicates that the fitness of hybrids species. This is in parallel with the observations is intermediate to that of B. campestris and B by Hauser et al. (in press) in the combination B. napus , and significantly higher than that of B. napus x B. campestris. They found that there was campestris. This is in contrast to the common a more pronounced selection against hybrids, generalization that species-hybrids suffer from a when oilseed rape was the mother. low fitness. Significant differences were detected between hybrids from different combinations of B. campestris populations and B. napus varieties for several of the fitness components and for the combined fitness estimate. The propensity to

48 Riso-R-972(EN) hybridize in the field may thus vary, depending fields, field edges and fallow (Hauser et at. in on the varieties used and the local populations of press). the weed. In the second crossing generation, we have intercrossed hybrids, originating from crossings • B. campestris between two of the populations and two of the ■ B. napus varieties from the first generation, B. campestris , 0.8 - and B. napus plants to obtain F2, reciprocal backcrosses, and pure B campestris and B. napus seeds. Offspring plants were grown in the field in 0.6 - the summer of 1996. The same fitness com­ ponents as described above were estimated, and statistical analyses are presently being performed. 0.4 -i Preliminary results show that F2 and backcross plants survive and reproduce well in the field.

Together with the results from the F, generation, 0.2 - and other results from our research group, this strongly suggests that introgression of genes from oilseed rape to weedy B. campestris is a likely process. The pollination environment may, in many Proportion of heterospecific pollen in mix ways, influence the fitness of hybrids in experi ­ Figure 3.10.2 Proportion of hybrid seeds that ments and in the field. In nature, stigmas of B. developed after the pollination of Brassica campestris and B napus will often receive a campestris and B. napus with different propor­ mixture of pollen from the two species. Competi ­ tions of heterospecific pollen. Observed pro­ tion between pollen of different parentage within portions of hybrids are shown by the data points the styles and/or competition between seeds (error bars indicate + 1 SF), and the proportions within pods could potentially diminish the fitness of hybrids predicted by a statistical model are of hybrids relative to when only pure B. indicated by the broken lines. The unbroken line campestris or B napus pollen is deposited, as in indicates the proportions of hybrids expected if most experiments. To study whether pollen or no selection takes place for or against hybrids. seed competition influences the relative fitness of hybrids, different mixtures of B. napus and B. Expression of Transgenes in a New Genetic campestris pollen were applied to stigmas of both Background species. By comparing the applied proportions of Hybrids were produced by controlled crosses heterospecific pollen to the proportions of between (as female) a transgenic variety of hybrids found in the seeds, determined by oilseed rape (male-sterile, BASTA-resistant) and isozymes, we detected a significant deficit of individuals from a wild Danish population of B. hybrids (Figure 3.10.2). Analysis of the data campestris. Such transgenic hybrids were com­ showed that B. campestris pollen is selected pared with transgenic oilseed rape plants with against in the styles of B napus, and that hybrids respect to the phenotypes produced by transgene survive less in pods of mixed parentage than in expression, i.e. male-sterility and BASTA pods from pure pollinations. Hybrids in B. tolerance. No differences were found between campestris pods survived better than hybrids in hybrids and oilseed rape. Growing plants at B. napus pods, which is in contrast to the elevated temperature - which have been reported common finding that hybrids are easier to to break down male sterility, also in transgenic produce on B. napus. Our results thus indicate plants - did not produce any effect, neither in that the likelihood of producing hybrids in the oilseed rape, nor in hybrids. Approximately 20 field may vary depending on the relative backcross plants, made by crossing transgenic abundances of the two species, e g. between male sterile hybrids with B. campestris. were also investigated with respect to male-sterility and

Ris0-R-972(EN) 49 BASTA tolerance. All plants carrying the environmental risk? A case study on oilseed rape transgene were BASTA tolerant and male sterile (Brassica napus) and weed) Brassica campestris. (Mikkelsen 1996). Ph.D. Thesis. Rise National Laboratory and University of Copenhagen, 96 p. Mikkelsen T.R., Andersen B.. Jorgensen R.B. (1996a) References Spread of transgenes. Nature 380: 31. Frello S., Hansen K.R., Jensen J.. Jorgensen R.B. Mikkelsen T.R., Jensen ./., Jorgensen R.B. (1996b) (1995) Inheritance of rapeseed (Brassica napus) Inheritance of oilseed rape (Brassica napus) RAPD specific RAPD markers and a transgene in the cross markers in a backcross progeny with Brassica B. juncea x (B. juncea x B. napus). Theor. Appl. campestris. Theor. Appl. Genet. 92: 492-497 Genet. 91: 236-241. Mikkelsen I, Jorgensen R.B. Kulturafgrodcrs niuligc Hauser T.P.. Jorgensen R.B., Ostergard II. Preferential krydsningspartnere i Danmark. Skov- og Natur- exclusion of hybrids in mixed pollinations between styrelsen. In press. oilseed rape (Brassica napus) and weedy B. campestris. Amer. J. Bot. In press. Jorgensen R.B. (1996). Spontaneous hybridization 3.11 Seeds for Educational Purposes (J. between oilseed rape (Brassica napus) and weedy H. Jorgensen) relatives. Acta Horticulturae 407: 193-200. Jorgensen R.B.. Hauser 7., I.andbo L., Mikkelsen T.R., The Gene Technology and Population Biology Ostergard //. (1996) Transfer of engineered genes from crop to wild plants. Trends in Plant Science Programme offers seed samples of barley and 10: .356-358. other plant species for teachers in biology. The Landbo I... Andersen B. Jorgensen R.B. Natural samples illustrate Mendelian gene-segregations hybridization between oilseed rape and a wild and the effect of ionizing radiation on various relative: hybrids among seeds from weedy B. plant species. In 1996, a total of 1228 individual campestris. Hcrcditas. In press. experiments were performed. Mikkelsen T.R. (1996) Flow of engineered genes from crops to wild or weedy relatives - is there an

50 Ris0-R-972(EN) 4 Plant Nutrition and Nutrient Cycling

Introduction (G.G. Nielsen) pounds, including fertilizers in plant production. With the objective of improving the utilization of The working environment was considerably im­ accumulated phosphorous in the soil, we have proved at the beginning of the year. A new 200 m2 started a project on the genetic background in barley section of the laboratory building was inaugurated in for P-uptake. January, and the old part of the building was In order to fertilize the different parts of fields in renovated to a high standard. This made increased accordance with the natural variation in topography activities in the laboratories possible, including the and fertility, a system has been developed to conti ­ employment of more postdocs, and Ph D. students. nuously measure the grain yield and the position of The objective of the research programme is to the machinery in the field, so called computer-aided control plant uptake processes to a greater extent. farming. We have the necessary equipment, and are Thereby, management of the nutrient uptake by involved in a national programme to study and plants can be optimized. Our contribution is to evaluate the system as a means to improved plant obtain a better understanding of the biological and production. chemical processes in the soil-plant-atmosphere A number of polycyclic, organic compounds system. This will lead to a reduced requirement for may have a negative impact on the environment. We energy -consuming fertilizers and to a reduced loss are continuing our studies on the effect of azaarenes of nutrients. on plants. The microbial biomass in the soil is of funda­ In general, the programme of plant nutrition mental importance to the mineralization of nutrients deals with fundamental processes in the turnover of in plant residues and animal manure. Special atten ­ nutrients in the soil-plant-air system. These tion is directed towards processes involved in the processes govern the utilization of the nutrients and cycling of nitrogen and turnover of organic matter. thereby the potential loss to the environment. Nitrogen is essential for plants, but also a risk to the Through close co-operation with colleagues at Rise environment through leaching of nitrate and and at other research institutes, covering research volatilization of some other nitrogen compounds. ranging from a very basic to a more applied level, The study of these processes will provide the basic our results contribute to the development of a sus­ information needed for economical, ecological and tainable plant production. sustainable plant production. Symbioses between plant roots and microorga­ nisms are directly involved in the nutrient uptake by Summary of Projects plants. Nitrogen is provided by Rhizobium bacteria fixing atmospheric nitrogen in symbiosis with legu­ 4.1 Genetical Background for Plant mes. Our research in this area is particularly aimed Uptake of Minerals and Organic at studies on the exchange of signal components between bacteria and host plant across the peri- Compounds bacteroid membrane. The soil-plant transport of mineral nutrients, especially phosphoms, is 4.1.1 Genetical Background for Uptake of mediated by arbuscular mycorrhizal fungi in sym­ Phosphorus biosis with most plant species. In our programme, we deal with a number of important aspects of There is an increasing political demand for a mycorrhiza for plant growth, such as the influence reduced input of fertilizers in plant production. of soil-P status on the mycorrhizal transport of Danish farmers have used a surplus of organic and phosphorous, and the efficiency of different fungi. inorganic P-fertilizers, especially during the latest 50 There is an increasing public demand to cut years, resulting in a considerable amount of strongly down drastically on the input of chemical com­ fixed soil-P.

Ris0-R-972(EN) 51 With the perspective of identifying genes in microbial biomass mediates the transformation of C barley for a better utilization of soil-P, we have and N in organic matter which has been added to started a series of experiments to evaluate the soils, e g. as crop residues, animal manure, organic importance of different plant compounds on P fertilizer and waste, into inorganic compounds such uptake. Genetically well-known genotypes of spring as NH4 and C02. The C and N, which are not and winter barley were grown for 1 -2 weeks under mineralized, end up in more and more recalcitrant sterile and non-sterile conditions, and the P-mono- organic compounds, humus. However, the pro­ and P-diesterase, associated with the roots and cesses of nutrient and C02 release versus SOM released to the nutrient solution, were determined. formation depend on many factors, including the The results showed considerable differences quantity and quality of the organic matter added to between genotypes (Asmar and Gissel-Nielsen, in the soil. press). These, and similar studies, will be continued The aim of our research is to improve the und­ and the results compared with P-uptake from soil. erstanding of the biogeochemical cycling of C and When the P-uptake related compounds are N in agro-ecosystems, with special emphasis on the identified, the relevant genes will be mapped in N supply of plants. An improved understanding of relation to well known marker genes. the interactions between microbiological, biological, chemical and physical factors, determining SOM Reference formation and nutrient release, is required to predict Asmar Ml-'., Gissel-Nielsen G. Extracellular phospho- the release of nutrients from the soil with crop niono- and phosphodiesterase associated with, and demand and minimize unwanted losses of nutrients released by the roots of barley genotypes: A non­ from agro-ecosystems. destructive method for the measurement of the extracellular enzymes of roots. Biology and Fertility of Symbiotic N2 fixation by leguminous crops Soils. In press. constitutes an important N source in sustainable cropping systems, especially those without animal 4.1.2 Effect of POM on Plants. husbandry and/or no use of industrially fixed N, e g. organic farming systems. However, symbiotic N2 The study of the effects of N-PAH compounds, fixation is limited by a number of environmental azaarenes, on plants started in 1995 (Gissel-Nielsen factors. Our research aims at improving the and Nielsen 1996) and was continued in 1996, using knowledge of the interactions between soil, two azaarenes, acridine and quinoline, in a pot- fertilizer/ manure and biologically fixed N as N experiment with Italian rye-grass, rape, navew, and sources in cropping systems. We shall investigate cornflower. The azaarenes were added to the soil in how interactions are influenced by environmental concentrations of 0.1, 1, and 10 ppm. The quinoline factors and the competition between legumes and seemed quite harmless to the plants, but even 1 ppm non-legumes in intercrops. acridine slightly inhibited the effect on the plants, while 10 ppm had a pronounced effect. 4.2.1 Plant Residue Decomposition and N Conservation in Agro- Ecosystems Reference Gissel-Nielsen (/., Nielsen T. (1996) Phytotoxicity of Roots, straw and other crop residues are the acridine, an important representative of a group of tar principal energy and nutrient source for microbial and creosote contaminants, N-PAC compounds. growth and activity in agricultural soils. Polycyclic Aromatic Compounds 8: 243-249. Consequently, crop residues are an important resource in a sustainable cropping system for the 4.2 Biogeochemical Cycling of Carbon maintenance of soil fertility and the supply of and Nitrogen in Agro-Ecosystems (KS. nutrients from the soil to future crops. Jensen, P. Sorensen, P. Ambus, F. V Jorgensen, Besides contributing to the nutrition of P.K. Kristensen) subsequent crops and SOM formation, crop residues incorporated in the autumn may conserve soil N and Soil organic matter, SOM, is an important factor in reduce N leaching, due to immobilization of N determining soil fertility, including the capacity of during residue decomposition. However, the soils to supply plants with nutrients. The soil efficiency of crop residues in reducing soil N

52 Ris0-R-972(EN) leaching is dependent on their quality, quantity and barley residues reduced the leaching. However, the the way they are managed. particle size had no significant effect on the N- A small-lysimeter experiment was carried out to conserving effect of crop residues. determine the effect of pea, low C/N, and barley, Conventional crop residue management in high C/N, residue particle size: ground (< 3mm) or autumn is done by harrowing and mouldboard cut (25mm) on nitrate leaching during the autumn ploughing. Plastic tracers (Figure 4.2.1.1 a) were and winter. We hypothesize that a greater surface used to determine the spatial distribution of barley area and a better contact between residues and soil straw in the soil. Conventional crop residue would lead to greater immobilization of soil N. The management and tillage resulted in the placement of results showed that more N was leached from residues in strings 10-20cm in diameter in the 10- previously pea-cropped soil than from barley 15cm depth with about 40cm between each string cropped soil, and incorporation of both pea and (Figure 4.2.1.1 b).

Figure 4.2.1.1 a) Plastic tracers are spread at the soil surface with the straw, h) 3 months later, soil was excavated to determine the spatial distribution of tracers.

An incubation experiment showed that barley cattle is in urine, which is collected in animal houses straw, cut in 3cm pieces and distributed and stored in tanks under anaerobic conditions. homogenously in soil, produced 25% more C02 Urine is either stored alone or mixed with dung as a than that of whole straw. The net immobilization of slurry. N per gram of whole straw C added, was only 55% We carried out experiments to determine the fate of the straw fragments. Both effects are probably of urine N when applied to ryegrass in the field, due to the fungi and bacteria having less access to using 15N-labelled urine obtained from a sheep fed the straw C when whole. If the straw is incorporated labelled ryegrass (Sorensen and Jensen, in press). In more homogenously in the soil, the soil N a sandy loam soil, about 60% of the soil conserving effect of straw is likely to be greater. incorporated urine N was recovered in three grass cuts during 5 months of growth, whereas about 80% 4.2.2 Cycling of Animal Manure in Plant-Soil of an equal amount of urea N was recovered. In a Systems sandy soil, about 50% of both urine and urea N was recovered in the ryegrass herbage. When urine and There is an urgent need to obtain knowledge, which urea were applied to the soil surface before sowing can contribute to an improved utilization of nitrogen the grass, 20-25% less N was recovered, and when a in animal manure, especially from the organic urination in the grass was simulated, the herbage fraction, which is not plant available when supplied recovered 70% of the urine-N in three cuts to crops. More than 60% of the N excreted from (Sorensen and Jensen, in press). More N was lost,

Ris0-R-972(EN) 53 probably by NH3 volatilization, from the sandy soil inorganic N concentration, or indirectly due to and when urine and urea were applied at the soil increased grass growth, which will reduce fixation surface. due to competition for other plant growth factors About 40% of the urine and 20% of urea-N were (Jensen et al. 1996). immobilized in the sandy loam, whereas there was Due to methodological problems, it has not yet no difference (13-16%) in the sandy soil. The been possible to determine the effect of a dung pat immobilization of urine N was similar whether the on N2 in grass/clover. Using cross-labelling urine was applied alone, or in a slurry consisting of techniques and 15N-labelled sheep faeces to simulate labelled urine and unlabelled faecal N. The results a cattle dung pat, we report the first data on the indicated that the net mineralization of urine N was short-term effect of a dung pat on N2 fixation and similar to that of urea in a sandy soil, but only 75% plant uptake of dung N, in the border area of the pat of the urine N was net mineralized in a sandy loam in ryegrass/white clover mixtures and pure stands, in soil, when urine was applied prior to sowing. Thus, a container experiment with a controlled environ ­ the fertilizer value of urine-N is expected to be ment (Jorgensen and Jensen 1996). White clover lower than that of urea on finer-textured soils. fixed from 88 to 99% of its N and the dung pat N In order to assess the relationship between did not influence the percentage of clover N2 quality parameters of feed stuffs, ruminant faeces fixation in the mixture; there was only a slight and the mineralization of C and N from faeces in reduction relative to the pure clover treatment. The soil, we carried out an incubation experiment at amount of N fixed was not influenced by the dung 15"C with 20 different faeces samples during 12 pat N. weeks of decomposition (Kristensen 1996). The Using this technique, it was also possible to C/N ratio and the log transformed N concentration determine the fate of dung pat N in the controlled of faeces showed the best correlation with the net environment and in the field. After 16 weeks of mineralization of N, and the apparent digestibility of growth, 10% of the dung N was taken up in the organic matter was the quality parameter of the feed, grass/clover mixture and 57% incorporated in the which had the highest correlation with the faeces N soil by earthworm activity. In the field, only 4% of net mineralization after 12 weeks. In addition, the the dung pat N was recovered in a grass/clover work showed that there was a correlation between mixture during 13 months. The lower recovery in feed N concentration/ apparent digestibility and the the herbage was probably due to dry conditions in N concentration of faeces. Consequently, it is the field. It is concluded that N released from a dung possible to obtain a reasonable estimate of the pat is not likely to reduce the N2 fixation in the relative mineralization of ruminant faeces N from border area in the short term. data of feed N concentration and digestibility (Kristensen 1996). References Jensen E.S., Hogh-Jensen //., Jorgensen F.F., Schjoerring References J.K., Vinther /•/’ (1996) Management of biological Kristensen I’.K. (1996) The mineralization of faeces nitrogen fixation in grass/clover leys. In proceedings of nitrogen in relation to the feed and faces composition. the 11th IFOAM scientific conference. Copenhagen 12- M.Sc. Thesis. 42 p. 15 August 1996. Sorensen Jensen K.S. The fate of fresh and stored 15N- Jorgensen F.I., Jensen F.S. (1996) Does dung influence labelled sheep urine and urea applied to a sandy and a N; fixation in grass/clovcr mixtures in the short sandy loam soil using different application strategies. term? In: Legumes in Sustainable Farming Systems. Plant and Soil. In press. D. Younie (ed.) Proceedings of Occasional Symposium No.3(). Organized by British Grasslands Society, Aberdeen, Scotland, 2-4 September 1996. 4.2.3 Nitrogen Cycling in Grazed Grass/Clover Pastures 4.2.4 The Role of Grain Legume N2 Fixation in the N Cycling of Agro-Ecosystems During grazing of a grass/clover pasture, more than 80% of the N in the feed may be returned to the Grain legumes, such as pea and beans, fix N in pasture in animal excreta, with the major part in symbiosis with Rhizobium. Crops also take up urine. The recycling of this N in the pasture may available soil N, especially when grown in pure inhibit/reduce N2 fixation either directly due to high

54 Ris0-R-972(EN) stands. It is to be expected that crops will reduce soil N in the autumn. Over the long-term, crop their N2 fixation when they can take up soil residues are an important factor in maintaining soil inorganic N. Thus, the full capacity of the legume fertility and supplying plant-available N via for fixation is not utilized and the soil N source is mineralization. used inefficiently. A field experiment was carried out to evaluate Reference the possibility of reducing the availability of soil N Jensen E.S. Nitrogen acquisition by pea and barley and the during the growth of pea, by no-till or high C/N effect of their crop residues on available nitrogen for straw incorporation and by using ,5N isotope subsequent crops. Biology and Fertility of Soils. In press. dilution techniques. The pea yield was not influenced by soil tillage or straw treatment. The amount of N was not significantly reduced by 4.3 Root-Microbe Symbioses tillage/straw treatment, being on average 132 kg N/ha. The no-till treatment did not reduce the Future sustainable agricultural resource manage ­ estimated amount of soil N uptake in pea, but it was ment will depend on an optimal utilization of more than halved by straw incorporation. This plant-microbe symbioses. An extended and care­ indicates that soil N can be conserved by straw fully controlled use of beneficial microorganisms incorporation and the subsequent N immobilization, which directly aid the supply of nutrients to without having a negative influence on the pea yield plants, provides a visionary and attractive and N2 fixation. alternative to the use of energy-consuming The nitrogen acquisition by field pea and spring industrial fertilizers. Rhizobium bacteria and barley grown on a sandy loam soil, and the mycorrhizal fungi are beneficial soil microbes, as availability of N in subsequent sequences of a they can supply nitrogen and phosphoms to cropping system were studied in an outdoor pot ex ­ plants through symbiotic associations. Optimal periment. The effect of crop residues on the N- exploitation of these microbes must be based on a availability was evaluated using I5N-labelled resi­ thorough knowledge of the complex interactions dues. Field pea fixed 75% of its N requirement and between the organisms and the environment. the N2 fixation almost balanced the N removed with seeds. The barley crop contained 80% of the 4.3.1 Symbiotic Nitrogen Fixation 15N-labelled fertilizer N supplied and the N in (L. Rosendahl, P. Mouritzen, A. C. W. Simonsen, barley seeds corresponded to 80% of the fertilizer A.M. Scharff) N taken up by the crop. The uptake of soil-derived N, by a test N-catch crop of white mustard grown The symbiosis between legumes and Rhizobium in the autumn, was higher after pea than after bacteria enables the plant to benefit from the barley. The N uptake in the test crop was reduced nitrogen fixed by the bacteria. The bacteria by 27 and 34% after pea and barley residue incor ­ inhabit characteristic root nodules that are formed poration, respectively, probably due to N in response to an intimate molecular communica ­ immobilization. The dry matter production and tion between the symbionts. The central unit of total N uptake of a spring barley crop succeeding all nitrogen fixing root nodules is the pea or barley, with a period of unplanted soil in the symbiosome, which comprises the bacteroid autumn/winter, were significantly higher after pea imbedded in the peribacteroid space that is than after barley. The barley crop succeeding pea surrounded by the plant-derived symbiosome and barley contained 11 % of the pea and 8% of the membrane. The membrane interface constitutes barley residue N. The pea and barley residue N the main point of symbiotic regulation, as any constituted only 2.5% and <1%, respectively, of exchange of compounds and signals between the total N in the N-fertilized barley. In the short-term, symbionts must proceed via this interface. The the incorporation of crop residues was not capacity to transport nutrients and phyto ­ important in terms of contributing N to the hormones across the membrane interface has subsequent crop compared to soil and fertilizer N been demonstrated by in vitro experiments with sources, but residues improved the conservation of isolated symbiosomes (Rosendahl el al. 1992,

Ris0-R-972(EN) 55 Rosendahl and Jochimsen 1995). Environmental It was demonstrated that the [14C]-labelled stress, e.g. drought, has an adverse effect on NH4 analog methylamine, MA, was taken up symbiotic nitrogen fixation. Recently, we have from the incubation buffer in a time dependent demonstrated that drought stress affects the manner, when a membrane potential as well as a transport properties of the symbiosome mem­ pH gradient, were present. Analysis of MA brane and that the protein composition of the uptake by vesicles in a concentration range from symbiosome membrane is altered in response to 25mM to 150mM suggested the existence of two stress, which may indicate degradation of specific transport mechanisms. Both mechanisms showed symbiosome membrane proteins or an altered increased transport when a membrane potential biosynthesis of symbiosome membrane poly ­ was imposed and this makes it difficult to peptides (Pedersen ct al. 1996). The overall perform separate analysis of the individual communication between the involved partners in transport mechanisms. However, the data suggest plant-microbe symbioses is far from understood. that one mechanism cannot be saturated and Our research effort in this field aims at providing operates with large capacity in high concen ­ extended knowledge on the transport mechanisms trations, > 5mM. The other mechanism operating in the symbiosome membrane, characterization of in low concentrations, < ImM, can be saturated the proteins in the membrane interface, and and concentrates MA 3 to 4 times inside vesicles. knowledge on plant response to microbes of Uptake also appeared to depend on the presence variable nutritional benefit. of a pH gradient across the symbiosome membrane, high pH inside, with a maximal Identification of Transport Mechanisms in the transport at a pH gradient of 2. The strong pH Symbiosome Membrane gradient dependency may be an indication for co­ It is generally agreed that dicarboxylic acids, transport of NH4 and H via a symport in the especially malate, are the primary reduced carbon symbiosome membrane. Such a mechanism is source supplied from the host plant to support known to exist in roots of higher plants, where K bacteroid activity. In contrast, the chemical form is known to be co-transported with H via a in which fixed nitrogen is exported from the symport which is also capable of transporting bacteroid to the host plant cytoplasm remains a NH4\ Saturation in the symbiosome membrane, matter of controversy (see Streeter 1995). Since which operates in the low concentration range, the exchange of all compounds occurs across the may share many of the properties of this symport, symbiosome membrane, identification and whereas the mechanism operating in the high characterization of transport mechanisms in this concentration range is likely to be identical to the membrane is a prerequisite for understanding the NH4 channel, which has recently been regulation of molecular compound exchange in characterized in the symbiosome membrane of symbioses. soybean by patch clamp technique (Tyerman cl The technique for isolation of inverted sym­ al. 1995). biosome membrane vesicles, as described by Christiansen ei al. (1995), has been used for in Import of Plant Proteins to Different vitro experiments to identify and characterize Compartments of the Symbiosome NH4 transport mechanisms in the symbiosome Symbiosomes may be compartmentalized into the membrane of pea. The generation of an electro­ bacteroid cytosol, bacteroid membrane, peri- chemical gradient is a prerequisite for in vitro ion bacteroid space, and the plant-derived sym­ transport into isolated membrane vesicles. We biosome membrane. Each compartment of the have adapted a method by which K -gradients symbiosome has a distinct pattern of proteins across the vesicle membrane, together with the (Rosendahl and Jochimsen 1993). These proteins K-ionophore valinomycin, are used to generate must originate from the plant and/or the membrane potentials in the range 0 to -150mV bacteroid, as there is no protein synthesis in the (Figure 4.3 .1). The size of the imposed potentials interface between the symbionts. An in vitro was confirmed by measuring uptake of [14C]- protein translocation assay, with symbiosomes tetraphenylphosphonium. and 35S-methionine labelled plant proteins, has been established to determine the localization of

56 Ris0-R-972(EN) Figure 4.3.1 The uptake of ammonium by symbiosome membrane vesicles is facilitated by the generation of an electric potential across the membrane. The potential is generated when the potassium transporter “valinomycin " is added to vesicles with a high internal concentration of potassium. Valinomycin carries positively charged potassium ions across the membrane along the potassium gradient leaving a negative net-charge behind.

plant originated proteins inside the symbiosome. special fluorographic reagent for treatment of The radiolabelled proteins residing in different polyacrylamide gels. In addition, a new type of compartments were identified by polyacrylamide film that provides maximum resolution, clarity, gel electrophoresis followed by fluorography. and sensitivity for detection of 35S-methionine The plant protein translocation assay was has been introduced. optimized in a number of steps. Increased levels The optimized translocation assay reveals that of adenosine triphosphate, ATP, were included in radiolabelled plant proteins are primarily located the incubation experiments, as phosphorylation in association with the bacteroid membrane and may be essential to protein import into the sym­ in the peribacteroid space. No radiolabelled plant biosome. In order to suppress de-novo synthesis proteins are detected in association with the of radiolabelled proteins of bacteroid origin, an symbiosome membrane by the present procedure. inhibitor of bacterial protein synthesis, chlor ­ This may indicate that plant proteins, intrinsic to amphenicol, was added to the assay. Additional the symbiosome membrane are either not purification steps were included in the protocol synthesized in the present in vitro translation for the separation of symbiosome compartments, system or may not be integrated into the to avoid cross contamination between com­ membrane in this experimental procedure. partment fractions. The detection limit of radio- labelled proteins was improved by the use of a

Ris0-R-972(EN) 57 Effect of Symbiotic Microorganisms on Rosendahl L., Dilworth M.J., Glenn Adi. (1992) Phytoalexin Accumulation in Soybean Roots Exchange of metabolites across the pcribactcroid Plants possess a broad spectrum of basic defence membrane in pea root nodules. J. Plant Physiol. 139: 635-638. mechanisms, pre-established or induced, which Rosendahl Jochimsen B.U. (1993) Characterization render them resistant to most potential colonizing of proteins in the pcribactcroid space of pea root microorganisms. One of these defence mechanisms nodules. In: New Horizons in Nitrogen Fixation. R. involves the inducible accumulation of low Palacios, J. Mora, W.E. Newton (cds.) Proceedings molecular weight, antimicrobial compounds known of the 9th International Congress on Nitrogen as phytoalexins. A plant-microbe symbiosis is Fixation. Cancun, Mexico, 6-12 December 1992. Kluwcr Acad. Publ. p. 368. possible if plant defence responses are modulated in Rosendahl /,., Jochimsen B.V. (1995) In vitro indole-3 - a way that permits the microorganism to colonize acetic acid uptake in symbiosomes from soybean the plant tissue, or if the microorganism can (Glycine max L.) root nodules. Symbiosis 19: 99- overcome the defence mechanisms. 110. We have investigated plant phytoalexin accumu­ Streeter J.G. (1995) Recent developments in carbon lation in response to different symbiotic micro­ transport and metabolism in symbiotic systems Symbiosis 19: 175-196. organisms to elucidate whether phytoalexins have a Tyerman S.D., Whitehead [.!■, Day D A. (1995) A role in determining compatibility, and whether the channel-like transporter for NH., on the symbiotic symbiotic performance of the microorganism is interface of N:-fixing plants. Nature 378: 629-632. reflected in plant phytoalexin levels. Soybean plants, in combination with different root associating microfungi, covering the spectrum from mutualistic 4.3.2 Mycorrhiza (I. Jakohsen, M. Gavito. A. to parasitic symbionts, were used as model Johansen, J. Larsen, S. Ravnskov, P. Schweiger. organisms in a six-week study. Phytoalexin accu­ I. Thingstrup) mulation in roots was analysed at five harvest times by HPLC. The parasitic fungus induced a massive Symbioses between arbuscular mycorrhizal (AM) phytoalexin response in plant roots, and the fungus fungi and plant roots are important in the only colonized a small part of the root tissue. The maintenance of adequate nutrient uptake rates by the three studied isolates of mutualistic, mycorrhiza- plant. The root ’s external phase of the AM fungi forming fungi all colonized a major part of the plant extends into the soil volume and gains direct access root system, but stimulated plant growth to very to those nutrients which are transported to the root different extents. The different degree of plant by slow diffusion processes. The overall nutrient growth stimulation by the mutualistic fungi was, transport by the mycorrhizal fungi is a result of however, not reflected in the root phytoalexin levels, hyphal nutrient uptake from the soil, nutrient which at all harvest times were statistically translocation through the hyphae and nutrient indistinguishable from plant basal levels. This indi ­ transfer across the symbiotic fungus-plant interface cates that phytoalexin accumulation is not a mecha­ in the cortex cells. The nutrient quantities nism by which plants may circumvent interactions transported are influenced by the isolate of AM with inefficient mutualistic microorganisms. fungus (Jakobsen et al. 1992), by the host fungus compatibility (Ravnskov and Jakobsen 1995) and References by the abiotic and biotic environment (Abbott et al. Christiansen J.U.. Rosendahl /.., Hi del I .S'. (1995) 1995). Our major goal is to understand mycorrhizal Preparation and characterization of sealed inside-out function in relation to these factors. Such under­ pcribactcroid membrane vesicles from Piston standing is a prerequisite for future management of sativum L. and Glycine max L. root nodules by aqueous polymer two-phase partitioning. J. Plant the mycorrhizal symbiosis as a means of enhancing Physiol. 147: 175-181. the sustainability of plant production systems. Pedersen AL, Feldner U.C., Rosendahl /.. (1996) Effect of prolinc on nitrogenase activity in sym- Utilization of Mycorrhizas in Sustainable biosomes from nodules of soybean (Glycine max L.) Agricultur subjected to drought stress. J. Exp. Bot. 47: 1533- We investigate how the indigenous populations of 1539 AM fungi contribute to the P uptake by crops grown at a range of soil P levels. This information is

58 Ris0-R-972(EN) required in order to predict to which degree AM (Figure 4.3.2 2). This suggests that mycorrhizal fungi may contribute to the maintenance of fungi contributed significantly to the P nutrition of reasonable crop yields after reductions in the rates of the plant in plots where no P had been applied for 20 P fertilizer application. The experiments are carried years, and that the mycorrhizal fungi were less out in long-term P fertilization trials under the important at high levels of P fertilizer. Plots Danish Institute of Plant and Soil Science, Lyngby. receiving no P contained 31 ppm NaHCOr Linseed was grown at three rates of P application extractabie P, which is only 5-10 ppm lower than (Figure 4.3.2.1) maintained over 20 years with half the average for Danish arable soils. These results of each plot fumigated with Basamid, a general confirm those from our previous field studies, where biocide, before planting, to prevent the development dry matter yield of mycorrhizal linseed was only of mycorrhizas. Mycorrhizal fungi colonized 42% marginally influenced by soil P levels ranging from of the root length in untreated soil when no P had 20-55ppm P (Jakobsen et al 1994). In conclusion, been applied and the colonization decreased with we have shown that indigenous populations of increasing P application (Figure 4.3.2.1). Basamid mycorrhizal fungi in Danish field soils can almost completely prevented mycorrhiza formation compensate for significant reductions in the (Figure 4.3.2.1). Mycorrhizal plants had a high P application of P fertilization. The mycorrhizal fungi uptake which was only slightly affected by rate of P are, therefore, an important biological resource application (Figure 4.3.2.2). In contrast, P uptake which deserve particular attention in the devel­ was markedly reduced in plants grown without opment of nutrient management strategies in mycorrhizas at the two lowest P application rates sustainable agriculture.

Phosphorus application (kg ha "1 yr"1) I - Basamid EH + Basamid Figure 4.3.2.1 Mycorrhiza formation in roots offield grown linseed at flowering is reduced by P fertilization and Basamidfimigation.

Riso-R-972(EN) 59 Phosphorus application (kg ha "1 yr"1) • -Basamid o —o +Basamid figure 4.3.2.2 Phosphorus content of field grown linseed at harvest: interactive effects of phosphorus fertilization and Basamid fumigation.

The Use of Excised Root-External Hyphae for can utilize amino acids as a source of nitrogen studying Metabolismand Nutrient Uptake in and the carbon skeleton in amino acids as a Mycorrhizal Fungi carbon source for respiration. The respiration rate The mechanisms underlying hyphal nutrient up ­ was constant during the two hours of measure­ take and metabolism are difficult to study in AM ment and this suggests that excised hyphae are fungi which cannot be grown in pure culture. suitable for short term in vitro studies. This obstacle can be partly overcome by using Evidence for the ability of AM fungi to utilize external hyphae extracted from the sand sur­ soil organic P is scarce and it has remained rounding mesh bags which enclose the unclear whether the external hyphae possess mycorrhizal roots (Johansen et at. 1996). external phosphatases to help in the mobilizing of An in-vitro incubation method was used to P from organic sources. To clarify this, p- study whether AM fungi can use amino acids as a nitrophenyl phosphate was used as a substrate for source of nitrogen and carbon. Excised external excised external hyphae of G. intraradices and G. hyphae of G. intraradices and G. claroideum claroideum in an in-vitro incubation experiment. accumulated l4C activity when incubated with The phosphatase assay was performed in a l4C-labelled aspartate or glutamate. The supply of modified universal buffer liquid medium with pH FCCP, which disrupts the membrane proton ranging from 4.0 to 8.8. Phosphatase activity, gradient, decreased the amino acid uptake by measured as the rate of substrate hydrolization, 95%, suggesting that amino acids are actively was detected at all pH levels and with both fungal transported across the fungal membrane. C02, isolates, but with a single maximum at pH 5-6 respired by the external hyphae of G claroideum, (Figure 4.3.2.3). The phosphatase activity in G. was collected in NaOH and 14C activity appeared intraradices was approximately twice as high as in the C02 traps a few minutes after the hyphae that observed for G. claroideum at all levels of were supplied with 14C-labelled aspartate. These pH. This is, therefore, a strong indication that results indicate that external hyphae of AM fungi external hyphae of AM fungi possess an external

60 Ris0-R-972(EN) acid phosphatase, whereas alkaline phosphatase culmorum did not affect hyphal growth and P seems to be absent, or at least not expressed, transport of G. intraradices. Phospholipid fatty under present experimental conditions. acid profiles were used to study mycelial interactions between G. intraradices and F. culmorum and their effects on soil bacteria. Apparently, the two fungi did not interact, but they both increased the total number of soil bacteria (acridin orange direct counting) and the G. intraradices amount of bacteria-specific phospholipid fatty G. claroidum acids. The mechanisms involved have not yet j 0.6 - been clarified, but the enhanced growth of the soil bacteria could have been caused by carbon compounds released by living or decomposing hyphae. A gene-transformed P. fluorescens isolate E 0.2 expressing luciferase under phosphorus starvation is used to study the effect of AM fungi on the phosphorus status of P. fluorescens. We also focus on the interactions between AM fungi and the saprophytic fungus Trichoderma harzianum which is often antagonistic towards other micro­ Figure 4.3.2.3 Activity of external phosphatase organisms. This study comprises a gene- in external hyphae of two mycorrhizal fungi as a transformed T. harzianum isolate, which consti- fmction of medium pH. tutively expresses glucoronidase and which can, therefore, be used as a marker of the presence and Interactions between AM Fungi and other Soil the physiological status of the fungus. Microorgan ism The root external phase of AM fungi co-exists Effects of Pesticides on Mycorrhizal with a broad range of other soil microorganisms Functioning and competition for inorganic nutrients and space Potential negative side-effects of pesticides on might occur. These interactions and their possible AM functioning are studied as part of a national influence on nutrient transport by the AM fungi cooperative research project entitled, Pesticide are studied in collaboration with RVAU, Effects on the Agricultural Ecosystem. The Copenhagen and Lund University, Sweden. Com- pesticide effects were tested by means of a model partmented growth systems with root-free soil growth system consisting of a root compartment compartments are used, such as interactions and a root-free hyphal compartment (Kling and between the external mycelium of AM fungi, and Jakobsen, in press). other soil microorganisms can be studied without Pea plants were grown in an agricultural soil interfering effects from roots. from an organic cropping system and their roots Previously, numbers of the plant growth pro ­ became colonized by the AM fungi naturally moting rhizobacteria Pseudomonas fluorescens occurring in the soil. The plants were grown for were found to increase in the presence of an five weeks, at which time external hyphae of the external mycelium of the AM fungi Glomus AM fungi had spread throughout the root-free intraradices and G caledonium. In that experi ­ hyphal compartment. Solutions of varying ment, the hyphal length of the two AM fungi was pesticide concentrations, ranging from well below unaffected by the presence of P. fluorescens. to well above the recommended field dosage, Recently, another study revealed that the P were applied to the hyphal compartment as a soil transport by the external mycelium of the same drench. An aqueous solution of 32P was applied two AM was unaffected by the presence of P. immediately after. The fungicides carbendazim, fluorescens. Similarly, in a third experiment, the propiconazole and fenpropimorph and the presence of the saprophytic fungus Fusarium insecticide dimethoate, all contained within their

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Johansen A., Finlay R.D., Olsson P.A. (1996) Nitrogen consisted of 13 slightly damaged trees, 30 trees metabolism of external hvphae of the arbuscular with midlevel damage and a needle loss of 26- mycorrtiizal fungus Glomus intraradices. New 60%, and 7 heavily damaged trees with a needle Phylologist 133: 705-712. loss of 61-99%. There were more than 10 dead Kling M, Jakobsen 1. Direct application of carbendazim and propiconazole at field rates to the external hyphae trees and markings had disappeared due to bark of three AMF species: Effects on 3"P transport and SDH loss. Needle samples from the three plot activity. Mvcorrhiza. In press. categories, taken from the 6th whorl at minimum Ravnskov S., Jakobsen /. (1995) Functional compatibility metabolism in December, showed arginine in arbuscular mveorrhizas measured as hyphal P contents of 0.16-0.3 gram kg"1, similar to that of transport to the plant. New Phylologist 129: 611-618. nondamaged trees and of the controls in pot experiments with balanced nutrition. This is very 4.4 Plant Constituents different from the situation in N-overloaded plants. Thus, there is no indication in the amino 4.4.1 Forest Decline, Ammonia Stress and acid composition that the diseased trees contain Amino Acids (K.C. Engvild, P.B. Holst) excessive levels of nitrogen.

The “ammonium hypothesis ” of Nihlgard (1985) postulates that certain types of forest decline are caused by an excess of ammonia from the air. There is one well-documented case in the Netherlands where Scots pine became strongly chlorotic (Van Dijk and Roelofs 1987). In 1989, a new disease “rode rodgraner ” or “red decline ” of Norway spruce became a serious problem in 100 - plantations in Jutland on poor, sandy soils. Some trees became red and lost all their needles over 2- 3 years with trees on the peripheries having the highest levels of damage; symptoms began at the tips of current year shoots. In some places, more than 10% of the trees have died, and much premature logging has been necessary. 10-25% 26-60% 61-99% 0 12 4 8 The only important pollutant in mainly rural NEEDLE LOSS ADDED N x NORMAL, NH4N03 Jutland is ammonia from the many farms, Figure 4.4. i Arginine content in spruce over­ resulting in N depositions of between 10-30 kg N loaded with N and in spruce with “Red ” decline ha"'year -1. Hence, the possibility that “rode rod­ symptoms. graner ” could be caused by nitrogen overload was investigated. Investigations of inorganic nutrients There have been many other hypotheses for in diseased trees did not show significant differ­ the “rode rodgraner ” disease: salt damage, ences (Saxe 1993). A more sensitive measure of nutrient leaching from needles or soil, acid rain, nitrogen status of the trees is the content of free drought, mild winters, and the combined effects urea cycle amino acids, arginine and ornithine. In of many stresses. No hypothesis has been proven pot experiments with a four times overload of or disproven, except that there is no indication of ammonium nitrate, the arginine content increased pathogens or pests. We felt that we had to more than 100 times (Figure 4.4.1). At the same abandon the ammonium hypothesis as an time, the needles reddened and fell off; the explanation. So what may be the cause of “rode pattern was, however, different from “rode rodgraner ”9 In the U K. there is a common rodgraner ” because the oldest needles fell off disease called “top dying”, which actually limits first. the planting of Norway spruce. This disease We investigated trees in an observation plot of shows “spectacular browning or reddening of the “rode rodgraner ” at Feldborg, which had also foliage ... the needles brown downwards from the been investigated by Saxe (1993). The plot shoot tip ”. It is believed that this disease is caused

Riso-R-972(EN) 63 by several mild windy winters in a row. In pea fertilized with nitrate. Experiments with 36C1 Denmark, we have had 8 mild winters in a row labelled rape seeds are also in progress. since 1987. The simplest hypothesis is to assume that “top dying ” and “rode rodgraner ” are the Reference same disease. If that is the case, then it should EngvildK.C. Herbicidal activity of 4-chloroindolcacctic become much less serious after the hard winter of acid and other auxins on pea. barley and mustard. 1995-96 and the conditions should improve even Physiol. Plant. 96: 333-337. more, if further cold winters follow. 4.4.3 Agrostemma githago as a Biomass Crop References Eng\’ild K.C. (1996). Sidste ar med "rode rodgraner” ? There is a need for an annual biomass crop which Skovcn 28 (8). 356-357. can be used for fuel. There are good permanent Sihlgdrd IS. (1985) The ammonium hypothesis - an crops, such as poplars and elephant grass. The additional explanation to the forest die-back in use of annual crops such as wheat, rye or triticale Europe. Ambio 14: 2-8. for burning meets with considerable political Phillips D.H., ISurdekin DA. (1982) Diseases of forest resistance, as does the use of cannabis. During and ornamental trees. MacMillan, London, pp. 134- 137. investigations of weed biology, Doll found quite Saxe II (1993) Triggering and predisposing factors in high yields of Com cockle (Agrostemma githago, the "Red” decline syndrome of Norway spmee klinte). Yield results of up to 15 ton of biomass iPicea ahies). Trees 8: 39-48. (at 67 kg N ha "’) were confirmed and we have van Dijk UFA',.. Roelojs J.G.M. (1988) Effects of seen yields up to 20 tons ha" 1 in single plots in excessive ammonium deposition on the nutritional Agrostemma sown in the fall. We continue to status and condition of pine needles. Phys. Plant. 73: experiment with fertilizer levels, sowing density, 494-501. varieties (from the Gatersleben gene bank) and 4.4.2 Natural Chlorinated Compounds and winter versus spring sowing, as well as experi ­ their Biology menting with minimum tillage, sowing Agrostemma directly in stubble. The natural chlorinated auxin in pea, 4-chloro- indoleacetic acid, was investigated for herbicide 4.5 Climate Change Experiment activity; it has a strength about one fifth of the strength of 2,4-D in pea and mustard. 4- 4.5.1 Climate Change Experiment, CLIMEX chloroindoleacetic acid is found in the immature (L. Rasmussen. C. Beier) pea seeds in concentrations corresponding to a 50% reduction in growth. This is consistent with In 1996, Riso joined the EU project CLIMEX, the idea that the chlorinated auxin may be a which is an integrated, whole-ecosystem research senescence factor of the developing seeds which project studying the response of entire forested weakens the mother plant. catchments to increased C02 and temperature in Experiments in labelling barley with radio­ relation to the foreseen global climate change active ,6C1 have confirmed that Cl is incorporated problems. Six research teams from different into organic compounds in barley. The very low countries, U.K., Norway, Sweden, The Nether ­ specific activity of 36C1 makes it necessary to lands and Denmark, work at the same field site in detect activity down to 10-100 counts min" 1 on southern Norway. Two experimental plots autoradiographs. This has been achieved by covered with large greenhouses, 1200m2 over the adapting the technique of preflashing the film, forest, are used to simulate atmospheric and exposing it at -80° C using hyperscreens. This conditions with increased C02 concentrations has reduced necessary exposure times from 2-3 and air temperatures. The objectives of the months to 3 days. Experiments are under way to project are to measure changes in plant C02 search for the chlorine-containing amino acid uptake and gas exchange, and to measure changes which should be formed in the nodulated roots of in forest growth and biogeochemical cycling of elements in the ecosystem. An ultimate purpose

64 Riso-R-972(EN) of the project is to develop and validate process- nitrogen was labelled with 15N tracer to oriented models linking terrestrial and aquatic investigate a possible preferential uptake of responses to global change, and up-scale the nitrogen. The effect of increased nitrogen load to results to an evaluation of the importance of the forest ecosystem is being studied by the global change for European forests. The project sampling of soil water and needles. started in 1992 and is planned to run until 1999. The N application has caused higher concen ­ trations of NOf and sometimes of NH4 in single samples of soil water. A general and significant 4.5.2. Nitrogen Circulation in Spruce Forests effect on soil water concentrations ofnitrogen was ((1 Beier) not seen (Figure 4.5 2.1). However, from the data, it is very difficult to judge the effect of the Forest ecosystems are very often growth limited N addition on the N leaching because of the very by the availability of nitrogen. However, dry weather conditions at the site in 1995 and increased nitrogen deposition may lead to 1996. The drought caused the sampling of soil leaching of nitrogen to ground and surface water to be very discontinuous. The sampling of waters. The potential problem of nitrogen soil water in the spring and summer of 1996, in saturation is being studied in a spruce forest at combination with the needle analyses from Klosterhede in Western Jutland. 50 kg N/ha/yr autumn 1996 and the l5N analyses, will hopefully (NH4N03) were applied manually to the two clear up some of the uncertainties. study plots in April-June 1995. The applied

0.03 Tree A1 0.03

0.02 0.02

■ \ ■ 0.01 ■ 0.01 ■ \ ■ \ -l_ ♦------A • ▲ w w w w WWWA W WW'i llW------A—A A------* —▲------apr- okt- apr- okt- apr- apr- okt- apr- okt- apr- 94 94 95 95 96 94 94 95 95 96

0.03 -----Tree N3- —- 0.02 .. ; 0.01 \ /• 0.00 j\WW WW'i ------♦—i— apr- okt- apr- okt- apr- 94 94 95 95 96 Figure 4.5.2.1 Soil solution concentration ofN03 in single samplers (3 samplers per tree) beneath the root zone at Klosterhede before and after application of 50 kg N/ha with labelled NH_, (upper two figures) or labelled N03 (lower two figures). The vertical dotted line indicates the application time.

Needle analyses have so far, not shown any The ultimate goal of the project is to significant differences between the control plot incorporate the results from this study, as well as and the N treated plots (Figure 4.5.2.2). Analyses those from other Nordic nitrogen studies, into a of ,5N in the needles will be performed in late nitrogen circulation model, SOILN. Based on the 1996. hydrological part of this modelling, a procedure

Ris0-R-972(EN) 65 has been developed to calculate element fluxes in there is still a need to improve microbial biomass forest soils, taking the spatial variability in input kinetics and N-cycling. fluxes into account (Beier, in press). The application of the SOILN model to the control References plot at Klosterhede was initiated and is still Beier C. (1996) Application of the SOILN model to a running (Beier 1996). So far, the parameterization Norway spruce plantation in Denmark. In: Work­ has focused on the microbial component in order shop reports 1996 from Annual meeting in Laugarvatn, Iceland. 24-28 August. to obtain reasonable development in the above- Beier C. Water and element fluxes calculated in a sandy and below-ground organic N and C pools. The forest soil taking spatial variability into account. model has given reasonably good results, but For. Ecol. Manage. In press.

' Current year needles

Cont A1 A2 N3 N4 Figure 4.5.2.2 Concentration of N in needles (mgN/gr) in Norway spruce trees at Klosterhede as a fiinction of treatments. Cont=controI; A l and A2=50 kgN/ha added with labelled ammonium; N3 and N4=50 kgN/ha added with labelled nitrate. White bars=1994, black bars=1995.

66 Ris0-R-972(EN) 5 Trace Analysis and Reduction of Pollution in the Geosphere

Introduction (V. Gundersen) Summary of Projects

The aim of our research programme has been 5.1 Chemical Analysis redirected towards analysis of agricultural pro ­ duction and conversion of biomass to useful 5.1.1 Measurement of Calcium Isotopic Ratios products, in a pertinent and effective way. in Human Urine using HRICPMS (S. Stump) Similarly, external sources now finance purely scientific programmes, rather than mixed Isotopic tracers, radioactive or stable, are often commercial activities. used to measure the bioavailability of elements of Trace elements in the human food chain and in nutritional importance, e.g. Ca, Fe and Zn from agricultural produce are investigated within the food or drink. The main advantage of using stable national programme in food technology, F0TEK. isotopes instead of radioactive ones, is that they Our new analytical facilities, the clean room and can be used without risk to subjects or the high resolution inductively coupled argon investigators. The analysis of stable isotopes is plasma mass spectrometer, are indispensable in difficult and requires an accurate and precise this work. We have investigated more than 60 mass spectrometric technique. Most often TIMS elements in 10 agricultural and gardening (Yergey et al. 1980, Yergey et al. 1987, Price et produce from 200 different farmers. More than al. 1990) or FABM (Smith 1983, Fairweather- 2,000 samples were analysed for more than 60 Tait et al. 1989) was used for the determination elements. For this purpose, it was necessary to of calcium isotopic ratios. The use of HRICPMS, develop a TIMS and an advanced computer which was installed at Rise in 1995, is another programme for data calculation from additional alternative. Quadrupole ICPMS was used for the calibrations. Sophisticated tools for sample determination of the 44Ca/43Ca ratio in urine after preparation and homogenization, without con ­ the precipitation of calcium with oxalate (Luten et tamination from metals, were developed. al. 1993). The 42Ca/3Ca ratio cannot however, be Statistical treatment and interpretation of the measured using Quadrupole ICPMS due to an analytical data will be completed and published overlap from 40ArH2+ and 40ArH on the 42Ca next year. peak. These interferences can be resolved by Our analytical equipment for organic analysis using HRICPMS and a resolution of 4000 m/Am has been extended with GC/MS/MS and (Figure 5.1.1). Interferences from doubly charged analytical procedures for analysis of triazines in strontium, Sf \ cannot be resolved even using sediments were developed. The use of municipal maximum resolution (RP= 10000). This overlap sludge in farming raises the problem of uptake of has, therefore, been mathematically corrected by compounds such as LAS and DEPH by plants. using the natural abundances of the strontium The development of analytical procedures for isotopes after the measurement of Sr++ at m/z = analysis of LAS and DEPH in sludge, soil and 43.5. Rubidium also has a doubly charged ion at plants have recently started. m/z = 43.5. But since the second ionization Soil and ground water contaminated with potential of rubidium is very high (28.5 eV), volatile organic compounds, eg. gasoline is a doubly charged ions are not formed in the argon serious problem at many sites. A development plasma (the first IP of argon is 15.8 eV). The project for soil and ground water remediation by second ionization potential of strontium is 11 eV bioventing and air sparging has just started. and doubly charged strontium is readily formed in the argon plasma. Approximately 5% of all strontium is seen as doubly charged ions. This HRICPMS method was employed for the measurement of isotopic ratios of calcium in urine following a double-label stable isotope

Ris0-R-972(EN) 67 technique, where volunteers were given 42Ca and approximately 0.15% was obtained, which is 44Ca orally and intravenously, respectively, with close to the theoretical value of 0.13%, calculated the purpose of determining the calcium from counting statistics. The calcium absorption absorption rates from milk and fish in humans. rates obtained, using HRICPMS, will be The 42Ca/43Ca and 44Ca/3Ca isotopic ratios were compared with results from a radiotracer study measured directly in the urine after a 100-fold using• 48 /~iLa. dilution with 0.14 M nitric acid. A precision of

25000

20000

O 10000

Mass Figure 5. 1.1 HRICPMS spectra of42Ca in urine.

References 5.1.2 Measurement of Total and Extracellular Yergey A.L., Vieira N.E., Hansen ./.IV (1980) Isotope Body Water from the Determination of ratio measurements of urinary calcium with a Deuterium and Bromine in Human Sera using thermal ionization probe in a quadrupole mass HRICPMS (S. Stiirup) spectrometer. Anal. Chern. 52: 1811-1814. Vergev A.L.. Vieira X V., Coveil D C1. (1987) Direct measurement of dietary fractional absorption using An accurate and rapid determination of total and calcium isotopic tracers. Biomedical and extracellular water is valuable in many clinical Environmental Mass Spectrometry 14: 603-607. situations. This includes an assessment of fluid Price R.I. et at. (1990) Kinetics of intestinal calcium status in patients with heart failure or renal absorption in humans using stable isotopes and high- failure. In addition, these measurements are precision thermal ionization mass spectrometry. necessary for the determination of body Biomedical and Environmental Mass Spectrometry 19: 353-359. composition in nutritional studies. Bioimpedance Smith D.I.. (1983) Determination of stable isotopes of is the most commonly used method for the calcium in biological fluids by fast atom bombard ­ determination of total and extracellular body ment mass spectrometry. Anal. Chem. 55: 2391- water since the apparatus is non-invasive, 2393. portable and reproducible. As bioimpendance is Fairweather-Tait S.J. et al. (1989) Studies on calcium prone to systematic errors, a reference method is absorption from milk using a double-label stable isotope technique. British Journal of Nutrition 62: needed. Labelling water with tritium or 379-388. deuterium, followed by beta emission detection Luten J.B.. Mays T., Dokkuni IV ran (1993) The or mass spectrometer (Schoeller et al. 1980, Patel determination of stable isotope ratio of zinc, copper, et al 1994) is normally used for the iron in faeces and calcium in urine by 1CPMS. determination of total body water, since both Bioavailability ‘93: Nutritional, chemical, and food deuterium and tritium distributes equally in all processing implications of nutrient availability 161- 168. body water. The use of deuterium is preferable, as it is nontoxic. Extracellular body water can be determined after ingestion of bromine, which is measured using fluorescent excitation. Bromine cannot pass through cell membranes and distributes in the extracellular water only and

68 Ris0-R-972(EN) therefore, the reference method is made up of two tration of deuterium even in the basis serum. The different analytical techniques. Using HRICPMS total body water is then calculated from the both deuterium and bromine can be measured difference in deuterium concentration originating with the same analytical technique, in the same from the ingestion of D20. Bromine is measured sample, at the same time. Furthermore, the at m/z = 81 in sera samples before and after sample preparation is a very simple 20-fold ingestion of NaBr. A resolution of 4000 was also dilution of the serum with deionized water. used for bromine to avoid interference from an Deuterium is measured at m/z = 2 using a unknown polyatomic substance. Preliminary resolution of 4000 m/Am to resolve the 2H peak results show very good agreement between the from the 1H2t peak (Figure 5.1.2). The deuterium HRICPMS and the bioimpedance method. At a content is measured in sera samples taken before later stage, the deuterium results will be and after ingestion of D20. Since 0.015% of compared with results from the mass hydrogen is deuterium, there is a high concen ­ spectrometric method.

Deuterium

12000

10000

Mass Figure 5. 1.2 HRICPMS spectra of deuterium in serum.

References geological and geochemical mapping, was Schoeller D A. et al. (1980) Total body water measure ­ therefore, initiated by the Geological Survey of ment in humans with lsO and "H labelled water. Norway and the University of Bergen to outline, Am. J. Clin. Nutr. 33: 2686-2693. on a three-dimensional base, the distribution of Patel R.V. et al. (1994) Estimation of total body and extracellular water using single- and multiple- contaminants, heavy metals and organic com­ frequency bioimpendance. Ann. Pharmacotlier 28: pounds. Some results of this work were presented 565-569. earlier and are now published in a special volume (Kunzendorf et al. 1996, Boe et at. 1996, 5.2 Trace Metals in the Geosphere Christiansen et al. 1996). Discussions of the signals reaching the Baltic Sea were presented at 5.2.1 Geochemical Mapping in the Skagerrak- some meetings (Kunzendorf and Christiansen Kattegat Area (H. Kunzendorj) 1996, Neumann et al. 1996) and similar cooperative work in another area was also The Skagerrak and the Kattegat are considered to presented (Nagender et al. 1996). be sedimentary basins which receive important inputs from the adjacent North Sea and Atlantic References Ocean, and from weathering of the agriculturally Kunzendorf H., Longva ().. Paetzel M. (1996) Recent sedimentation rates across the Norwegian Trough. and industrially exploited land of Scandinavia. NGU Bull 430. 67-74. Both sea areas are connected to the Baltic Sea and Boe R., Rise L., Thorsnes T.. de Haas II. Saelher O.M its problems of industrial pollution (Kunzendorf Kunzendorf II. (1996) Sea-bed sediments and et al. 1994). Systematic work, including

Ris«-R-972(EN) 69 sediment accumulation in the Norw egian part of the Late-Quaternary deposits of the Bornholm Basin. In: Skagerrak. NGU Bull. 430: 75-84. E. Emelyanov, C. Christiansen, O. Michelsen (eds ). Christiansen C, Kunzendorf //., Laima M.J.C., Lund - Geology of the Bornholm Basin, Aarhus Geoscience Hansen L.C., Petersen AM. (1996) Recent changes 5: 175-188. in environmental conditions in the southwestern Baturin G.N., Emelyanov EM., Kunzendorf II. (1996) Kattegat. Scandinavia. NGU Bull 430: 137-144. Authigeneous deposits in the Bornholm Basin. In: E. Kunzendorf II, Kuijpers A., Dennegard B., I.ongvaa (). Emelyanov, C Christiansen, O. Michelsen (eds ), (1994) Marine geochemical studies at the entrance Geology of the Bornholm Basin, Aarhus Geoscience to the Baltic Sea. In: J.C. Alexander, J. Menon 5: 189-194. (eds), Trends in Chemical Geology, Vol.l Research Trends. Trivandrum, India, p. 77-89. 5.2.3 Marine Geochemistry of Gotland Basin Kunzendorf II., Christiansen C. (1996) Tracing and use Sediments (H. Kunzendorf) of radiometric signals in Baltic Sea surface sediments. Journal of Conference Abstracts, 1996 V.M. Goldschmidt Conference, 31 March-4 April. The Gotland Basin Experiment (GOBEX), a pilot Heidelberg, Germany, pp. 340. study for a MAST-3 Regional Seas project, was Neumann /’., (Kristiansen ('., Emeis K.-E., Kunzendorf terminated in 1996. The geological/geochemical II. (1996) Storre indstromiring til 0stersoen registre- results were compiled in a report (Christiansen rcs i sedimentet. 9. Danske Havforskermode. DTU. and Kunzendorf 1996). Some of the scientific Lyngby. 16-18 January , poster spin-off from the project, such as the chemical Nagender Nath II, Kunzendorf II., Pliiger W.L. (1996) Geochemical signatures of extreme chemical environment in the Gotland Basin during the last weathering in southwest coastal sediments of India. 300 years (Endler et al. 1996) and the variability Int. Symp. Geology and Geophysics of the Indian of sedimentation rates in the Gotland Basin Ocean. Goa, India, 21-25 October. Abstract p. 59. (Christiansen et al. 1996) was presented at the Baltic Marine Science Conference in Ronne in 5.2.2 Geology and Geochemistry of the 1996. Bornholm Basin (H. Kunzendorf) References At the end of the 1980s, a scientific co-operation Christiansen ( ., Kunzendorf II. (1996) Signals of between several Russian and Danish institutes in inflow to the Gotland Basin in the sedimentary the Bornholm Basin resulted in a need to edit this record - Preliminary results. In: E. Hagen (cd ), work as a basis for future more detailed GOBEX summary report. Marine Science Reports investigations in the area. The publication of No. 19, IOW, Wamemiinde, p. 10-14. Endler R, Struck V., Benesch I)., Neumann I’., these data was accomplished by collaboration Kunzendorf II., Christiansen C. (1996) The between Russian and Danish authors on several chemical environment of the Gotland Basin over the topics. The Riso group was chosen to edit the last 300 years - Results of GOBEX sediment studies. marine geochemistry section in co-operation with Baltic Marine Science Conference, Ronnc, 22-26 scientists from Kaliningrad and Moscow. Both October. Abstract p. 56-57. trace metals in seawater and in the pore waters of Christiansen ('., Kunzendorf II, Emeis K.-C., Endler R., Struck {/., Benesch /)., Neumann /'. (1996) the sediments (Emelyanov et al. 1996a) and the Sedimentation rate variabilities in the Gotland Basin general geochemistry of the recent sediments in (GOBEX). Baltic Marine Science Conference. the Bornholm Basin (Emelyanov et al. 1996b) Ronne, 22-26 October. Abstract p. 42-43. were addressed. Finally, some authigeneous de­ posits, such as ferromanganese nodules were also 5.2.4 Analytical Work in the National Gamma described (Baturin et al. 1996). Dating Center (H. Kunzendorf)

References The national Gamma Dating Center (GDC) is Emelyanov EM, Kravtzov V.A., Kunzendorf II. (1996a) operational, but performance parameters for the Trace metals in sea and pore waters from the new location have not yet been studied suffi­ Bornholm Basin. In: E. Emelyanov. C. Christiansen, ciently. Earlier work on the dating method (lead- O. Michelsen (eds.), Geology of the Bornholm Basin, Aarhus Geoscience 5: 157-174. 210), involving the possible correction of self- Emelyanov EM., Baturin G.N., Kunzendorf II. (1996b) absoprtion in the sediment, was published Distribution of chemical elements in recent and (Kunzendorf 1996b) and the use of a gamma

70 Ris0-R-972(EN) probe for seafloor inspection was presented at an ground waters in several regions of Europe and invited lecture (Kunzendorf 1996c). the US. High NOM is undesirable due to a variety of reasons: the yellow to brown colour of References the NOM is unsavoury, removal of dissolved iron Kunzendorf II. (1996a) Frelies center for datering af and manganese during treatment is impeded, recente sedimenter (gammadatering). 9. Danske microbial regrowth is enhanced in the supply Havforskcrmode, DTU, Lyngby. 16-18 January. system, and NOM is the precursor of Abstract p. 73-74. trihalomethanes and other harmful compounds Kunzendorf II., Jensen H.S., Odgaard B., Pejrup A/, Christiansen C. (1996) Gamma-dateringscentrets during chlorination of the drinking water. grundlag og opgave. 9. Danske Havforskermode, In order to enable efficient, simple treatment DTU. Lyngby. 16-18 January 1. Abstract p. 75. of humic raw water, operating parameters for Kunzendorf II. (1996c) Marine mineral exploration - nanofiltration were developed for the removal of Realities and strategies at the end of the nineties. NOM, maintenance procedures were established, Int. Symp. Geology and Geophysics of the Indian and the performance and economy were Ocean, Goa. India. 21-25 October, p. 189. Kunzendorf II. (1996b) A practical approach for self­ evaluated at three sites with humic ground water. absorption correction in 2l0Pb gamma-spectrometric The NOM found in the ground water at the dating. J. Radioanal. Nucl. Chem. 204 (1): 23-31. Kisserup site in Zealand and the Skagen site in northern Jutland had similar structural properties 5.2.5 Review of Processes Operating at the to the NOM of many surface waters, whereas the K/T Boundary (H. Kunzendorf) NOM at the Fjand site was similar to the ground water organic matter found in Western Germany, Participation in a project, studying the geological France and the “Trona waters” reported from the and geochemical processes that led to the US (Gran et al. 1996). extinction of e g. the dinosaurs at the Cretaceous/ Tertiary (K/T) boundary, resulted in a review The Technique article (Glasby and Kunzendorf 1996). As a result In nanofiltration, water and small dissolved of a thorough literature search, it was concluded molecules, such as inorganic salts, are forced by that the generally gradual mass extinctions pressure through a membrane of specified pore observed at the boundary, most probably were size. The larger NOM molecules are retained by caused by volcanic activities, Deccan Traps, the membrane. In the removal of NOM, the prevailing at the end of Cretaceous times. This filtrate or permeate, is used as drinking water, and theory does not conflict with the asteroid impact the retained water and its solutes, the retenate, theory, but merely suggests that the impact traced must be disposed of. In order to reduce plugging at the boundary was not responsible for the of the membrane pores during operation, the gradual extinctions. water is pumped across the membrane during filtration in a cross flow operation. Reference Glasby G.P., Kunzendorf H. (1996) Multiple factors in The Operating Parameters the origin of the Cretaceous/Tertiary boundary: the The membranes were operated as spiral wound role of the environmental stress and Deccan Trap units on site in mobile pilot plant units. volcanism. Geol. Rundschau 85: 191-210. Membranes with larger pore sizes gave higher productivity, but with the more open membranes, 5.3 Organic Matter and Pollution in the NOM removal decreased, depending upon the the Geosphere (C. (Iron) NOM molecular size (Figure 5.3.1). Higher cross flow, lower pressure and higher distance between 5.3.1 Removal of NOM from Humic Ground membrane layers known as spacing, reduced the Waters by Nanofiltration productivity decline during operation. With optimized operational parameters, 90% High concentrations of naturally occurring of the raw water could be used as drinking water, organic matter, NOM, are found abundantly in whereas the remaining 10% with high concen ­ surface waters all over the world and even in trations of NOM would have to be discarded. The

Ris0-R-972(EN) 71 membrane filtration treatment technique proved using several membrane units resembling the simple and stable to operate, once operational ones used in this present project, the operational parameters matching the individual water quality parameters are directly applicable for large scale were selected. As the process can be upscaled for water treatment. use in drinking water treatment plants, simply by

NOM in treated water Productivity 300

200-300 D 2000 D 6000 D 200-300 D 2000 D 6000 D Pore size as molecular weight cut off Pore size as molecular weight cut off

Raw water with Raw water with fP|flarge NOM t®*! small NOM ■llarae NOM 1 Ismail NOM ■■molecules isilmolecules HIS molecules 1_J molecules

Figure 5.3.1 Effect of membrane pore size on productivity and efficiency for NOM of two different molecular size distributions.

Maintenance carbon NPOC, could be removed for the water Even with optimized operational parameters, a types tested (Table 5.3.1). Furthermore, dissolved productivity decline was observed due to mem­ iron removal was improved and, for calcareous brane fouling. From highly calcareous raw water waters, a softening effect was obtained. with low molecular weight NOM (fulvic acids), Generally, the produced water complied with the calcite crystals precipitated and plugged the mem­ Danish drinking water regulations. brane. Periodical cleaning with an acidic deter­ gent, followed by an alkaline detergent, could re­ Conclusions and Perspectives store the initial productivity completely and thus Nanofiltration has been developed to an efficient, enable a long membrane life. At a site with low inexpensive and simple technique for the removal calcium content and higher molecular weights of of NOM in drinking water treatment of humic the NOM (humic acids), the membrane fouling ground water, and a rationale for identifying the was caused by organic matter which could be best operational parameters from data on removed almost completely using the alkaline inorganic hydrochemistry and NOM structures detergent only. was established. The results obtained can be extended to treat humic surface waters, and in the Water Quality future, the technique may even be employed for With the selection of suitable membranes and the removal of low molecular weight organic optimized operational parameters, more than 95% contaminants, such as pesticides. of NOM, measured as non purgeable organic

72 Ris0-R-972(EN) Table 5.3.1 Selected data on composition of raw water and treated water at two sites.

NOM Fration of NOM Sodium Calcium Total iron (mg C/L) with molecular weight above 5 000 D (mg Na +/L) (mg Ca ++/L) (mg Fe/L) (%) Kisserup Raw water 11 <5 21 107 4,0 Treated water 0,6 - 20 82 <0,1 Fjand Raw water 22 22 87 12 <0,1 Treated water 0,4 - 52 13 <0,1

References pretreatment processes to overcome this are wet Alhorzfar A/., (iron ('., Jonsson G. (1994) Ultrafiltration oxidation (aqueous, elevated temperature, oxygen) in producing drinking water from humic ground (Bjerre et al. 1996a) and steaming (Schmidt et al. waters. Environ. Int. 20(3): 411-417. 1996). They partly solubilized the hemicellulose Alhorzfar M. (1996) Membrane filtration and artificial recharge of humic ground water. Ph D. Thesis. and partly degraded the lignin, facilitating a Technical University of Denmark. fractionation of the constituents and making the Alhorzfar M., Jonsson G., I'illumsen A., Gron C. (1996) polysaccharides accessible for bioconversion. Treatment of “brown water ” using membrane- The wet oxidation under alkaline conditions was filtration (in Danish). Vandlcknik 64(6): 311-316. previously found (Ahring et at. in press and (iron 1 Vassenaar /,., Krog M. (1996) Origin and 1996, Bjerre et al. 1996a, 1996b) to be very stmclurcs of groundwater humic substances from efficient for solubilization of the hemicellulose three Danish aquifers. Environ. Int. (22(5): 519-534. fraction in wheat straw, producing fewer toxic 5.4 Pretreatment and Conversion compounds from the lignocellulosic carbohydrates, e.g. furfural and 5-hydroxymethylfurfural, than Processes of Agricultural and Industrial steam explosion (Schmidt et al. 1996). Waste Birch wood and wheat straw represent lignocellulosic material of high and low acetyl 5.4.1 Bioethanol Production (A ll Bjerre. content, respectively. The optimum steaming con ­ A S. Schmidt , A. Woidetnann, A. Ploger) ditions for hemicellulose solubilization and recovery from birch wood were not optimal for Lignocellulosic biomass, such as wood and residues the treatment of wheat straw and vice versa , as from agricultural crops, is potential raw material for seen in Figure 5.4.1 (Schmidt et al. 1996). the production of fuel ethanol. In contrast to Steaming looked more promising for pre ­ petroleum, fermentation ethanol does not contribute treatment of birch wood, whereas the wet to the green-house effect as it is a C02 neutral fuel oxidation process looked more efficient for (Bjerre 1996). Lignocellulose consists of three pretreatment of wheat straw. The formation of major constituents i.e. cellulose, hemicellulose furfurals could be minimized by the addition of and lignin. Cellulose and hemicellulose are most carbonate to the hydrothermal process (Schmidt interesting for the production of a variety of et al. 1996). In the case of birch wood, this potential valuable products such as ethanol, resulted in very low hemicellulose solubilization furfural, enzymes, xylitol, carboxylic acids and probably due to an interruption of the oligosaccharides (Schmidt and Bjerre, in press). autohydrolytic action of acetic acid (Schmidt et These polysaccharides are not directly available al. 1996). However, a high hemicellulose for bioconversion, as the lignin component acts solubilization at alkaline conditions was achieved as a physical barrier. Two of the most promising

Ris0-R-972(EN) 73 for wheat straw, which has a smaller content of extracellular enzyme activities were highest after acetyl groups in the raw material. about 3-4 days, as the activity in the pellets was decreasing. In addition to the 3 enzymes mentioned 5.4.2 Enzyme Production and Hydrolysis above, other extracellular enzymes were produced (A.B. Bjerre, A.S. Schmidt , A. Woidemann) including a-arabinofuranosidase (EC 3.2.1.55) and several esterases. Hemicellulolytic enzymes by an Aspergillus niger The enzymatic hydrolysis of pretreated ligno- isolate were produced on the hemicellulose-rich cellulose was studied to see whether it was fraction of wet-oxidized wheat straw. In wheat advisable to replace the acid hydrolysis (Bjerre el al. straw, the hemicellulose is an arabino-4-0-methyl- 1996c) with enzymatic hydrolysis. Rise’s own glucuronoxylan. This solubilized hemicellulose enzyme mixture produced by A. niger on pretreated proved to be a direct substrate source for A. niger wheat straw, proved to be a better enzyme mixture producing hemicellulases. Attention was focused on than both Celluclast and Viscozyme from Novo production of the intracellular and extracellular p* Nordisk, as it released the same monosaccharides xylosidase (EC 3.2.1.37), and the endo-p-(1.4)- produced by acid hydrolysis. An enzyme mixture xylanase (EC 3.2.1.8) activities. These enzymes from Trichoderma reesei, supplied by VTT, have a key role in the total enzymatic hydrolysis of Finland, was also able to hydrolyze the solubilized hemicellulose into xylose. Pellet-shaped mycelia hemicellulose to 85% of that found by acid were formed after 3-4 days growth, indicating no hydrolysis. As the hydrolysis conditions have been inhibition of A niger in wet-oxidation substrates and optimized, the lower yield of sugars obtained might about 13 U intracellular P-xylosidase/g dry mycelia be due to inaccessible hemicellulose or missing was produced (Schmidt and Bjerre 1995). The enzymes.

60 Birch Wood Wheat Straw „ . ■ o...... o O' 50 o

A . 40 0) o co o >> A A X 30 A o A >1 ■ a> A > 20 o o o o 0) - CC A' 10 ---A--- Wet Oxidation -o Steaming O

0 ___ i —1----1---- 1___i___ 1___i___ 1----1___ 1___i___ 1___i__ 170 180 190 200 210 150 160 170 180 190 200 210 Temperature (°C) Figure 5.4.1 The recovery of the xylose from wheat straw and birch wood pretreated by wet oxidation (60 g/L material; 6.5 g/L Na 2C03; 12 bar 02; 15 minutes) and steaming (10 minutes). Values in % of the xylose in the raw materials.

74 Ris0-R-972(EN) 5.4.3 Detoxification of Tar Containing Pyrolysis involving hydroxyl radicals and possibly also the Wastewater (A.B. Bjerre, A. Ploger, H.H. Kilen, reactor material (Bjerre, submitted). A. Woidemann) 5.4.5 Fibres for Composite Materials The tar-containing wastewater, derived from the gas (A.B. Bjerre, A S. Schmidt) pyrolysis of straw and wood chip damages con­ ventional biological treatment. Purification methods Lignocellulosic biomass is also a potential fibre for for detoxification of this waste stream were industrial application as new environmentally investigated by wet oxidation (WO) and wood chip friendly materials and products. These fibres have columns. The toxicity was evaluated biologically by received a lot of interest due to their properties Nitrosomonas and Nitrobacter as well as by the for replacing traditional fibres, such as asbestos inherent biodegradability test, the modified Zahn- and glass in composite applications, e g. thermo ­ Wellens test. The reaction products form the WO plastics or cement. The low density and high treatment were identified and quantified by a com­ strength of annual grown lignocellulosic fibres, bination of GC/MS, HPLC and an application of e.g. flax and hemp, provide structural superiority deuterium-labelled compounds (Bjerre et al. over traditional fibres (Schmidt and Bjerre, in submitted). The preliminary results show that WO is press). Riso is part of a newly formed Center for a very promising method for the detoxification of Plant Technology together with RVAU, DTU, waste water, as well as for the removal of the DIPPS and Novo Nordisk. The aim of the Center is dominating tar compounds. In addition, the WO to promote the use of lignocellulosic fibres for products could be transformed to biogas by means industrial applications. of methane producing bacteria. References 5.4.4 Reaction Mechanisms of Tar Compounds Ahring B.K., Jensen K., Nielsen P., Bjerre A.B., Schmidt in Wet Oxidation (A.B. Bjerrej A.S. Pretreatment of wheat straw and conversion of xylose and xylan to ethanol by thermophilic anaerobic bacteria. Bioresource Technol. In press. Tar products, coming from gasification and oil shale Ahring B.K., Licht D., Nielsen P., Schmidt A.S., Bjerre A.B. processing, are of environmental concern as they are (1996) Conversion of wheat straw to ethanol: Wet harmful to biological treatment containing toxic oxidation pretreatment followed by anaerobic compounds. Quinoline represents a group of PNA- thermophilic fermentation of the hcmicellulose compounds, Poly Nuclear Aromatics, which are fraction. 9th European Bioenergv Conference, closely related to the PAHs. Hence, quinoline could Copenhagen, Denmark, 24-27 June 1996. Bjerre A.B. Degradation of quinoline by wet oxidation - be used in this fundamental study of the kinetics and kinetic aspects and reaction mechanisms. Water Res. reaction mechanisms in WO of tar compounds. WO Submitted. of quinoline was studied as a function of initial Bjerre A.B. (1996) Bioteknologi forvandler plantcrestcr til concentration, pH and reaction temperature. At flydende bnendstof. Riso Nyt March (1) p. 14-15. neutral and acidic conditions, quinoline was Bjerre A.B, Femq\>ist T., Ploger A., Schmidt A.S. (1996a) efficiently oxidized at WO temperature at 240°C or Delignification of wheat straw by wet oxidation resulting in bioconvertible cellulose and hemicellulose. above, whereas at alkaline conditions the oxidation In: Management of Urban Biodegradable Wastes J.-A. was considerably slower. The reaction is most likely Hansen (ed.) James and James Science Publishers, an auto-catalyzed, free radical chain reaction. London, p. 209-217. Depending on the initial quinoline concentration, up Bjerre A.B., Olesen A.B., Femqvist 7!, Ploger A., Schmidt to 30-50% was oxidized to C02 and water. Fourteen A S. (1996b) Pretreatment of wheat straw using other reaction products were identified and combined wet oxidation and alkaline hydrolysis resulting in convertible cellulose and hemicellulose. quantified by a combination of GC/MS, HPLC and Biotechnol. Bioeng. 49: 568-577. an application of deuterium-labelled quinoline Bjerre A.B., Ploger A., Simonsen T., Woidemann A., (Bjerre et al. submitted). Nicotinic acid was the Schmidt A.S. (1996c) Quantification of solubilized main reaction product, accounting for up to 35% of hemicellulose from pretreated lignocellulose by acid the initial quinoline. A reaction mechanism was hydrolysis and high performance liquid proposed (Figure 5.4.4, shown at end of Chapter 5) chromatography. Riso-R-855(EN). for the degradation of quinoline by wet oxidation

Ris0-R-972(EN) 75 Bjerre A.B., Fgsgaard II., Nielsen I'., Plager A.. Analytical Schmidt A S., Puls J., Bjerre A.B. (1996) Comparison of chemistry directed to reconditioning of contaminated wet oxidation and steaming for solubilization of the soil by wet oxidation. Submitted. hemiccllulosc traction in wheat straw and birchwood. Schmidt A S., Bjerre A.B. (1995) Optimization of wet 9th European Bioenergv Conference. Copenhagen. oxidation of wheat straw for the enzyme production by Denmark, 24-27 June 1996. Aspergillus niger. GIAM X - 10th International Schmidt A.S., Bjerre A.B. Pretreatment of agricultural crop Conference on Global Impacts of Applied residues for conversion to high-value products. In: Microbiology and Biotechnology, Elsinore, Denmark. Cereals: Novel Uses and Processes. G.M. Campbell. C. 6-12 August 1995. Webb, S. McKee (eds.) Plenum Press. In press.

Abstraction

b

Ring oxidation/opening

2 acetyl pyridine H+ | H2O -2C02 O CHO

cx:N' ''CHO O, /CHO Cx'N' ''COOH

I- C02 cx CHO 3-pyridine aldehyde

GrN c nicotinic acid Figure 5.4.4 Proposed reaction mechanism of quinoline in wet oxidation.

76 Ris0-R-972(EN) 6 Ecology

Introduction (A. Aarkrog) encouraging that the Emergency Management Agency of the Danish Ministry of the Interior The aim of the ecology programme is to improve continued to support the Ecology Programme in our understanding of the bio-geochemical 1996. Without this support, radioecological behaviour of stable, as well as radioactive research in Denmark might have been injuriously elements. Furthermore, the purpose is to develop affected. methods which can reduce environmental and The biosphere-atmosphere exchange studies in radiological impacts from pollutants, and utilize the Ecology programme include the deposition of radioecological principles and measuring me­ stable elements in the ecosystems in addition to thods to solve general environmental problems. biological emissions. Deposition of airborne The determination of the generally very low pollutants may influence plant production, concentrations of environmental pollutants re­ specially vulnerable ecosystems. In a broader quires advanced measuring techniques. Forty environmental perspective, the study of biological years experience in the measurements of emissions may identify those substances which environmental radioactivity has placed the pro ­ contribute to crucial global processes, such as the gramme in a favourable position with respect to greenhouse effect and the depletion of the ozone low-level measurements and radiochemical layer. analysis. After the installation of an ICPMS in the Department, new possibilities for low-level analysis have appeared. A major effort is now Summary of Projects directed towards an application of ICPMS for the determination of very long-lived radionuclides, 6.1 Biosphere Atmosphere Exchange e g. 99 Tc and 2,y-240p u in environmental samples. The aim is to improve the sensitivity, so far obtained by radiochemical analysis, by one to two 6.1 1 Exchange of Oj and NOx between the orders of magnitude. If this effort is successful, Atmosphere and Forested Ecosystems the size of many environmental samples can be (K. Pilegaard) reduced substantially. This will be a great advantage for the programme, because our International Biosphere-Atmosphere Exchange activities are global and shipments of large Experiment samples create problems. In June 1996, the exchange of nitrogen oxides The activities in the ecology programme not and ozone between forest and atmosphere was only improve general knowledge of measured. TNO (The Netherlands) and UMIST environmental problems, but the programme also (United Kingdom) participated in the experiment. contributes to the nuclear preparedness nationally The site was an 80-year-old beech forest near and internationally. Our recent participation in Sore, where a 57m high meteorological mast and IAEA’s Mururoa programme exemplifies this. a 24m high tower were available. The objectives In 1996, the 10-year anniversary of the were to provide better estimates of the exchange Chernobyl accident was commemorated by a rates of ozone and nitrogen oxides. Therefore, the number of international scientific meetings sum­ fluxes were measured at several heights above marizing the knowledge achieved since the and below the canopy, in addition to the biogenic accident. It is thought-provoking that the present emission of nitric oxide from the soil. Preliminary general tendency, at least outside the former results show that under some circumstances, the Soviet Union, is for a substantial reduction of forest acts as a source of atmospheric nitrogen radioecological research. This could, in the case oxides and under other circumstances, as a sink. of a new nuclear accident, have serious The exchange processes are, therefore, important consequences, because the radioecological for the oxidizing capacity of the atmosphere. expertise may rapidly disappear. It is, therefore,

Ris0-R-972(EN) 77 Vd (Ozone)

un 94 Sep 95

.+ ®d"

Figure 6.1.1.1 The tower and the mast in the forest.

Ozone Fluxes over Forested Ecosystems and summer, when light is not limiting and there is no Mechanisms of Removal water stress. This indicates that stomatal uptake is The seasonal flux of 03 was measured over a an important mechanism of removal. However, a coniferous forest during several field experi ­ substantial deposition occurs during the night, ments. Generally, there is a characteristic diurnal when stomata are probably closed. The pattern in the deposition velocity with the highest deposition velocity of 03 during the night is values occurring during daytime (Figure 6.1.1.2). almost half of that during the day. Therefore, There is also a seasonal pattern with the highest there must be other important sinks for 03 other deposition velocity during the early and late than stomatal uptake.

Figure 6. 1.1.2 Diurnal variation of deposition velocity of ()< in June 1994. April and September 1995.

78 Ris0-R-972(EN) The following sinks for 03, other than stomatal measured by the eddy correlation method and the uptake, must be considered: NO emissions were measured by the steady-state 1) destruction at external plant and soil surfaces chamber techniques. Further details are given in 2) chemical reactions with isoprene and mono- Pilegaard et al. (1995) and Pilegaard et al. (in terpenes emitted from the trees press). 3) chemical reactions with NO emitted from the The concentrations of 03 were 20-40 ppbv forest floor. with the lowest values found during the late night and the highest during late afternoon (Figure The objective of the following experiment was 6.1.1.3). Fluxes ranged from -0.2 to-0.5 Tg m'2 s' to measure the effect of 03 removal by NO on the 1 with the highest values around noon. The total flux of 03 above a spruce forest. The deposition velocity peaked before noon with an measurements were carried out in a Norway average maximum of 8mm s"1 and a night-time spruce (Picea abies) plantation near Ulfborg in a minimum of 3mm s'1. The time shift in the remote rural area of western Jutland. The trees occurrence of the maxima of concentrations, had a height of approximately 12m and there was fluxes and deposition velocities, indicates that the a good fetch in most directions, except from a underlying surfaces regulate the deposition. small sector towards SW. The 03 fluxes were

Figure 6.1.1.3 Average diurnal variation of ozone concentration (C), flux (FJ and deposition velocity (Vj over a Norway spruce stand during 7-15 September 1995. The plots show mean +- s.e. and a smoothed line.

In September 1995, there was a substantial the variation within a single plot was small, soil NO emission with an average of 21 ng N m"2 whereas the variation between plots was large. s"1 and a range of 0-82 ng N m"2 s'1. Generally, There was a high correlation between C02-

Ris0-R-972(EN) 79 emission and NO-emission confirming that the the soil following a long drought period during NO was bacterially produced. the summer. When sufficient 03 is available, it Compared to most of the previous measure­ can be assumed that all the soil emitted NO reacts ments of NO-emission from temperate forest with 03 within the canopy. The resulting 03 flux soils, the values presented here are high. This is shown in Figure 6.1.1.4. might be due to “pulsing ” caused by re-wetting of Ozone flux, Sep 95

09/08 09/10 09/12 09/14

Figure 6.1.1.4 Flux of 03 7-15 September 1995 (line) and the corresponding flux as a result of the reaction of O, with soil emittet NO (dots); mean value (-0.036ppm m s!) indicated by dotted line.

It is concluded, that at certain periods of the 6.1.2 Characterization of Airborne Particles year, the soil emission of NO and subsequent from Rubber Tires and Asphalts (P. Fauser) reaction with 03 can account for the removal of a substantial amount of the total 03 flux to a spruce A method for identifying airborne particles forest, especially at night. If all the emitted NO generated from the wear of automobile tires and reacts with 03 within the forest, this reaction can roadway asphalt, bitumen, has been developed. account for about half of the 03 flux measured Organic zinc and the rubber monomer styrene during night-time or about 20% of the total flux. butadiene rubber, SBR, are used as tire indicators and are measured by heated graphite furnace References atomic absorption spectroscopy and infrared Pilegaard K., Jensen N.O., llummelshoj P. (1995) spectrometry respectively. Large organic mole­ Seasonal and diurnal variation in the deposition cules are used as bitumen indicators and these are velocity of ozone over a spruce forest in Denmark. quantified by high performance gel permeation Water, Air and Soil Pollution 85: 2223-2228. Pilegaard A'., llummelshoj P.. Jensen N O. Nitric oxide chromatography. emission from forest soil. In: P.M. Borrell, P. Air samples in heavily trafficated areas, as Borrell, T. Cvitas, K. Kelly. W. Seiler (eds.) well as in rural sites, are collected with high- EUROTRAC’96, Computational Mechanics Publi­ volume samplers and cascade impactors. Particle cations. Southampton. In press. fractions down to 0.01 pm are collected and examined for asphalt and tire debris. Figure 6.1.2.1 shows the total suspended particulate

80 Ris0-R-972(EN) matter, TSP, and the bitumen- and tireconcen- Copenhagen air in January 1996 during moist and trations in the different particle fractions found in cold weather. The total airflow is 432.82 m3.

Particulate concentrations in air

0 TSP

■ Bitumen

□ Tire

particle aerodynamic diameter fjimj

Figure 6.1.2.1 Particulate concentrations in air.

It is surprising that 92% of the airborne tire-tread Soil samples taken in June in dry weather, at debris has an aerodynamic diameter smaller than right angles to a north-south highway near lpm. This fraction can reach the alveoli in the Copenhagen, had an almost exponential decrease human respiratory system and has the potential to in bitumen concentrations with the distance from spread over long distances in the environment. the road. The results indicate a predominant 5% of the total TSP is found to be tire debris. easterly wind in the period.

Particulate concentrations in soil

- -i-

E3 bitumen

■ tire

Distance from road [mj

Figure 6.1.2.2 Particulate concentrations in soil.

Leaf samples were taken in highly trafficated late adsorption was measured in dry periods and areas and at rural sites. Both hairy leaves and after rainfall for both upper and lower leaf smooth surfaced leaves were examined. Particu­ surfaces. Some preliminary results are:

Ris0-R-972(EN) 81 Hairy surface:

(l limits glabra) Elm leaves untreated dry weather 12 m from highway: 5.92 pg tire pr. cm2 Elm leaves upperside dry weather 12 m from highway: 2.92 pg tire pr. cm2 Elm leaves lowerside dry weather 12 m from highway: 2.42 pg tire pr. cm2

Elm leaves untreated after rain inner city 6.76 pg tire pr. cm2 Elm leaves upperside after rain inner city 1.32 pg tire pr. cm2

Smooth surface:

(Primus cerasifera) Cherry plum leaves untreated dry weather 5 m from highway 8.13 pg tire pr. cm2 Cherry plum leaves upper side dry weather 5 m from highway 5.56 pg tire pr. cm2

No firm conclusions can be made from the currents. The main pathways from the two preliminary results of the plant samples. There is, discharge points to the present sampling points however, a tendency for the upper and lower are shown in Figure 6.2.1. In this study, brown sides of the leaves >12 m from the highway to algae, Fucus vesiculosus, were used as have equal concentrations of tire residues. This is bioindicators, as these plants concentrate and a consequence of the particulate size, since sub ­ time-integrate the weak radionuclide signals in micron particle transport is dominated by diffu­ water (Dahlgaard and Boelskifte 1992). This is sion and not by gravitational settling. especially the case for technetium, which is Only 5 m from the road, the adsorbed amount concentrated to high levels in Fucus. on the upper leaf side is larger, due to the In the present project, the Institute for Energy gravitational settling of the larger particles, Technology in Kjeller, Norway, made seaweed >10pm. samples available for our analysis of 99 Tc (Chen et al. 1994). The samples are part of a larger 6.2 Marine Tracers (H. Dahlgaard) sampling programme covering the entire Norwegian coast (Christensen and Selnaes 1995). Marine Brown Algae Fucus vesiculosus and In the following calculations, we compare data Chemical Oceanography from the Utsira seaweed samples with average Controlled radioactive releases to coastal waters annual discharge data 1-2 years earlier from La from the nuclear reprocessing plants in Sellafield Hague and 3-4 years earlier from Sellafield. We (United Kingdom) and La Hague (France) have will thus assume that the transit time to Utsira - as been used as tracers for long-distance transport well as to the other locations further north along and dispersion of coastal pollution for several the Norwegian coast - is 2 years longer from years (Dahlgaard 1995). The dispersion of these Sellafield than from La Hague. For the two discharges follows the general circulation of the sampling locations in north Norway, 4) Ingoy and ocean currents, and the radionuclides are, 3) Indre Kiberg at Vardo, a one year extra therefore, excellent tracers for studying these transport time was stipulated.

82 Ris0-R-972(EN) 80' 80' 70'

Barents Sea

Return \ Atlantic C. (subm) Greenland Greenland Sea v

1 Jan " Mayen

Norwegian Sea

Iceland

North Sea

40° W 20°W Figure 6.2. 1 Sampling locations for Fucus vesiculosus on the Norwegian coast and main currents carrying activity from the reprocessing plants Sellafield and La Hague to Norway.

| Sellafield Tc-99 Cs-137 n 200 r 5000 r- '////A La Hague

4000 I- 150 l-

1000 -

Figure 6.2.2 Controlled discharges of99Tc and L' Cs from Sellafield and La Hague.

Ris0-R-972(EN) 83 A Transfer Factor, TF, is calculated as the are given in Table 6.2.1. By comparing measured quotient between observed concentrations in values from the two north Norwegian stations water (Bq m"3) at the sampling site and an with the values from Utsira, an average correction average discharge rate (PBq yr"1) t years earlier, factor of 0.45, as compared to Utsira, was found. where t is the transit time (Dahlgaard 1995). The Given the information above, it is easy to make a unit for the transfer factor, Bq m"3/PBq yr"1, is “back of an envelope ” model calculating expected equal to ng m"3/ton yr"1. The calculations make radionuclide concentrations in the seaweed further use of a concentration factor, CF, defined samples. These calculations (bars) are compared as the quotient at an assumed steady state with measured values (crosses) in Figures 6.2.3 between radionuclide concentrations in Fuats and 6.2.4. vesiculosus and in water, Bq kg"1 dry Fuats/Bq kg"1 water. The actually chosen TF and CF values

Table 6.2.1 Transfer factors, TF = Bq m"3/PBq yr"1 and concentration factors, CF = Bq kg"1 dry Fitctts/Bq kg"1 water used for the calculated values, bars in figure 6.2.3. The Cs value from La Hague is in parenthesis, as it cannot be evaluated due to insignificant discharges from La Hague.

99^-r 137Cs Tc TF, Sellafield to Utsira 16.5 24 TF, La Hague to Utisra (47) 55 CF from water to Fucus vesiculosus 180 105

Tc-99, Utsira (10) Cs-137, Utsira (10)

Sellafield

La Hague

Measured, 10

10 -

too -

Year Figure 6.2.3 99Tc and 13 Cs in Fuats vesiculosus from Utsira , S’IV Norway. Calatlated (modelled) contributions from Sellafield and La Hague (bars) and measured values (marks).

84 Ris0-R-972(EN) Sellafield o Tc-99 V///\ La Hague Cs-137 Measured, 4

Measured, 3

O O

Figure 6.2.4 l'c-99and Cs-13'7 in Fucus vesiculosus from the two north-Norwegian locations 4) Ingoy and 3) Indre Kiberg at Vardo. Calculated (modelled) contributions from Sellafield and La Hague (bars) and measured values (marks).

It is expected that a higher TF value for 99 Tc sampling locations, we expect to get a better than for the less conservative element Cs will be estimate of the TF value for technetium from found. In its list of Kd values, IAEA (1985) gives Sellafield. It will then be possible to give a better a value of 100 for Tc in coastal sediments, estimate of the differences in TF values between whereas the value for Cs is given as 3000. Kd is caesium and technetium due to differences in Kd, the distribution coefficient defined as the quotient i.e. different degrees of conservativity, and thus between element concentrations in sediment and compare the different dilution factors from water, kg kg'1 dry sediment/kg kg'1 water. Sellafield and La Hague to the Norwegian coast The effect of the 1986 Chernobyl discharge with the almost completely conservative tracer and the following 137Cs enrichment of the Baltic 99 Tc. A further discussion on the chosen para ­ outflow is evident in Utsira, as well as at the two meter values is given in Dahlgaard et al. (1996). north Norwegian locations. For 99 Tc, the corre­ lation seems to be best for the period where La References Hague discharges were dominant. As technetium Dahlgaard If. (1995) Marine Pollution Bulletin 31: 3-7. is more conservative than caesium, it is not likely Dahlgaard H., Boelskifte S. (1992) J. Environ. Radio ­ that the TF value for 99 Tc from Sellafield should activity 16: 49-63. Dahlgaard II , Bergart T.D.S., Christensen G.C. (1996) be reduced further. It is therefore suggested, that Technetium-99 and Caesium- 137 time series at the the reported 1978 value for the Sellafield Norwegian coast monitored by the brown alga discharge may be overestimated. It was in fact the Fucus vesiculosus. RADOC-96: Radionuclides in first reported value for 99 Tc - the earlier values the Oceans, 7-11 October 1996, Cherbourg. France are all estimated from environmental data. When Christensen G.C., Selnaes T.D. (1995) Study of marine the new discharges from 1994-1995 reach the radioactivity along the Norwegian coast. 1980 - 994.

Ris0-R-972(EN) 85 Environmental impact of radioactive releases. Pro ­ Materials and Methods ceedings of an International Symposium. Interna ­ tional Atomic Energy Agency. Vienna, Austria: Model Description 618-622. The marine dispersion model covers the Arctic Chen O.J., Dahlgaard //., Nielsen S.P. (1994) Determi­ nation of 99Tc in sea water at ultra low levels. Ocean and the North Atlantic Ocean. Box-model Analytica Chi mica Acta 285: 177-180. analysis is used to simulate the movement of IAEA (1985) Sediment Kds and Concentration Factors radioactive material between the different boxes into for Radionuclides in the Marine Environment. which the marine environment is subdivided. This type of modelling assumes instantaneous uniform mixing within each box, with rates of transfer being 6.3 Radioecological Models proportional to the inventories of material in the (S.P. Nielsen) source boxes. Box models have been used in connection with 6.3.1 A Sensitivity Analysis of a Radiological studies of the dispersion of radioactive discharges Assessment Model for Arctic Waters from European civil nuclear installations (CEC 1990). The present model is a combination of two Introduction models: 1) an adjusted version (Nielsen 1995) of a Information on the disposal of radioactive waste in regional box model used for radiological the shallow waters of the Arctic Seas by the former assessments in north-west European coastal areas Soviet Union became available to the international (European Commission 1995) and 2) a larger box community from official Russian sources in 1993 in model covering the Arctic Ocean and the North the White Book (Yablokov et al. 1993). Since then, Atlantic (Chartier 1993). The larger box model is several initiatives have been taken to assess the derived from a world ocean General Circulation potential impact of this waste on man and the Model (Marti 1992) from which the results were environment. Dumping has taken place in the used for the design of a box structure in the Arctic Barents Sea west of Novaya Zemlya and in the Kara Ocean and surrounding waters. Sea east of Novaya Zemlya during the period 1960- The box-model analysis uses first order 1991. differential equations to describe the transfer of The countries bordering the Arctic Seas are contaminants between the boxes. The equations are particularly concerned about the possible con­ of the form: tamination of marine produce from these waters. dA { Norway and the Russian Federation have entered dt into close co-operation to investigate the present j-1 levels of artificial radionuclides in the environment where kii=0 for all i, A, and Aj are activities (Bq) at time t in boxes i and j, ky and kj; are rates of transfer near the sites where solid reactor waste and spent (y"1) between boxes i and j, k; is an effective rate of nuclear fuel were dumped in the Kara Sea. In 1993, transfer of activity (y'1) from box i taking into the International Atomic Energy Agency (IAEA) launched the International Arctic Seas Assessment account loss of material from the compartment without Project (IASAP), which is planned to last for four transfer to another, for example radioactive years. The programme is organized by IAEA in co­ decay, Q is a continuous source of input into box i (Bq y"1) and n is the number of boxes in the system. operation with Norway and Russia. Several projects supported by the European Union include The rates of transfer between the aquatic boxes, ky are related to the volume exchanges, R,p supplementary investigations of the potential impact. This section describes a parameter sensi­ according to: tivity analysis of a radiological assessment model for the Arctic waters, for the purpose of identifying where V; is the volume of water represented by important contributions to the predictive accuracy of box i. the model.

86 Ris0-R-972(EN) Figure 6.3.1.1 shows the regions used in the exchange rates may be found in Nielsen et al. marine box model in the Arctic Ocean. Each of the (1995). The suspended sediment loads and water compartments contains associated suspended sedimentation rates are default values from the sediment and has underlying seabed sediment com­ regional model mentioned above (European partments. Details of volumes, depths, suspended Commission 1995). sediment loads, sedimentation rates and volume

Figure 6.3.1. 1 Regions in the Arctic Ocean covered by the model. The numbers refer to surface water boxes.

At any given time, the activity in the water k - ^ column is partitioned between the water phase and 9 df\ + KdSSf ’ the suspended sediment material. The fraction of the where dj is the mean water depth of the water activity (Fw) in the water column, which is in column and SR the mass sedimentation rate. solution, is given by: Transfer between the sediment box and the water F„, = 1 , column is parameterized by diffusivity through the \ + KdSSL pore water and diffusivity due to bioturbation which where Kd is the sediment-water distribution gives the following transfer across the boundary: coefficient and SSL the suspended sediment load. - = (8 + 000)8^-^, Activity on suspended sediments is lost to the dt hB underlying boxes when particulates settle out. The where A is the radionuclide inventory in the water fractional transfer from a water column (box i) to the column, B the bioturbation coefficient, or the sedi­ sediments (box j) due to sedimentation is given by: ment porosity, D the pore water diffusion coefficient, S is the surface area of the interface, Cs the

Ris0-R-972(EN) 87 radionuclide concentration in sediment pore water, concentrations of 0.25 pg m"3 and 10 g m"3, Cw the radionuclide concentration in the water respectively. Furthermore, a hypothetical critical column, and h B the thickness of the bioturbated layer. group of military personnel, staying at the bays with Furthermore, removal of activity from the top dumped nuclear waste was included. The exposure surface sediment to lower sediment layers is taken pathways of this group, however, cover inhalation into account by assuming that the burial rate is equal and external exposure from beach occupancy only. to the flux of particles, which settle from the overlying waters. Radioactive decay is included in Source Terms all the boxes. The source terms used for the calculations were The contamination of fish, crustaceans and obtained from the IASAP project. The best- molluscs is calculated from the radionuclide concen­ estimate release scenario was selected, trations in filtered seawater in the different water comprising release rates of radionuclides from regions. Concentrating factors for biological shallow bays on the east coast of Novaya Zemlya, material are used, for this purpose. Data for the Abrosimov Bay and Tsivolki Bay, and from the world catch of seafood in the various regions were deeper waters of the Novaya Zemlya Trough. The compiled from the Marina Study (CEC 1990) and radionuclide specific releases considered, are FAO (FAO 1992). The following assumptions for shown in Table 6.3.1.1, and Figure 6.3.1.2 shows the edible fractions of seafood catch used are: 50% the release rate assumed for the Tsivolki Bay, for fish, 35% for Crustacea and 15% for molluscs. where the reactor compartment and spent nuclear Collective doses to the world population are based fuel from the Lenin icebreaker were dumped in on the ingestion of seafood only. Representation 1967. The graph shows that the best-estimate groups are assumed to be located at various scenario assumes release of the activation distances from the dump sites, on the Yamal products only (Co-60, Ni-59, Ni-63) until the peninsula at the Kara Sea, on the Kola peninsula at year 2300 due to corrosion of the protective the Barents Sea and in northern Norway. The barriers. At that time, contact is assumed between following assumptions were made of their habits to the seawater and the spent nuclear fuel, and a estimate radiation doses: ingestion of fish (200 kg peaked release of fission products (Cs-137 and y"1), crustaceans (50 kg y"1) and molluscs (50 kg y" Sr-90) and transuranics (Pu-239, Pu-240, Am- '); exposure to external gamma radiation for 250 h 241) is assumed followed by a steady release due y"1 in coastal areas; and inhalation for 2000 h y"1 of to corrosion of the nuclear fuel. resuspended beach sediment and seaspray at

Table 6.3.1.1 Integrated release of radionuclides (TBq) dumped in the Kara Sea, IASAP best-estimate scenario. Tsivolki Abrosimov Novaya Zemlya Bay Bay Trough Pu-239 4.9 0.73 0.09 Pu-240 2.1 0.32 0.04 Am- 0.41 0.12 0.01 241 Co-60 0.03 2.9 0.08 Ni-63 370 110 1.9 Ni-59 40 110 2.3 Cs-137 0.10 41 8.7 Sr-90 0.07 34 7.4

88 Ris0-R-972(EN) TSIVOLKI BAY

1 OOE*-13

1.00B-12

1 OOE+10 ™ -RJ240 - - -AM241 i . - CO60 ™ - NI63 1 00B-08 ■ N159

1 00B-07 ...... ~SR90

1 00B-06

1 00B-05

1 00B-04 2400 3400 4900 YEAR

Figure 6.3.1.2 Radionuclide release rates (TBq y') for Tsivolki Bay estimated by the lASAP Source Perm Group.

Parameters investigated all the above mentioned parameters and running The parameter sensitivity of the doses to indi ­ the model repeatedly, about 500 times. viduals and populations was investigated con ­ Correlation coefficients were calculated between cerning three main processes: water movement parameter values and dose values, and the square and mixing, sediment-water interaction, and of the correlation coefficients (R2) indicated how biological transfer. For the hydro dynamical much of the dose variability is explained by the processes, four parameters were investigated: the linear relationship to the parameters. The advection and mixing between the Kara Sea and parameter sensitivities are thus expressed as a the Laptev Sea to the east, the advection and percentage of the total variability. mixing between the Kara Sea and the Barents Sea to the west, the vertical mixing between surface Results and deeper waters over the Novaya Zemlya Trough, and the rates of exchange of water Collective Doses between the bays containing the dumped waste The results of the parameter sensitivity analysis and the open Kara Sea. The sediment-related for the collective doses are shown in Table parameters were the following: sedimentation 6.3.1.2, which summarizes the integrated rates, sediment distribution coefficients (Kd), releases, the collective doses and the main suspended sediment loads, depth of the mixed parameter sensitivities, >1%. The total collective surface sediment layer, and the mixing rates in dose is calculated to 0.4 manSv (truncated at the surface sediments. The three parameters 1000 y) with dominating contributions from representing biological concentrating factors for plutonium isotopes of 57% and from Cs-137 of fish, crustaceans and molluscs were included for 37%. The parametersensitivities are seen to vary the biological transfer processes. across the radionuclides from Sr-90, which has a The parameter sensitivity analysis was carried low Kd to the transuranics and activation out by assigning identical variabilities of 10% to products, which have high Kds. For the total

Ris0-R-972(EN) 89 collective dose, the main parameter sensitivities biological transfer processes (concentrating factor are due to sedimentation processes (sedimen ­ for fish). Furthermore, it is noted that the tation rate, Ka, suspended sediment load) and collective dose is very small.

Table 6.3.1.2 Integrated releases (TBq), collective doses (manSv) and main parameter sensitivities (%).

Nuclide Cs-137 Sr-90 Pu-239 Pu-240 Co-60 Am-241 Ni-63 Ni-59 Total

Total release (TBq) 49 42 6 2.4 3 0.5 480 150 Coll, dose (manSv) 0.14 0.02 0.16 0.06 7E-05 0.001 0.002 0.001 0.4 Coll, dose (%) 37 5 41 16 0.02 0.4 0.6 0.3 100 Sedimentation rate, (%) 19 66 66 54 61 53 53 45 Cone, factor fish (%) 65 90 13 4 4 28 K,( (%) 11 13 13 25 7 24 24 12 Susp. sediment load, (%) 16 16 4 14 1 1 9 Cone, factor molluscs (%) 9 9 5 1 1 5 Adv., Kara Sea, west (%) 4 1 4 4 2

Individual Doses 137 and Sr-90 will have decayed and that the The results of the parameter sensitivity analysis, transuranics dominate the release. These for the maximum annual doses to individuals in radionuclides associate readily with sediments; the critical groups, are shown in the Tables for this reason the main parameter sensitivities 6.3.1.3 to 6 3.1.6 which summarize the dose rates for long range transport are related to sediment from each nuclide and the main parameter dynamics, e.g. sedimentation rate, Kd, suspended sensitivities, > 1%. The time for occurrence of the sediment load. The advection and mixing of maximum dose rate for all the critical groups is water from the Kara Sea to the Barents Sea also related to the peak release of radionuclides from has some influence on the maximum dose rates to Tsivolki Bay in the year 2300, when the the critical groups in northern Norway and on the protective barriers of the spent nuclear fuel from Kola peninsula. The maximum dose rates to these the Lenin icebreaker will be penetrated due to groups are very low, of the order of one corrosion. The late occurrence of that event nanosievert per year. means that the shorter-lived fission products Cs-

Table 6.3.1.3 Maximum dose rates (Sv y’1) in year 2300 to a critical group in northern Norway and main parameter sensitivities (%).

Nuclide Cs-137 Sr-90 Pu-239 Pu-240 Co-60 Am-241 Ni-63 Ni-59 Total

Dose rale (Sv/y) 7E-13 7E-14 2E-10 8E-11 0E+00 2E-11 5E-14 5E-15 3E-10 Dose rate (%) 0.2 0.0 64 28 0.0 7 0.0 0.0 100 Sedimentation rate (%) 43 Adv., Kara Sea, west (%) 15 Kd (%) 14 Cone, factor molluscs (%) 10 Bay flushing rate (%) 7 Susp. sediment load (%) 5

90 Ris0-R-972(EN) Table 6.3.1.4 Maximum dose rates (Sv y'1) in year 2300 to a critical group on the Kola peninsula at the Barents Sea and main parameter sensitivities (%).

Nuclide Cs-137 Sr-90 Pu-239 Pu-240 Co-60 Am-241 Ni-63 Ni-59 Total Dose rate (Sv/y) 7E-12 6E-13 2E-09 7E-10 0E+00 IE-10 4E-13 5E-14 3E-09 Dose rate (%) 0.3 0.0 66 29 0.0 4 0.0 0.0 100 Sedimentation rate (%) 34 Kd (%) 21 Adv., Kara Sea, west (%) 20 Cone, factor molluscs (%) 11 Bay flushing rate 9

For the critical group on the Yamal peninsula first recipient of the radioactivity from the at the Kara Sea, the concentration factor for Tsivolki Bay. The maximum dose rate to the molluscs and the bay flushing rate have higher critical group is very low, of the order of one sensitivities than those of the sediment-related microsievert per year. parameters. This is because the Kara Sea is the

Table 6 3.1.5 Maximum dose rates (Sv y"1) in year 2300 to a critical group on the Yamal peninsula at the Kara Sea and main parameter sensitivities (%).

Nuclide Cs-137 Sr-90 Pu-239 Pu-240 Co-60 Am-241 Ni-63 Ni-59 Total Dose rate (Sv/y) 2E-09 IE-10 7E-07 3E-07 0E+00 IE-07 2E-10 2E-11 IE-06 Dose rate (%) 0.1 0.0 63 28 0.0 9 0.0 0.0 100 Cone, factor molluscs(%) 32 Bay flushing rate (%) 31 Sedimentation rate (%) 20 Kd (%) 15 Cone, factor crastac. (%) 3

Only external exposure and inhalation pathways to the sediment where the concentration are considered for the critical group located at the (activity/volume) depends on the value of the Tsivolki Bay. Inhalation dominates due to the sediment mixing depth. A small sediment mixing transuranic nuclides. The most sensitive para ­ depth gives a small sediment volume and a high meter is the sediment mixing depth, which shows sediment concentration from which resuspension a strong negative correlation with the dose rates causes high doses due to inhalation. The and a high sensitivity, 56%. This is due to the maximum exposure of the critical group is association between sediments and transuranic estimated to be of the order of ten millisieverts elements, whereby the transuranic elements are per year. removed by sedimentation from the water column

Table 6.3.1.6 Maximum dose rates (Sv y'1) in year 2300 to a hypothetical critical group located at the Tsivolki Bay and main parameter sensitivities (%).

Nuclide Cs-137 Sr-90 Pu-239 Pu-240 Co-60 Am-241 Ni-63 Ni-59 Total Dose rate (Sv/y) IE-07 4E-07 5E-03 2E-03 0E+00 6E-04 7E-09 6E-10 7E-0.3 Dose rate (%) 0.0 0.0 64 28 0.0 8 0.0 0.0 100 Sedim. mixing depth (%) 56 Kd (%) 18 Bay flushing rate (%) 10 Sedimentation rate (%) 9 Susp. sediment load (%) 3

Ris0-R-972(EN) 91 Conclusions radionuclides to the environment, EUR 15760EN A parameter sensitivity analysis was carried out European Commission, Luxembourg. for a radiological assessment model used for a Chartier M.( 1993) Radiological assessment of dumping in tire Kara and Barents Seas: Design of a conrpartmcnlal prediction of doses to man from dumping of structure for the Arctic Ocean and surrounding oceans. radioactive waste in the Kara Sea. The model Project Report, CETTIS. Ivry sur Seine, France. covers the Arctic and Atlantic Oceans and Marti O. (1992) Etude dc l'Occan Mondial: Modelisation calculates doses to the world population, as well de la circulation cl du transport dcs traccurs as to critical groups from a range of exposure anthropiques. Doctor thesis from University Pierre ;tnd pathways. Release rates of radionuclides from Madame Curie, Paris. Nielsen S.P., Iosjpe M., Strand P. (1995) A preliminary waste dumped in the Kara Sea were used as assessment of potential doses to man from radioactive source terms. These data were made available waste dumped in the Arctic Sea. Riso-R-841(EN). Rise from the International Arctic Seas Assessment National Laboratory; Project of the International Atomic Energy Food and Agriculture Organisation of UN (1992). Fishery Agency. The sensitivity analysis has focused on Statistics, Vol. 79, 71. FAO, Rome. three areas: 1) hydrodynamical processes 2) water-sediment interaction processes, and 3) 6.3.2 An Intercomparison Exercise on biological transfer processes. The doses to man Radionuclides in Sediment Samples are generally dominated by contributions from long-lived transuranic radionuclides, plutonium Introduction and americium isotopes, which associate readily Under the EKO-1 Project on marine radio - with sediments. For that reason, the sediment- ecology, which is organized by the Nordic related processes and parameters generally show Nuclear Safety Research Programme (NKS) for high sensitivities, particularly where doses are the period 1994-97, an intercomparison exercise delivered some distance from the dump sites, e g. on radionuclides in sediments was initiated in in the Barents Sea. At closer distances, in the 1995 by Riso National Laboratory. A total of 21 Kara Sea and in the Novaya Zemlya bays, laboratories have participated in the present biological transfer and hydrodynamical processes exercise: 14 from the Nordic countries, 6 from show high sensitivities. the Baltic republics, and one from Japan. This study illustrates that more detailed information on sediment-related processes, biolo ­ Material and Methods gical transfer processes and hydrodynamical pro ­ cesses in the Arctic waters will help reduce the Sediment Samples predictive uncertainty of the dose assessment. Two sediment samples were obtained: one from the Baltic Sea and one from the Kattegat. The References Baltic Sea sediment sample was obtained from Yablokov A.l'., Karasev I'.K., Rumyantsev l ’.A/, Kokeev ME, Petrov O.J. (1993) Facts and problems related to the German Federal Maritime and Hydrographic radioactive waste disposal in seas adjacent to the Agency, who collected the sample in the western territory of the Russian Federation. Materials for a Baltic Sea. The Kattegat sample was obtained Report by the Government Commission on Matters from the Danish National Environmental Related to Radioactiv e Waste Disposal at Sea. Created Research Institute, who collected it in the central by Decree No.613 of the Russian Federation President, Kattegat. Both samples were homogenized and 24 October 1992; Small World Publishers, Moscow. CEC (1990) The radiological exposure of the population of tested for homogeneity by gamma-spectrometric the European community from radioactivity in North measurements on 9 sub-samples. Homogeneity of European marine waters. Project 'Marina'. Commission the sediment samples was demonstrated for Cs- of tire European Communities. Bruxelles, EUR 12483. 137 and natural radionuclides to within a few Nielsen S.P. (1995) A box model for North-East Atlantic percent. coastal waters compared with radioactive tracers. J. The sample material was distributed in June Marine Systems 6: 545-560. 1995, by sending 100 g Baltic Sea sediment and European Commission (1995) Methodology for assessing tire radiological consequences of routine releases of 200 g Kattegat sediment to the participating laboratories. Some laboratories shared the same

92 Ris0-R-972(EN) samples. One participant received a Kattegat Values of z-scores may furthermore, be sediment sample only. During the intercompa ­ combined into a sum of squares of z-scores, SSZ, rison, two of the participants submitted an extra which are calculated according to set of results, so the number of results received per radionuclide varied from 1 to 23. The data W = I>,2 sets were identified by numbers because of i I This value may be used for a test in the %- anonymity. distribution with n degrees of freedom. The SSZ- value is an indicator of analytical accuracy. Statistics Two-sided tests were made for the normal The data were evaluated, by comparing the distribution and the x 2-distribution. The following laboratory mean results of radionuclide concen ­ levels and symbols of statistical significance were trations in the sediment samples, to the used: probably significant (P>95%, *), significant corresponding overall mean concentrations. The (P>99%, **) and highly significant (P>99.9%, procedures were adopted from those used in a ***\ recent proficiency testing exercise on radio­ The scores were combined across the two nuclides in milk powder carried out by the sediment samples for each set of results to allow Central Science Laboratory (CSL 1996). The an evaluation of the laboratory performances for overall mean concentrations were determined for each radionuclide to be made. Additionally, the those radionuclides where six or more sets of data scores were combined across all radionuclides to were received, and after exclusion of outliers, permit an evaluation of the overall analytical determined on a subjective basis. No evaluation performance for each participant. was carried out for radionuclides where less than It is important to note the elements of six sets of data were received. subjectivity in the statistical tests. The overall For each laboratory mean result a z-score was mean values are, in several cases, determined calculated according to after elimination of subjectively identified z=x~ xom outliers, and the target values of the standard deviations (ct,) around the overall mean values are for the standard deviation around the overall chosen on a subjective basis. mean value for the analysis in question. The target values were chosen to be 5%, 10%, 15% or Results 20% depending on the where x is the laboratory Results were received on a wide range of mean value, xom the overall mean value and a, a radionuclides. The artificial radionuclides target value radionuclide and represented comprise Cs-137, Cs-134, Co-60, Sb- 125, Pu- uncertainties typical of those reported by the 239,240, Pu-238, Am-241 and Sr-90. The natural participants. The z-score may be used as a test- radionuclides comprise Ra-226, Th-232, K-40, value in the normal distribution such as values Pb-210, Po-210 and U-235. We present results between -2.0 and 2.0 would be acceptable at the for some of the radionuclide and sediment 5% level. samples where six or more data sets were Values of z-scores may be combined into a received. The graphs show the sorted results and rescaled sum of z-scores, a RSZ-value which is include an indication of the overall mean value calculated according to and the 95% confidence interval for the data on which the overall mean is based. The results are shown with error bars that represent either standard errors of the mean for multiple where n is the number of z-scores being determinations, or reported uncertainties for combined. The RSZ-value may be used for a test single values. Furthermore, an evaluation of the in the normal distribution. The value is an indi ­ laboratory scores is shown. The results are cator of analytical bias. reported in detail in Nielsen (1996).

Ris0-R-972(EN) 93 94

CS-137 (BQ/KG) | CS-137 (BQ/KG) BALTIC KATTEGAT RESULT

Cs-137 SEA

NO.

SEDIMENT SEDIMENT Ris0-R-972(EN) Table 6.3.2.1 Summary of combined z-scores for Cs-137 results showing rescaled sum of scores (RSZ), sum of squares of z-scores (SSZ) and the corresponding levels of statistical significance.

Res. No. RSZ Sign SSZ Sign Res. No. RSZ Sign SSZ Sign 1 1.9 ns 5.6 ns 13 -3.3 ** 10.7 ** 2 -0.1 ns 0.3 ns 14 1.9 ns 3.8 ns 3 -5.1 *** 28.7 *** 15 1.8 ns 3.8 ns 4 -2.4 ** 5.8 ns 16 0.4 ns 7.3 ns 5 1.6 ns 2.7 ns 17 0.3 ns 4.5 ns 6 -0.1 ns 0.0 ns 18 -1.7 ns 3.0 ns 7 -1.2 ns 8.2 * 19 10.0 *** 125.1 *** 8 4.8 *** 23.2 *** 20 0.7 ns 1.0 ns 9 -8.5 *** 84.4 *** 21 3.7 *** 16.4 *** 10 -1.1 ns 1.4 ns 22 -3.3 *** 11.8 ** 11 -0.5 ns 0.8 ns 23 2.1 * 4.4 ns 12 13.4 *** 285.9 ***

The statistical tests in Table 6.3.2.1 show that Pu-239,240 12 of the 23 data sets received on Cs-137, agree The statistical tests show that 5 of the 8 data sets, within 5% of the two overall mean values and received on Pu-239,240, agree within 10% of the that 9 of the data sets disagree significantly. two overall mean values and that one of the data sets disagrees significantly.

BALTIC SEA SEDIMENT

*o md

D Q_

KATTEGAT SEDIMENT

(L3

17 18

RESULT NO.

Ris0-R-972(EN) 95 Ra-226

BALTIC SEA SEDIMENT

19 13 22 16 12 18

RESULT NO.

KATTEGAT SEDIMENT

45

40

35

30

25

20

15

10

5 0 13 22 16 18 8 19

RESULT NO.

The statistical tests show that 3 of the 12 data sets received on Ra-226 agree within 10% of the two overall mean values and that 5 of the data sets disagree significantly.

96 Ris0-R-972(EN) Th-232 f BALTIC SEA SEDIMENT

60

50 ; f < 40 *o d i Te m m 1 30 . i ■ i

20

10

0 ------1 12 16 3 22 13 18 19 5 7 8 1

RESULT NO.

i KATTEGAT SEDIMENT

50

45

40 i

35

30

25 i

20

15

10

5

0 h------1------1------1------—i------1------1------1------1------i 16 13 3 12 22 18 7 5 1 19 8 6

RESULT NO.

The statistical tests show that 6 of the 12 data sets received on Th-232 agree within 10% of the two overall mean values and that 5 of the data sets disagree significantly.

Ris0-R-972(EN) 97 K-40

BALTIC SEA SEDIMENT

1000

900 l { 800 : . . I ftt ii 1 *CD 700 i «- x -c idT 600 1 [

500

400 ----1---1---1---1....1---1---1--- 'T cd m g) co o co r- RESULT NO.

KATTEGAT SEDIMENT

XO d 800 -44- "fi"" • rr 700

600

500

400 15 14 22 11 18 17 4 13 23 6 5 21 3 20 1 7 8 12 19 16 RESULT NO.

The statistical tests show that 16 of the 20 data sets received on K-40 agree within 10% of the two overall mean values and that 4 of the data sets disagree significantly.

98 Ris0-R-972(EN) Radionuclides combined agree within target standard deviations of the The overall analytical performance for Cs-137, overall mean values and that 14 of the data sets Cs-134, Co-60, Pu-239,240, Pu-238, Ra-226, Th- disagree significantly. It should be noted, how ­ 232 and K-40 of each participant may be ever, that the number of results submitted by the evaluated from the data in Table 6.3.2.2. The participants vary considerably, from 2 to 14. statistical tests show that 4 of the 23 data sets

Table 6.3.2.2 Summary of combined z-scores for Cs-137, Cs-134, Co-60, Pu-239,240, Pu-238, Ra-226, Th-232 and K-40 showing rescaled sum of scores (RSZ), sum of squares of z-scores (SSZ) and the corresponding levels of statistical significance. The number of z-scores combined is given as n.

Res. No. n RSZ Sign. SSZ Sign. Res. No. n RSZ Sign. SSZ Sign.

1 8 2.3 * 28.8 *** 13 9 -2.4 * 25.1 ** 2 2 -0.1 ns 0.3 ns 14 4 -0.8 ns 13.0 * 3 11 -3.2 ** 50.2 *** 15 8 25.8 *** 3044.7 *** 4 4 -2.0 * 6.0 ns 16 8 0.3 ns 41.8 *** 5 9 2.9 ** 18.5 ns 17 10 0.8 ns 8.8 ns 6 6 4.7 *** 111.0 *** 18 13 0.2 ns 16.0 ns 7 8 -1.4 ns 21.0 * 19 8 6.2 *** 153.3 *** 8 9 6.5 *** 59.0 *** 20 5 13.7 *** 821.6 *** 9 2 -8.5 *** 84.4 *** 21 7 3.2 ** 23.9 ** 10 2 -1.1 ns 1.4 ns 22 14 -3.2 ** 26.1 * 11 8 -2.3 * 9.1 ns 23 6 -0.6 ns 15.0 * 12 8 9.7 *** 416.0 ***

Conclusions Sb-125, Pu-239,240, Pu-238, Am-241, Sr-90, Ra- An intercomparison exercise was carried out 226, Th-232 and K-40, Pb-210, Po-210 and U- under the EKO-1 project of the Nordic Safety 235. The analytical performance of the partici ­ Research Programme, NKS, for the period 1996- pants was evaluated for those radionuclides 97. The exercise has dealt with the analysis of where six or more data sets were received. artificial and natural radionuclides in two Statistical tests were made to see if individual sediment samples, one from the Baltic Sea and results agreed with overall average radionuclide one from the Kattegat. The measurement concentrations in the two sediment materials techniques have included direct gamma-ray within target standard deviations. The results of spectrometry with Ge and Nal detectors, and these tests are summarised in Table 6.3.2.3 which radiochemical procedures followed by beta shows the radionuclides, the selected target counting and alpha spectrometry. The participants standard deviations, the number of data sets have comprised 21 laboratories of which 14 are submitted and the number of data sets which from the Nordic countries, 6 from the Baltic disagree significantly with the overall mean republics and one from Japan. values. Results were submitted for a large number of radionuclides comprising Cs-137, Cs-134, Co-60,

Ris0-R-972(EN) 99 Table 6.3.2.3 Summary of analytical performance across all data sets.

Nuclide Target standard deviation Number of data sets Number of data sets that disagree significantly with (%) performance criteria Cs-137 5 23 9 (39%) Cs-134 20 6 2 (33%) Co-60 15 6 0 (0%) Pu-239,240 10 8 1 (13%) Pu-238 20 8 1 (13%) Ra-226 10 12 5 (42%) Th-232 10 12 5 (42%) K-40 10 20 4 (20%) Nuclides combined 23 14 (61%)

It is noteworthy, that with the exception of Co- 6.4 Contamination Physics 60, the performance criteria are not met in about 20-40% of the data sets for the relatively straight 6.4.1 Decontamination in a Russian forward gamma-spectrometric analyses of Cs- Settlement (J. Roed) 137, Cs-134, Ra-226, Th-232 and K-40. For the more complicated radiochemical analyses of Due to governmental restrictions, implemented to plutonium isotopes, only 13% of the data sets do reduce the radiation level from radionuclides not meet the performance criteria. The test of the deposited on various types of surface after the overall analytical performance shows that 61% of Chernobyl accident in 1986, large previously the data sets do not meet the combined per ­ inhabited areas in Russia are now deserted. formance criteria. This shows that there is room Almost a decade after the accident, investiga ­ for considerable improvement in analytical tions, supported by the Emergency Management quality for most of the laboratories which have Agency under the Danish Ministry of the Interior, participated in this intercomparison. were made on the effect of different feasible The intercomparison exercise has, further ­ methods to reduce this level of contamination. more, demonstrated several elementary problems The ultimate goal of the work was to increase the in analytical work such as interchange of interest for decontamination possibilities of samples, mistakes in calculations and correction residential areas, thereby supporting the process of background levels. High-quality analytical towards a stoppage of migration from the performance requires careful and dedicated staff contaminated areas, and instead facilitate a using well-established laboratory procedures and resettlement of the deserted areas. frequent participation in international intercompa ­ A mobile laboratory travelled from Denmark rison exercises. through Poland and Belarussia packed with equipment for the dose-reducing treatment of the References area and for measurements and mapping of the CSL (1996) Gems/Food-Euro Proficiency Testing contamination levels. The Novozybkov Branch of Exercise 95/01. Central Science Laboratory, Norwich, U.K. the Russian Institute for Radiation Hygiene Nielsen S.P. (1996) An intercomparison exercise on located a suitable undisturbed area in Russia, radionuclides in sediment samples. Riso-R-914(EN), where the work could take place - a settlement Rise National Laboratory. called Novo Bobivichi, by the bank of the Iput river, approximately 25km north-north-east of Novozybkov. This settlement was not permanent ­ ly inhabited and had a sufficiently high level of contamination, about 1 MBq/m2, to enable the

100 Ris0-R-972(EN) effectiveness of each step of the decontamination contamination in the decontaminated soil area. strategy to be accurately measured. Other Russian The strategy, therefore, comprised the following institutes also participated in the collaborative steps: work: St. Petersburg Institute of Radiation Hygiene, the RECOM Institute in Moscow, the 1) Removal of the major part of the soil Typhoon Research Institute at Obninsk and the contamination in an area of about 20m by 20m Obninsk Institute of Nuclear Power Engineering. around the three wooden houses soil by A total of 18 people participated in the work over skimming off the top 5 to 10 cm layer. a period of three weeks. 2) Cleaning of the roofs of the three wooden The strategy for decontamination of the area houses in the central part of the area. was worked out as follows: 3) Application of a clean soil/sand mixture to the After the mapping of the site, it was clear that decontaminated soil area. the recorded deposition pattern on the different surfaces in the area was in good agreement with The tools applied for the soil removal were previous observations after a wet deposition of spades, shovels and wheel barrows. The deconta ­ B7Cs. mination of the roofs included two steps: The main contributor to the dose rate inside the wooden houses in the area was found to be 1) Removal of organic litter from the roofs, soil contamination. The radiocaesium on the needles, leaves, etc. roofs was also found to give a significant contri ­ 2) Cleaning of the asbestos sheets. bution to the dose rate, while the contamination level on the walls was found to be negligible. The For the cleaning of the asbestos sheets, a many large pine trees in the area had changed specially constructed roof cleaner was brought to their needles several times since the accident and the area. This specially constructed device the present level of contamination was fairly low, consists of a rotating brush with a shape that compared to the contamination level of the soil. follows the curves of the roof. The device is The road pavements, in and around the area, had driven by pressurized air at 700 1/min., 7-8 bar, a very low contamination level. Due to traffic and and uses water at ordinary tap water pressure. weathering, the level of contamination on such The fact that the cleaning is carried out at low surfaces usually decreases by at least an order of pressure and in a wet medium makes the method magnitude over the first decade following especially suitable for work in contaminated deposition. areas, as the resuspension hazard is minimized. It was decided to focus on soil and roofs. In a Further, this relation satisfies the existing legal real case of decontamination it would be natural demands in Europe regarding treatment of to begin with the roof, followed by the removal asbestos surfaces. The brushing pressure and of the top soil layer, as water from the roof might cleaning function were appropriately adjusted to raise the contamination level of the soil around ensure that only coatings, such as moss and algae, the house. In this case, however, it was decided to were removed, while the abrasive effect on the reverse the order of the implementation of these roof material was negligible. An earlier experi ­ procedures in order to measure the effect of the ment showed that very little caesium conta ­ roof cleaning with better accuracy. The removal mination would be found in the liquid phase of of the soil was expected to give a highly the washed off material from such operations, < significant dose rate reduction and the measure­ 1%, while the rest would be associated with the ments of the dose rate, before and after the roof solid fraction, moss, algae, etc. was cleaned, could then be made with less A novel technique was applied for the influence from contamination on other surfaces. management of the waste from the roof cleaning Consequently, the uncertainty of these measure­ operation. As the radioactive matter is almost ments would be smaller. Finally, it was decided entirely associated with the solid waste, the that a mixture of clean sand and soil was to be amount of waste can be greatly reduced by added to the cleaned area after the soil removal, filtration. The waste is collected in the roof gutter to obtain a shielding effect against any remaining and led through a down pipe into a large vessel,

Ris0-R-972(EN) 101 where a filter material coating on a plastic area than would be treated, and the real effect covered metal net filtrates the solution so that would certainly be greater. only the liquid fraction can penetrate. On the 2) Direct radiation is not the only significant other side of the filter, the water is pumped to source. Sky-shine, the dose increasing effect another vessel from which it can be applied again on radiation scattered in the air, may also be of for the decontamination operation. The filtering great importance, as sky-shine radiation may material is a commercially available polymer travel over considerable distances. It is pos ­ fibre textile called Typar, with a pore size of sible that this widespread radiation may 0.14mm. The cost of this material is only about penetrate poorly shielded houses through thin 0.50 ECU/m2. Using this method, it is possible to roofs and walls. recycle the water applied for decontamination without risk of redistributing the removed 3) A part of the residual dose rate contribution material. This also has a beneficial effect on the from Chernobyl fallout could also come from cost of carrying out the procedure, which could the residual soil contamination in the treated be kept well under 1 ECU per square meter area. However, the shielding effect of the decontaminated. application of a 4 cm layer of gravel or sand Looking at all the information obtained from after the soil removal was found to be the decontamination exercise, it is possible to considerable, and the shielding layer has account for the fraction by which the initial dose undoubtedly reduced this dose rate contribu ­ rate was reduced, by each of the different tion by at least a factor of 2. decontamination steps. It is also possible to 4) Contamination of indoor surfaces may also identify the likely sources of the residual have contributed to the dose rate inside radiation. Figure 6.4.1 shows the fractions by buildings. Only small radiocontaminant which the dose rate was reduced by imple ­ concentrations would be required on indoor mentation of the different countermeasures. The surfaces to give a significant dose contri ­ dose rate pies, refer to measurements of the dose bution, as the indoor shielding is often rate made inside the three different houses after negligible. The indoor surfaces may have been each step of the decontamination strategy. As can exposed to airborne contamination, but as the be seen, the total effect of the strategy is a contamination took place with rain, the dose reduction of the dose rate due to the Chernobyl contributions from deposition on walls, indoor fallout by up to 64%. Several sources in the or outdoor, are negligible. However, contami ­ environment may contribute significantly to the nation from the outdoor environment may residual fallout component dose rate: have accidentally been brought into the houses 1) The residual dose rate is greatest in house 4, by their occupants, for instance under the which lies closest to the untreated soil areas shoe-soles. and smallest in house 6, which is surrounded 5) The roof of house 6 was actually replaced. The by decontaminated soil and shielding neigh ­ other two roofs were cleaned, see above, and bouring walls. Therefore, it seems evident that still carried some contamination. In all 3 cases, the dose rate contribution from surrounding contamination may also have reached the soil areas is relatively large. Recent Monte construction materials directly under the roof, Carlo calculations, based on the recorded and there may be a dose rate contribution from vertical distribution of the contamination in this, even after the decontamination strategy the soil, have shown that the dose rate in a was carried out. field, probably comes from a much greater

102 Ris0-R-972(EN) Figure 6.4.1 Pie charts showing the contributions to dose rate inside houses 4, 5 and 6 from the different contaminated surfaces in the environment. The effects of the different cleaning operations are indicated. After the operation, a cosmic, a natural and a residual fallout contribution remain.

Ris0-R-972(EN) 103 The overall conclusion of the decontamination logue existed at Riso, which was therefore given work at Novo Bobovichi is that, contrary to this role. previous claims, it is still possible to obtain a Throughout the decade that has passed since significant reduction of the external radiation the Chernobyl accident in 1986, the impact of a level, although 10 years have now passed since contamination of different types of inhabited the Chernobyl accident contaminated the area. In areas has been investigated carefully. For house 6, a reduction of the dose rate component instance, the contamination levels on different originating from the Chernobyl accident by a surfaces in and around the town of Gavle, factor of 3 was achieved; if the procedure were to Sweden, which received a relatively high level of be applied over a large area, as it would be in contamination from the Chernobyl accident, have practice, calculations have shown that a reduction been followed through the years in seven Riso by a factor of about 5 is anticipated. It has measurement campaigns. previously been claimed that deposition of The URGENT model, which was developed at resuspended contaminated soil from outside a Riso mainly on the basis of this in situ cleaned site would rapidly lead to a re­ measurement data, was one of the very first contamination, but subsequent measurements of dynamic external dose models to comprise the the radiation levels have shown the effect of whole urban environment in the event of an resuspension to be without importance in this accidental contamination with radiocaesium. It context. was, for instance, the first extensive model to take into account the contamination on trees and Reference vegetation, which has proved to be a potentially Roed J., Lange C., Andersson K.G., Prip //., Olsen S., important factor in dry deposition scenarios. The Ramzaev V.P., Ponomarjov A.V., Borkovsky A.N., model uses the results of Monte Carlo photon Mishina A.S., Vorobiev B.F., Chesnokov A.V., transport calculations to link the time-dependent, Potapov V.N., Shcherbak SB. (1996) Decontamina ­ tion in a Russian settlement. Riso-R-870(EN), ISBN radio-contaminant concentrations to dose rates to 87-550-2152-2. persons staying at different locations in different housing environments of varying population 6.4.2 Evaluation of Decontamination density. Procedures (K.G. Andersson) The URGENT model was used, together with practical experience from semi-large scale A specific request for an identification of a decontamination testing (e g. Roed et al. 1995, preparedness strategy for early phase clean-up, in 1996, Roed and Andersson 1996), to evaluate the the event of a major accidental contamination of feasible remedial countermeasures when applied inhabited areas, has recently been presented by a in the early phase. The modelled scenarios reflect special law in Sweden. Due to obvious common what is considered to be typical situations in the interests between the different Nordic countries, Nordic countries with areas of varying population it was decided to make the investigation the density, from small single-family houses, made responsibility of the Nordic Committee of of wood or brick, to multi-storey house blocks, Nuclear Safety Research, NKS. and varying weather conditions. The need for a catalogue, which could The resulting catalogue consists of 66 facilitate the local decision-making process was individual data sheets. Each data sheet describes required, stating the advantages and dis­ the advantages and disadvantages of the intro ­ advantages of each method considered to be duction of a specific clean-up procedure in the feasible for cleaning of each type of contaminated early phase, after a contamination of a specific surfaces in a few representative Nordic housing type of area. environment scenarios. Only external doses were The relative contribution of the surface to be to be considered. treated, e g. grass, walls, roofs, etc., to the dose After the first meeting of the Nordic working rate in the early phase is estimated. Also, the group, it became apparent to NKS that the lifetime dose, relative to the initial contamination expertise and experience to make such a cata­ level, to people staying in a particular type of area, assuming that no dose-reducing measures

104 Ris0-R-972(EN) are introduced, is treated. Further, estimates are account the contamination, not just on the given of the achievable immediate dose rate ground, but also that on other outdoor surfaces reduction, if the clean-up procedure is carried out such as roofs, walls and trees. Furthermore, we in the early phase. Finally, the achievable reduc­ have considered the potential contribution to dose tion of the lifetime dose to people staying in a from contamination on indoor surfaces which, type of area is estimated. prior to the investigations at Imperial College On the background of a general evaluation of London and Riso, was considered by several the current situation, this information can be used countries at the forefront of radiological protec ­ to estimate the potential beneficial effects of the tion, e g. Germany, the U K. and Sweden, to be implementation of a countermeasure. of little or no consequence. Also stated on each data sheet is a short The Contamination Physics Group at Rise is description of the method, with e g. indications now addressing the potential problem of dose on the personnel requirements and costs, from beta-emitters deposited indoors on clothing overheads, costs of necessary equipment, and on human skin. This is the subject of the amounts and costs of disposal of waste, etc. following experimental investigation. Likewise, additional instructive chapters were supplied by Riso on, e.g. waste disposal, which is Method a problem of general nature, on indoor conta ­ In order to measure very small amounts of mination, which in some cases needs careful particulate, deliberately deposited on surfaces in consideration, and on methods to actually form a experiments, we use particles labelled with clean-up strategy from the data sheets. dysprosium and/or indium, which on neutron activation yield the radionuclides 165Dy and U5ln References respectively. For the particle size range 2-8pm RoedJ., Andersson K.G., Prip H. (eds.) (1995) Practical diameter, we use dysprosium-labelled porous means for decontamination 9 years after a nuclear spherical silicon-oxide, and for producing accident. Riso-R-828(EN), ISBN 87-550-2080-1. particles in the sub-micron range, about 0.5pm RoedLange C., Andersson K.G., Prip II., Olsen S., Ramzaev IP., Pononiarjov A A'., Borkovsky A. N., AMAD, we nebulize a suspension of indium Mi shine A.S., Vorobiev B.F., Chesnokov A. V., acetyl acetonate in alcohol. Following deposition Potapov (’.A., Shcherbak SB. (1996) Decontami ­ on surfaces, the particles are measured by nation in a Russian settlement. Riso-R-870(EN), neutron-activation/ gamma-ray spectrometry. ISBN 87-550-2152-2. Roed ,/., Andersson K.G. (1996) Clean-up of urban Experiments areas in the CIS countries contaminated by Six human adults were placed in a room. Five sat Chernobyl fallout. J. Environ. Radioactivity 33(2): 107-116. around a table playing cards and the sixth, a member of the technical team, moved around the 6.4.3 Deposition on Skin (J. Roed) room as necessary. Three of the card-players were naked to the waist so that deposition to the Introduction chest, back, and shoulders could take place. The For many years, the gamma radiation dose to other three persons wore white cotton coats. people inside buildings from radioactive fallout Filter papers, Whatman 542, were fixed to parts was calculated from measurements of radiation of the ceiling, walls, window and floor to on the ground. The standard procedure is to measure deposition on these surfaces. Deposition measure the dose rate at 1m above the ground and on the cotton coats was measured by cutting out employ a shielding factor to allow for attenuation the appropriate areas and analysing them by by the building itself. This approach is now neutron activation/gamma spectrometry. The known to be too simplistic for predicting dose filter papers were analysed by the same method rate to people in multi-storey buildings and is The mass of particulate deposited on skin was inadequate even for two-storey houses. determined by three times cleaning by filter paper In order to improve the current situation, by and neutron activation/gamma spectrometry. making more accurate assessments and dose The concentration of particles in the air inside predictions, our approach was to take into the room was measured by analysing filters

Ris0-R-972(EN) 105 through which known volumes of air had been subsequently transferred to either clean BW, drawn. clean fresh water (FW) or clean seawater (SW) at 16°C. The fish were labelled with 32P-phosphate Results and 14C-acetate. Depending on previous exposure Deposition velocities to skin were found to range to Cu, 32P- and 14C-labelled gill membrane from 0.05 to 0.4cm per second for particles of phospholipids degraded if the fish were diameter 2.5pm, and 0.002-0.02cm per second transferred to SW or remained in BW, but not for 0.5pm particles. These values are higher than when they were transferred to FW. Corre­ those found for deposition to walls inside houses, sponding experiments, where the fish were where the figures were 0.05-0.12cm per second exposed to Cu in BW for 12 days, showed a for 2.5pm and 0.001-0.005cm per second for similar subsequent degradation in SW and BW of particles of about 0.5pm AM AD. Although some both gill and esophagus membrane phospho ­ samples have yet to be analysed, we have already lipids, but to a much lesser degree in gill tissue seen some surprising results and there is no doubt than after 4 days Cu exposure. Plasma Na was that this approach will lead to a much improved similarly reduced by up to 8%, depending on model for predicting indoor dose from fallout. previous ambient Cu, but in this case only after transfer to FW and only after 4 days of exposure. Both the effect on membrane lipid metabolism 6.5 Ecophysiology and on plasma Na+ thus showed acclimation to (H.J.M. Hansen, M. Groseil) ambient Cu, but there was apparently no direct correlation between the two different types of 6.5.1 The Effect of Cu on Gill and Esophagus observed changes in membrane function. Lipid Metabolism in the Rainbow Trout (Oncorhynchus my kiss) Reference Hansen H.J.M, Olsen A.G., Rosenkilde P. (1996) The effect of Cu on gill and esophagus lipid Rainbow trout (Oncorhynchus mykiss) were metabolism in the rainbow trout. (Oncorhynchus exposed to Cu at 0, 100, 300 and 800 pgl" 1 in mykiss). Comp. Biochem. Physiol. 111C: p. 2.1-29. brackish water (BW) for 4 days at 13°C and

Figure 6.5.1 Degradation of gill membrane phospholipids after exposure to ambient Cu. Distribution patterns (%) of,4C- and 32P- activities among various trout gill phospholipids. Exposure to 0, 300 and 800 pgr 1 Cu in brackish water, BW, for 4 days and subsequent incubation with (l4C) acetate and (i2P) phosphate in seawater, SW. P, O summarizes the activities found in phosphatidylinositol, phosphatidyl- serine, sphingomyelin, lysophosphatidylcholine and phosphatidic acid; PC, phosphatidylcholine ; PE, phosphatidylethanolamine; P, 1 summarizes cardiolipin and various unidentified degradation products.

106 Ris0-R-972(EN) 6.5.2 Acclimation to Cu by the Highly involves an increased clearance of plasma Cu, Tolerant European Eel (Anguilla anguilla) especially in the rainbow trout, leading to a rapid and the More Sensitive Rainbow Trout normalization of plasma Cu, even during chronic (Onchorhynchus my kiss) exposure. The increased clearance of plasma Cu does not involve an increased accumulation in Preliminary studies have shown the European eel any of the investigated tissues, but in the to be highly tolerant to Cu (96h LC50>1500pg European eel rather a decreased hepatic Cu V1) compared to the rainbow trout. accumulation. The exchangeable Cu pools in the We have followed the ongoing uptake, liver of both species and in the kidney of rainbow turnover and excretion of Cu during chronic trout were found to be high, which leads us to exposure to waterborne copper in the European focus on Cu excretion; we found that Cu eel and the rainbow trout, using the radioactive acclimation involves an increased renal and isotope 64Cu, and found that Cu exposure to hepatic Cu excretion, to a different extent in the lOOpg Cu.I'1 for 28 days did not result in a two species. reduced branchial Cu uptake in the European eel. The acclimation did not lead to any significant Reference changes in branchial Cu affinity (K^) or binding Grosell M, Boetius /., Hansen Rosenkilde P. capacity (Vmax ). Furthermore, we have shown that (1996) Influence of preexposure to sublethal levels of copper on 64Cu uptake and distribution among >90% of the accumulated Cu is found in the tissues of the European eel (Anguilla anguilla). plasma and that Cu exposure leads to an initially Comp. Biochem. Physiol. 114C: p. 229-235. increased plasma Cu. The acclimation clearly

Newly accumulated Cu, bile starved fish

12 pg Cu l

lCu exposure (hours) Figure 6.5.2 Excretion of newly accumulated Cu via the bile is enhanced in eels adapted to 94pg Cu X1 for 6 (A or 28 (a) days, relative to controls (A); it is only slightly enhanced in eels equivalently adapted to I2pg Cu X'1 (•).

6.6 Radioecology (A. Aarkrog) AAR. The report will be published in June 1997, and the authors are not permitted to present any 6.6.1 The Arctic Monitoring and Assessment data or conclusions from the report before the Programme, A MAP official publication is available. The chapter on radioactivity will comprise Rise has participated as a key expert in the approximately 200 pages and after an intro ­ AMAP drafting group for the chapter on duction, will deal with fundamentals and radioactivity to the AMAP assessment report, definitions relating to radioactivity. This is

Ris0-R-972(EN) 107 followed by a description of past and present will be given. Thereafter follows a radiological contamination in the Arctic, dealing with wide­ assessment of potential releases, for example a spread contamination on land and sea, as well as hypothetical release from one of the Russian with localized contamination, for instance that nuclear vessels or storage sites located in the from the Thule accident. The next section Arctic. The section on spatial analysis of the describes the individual doses to man from vulnerability of Arctic ecosystems, compares natural and man-made contamination in the 8 radioecological sensitivity of the various Arctic Arctic countries, Alaska, Canada, Greenland, countries. This is based on the individual doses Iceland, the Arctic parts of Finland, Norway, assessed for the various countries and on the Russia and Sweden. In the section on Radio- production of Arctic key food products (e g. ecological assessment of previous releases, the reindeer). The chapter ends with conclusions and radiological impact in the Arctic of nuclear recommendations, where a number of defi­ weapons testing, liquid discharges from nuclear ciencies in present knowledge and abilities reprocessing plants, e g. Sellafield in the U.K., relevant to radioactivity in the Arctic are accidental releases, e g. the Chernobyl accident, identified.

I'lgure 6.6.1 Landscape in Greenland

6.6.2 South Ural Contamination Studies, our participants from Russia (IPAE in the Urals) SUCON and Ukraine (IBSS in the Crimea). The project was launched by a planning Through the Fourth Framework Programme from meeting at Riso in March, where a group of US 1996, the EU has supported a project on: scientists, working in the Ob river system, also Radiological assessment of past, present and participated. The aim was to co-ordinate the potential sources to environmental contamination European and American activities and to in the southern Urals and strategies from remedial exchange samples from our respective pro ­ measures, with the acronym SUCON. grammes. Two environments are studied: Rise is co-ordinator of the project with 1) The area in the Urals contaminated by the so- participants from the UK, Norway, Russia and called Kyshtym accident in 1957, Ukraine. The project is finansed through INTAS 2) The Ob river system, which was contaminated and MIKA, which makes it possible to support around 1950 by liquid radioactive releases to

108 Ris0-R-972(EN) the Techa river from the nuclear facility and one was received from our US colleagues, MAYAK in the South Urals. who were more successful than the Rise With regard to the Kyshtym contamination, a scientists in their sediment sampling efforts. The large number of soil samples were analysed for sediment samples analysed (Figures 6.6.2.1-4) 90 Sr, 137Cs and the transuranics, Pu and Am. (see show two peaks of 137Cs. The upper one may be Figures). due to the Chernobyl accident in 1986 and the The preliminary results indicate that approxi ­ lower is ascribed to the peak in global fallout mately 1 kg Pu might have been released by the around 1963. The samples analysed give no Kyshtym accident to the environment. Russian indication of a possible 137Cs contamination of reports have given figures nearly two orders of the sediments due to the above mentioned magnitude less than this estimate. releases from MAYAK around 1950. Such a The studies in the Ob river system have peak is observable in sediments from the Techa focused on river sediments. The aim has been to river in the upper part of the Ob river system, but obtain undisturbed sediment columns, which is apparently not present in the lower parts. The make it possible to date the sediment layers by question is whether a more mobile radionuclide, 210Pb. Undisturbed columns are difficult to such as 90 Sr, is detectable further down the river. obtain, but so far 3 have been obtained by Riso This will be studied in 1997.

57°N

EKATERINBURG, f2*- 24 Jfe > x 19

12 ** 18 ♦ -14 V '■ , 0' ■J' 7 o — io i1-1 " 7 ik 8 v 11 - & V? ‘ , 0 0 \\ ; ^9 A O' ' 56°N

55°N

o Ch m Figure 6.6.2.1 Sampling locations for soil 1990-1995 in the South Urals

Ris0-R-972(EN) 109 Sr-90

0.30 kBq/m2

global fallout Kyshtym Karachay

Figure 6.6.2.2 Relative mean distribution between sources of contamination for 90Sr, I37Cs and 239,240Pu in areasd contaminated both by the Kyshtym and Karachay accidents. The example shown is location No. 13, 73 km NE ofMAYAK.

Cs-137 in sediments from Ob-river 1995. Location 5 core C.

Depth (cm) Figure 6.6.2.3 Cs-137 profiles in Ob river

110 Ris0-R-972(EN) 7 Other Activities Photochemistry and Photophysics of Macrocyclic Compounds The work in this field is related to the general 7.1 Molecular Dynamics and area of material science. This refers in Photochemistry (R. Wilbrandt) particular, to optical switching on a molecular level, photochromism, and new optical mate­ rials, ferromagnetism, non-linear properties, and Ions and Free Radicals organic superconductors. In collaboration with The activity in this area was concentrated mainly the University of Copenhagen, we have been on two projects: 1) studies of radical cations and working on the photophysical properties of anions of importance for conducting polymers truxene. It is a large molecule, suggested and under stable conditions by resonance Raman studied previously as a possible ferromagnetic spectroscopy in low temperature glasses, and 2) material in the context of organic supraconduc- the structure of radical cations of amine-based tivity. In particular, fluorescence and phos ­ non-aromatic ring compounds, i.e. of electron phorescence properties, but also time-resolved donors important in supramolecular donor- absorption and Raman investigations, were acceptor complexes. carried out. This work is continuing in ongoing Short-lived species of importance in the studies of triphenylene and some of its photoconductivity of conducting polymers such derivatives as polyacetylene were studied. Such species include the butadiene and hexatriene radical References cations, but also other radical anions, cations Keszthelyi T., Wilbrandt R. (1996) The radical cation of and free radicals of conjugated polyenes. In trans- 1,3 -pentadiene: Resonance Raman spectrum particular, we have carried out a detailed study and ab initio calculations. J. Mol. Struct. 379: 211- of the butadiene radical cation and some of its 218. isotopomers in collaboration with the Institute of Keszthelyi !., Wilbrandt R., Bally T. (1996) The radical cation of 1,3-butadiene: Resonance Raman spectrum Physical Chemistry, University of Fribourg, of the hfi isotopomer. J. Phys. Chem. 100: 16843- Switzerland. 16849. In collaboration with the University of Keszthelyi T., Wilbrandt R., Bally T., Roulin J.-L. Amsterdam, The Netherlands, we continue a (1996) The radical cation of 1,3-butadiene: Reson ­ time-resolved resonance Raman and theoretical ance Raman spectrum of deuterated derivatives and ab initio study of a large number of radical improved force field. J. Phys. Chem. 100: 16850- cations of piperidine and piperazine based 16855. Keszthelyi T., Wilbrandt R. (1996) The radical cations chemical systems. A mainly theoretical study of of monomethyl substituted 1,3-butadienes: Reson ­ the dimethylaniline radical cation has appeared ance Raman spectra and molecular orbital calcula ­ in 1996. These radical cations can easily be tions. J. Phys. Chem. 100: 15785-15793. generated as transient species in polar solution Wilbrandt R. (1996) Time-resolved resonance Raman by laser photolysis. The resonance Raman spectroscopy: A retrospect on the early days. spectra of these compounds will allow us to gain Biospectroscopy 2: 263-275. Brouwer A.M., Wilbrandt R. (1996) Vibrational spectra a deeper understanding of intramolecular of N,N-dimethylaniline and its radical cation. An electronic charge distribution and interactions in interpretation based on quantum-chemical calcula ­ these species, which from a structural point of tions. J. Phys. Chem. 100: 9678-9688. view, are largely unknown. As these molecules Brouwer AM., Wiering P.G., Zwier J.M., Langkilde form part of many supramolecular systems in F.W., Wilbrandt R. Localization versus delocaliza ­ the context of electron transfer, we expect that tion in diamine radical cations. Acta Chem, Scand. Accepted. our results will be of general interest to the large Ojfersgaard J.F., Veng T., Skettrup T. (1996) Accurate community interested in electron transfer, method for determining the refractive-index profiles molecular switching, molecular computers, of planar waveguides in uniaxial media with the supramolecular photochemical devices, etc. optical axis normal to the surface. Appl. Opt. 35: 2602-2609.

Ris0-R-972(EN) 111 7.2 Dosimetry and Industrial Consulting activities were carried out in Irradiation (A. Miller) connection with maintenance and testing of irra­ diation of facilities and the establishment of new facilities. 7.2.1 Rise Industrial Irradiation One training course in Validation and Routine Monitoring of Electron Beam Sterilization with The volume of products irradiated at the 10 8 participants was arranged. This was the 7th MeV electron accelerator increased during 1996, arranged course within the past 4 years. A mainly due to irradiations for one of the meeting on Documentation Requirements for the irradiation facilities with whom we have mutual Radiation Sterilization with 30 participants was back-up agreements. The Rise accelerator has arranged in co-operation with DMDA. functioned without major problems, and scheduled repairs of the conveyor were carried 7.2.3 Dosimetry for Radiation Processing out during the summer. As a function of the newly approved EN and Calorimeters for the measurement of nominal ISO standards, the documentation requirements dose and also for the calibration of other for the irradiation process have been increased. dosimeters by irradiation at electron accelerators We were able to meet these requirements with energies down to approx. 1 MeV are being through our ISO-9002 certification, which was developed. This work is carried out under a reconfirmed by Danish Standard Association at research agreement with the IAEA. semi-annual audits. We have accepted IAEA fellows for visits here, in connection with the establishment of 7.2.2 High Dose Reference Laboratory traceable dosimetry in the fellow’s home country. DANAK has approved the accreditation for a We are engaged in two EU-contracts: one new 5-year period. During the approval process, under the programme for Standards Measure­ the alanine/EPR dosimeter was accepted under ment and Testing SMT and the other one in the the accreditation in the range 1-60 kGy, but Inco-Copernicus programme for collaboration measurements down to 50 Gy can be calculated with Eastern Europe. The SMT contract, with good precision. We have carried out the Dosimetry for Radiation Sterilization, concerns following tasks under the accreditation: the establishment of standard irradiation condi ­ tions for dosimetry calibrations and intercompa ­ rison. The project involves intercomparisons Dose map reports 50 with approximately 30 European industrial Irradiation at cobalt-60 gamma facilities. cell of dosimeters or samples 2 The Inco-Copernicus project, Accreditation Irradiation at 10 MeV electron accelerator for High Dose Measurements, is aimed at of dosimeters or samples 2 establishing documentation for the high dose Issue of calorimeters 10 measurements which may lead to accreditation at Issue of dichromate reference dosimeters 7 the involved laboratories: Institute of Isotopes, Issue of FWT-60 reference dosimeters 4 Hungary, Institute of Nuclear Chemistry, Issue of alanine reference dosimeters 2 Poland, Institute of Nuclear Physics, Albania. Measurement of irradiation We participate in the standardization work in facility parameters 5 DS, CEN, ISO and ASTM where we take part Measurement of dose distribution in the writing of the new standards and in irradiated products 55 guidelines for radiation processing.

112 Ris0-R-972(EN) 8 Special Facilities

8.1 The RIMI Field Station summer were normal regarding rainfall and tempe ­ (K. Pilegaard) rature, thus favouring a good yield of plant pro ­ duction. Rise's Integrated Environmental Project (RIMI) Yield mapping, with a Dronningborg combine is an interdisciplinary project studying pathways, equipped with GPS and a yield metering device, processes and effects on terrestrial ecosystems of was again used. The demonstration field, with Site nitrogen compounds, derived from human activi­ Specific application of N-fertilizer, did not show any ties. RIMI is managed by the Ecology section and differences in yield, presumably due to the very high involves scientists from the Environmental content of N-mineralization in the spring. Science and Technology Department, and scien ­ tists from the Meteorology and Wind Energy Department. The RIMI field station is a part of 8.3 Open Top Chamber Facility the project. It is used in the Danish contributions (V. Haahr) to the EUROTRAC projects BIATEX (managed by the Ecology section) and TOR (managed by In collaboration with the National Environmental the Chemistry section). A number of projects Research Institute, the technical performance of the within the SMP Center for Air Pollution Pro­ chambers was modified for experiments using cesses and Models make use of the field station. different concentrations of carbon dioxide. The One of these projects, concerning the atmospheric influence of different levels of C02 on spring wheat composition of nitrogen compounds, is managed was investigated in 1996. by the Chemistry section and performed in collaboration with the National Environmental Research Institute. 8.4 Rise Environmental Risk Apart from the continuing meteorological Assessment Facility, RERAF (K. Nilsson) observations carried out at the site, a special experiment was carried out in 1996. The purpose The RERAF-facility consists of two identical units, was to evaluate new equipment (ID Sonics) for RERAF 1 and RERAF2. The former was built the measurement of vertical wind speed. The during 1995, and was expected to be ready for use station has also been used as a test bed for a in the spring of 1996. However, due to a number of instruments and experimental designs. replacement of one of the original engineering The RIMI station serves as a reference station consultants, the starting date for RERAF 1 was for air pollution monitoring in Copenhagen by the postponed until the end of 1996. National Environmental Research Institute. RERAF 1 consists of six 24 m2 greenhouses with a height of 3 meters to the glass roof. Directly underneath each greenhouse is a 3-meter-deep root 8.2 Dyskaergard, the Experimental chamber, also 24 m2 in size. The greenhouses face Farm (V Haahr) north, and are equipped with a type of glass which allows the maximum amount of external daylight to enter. The greenhouse roofs have mobile high- The climate during the autumn and winter 1995-96 pressure Hg lamps, giving a maximum foton flux of was characterized by an extremely low amount of approximately 1000 mmol m"2 s'1 within the range rain and snow. This resulted in no drainage and of 400-700 nm. In the greenhouses, it is possible to consequently, in a very high amount of N-minerali- mix sunlight and artificial light in a controlled zation in the soil during the spring period. For that manner, since the lights can be controlled auto­ reason, the nitrogen fertilization was reduced by matically and daylight used whenever possible. 20%. The weather conditions during the spring and Mobile lamellar shutter systems are located above the lights. These allow individual simulation and

Ris0-R-972(EN) 113 variation of the daily lighting rhythm in the green ­ houses. Each of the greenhouse cubicles is equipped with its own individual climate control system, which controls light, temperature, humidity and composi­ tion of the air inside the growth chambers, and in addition, can simulate the diurnal cycle for all three parameters in a near natural manner. In the RERAF, it is possible to establish climatic conditions ranging from sub-arctic to tropical. Gasses such as nitrogen, oxygen, carbon dioxide, as well as air pollutants such as ozone, NOx and SOx can be added. The chambers can, when required, be certified for experiments with radioisotopes and with transgenic organisms. As a first performance test, tomato plants were placed in one of the climate chambers. Despite less than optimal conditions in this preliminary test, due to the conditioning systems not being completely under control, the tomato plants fared well and produced well-developed and tasty tomatoes. Later, maize and rapeseed, both of which are quite demanding with respect to the climatic conditions, were also tested in the greenhouse chambers with good results. However, the technical tests of the climatic chambers are still in progress and there have been some problems. These are currently being rectified. RERAF 1, and in the future also RERAF2, embody advanced plant growth facilities with separate root and shoot compartments, so that it will be possible to operate, and independently control, experimental conditions of both above and below ground parts of integral ecosystems. The climate and nutrition can be controlled by models based on empirical data or by feedback from analytical instruments. Using the RERAF-facility, it will be possible to conduct fully integrated multi-parameter experi ­ ments, where whole plants or plant materials are cultured under climatic and nutritional control over long periods of time. The biological material can be tested under steady state or transient conditions, or under conditions simulating the fluctuating natural environment.

114 Ris0-R-972(EN) h

Ris0-R-972(EN) 115 9 Publications

9.1 Refereed Journals Christiansen (.. Kunzendorf 11., Laima M.J.C., Lund- Hansen L.C., Pedersen A.M. (1996). Recent changes in environmental conditions in the southwestern Aarkrog A., Baxter MS., Bettencourt A.O., Bojanowski R., Kattegat, Scandinavia. NGU Bull. no. 430. 137-144. Botoga A., Charmasson .S', Chunha /., Delfanti R., Dahl S.W., Rasmussen S.K., Hejgaard J. (1996) Duran hi., Holm II., Jeffree R. Livingston H.D., Heterologous expression of three plant serpins with Mahapanyawong S., Nies II., Osvath /., Li Pingyu, distinct inhibitory- specificities. J. Biol. Chem. 271: Povinec P.P., Sanchez, l., Smith J.N., Swift D. (1997) A Comparison of Doses from 11 Cs and ‘to Po in Marine 25083-25088. Dahl S. tV„ Rasmussen S.K., Petersen L.C., Hejgaard J. Food. A Major International Study. J. Environ. (1996) Inhibition of coagulation factors by Radioactivity 34(1): 69-90. recombinant barley serpin BSZx. FEES Lett. 394: Abdel-Fatlah A.A., Miller A. (1996) Temperature, 165-168. humidity and lime. Combined effects on radio- Dahlgaard II. (1996) Polonium-210 in Mussels and Fish chromic film dosimeters. Radial. Phys. Chem. 47: from the Baltic-North Sea Esturay. J. Environment 611-621. Radioactivity 32(1-2): 91-96. Ahring B.K., Jensen K., Bjerre A.B., Schmidt A S. Damgaard C. (1996) Fixation probabilities of selfrng (1996) Pretreatment of wheat straw and concersion rate modifiers in simulations with several of xylose and xylan to ethanol by thermophilic deleterious alleles with linkage. Ev olution 50: 1425- anaerobic bacteria. Bioresourcc Technol. 58: 107- 1431. 113. Damgaard C. (1996) The rate of evolution in growing Ambus /’.(1996) Production of N20 in soil during populations. Trends in Ecology and Evolution 11 decomposition of dead yeast cells with different spatial (News & Comment): 107-108. distributions. Plant and Soil 181: 7-12. Damgaard Giese II. (1996) Genetic variation in Baturin G\ : Emelyanov EM. Kunzendorf II. (1996) Danish populations of Ervsiphe graminis f.sp. Authigcncous deposits in the Bornholm Basin. horded, estimation of gene diversity and effective Geology of the Bornholm Basin. Aarhus University. Department of Earth Sciences. Arhus, 1995. Aarhus population size using RFLP data. Plant Pathology 45: 691-696. Geoscience 5: 189-194. Damgaard C, Ostergdrd II. (1996) Genetic variation in Bilde A/., Sehested J., Magelherg T.E., Wallington T.J., the gene-for-gene system. Cereal Rusts and Powdery Xielsen O.J. (1996) Atmospheric Chemistry of Mildew Bulletin 24 (Supplement): 284-286. CF2BrH: Kinetics and Mechanism of Reaction with Egsgaard H. (1996) Investigation of The Initial F and Cl Atoms and Fate of CF2BrO Radicals. J. Reactions of The Calcolc Mechanism for Soot Phys. Chem. 100: 7050-7059. Formation. I. Am. Soc. Mass Spectrom. 7: 559-564. Bjerre .1/1., Bjerring Olesen A., Fernqvist T., Ploger Emelyanov E.M., Baturin (i. V. Kunzendorf II. (1996). A., Schmidt A.S. (1996) Pretreatment of wheat straw Distribution of chemical elements in recent and late- using combined wet oxidation and alkaline hydro ­ quaternary deposits of the Bornholm Basin. Geology lysis resulting in convertible cellulose and hemi- of the Bornholm Basin. Aarhus Geoscience 5: 175- ccllulose. Biotechnol. Bio eng. 49. 568-577. 188. Bjerghakke F. Sillesen A., Pagsherg P. (1996). UV Emelyanov E.M., Kravtzov V.A., Kunzendorf II. (1996). Spectrum and Kinetics of Hydroxycyclohcxadienyl Trace metals in sea and pore waters from the Radicals. J. Phys. Chem. 100: 5729-5736. Bornholm Basin. Geology of the Bornholm Basin. Brouwer A.M, Wilbrandt R. (1996) Vibrational Spectra Aarhus Geoscience 5: 157-174. of :V, A-Dimclhy laniline and Its Radical Cation. An Eng\’ild EC. (1996) Herbicidal activity af 4-chloro- Interpretation Based on Quantum Chemical Calcula ­ indoleacetic acid and other auxins on pea, barley and tions. J. Phys. Chem. 100: 9678-9688. mustard. Physiologia Plantarum 96: 333-337. Boe R.. Rise I... Thorsnes 7., Haas II. de, SaAher O.M., Gabert W.M., Jorgensen //., Brunsgaard G. Eggum Kunzendorf H. (1996). Sea-bed sediments and sedi­ B.O., Jensen J. (1996) The nutritional value of new ment accumulation rates in the Norwegian part of high-lysine barley varieties determined with rats and the Skagcrrak. NGU Bull. no. 430: 75-84. young pigs. Can. J. Anini. Sci. 76. 443-450. Christiansen C, Kunzendorf II. (1996). Signals of Giessing A.M.B., Feilherg A., Mogelberg T.E., Sehested inflows to the Gotland Basin in the sedimentary' ./, Bilde iVI.. Wallington T.J., Nielsen O.J. (1996) record. Preliminary results. GOBEX - summery Atmospheric chemistry of HFC-227ca: Spcctro- report. Institut fur Ostsecforschung, Wamemundc. kinetic investigation of the CF3CF2CF;02 radical, its Mecrcswisscnschaftliche Bcrichtc 19: 10-14. reactions with NO and NO: and the atmospheric fate

116 Ris0-R-972(EN) of CF3CF2CF20 radical. J. Phys. Chem. 100: 6572- Jensen E.S. (1996) Nitrogen and grain legumes: advantages 6579. and problems to balance. Special Report, Nitrogen & Gissel-Nielsen G., Nielsen, T. (1996) Phylotoxicity of Environment, Grain Legumes 14: 22. Acridine, an important representative of a group of Tar Jensen E S. (1996) Rhizodeposition of N by pea and barley and Creosote containments, N-PAC compounds. Poly ­ and its effect on soil N dynamics. Soil Biol. Biochem. cyclic Aromatic Compounds 8 : 243-249. 28:65-71. Glasby G.P., Kunzendorf H. (1996). Multiple factors in Jensen E.S.( 1996) Grain yield, symbiotic N2 fixation and the origin of the Cretaceous/Tertiary' boundary . The interspecific competition for inorganic N in pea-barley role of environmental stress and Deccan Trap intercrops. Plant and Soil 182: 23-38. volcanism. Geol. Rundsch. 85: 191-210. Johansen A., Jensen E.S. (1996) Transfer of N and P from Goddard A. J. 11, Byrne M A., Lange C., Roed J., Lai C.K. intact or decomposing roots of pea to barley (1996) Tracer-labelled aerosol deposition measure ­ interconnected by an arbuscular mycorrhizal fungus. ments on internal building surfaces: implications for Soil Biol. Biochem. 28: 73-81. indoor air quality . Indoor Air 3: 839-845. Johansen A., Finlay R.D., Olsson P.A. (1996) Nitrogen Grosell A/, Boetius /., Hansen Rosenkilde P. metabolism of external hyphae of the arbuscular (1996) influence of Preexposure to Sublethal Levels of mycorrhizal fungus Glomus intraradices. New Phvtol. Copper on MCu Uptake and Distribution Among 133: 705-712. Tissues on the European Eel {Anguilla anguilla) Comp. Justesen A., Somerville .S’, Christiansen S., Giese //. Biochem.Physiol. 114C(3): 229-235. (1996) Isolation of two novel genes expressed in Gron C., Dybdahl LLP. (1996). Determination of Total germinating conidia of the obligate biotroph Organic Halogens (TOX); Bias from a Non- Erysiphe graminis f.sp. hordei. Gene 170: 131-135, Halogenated Organic Compound. Environ.Int. 22: Jorgensen J.H. (1996) Effect of three suppressors on the 325-329. expression of powdery- mildew resistance genes in Gron C., Madsen. J.O., Simonsen S.. Boren H. (1996) barley. Genome 39: 492-498. Contamination of ground water samples from well Jorgensen ()., Egsgaard H., Larsen LI. (1996) Synthesis installations. Environ. Tech. 17: 613-619. of deuterated clenbuterol. J. Labelled Comp, and Gron (’., Wassenaar Krog M. (1996). Origin and Radiopharm. 38: 1007-1014. structures of groundwater humic substances from Jorgensen R.B. (1996) Spontaneous hybridization three Danish aquifers. Environ.hit. 22: 519-534. between oilseed rape (Brassica napus) and weedy Hansen J.M.H., Olsen A.G.,Rosenkilde P. (1996) The relatives. Acta Horticulturae 407: 193-200. effect of Cu on Gill and Esophagus Lipid Metabolism Jorgensen R.B., Hauser T., Mikkelsen T.R., Ostergdrd in the Rainbow Trout (Oncorhynchus mykiss). Comp. H. (1996) Transfer of engineered genes from crop to Biochem. Physical 113C( 1): 23-29. wild plants. Trends in Plant Science 1: 356-358. Hauser 71, Loeschcke V. (1996) Drougt stress and Jorgensen R.B., Chen B.Y., Cheng B.F., Heneen W.K inbreeding depression in Lychnis flos-cuculi Simonsen V. (1996) Random amplified polymorphic (Caryophyllaceae). Evolution 50: 1119-1126. DNA markers of the Brassica alboglabra chromo ­ Hickel B., Corfitzen //., Sehested K. (1996) Measure ­ some of a B. campestris-alboglabra addition line. ment of the Rate Constants of the Reactions OH + Chromosome Res. 4: 111-114 OH <-> O + H20 and OD + OD <-> + D20 in Keszthelyi T., Wilbrandt R. (1996) Radical cations of Forward and Reverse Directions. Kinetic Deter­ monomethyl-substituted 1,3-butadienes: Resonance mination of the pATs of OH and OD Radicals. 1. Raman spectra and molecular orbital calculations. J. Phys. Chcm. 100: 17186-17190. Phys. Chem. 100: 15785-15793. lkram A., Sudin AIN., Jensen LIS. (1996) Estimating N2- Keszthelyi T, Wilbrandt R. (1996) The radical cation of fixation bv Pueraria phaseoloides in rubber inter- trans- 1,3-pentadiene: Resonance Raman spectrum rowsusing the 1SN dilution technique. Journal of Natural and ab initio calculations. J. of Mol. Struct. 379, Rubber Research 10: 199-208. 211-218. Jensen E.S. (1996) Barley uptake of N deposited in the rhi- Keszthelyi T., Wilbrandt R., Bally T. (1996) Radical zosphcrc of associated field pea. Soil Biol. Biochem. cation of 1,3-butadiene: Resonance Raman spectrum 28: 159-168. of the h6 isotopomer. J. Phys. Chem. 100: 16843- Jensen E.S. (1996) Compared cycling in a soil-plant system 16849. of pea and barley residue nitrogen. Plant and Soil 182: Keszthelyi T., Wilbrandt R., Bally I., RoulinJ.L. (1996) 13-23. Radical cation of 1,3-butadiene: Resonanace Raman Jensen E.S. (1996) Effect of pea cultivation on the soil N spectrum of deuterated derivatives and improved balance. Special Report, Nitrogen & Environment, force field. J. Phys. Chem. 100: 16850-16855. Grain Legumes 14: 16-17. Kjcer B.. Jensen J. (1996) Quantitative trait loci for Jensen E.S. (1996) Nitrogen acquisition by pea and barley grain yield and yield components in a cross between and the effect of their crop residues on available a six-rowed and a two-rowed barley. Euphylica 90: nitrogen for subsequent crops. Biol Fertil Soils 23: 459- 39-48. 464.

Ris0-R-972(EN) 117 Knudsen E, Sandstrom B , Solgaard P. (1996) Zinc, Mogelberg T.E., Sehested J., Bilde M, Wallington T.J., copper and magnesium absorption from a fibre-rich Nielsen O.J. (1996). Atmospheric chemistry of diet. J. Trace Elements Med.Biol. 10: 68-76. CF3CFHCF3 (HFC-227ea): Spectrokinetic investiga ­ Kunzendorf H. (1996). A practical approach for self­ tion of the CF3CF02CF3 radical, its reactions with absorption correction in Pb-210 gamma-spectro- NO and N02, and fate of the CF3CFOCF3 Radical. metric dating. J. Radioanal. Nucl. Chem. Art. 204: J. Phys. Chem. 100: 8882-8889. 23-31. Mogelberg T.E., Bilde M., Sehested, J., Wallington T.J., Kunzendorf II., Longva ()., Paelzel M. (1996). Recent Nielsen, O.J. (1996) Atmospheric Chemistry of sedimentation rates across the Norwegian Trough. 1,1,1,2-Tetrachloroethanc (CC13CH2C1): Spectro ­ NGU Bull. no. 430: 67-74. kinetic Investigation of the CC13CC1H02 Radical, Its Longer S., Ljungstrom E., Wangberg I, Wallington T.J., Reactions with NO and N02, and Atmospheric Fate Hurley MI)., Nielsen O.J. (1996) Atmospheric of the CC13CC1H0 Radical. J. Phys. Chem. 100: chemistry of di-lert butyl ether: Rates and products 18399-18407. of the reactions with chlorine atoms, hydroxyl and Nielsen P.S., Rasmussen S.K (1996) A barley nitrate radicals. Int. J. Chem. Kinet. 28: 299-306. photosystem II associated 10 kD protein (Accession Larsen, ,/., Jakob sen I. (1996) Effects of a mycophagous No. X97771) shows that at least two forms arc Collembola on the symbiosis between Trifolium present in monocot plants (PGR96-108). Plant subterraneum and three arbuscular mycorrhizal fungi. Physiol. 112: 1398. New Phytol. 133: 295-302. Nielsen T. (1996). Traffic contribution of polycyclic Larsen J., Jakobsen I. (1996) Interactions between a aromatic hydrocarbons in the center of a large city. mycophagous Collembola dry yeast and the external Atmos. Environ. 30: 3481-3490. mycelium of an arbuscular mycorrhizal fungus. Nielsen Jorgensen HE., Larsen J.C., Poulsen M. Mycorthiza 6: 259-264. (1996) City air pollution of polycyclic aromatic Larsen J., Ihingstrup /., Jakobsen L, Rosendahl S. 1996: hydrocarbons and other mutagens: occurrence, Benomyl inhibits phosphorus transport but not fungal sources and health effects. Sci. Total Environ. alkaline phosphatase activity in a G/owMx-cucumber 189/190: 41-49. symbiosis. New Phytol. 132: 127-133. Nielsen T., Pilegaard K, Egelov A.II., Granby K, Linde-Laursen L, Seberg ()., Frederiksen -S’., Baden C. Hummelshoj P., Jensen N.O., Skov H. (1996) (1996) The karyotype of Festucopsis serpentini Atmospheric nitrogen compounds: occurrence (Poaceae Triticeae) from Albania studied by composition and deposition. Sci. Total Environ. banding techniques and in situ hybridization. PI. 189/190: 459-465. Syst. Evol. 201: 75-82. Offersgaard J.E., Veng T., Skettrup T. (1996) Accurate Madsen L., Grahl-Madsen L., (iron C, Lind L, Engell method for determining the refractive-index profiles J. (1996) Adsorption of Polar Aromatic Hydro ­ of planar waveguides in uniaxial media with the carbons on Synthetic Calcite. Org. Geochem. 24: optical axis normal to the surface. Appl. Opt. 35: 1151-1155. 2602-2609. McLaughlin W.L., Puhl J.M., Al-Sheikhly M., Christou Olsson P.A., Baath, E., Jakobsen /., Sonderstrom B. (1996) C.A., Miller A., Kovacs A., Wojnarovits L, Lewis Soil bacteria respond to presence of roots but not to I). /■' (1996) Novel Radiochromic Films for Clinical mycelium of arbuscular mycorrhizal fungi. Soil Biol. Dosimetry. Radiation Protection Dosimetry 66: 263- Biochem. 28 (4/5): 463-470. 268. Pagsberg P., Sillesen A., Jodkowski J.T., Ratajczak E Meunier II, Pagsberg P , Sillesen A. (1996) Kinetics (1996) Kinetics of the reaction F + NO + M —> FNO and branching ratios of the reactions NH2 + N02 -> + M studied by pulse radiolysis combined with time- N20 + H20 and NH2 + N02 H2NO + NO studied resolved IR and UV spectroscopy. Chem. Phys. Lett. by pulse radiolysis combined with time-resolved 249: 358-364. infrared diode laser spectroscopy. Chem. Phys. Lett. Pagsberg P., Sillesen A., Jodkowski J.T., Ratajczak E. 261: 277-282. (1996) Kinetics of the F + N02 + M -> FNO; + M Mikkelsen T.R., Andersen B., Jorgensen R.B. (1996) reaction studied by pulse radiolysis combined with The risk of crop transgenes spread. Nature 380: 31. time-resolved IR and UV spectroscopy. Chem. Phys. Mikkelsen T.R., Jensen J., Jorgensen R.B. (1996) Let. 252: 165-171. Inheritance of oilseed rape (Brassica napus) RAPD Pedersen A.L, Feldner H.C., Rosendahl L. (1996) Effect of markers in a backcross progeny with Brassica proline on nitrogenase activity in symbiosomes from campestris. Theor. Appl. Genet. 92: 492-497. root nodules of soybean (Glycine max L.) subjected to Miller A. (1996) Uncertainty of dose measurement in drought stress. Journal of Experimental Botany, radiation processing. Radial. Phys. Chem. 47: 479- 47(303): 1533-1539. 482.

118 Ris0-R-972(EN) Pedersen C., Rasmussen S.K., Linde-Laursen I. (1996) Contaminated Solid Waste and Sludge. J. Environ. Genome and chromosome identification in Sci. Health. A31(l): 13-29. cultivated barley and related species of the Triticeae Sorensen P., Jensen E.S. (1996) The fate of fresh and (Poaceae) by in situ hybridization with the GAA- stored 1 ^-labelled sheep urine and urea applied to a satellile sequence. Genome 39: 93-104. sandy and a sandy loam soil using different appligation Rasmussen S.K., Dahl S.W., Norgdrd A., Hejgaard J. strategies. Plant and Soil 183: 213-220.. (1996) A recombinant wheat serpin with inhibitory Sorensen P . Ladd J.N., Amato M. (1996) Microbial activity. Plant Mol. Biol. 30: 673-677. assimilation of 14C of ground and unground plant Rasmussen S.K., Klausen J. (1996) Ribosomal protein materials decomposing in a loamy sand and a clay soil. L24E homolog (Accession No. X94296) is Soil Biol. Biochem. 28: 1425-1434. expressed in barley endosperms. (PRG96-011) Plant Turgeon B.G., Sharon A., Wirsel S., Christiansen S.K., Physiol. 110: 1048. Yoder O. (1996) Structure and function of mating Rasmussen S.K., Klausen J., Hejgaard J., Svensson B., type genes in Cochliobolus spp. and asexual fungi. Svendsen I. (1996) Primary structure of the plant Can. J. Bot. 73: 778-783. serpin BSZ7 having the capacity of chymotrypsin Tyndall G.S., Orlando J. II., Wallington 1,11, Sehested J. inhibition. Biochim. Biophys. Acta. 1297: 127-130. Nielsen O.J. (1996) Kinetics of the reactions of RoedJ., Andersson K.G., (1996) Clean-up of rnban areas in acetonitrile with chlorine and fluorine atoms. J. the CIS countries contaminated by Chernobyl fallout. J. Phys. Chem. 100: 660-668. Environ. Radioactivity 33(2). 107-116. Wallington T.J., Bilde M., Mogelberg T.E., Sehested J., Rued J., Andersson K.G., Prip II. (1996) The skim and Nielsen O.J. (1996) Atmospheric chemistry of 1,2- burial plough: A new implement for reclamation of Dichloroethane: UV spectra of CH2C1CHC1 and radioactively contaminated land. J. Environ. Radio­ CH2C1CHC102 radicals, kinetics of the reactions of activity 33(2): 117-128. CH2C1CHC1 radicals with 02 and CH2C1CHC102 Rvsgaard .S', Piaster K., Dahlgaard II (1996) Primary radicals with NO and N02, and fate of the alkoxy production, nutrient dynamics and mineralisation in a radical CH2C1CHC10. J. Phys. Chem., 100: 5751- north-eastern Greenland fjord during the summer thaw'. 5760. Polar Biol 16: 497-506 Wallington T.J., Hurley M.D., Fracheboud J.M., Schmidt A.S., Andrews B.A.. Asenjo J.A. (1996). Orlando J.J., Tyndall G.S., Sehested J., Mogelberg Correlations for the partition behaviour of proteins T.E., Nielsen O.J. (1996) Role of excited alkoxy in aqueous two-phase systems: Effect of overall radicals in trifluoroactic acid formation during protein concentration. Biotechnology and Bio ­ atmospheric degradation of HFC-134 J. Phys. engineering 50: 617-626. Chem. 100: 18116-18122. Seherg ()., Linde-Laursen I. (1996) Premium, a new Wilbrandt R (1996) Time-resolved resonance Raman genus of the Triticeae (Poaceae) from Argentina. spectroscopy: A retrospect on the early days. Systematic Botany 21: 3-15. Biospectroscopy 2: 263-275. Sehested J., Bilde M, Magelberg T., Wallington T.J., Ostergard H., Shaw M.W. (1996) Linear models are Nielsen O.J. (1996) Kinetics and mechanism of the inappropriate to estimate relative parasitic fitness of reaction of F atoms with CH3Br. J. Phys. Chem. pathogens in heterogeneous host populations. 100: 10989-10998. Phytopathology 86 : 561-562. Sehested ./., Magelberg T, Wallington T.J., Kaiser E.W., Nielsen O.J. (1996) Dimethyl ether oxidation: Kinetics and mechanism of the CH3OCH2 + 02 reac ­ 9.2 Books tion at 296 K and 0.38-940 torr total pressure. J. Phys. Chem. 100: 17218-17225. Sehested J., Nielsen O.J., Rinaldi C.A., Lane S.I., Boesen B., Hovmoller M.S., Jorgensen J.H. (1996) Perrero J.C. (1996) Kinetics and mechanism of the Designation of barley and wheat powdery mildew reaction of CF3 radicals with N02. Int. J. Chem. resistance and virulence in Europe. In: E. Limpert, Kinet. 28: 579-588. M R. Finckh and M S.Wolfe (eds). Integrated Sehested K . Holcman J. (1996) A pulse radiolysis study control of cereal mildews and rusts: Towards co ­ of the OH radical induced autoxidation of ordination of research across Europe. COST 817, methanesulfinic acid. Radial. Phys. Chem. 47: 357- European Commission: 2-12. 360. Damgaard C, Ostergard H. (1996) Density dependent Sharon A., Yamaguchi K, Christiansen S.K., Horwitz growth of powdery mildew (Erysiphe graminis f.sp. B.A., Yoder O.C., Turgeon B.G. (1996) An asexual hordei) on a partial resistant barley variety: fungus has the potential for sexual development. Infection efficiency and spore production. In: E. Mol. Gen. Genet. 251: 60-68. Limpert, MR. Finckh and MS. Wolfe (eds). Sorensen P., Fernqvist T. (1996) Some Distinctive Integrated control of cereal mildews and rusts: Reactions of Mercury Involved in its Removal from Towards co-ordination of research across Europe. COST 817, European Commission: 241-245.

Ris0-R-972(EN) 119 Gallegher, M, Fontan, J., Wyers, F, Ruijgrok, W., de Protection et de Surete Nuclaire. ISBN: 2-86883- Duyzer, J., Hmtimelshoj, Filegaard, K. and 285-7, Les Editions de Physics: 95-119. Fowler, D. 1996. Atmospheric particles and their Walton, S., Gallagher, M, Choularton, T.W., Duyzer, ,/. interactions with natural surfaces. In: J. Slanina (ed): and Filegaard, K. 1996. The Speulderbos experi­ Biosphere-Atmosphere Exchange of Pollutants and ment 1993 - experimental and modelling results. In: Trace Substances, Springer, Heidelberg: 45-92. J. Slanina (ed): Biosphere-Atmosphere Exchange of Giese 11, Justesen A.F., Christiansen S.K. (1996) Pollutants and Trace Substmices, Springer, Towards the cloning of an avirulence gene from Heidelberg: 292-300. Frysiphe graminis f.sp. hordei. In: E. Limpert. M R. Welinder K.G., Jespersen H.M., Kjtersgard I.V.H., Finckh mid M S. Wolfe (cds). Integrated control of Ostergaard L, Abetskov A.K., Hansen L.N., cereal mildews and rusts. Towards co-ordination of Rasmussen S.K. (1996) What can we learn from research across Europe. COST 817, European Arabidopsis peroxidases? In: C. Obingcr, U. Burner. Commission: 217-222. R. Ebermann, C. Penel, H. Greppin (eds) Plant Per­ Kollist If., Moldau H., Mortensen L., Rasmussen S.K., oxidases: Biochemistry and Physiology. University llolt-Jorgcnsen /.. (1996) Ascorbate levels and of Geneva Press, Geneva. Switzerland. 173-178. ozone decay in cell walls of barley and wheal Ostergard If. (1996) Model for growth of a mixture of seedling. In: C. Obingcr. U. Burner. R. Ebermann. pathogen isolates. In: E. Limpert. M R. Finckh mid C. Penel, H. Greppin (eds) Plant Peroxidases: Bio ­ M S. Wolfe (cds). Integrated control of cereal chemistry and Physiology. University of Geneva mildews and rusts: Towards co-ordination of Press, Geneva. Switzerland: 558-362. research across Europe. COST 817, European Kristensen B.K., Rasmussen S.K. (1996) Expression of Commission: 237-240. the stress-related intercellular barley peroxidase Prx8 in barley mid transgenic tobacco. In. C. Obingcr, U. Burner, R. Ebermann, C. Penel, H. 9.3 Printed Abstracts and Proceedings Greppin (eds) Plant Peroxidases: Biochemistry and Physiology. University of Geneva Press, Geneva, Switzerland: 317-321. Aarkrog A. (1996) Inventory of nuclear releases in the Lyngkjier M.F., Ostergard II, Munk L. (1996) Papilla world. In: Radioecology and the Restoration of formation in leaves of Mlo-resistant barley attacked Radioactive-Contaminated Sites. NATO conference on by Mlo-virulent powdery mildew. In: E. Limpert, radioactive-contaminated site restoration. Zarechny. M R. Finckh mid M S. Wolfe (eds). Integrated Sverdlovsk (RU), 19-28 Jun 1995. Luykx. F.F.. Frissel. control of cereal mildews and rusts: Towards co ­ M.J. (eds ), (Kluwer Academic Publishers, Dordrecht. ordination of research across Europe. COST 817, 1996): 31-43.. Europemi Commission: 227-230. Aarkrog A. (1996) Up-to-date studies of radionuclides Nies If., Nielsen S.F. (1996) Radioactivity in the Baltic transfer in the river sy stem Tccha-Isct-Tobol-Irty sh-Ob Sea. (Co-ordinators: P. Gucgueniat. P Germain and H. to Kara Sea by a joint international group. In: Radio­ Metivier) Radionuclides in the Oceans, Inputs and ecology mid the Restoration of Radioactive- Inventories, Institut dc Protection et de Surete Nuclaire. Contaminated Sites. NATO conference on radioactive- ISBN: 2-86883-285-7, Les Editions de Physics. 220- contaminated site restoration. Zarechny, Sverdlovsk 231 (RU), 19-28 Jun 1995. Luykx. F.F., Frissel, M.J. (cds ). Nyjjeler F„ Cigna A.A., Dahlgaard If, Uvingston H.D. (Kluwer Academic Publishers, Dordrecht, 1996): 187- (1996) Radionuclides in the Atlantic Ocean: A Survey . 201. (Co-ordinators: P. Gucgueniat P. Germain and H. Aarkrog A., Folikarpov (!., (1996) Development of Metivier) Radionuclides in the Oceans, Inputs and Radioecology in East and West. In: Radioecology and Inventories, Institut de Protection ct de Surete Nuclaire. the Restoration of Radioactive-Contaminated Sites. ISBN: 2-86883-285-7. Les Editions de Physics: 1-28. NATO conference on radioactive-contaminated site Filegaard, K, Hummelshoj, F, Jensen, N.O. and restoration. Zarechny, Sverdlovsk (RU), 19-28 Jun Kristensen, L 1996. Measurements of fluxes of 1995. Luvkx, F.F., Frissel. M.J. (eds). (Kluwer particles, ozone, nitrogen ocides mid carbon dioxide Academic Publishers, Dordrecht, 1996): 17-29. to coniferous and deciduous forests. In: J. Slanina Andersson K. G. (1996) Modelling external radiation doses (cd): Biosphere-Atmosphere Exchange of Pollutants in contaminated urban areas: Implications for and Trace Substances, Springer. Heidelberg: 391- development of decontamination strategics. In: 1RPA 396 International congress on radiation protection. StrandF., SickelM, AarkrogA., Bewers J.M., Tsaturov T, Proceedings. Vol. 3.9. International congress on Magnusson S. (1996) Radioactive Contamination of the radiation protection Association. Vienna (AT). 14-19 Arctic Marine Environment. A Survey. (Co-ordinators. Apr. (IRPA. Seibersdorf): 265-267. P. Gucgueniat. P. Germain and H. Metivier) Radio­ Bilde A-/, Sehested J., Mogelberg T.F., Nielsen O.J. nuclides in the Oceans, Inputs and Inventories. Institut (1996) Chlorine mid fluorine atom adducts with halogcnatcd compounds in the gas phase.

120 Ris0-R-972(EN) Proceedings of The NOSA/NORSAC Symposium, Riso National Laboratory, Roskilde, Denmark: 96- LO-Hojskolen. Elsinore. Denmark. Nov 15-17. 98. 1996. Ed. by Poul Hummelshoj. Riso-R-934(EN). Granby K., Skov H., Hertel ()., Christensen C.S., Riso National Laboratory, Roskildc, Denmark, Egelov A.H., Lohse C, Nielsen 71, Gryning S.-E., 1996. 30 p. Hummelshoj P Platz J. (1996) Oxidants. Center Christensen L.K, Nicolaisen KM (1996) FT1R- News. The Environmental Research Programme. spectroscopy of atmospheric greenhouse gases. The Danish Centre of Air Pollution Processes and Proceedings of the NOSA/NORSAC symposium Models 4 (2): 9-10. 1996. NOSA/NORSAC symposium, LO-Hojskolen, Grosell M.H., Rosenkilde /’., Hansen II.J..M, H ood CM. Elsinore, Denmark, 15-17 Nov 1996. Ed. by P. (1996) Acclimatiion to Cu by the highly tolerant Hummelshoj. Riso-R-934(EN). Riso National European eel (Anguilla anguilla) and the more sensitive Laboratory , Denmark: 78. rainbow trount (Onchorhynchus mykiss). In: Program Dreiseitl, A., Steffenson, B.J., Jorgensen, J.H. (1996) and Abstracts, Adaptation to Stress in Aquatic and Diversity of the Czech and Slovak spring barley Terrestrial Ecosystems, Physiology, Biochemistry. cultivars to powdery mildew and leaf rust. Proceedings, Toxicology. Europaen Society' for comparative 7th Intern. Barley Genetics Symposium, Saskatoon, Phy siology and Biochemistry 17th Annual conference. Canada. 30 Julv-6 August 1996: 714-716. University of Antwerp (RUCA). Belgium, 27-31 Kgsgaard II, Larsen E. (1996) Direct sampling tandem August. Session E: 71. mass spectrometry of premixed flames. Proceedings Hansen 71, Jensen P.W., Rasmussen S.K. (1996) of the 44th ASMS Conf. Mass Spectrom. Allied Cellulter lokalisering af den stressrelatcrcde byg- Top., Portland, 12-16 May: 420a-b. peroxidase Prx7. (Abstract) SP rapport 2: 22 (in Kgsgaard II., Larsen E., Sehested J., Nielsen O.J. Danish). (1996) Intermediates in dimethy l ether flames. Joint Hauser T.P., Landho L., Qstergard II., Jorgensen R.B. meeting of the Portuguese, British, Spanish and (1996) Spontan overforsel af egenskaber mellcm Swedish sections of the Combustion Institute, af grade og ukrudtsbestande: raps (Brassica napus) Funchal, Madeira, Portugal, 1-4 Apr 1996: 18.3.1- og agerkal (B. campestris). 13. Danske Plante- 3.4. vaemskonference 1996. SP rapport 3: 169-177. (in Kauser P. (1996) Particulate adsorption on plant surfaces. Danish). NOSA/NORSAC meeting, 15-17 Nov.. Elsinore, Helweg C, Nielsen 71, Hansen P.E. (1996) QSPRs Denmark. Riso-R-934(EN): 56-58. compared with HPLC experiments for the Keilherg A., Holcman J., Lohse C., Nielsen T., Sehested estimation of log Kow for N-PAH compounds. QSAR K. Reactions of N-heteroaromatics with HO in rain, 96. 7th International Workshop on QSAR's in cloud-water and fog droplets. EUROTRAC Sympo ­ Environmental Sciences "Past, Present and Future". sium '96 on Transport and transformation of Book of Abstracts. National Environmental pollutants in the troposphere. EUROTRAC, Inter­ Research Institute, Roskilde. Danmark: 73-74. national Scientific Secretariat. Garmisch-Parten- Hovmoller M, Qstergard H. (1996) Prediction of kirchcn.: 2-8 . durability of powdery mildew resistance in autumn Keilherg A., Holcman J., Lohse Ch., Nielsen T., sown and spring sown barley varieties. In: G.H.J. Sehested K. (1996) Reactions of quinoline and 5- Kema, RE. Niks and R.A. Daamen (eds). nitroquinoline with OH and 02 in the aqueous phase. Proceedings of the 9th European and Mediterranean Proceedings of The NOSA/NORSAC Symposium, Cereal Rusts and Powdery' Mildews Conference. LO-Hojskolen, Elsinore, Denmark, Nov 15-17, COST. European Commission IPO-DLO, Wage- 1996. Ed. by Poul Hummelshoj. Riso-R-934(EN). ningen: 105-108. Riso National Laboratory, Roskilde, Denmark, Hummelshoj P., Sempreviva A.M., Jensen NO., Pilegaard 1996: 84-86. K. Nordstrom C., Sogaard II., (1996) Danish H:OZCO: Giese, II., Justesen, A.K., Christiansen, S.K. (1996) flux and concentration measurements intercomparison - Potential for the use of transformation in studies of RIMI’95. 21. General assembly of the European the obligate biotroph Erysiphe graminis f.sp. hordei. Geophysical Society, The Hague (NL), 6-10 May . Ami. In: G.H.J. Kema, R E. Niks and R.A. Daamen (eds). Geophys. Suppl. 2, v. 14: C470. Proceedings of the 9th European and Mediterranean Jorgensen K. 11, Jensen, E.S. (1996). Do dung patches Cereal Rusts and Powdery Mildews Conference. influence N2 fixation in grass/clover mixtures in the COST, European Commission IPO-DLO, Wage- short term. In: Legumes in sustainable farming sy stems ningen: 31-33. (ed. D. Younie), Proceedings of Occasional Granby K, Nielsen 71, Egelov A.II. (1996) Indications Symposium No. 30. Organised by British Grasslands of sources of atmospheric formic and acetic acid. Society, Aberdeen 2-4 September 1996, Scotland: 44- Proceedings of The NOSA/NORSAC Symposium, 45. LO-Hojskolen. Elsinore, Denmark, 15-17 Nov. Kristensen B.K., Rasmussen S.K. (1996) Promotor 1996. Ed. by Poul Hummelshoj. Riso-R-934(EN). analy se af to stress-rclaterede bygperoxidasc gencr

Ris0-R-972(EN) 121 og belysning af deres funktion i planten. (Abstract) Nielsen T., Hertel O. Christensen C.S., Fgelov A IL, SP rapport 2: 20 (in Danish). Granby K, Hansen A.B., Platz J., Skov II. (1996) Kunzendorf H. (1996) Marine mineral exploration. Evaluation of the Danish ACDEP model to simulate Realities and strategies at the end of the nineties. formation of tropospheric ozone, other photo ­ International symposium on geology and geophysics chemical oxidants and atmospheric reaction of the Indian Ocean - 1996. Abstracts. GIO - 1996, products. The Oxidizing Capacity of the Goa, India, 21-25 Oct 1996. National Institute of Troposphere. Abstracts of the 7th European Svmp. Oceanography, Goa: 189. on Physico-Chemical Behaviour of Atmospheric Kunzendorf II, Christiansen C. (1996) Tracing and use Pollutants. European Commission: 0-47. of radiometric signals in Baltic Sea surface Nielsen T., Hertel O., Granby K., Hansen A.B.. Platz J.. sediments. Sixth V.M. Goldschmidt conference. Skov II., Stenholt C., Fgelov AIL (1996) Extended abstracts. 6. V.M. Goldschmidt con ­ Comparison of field measurements and a trajectory ference, Heidelberg, Germany, 31 Mar-4 Apr 1996. model. Proceedings of the NOSA/NORSAC Sym­ Cambridge Publications, Cambridge, 1966. Journal posium, LO-Hojskolen, Elsinore, Denmark, 15-17 of Conference Abstracts 1(1): 340. Nov 15-17. Ed. by Poul Hummelshej. Riso-R- Lange C. (1996) Size distribution of atmospheric pollutants 934(EN). Rise National Laboratory', Roskilde. sampled with a BLPI and measured by neutron Denmark: 121-123. activation. NOSA/NORSAC meeting, 15-17 Nov., Nielsen /., Hertel ()., Granby K., Hansen A.B., Platz J., Elsinore, Denmark: 53-55. Skov II., Stenholt C, Fgelov .(.//.(1996) Chemical Lyngkjcer M.F., Jorgensen J.H., Ostergard, II. (1996) characterization and modelling of ozone, other Er Mlo-resistens i byg holdbar? 13. Danske Plante- photochemical oxidants, precursors and atmospheric vamskonference 1996. SP rapport 4: 145-155. (in reaction products during a summer high pressure Danish) episode in Denmark. EUROTRAC Symposium ’96 Lyngkjcer M.F., Carver T.L.W., Zeyen R.J. (1996) on Transport and transformation of pollutants in the Suppression of mlo powdery mildew resistance in troposphere, Garmisch-Partenkirchen, Germany , 25- barley. In: G.H.J. Kema, R.E. Niks and R.A. 29 Mar 1996. Poster 4-20. Daamen (eds). Proceedings of the 9th European and Nielsen 7., Hertel ()., Granby K., Hansen A. B., Platz J., Mediterranean Cereal Rusts and Powdery Mildews Skov //., Stenholt, C., Iigelov A.II. (1996) Conference. COST, European Commission IPO- Comparison of night to day ratios and dry deposition DLO, Wageningen: 67-68. velocities of different pollutants. In: Transport and Lyngkjcer M.F., Ostergard II (1996) Powdery mildew transformation of pollutants in the troposphere. infection of barley with the mlo resistance gene. In: Abstracts of lectures and posters. EUROTRAC G.H.J. Kema, R.E. Niks and R.A. Daamen (eds). symposium 96, Garmisch-Partenkirchen, Germany, Proceedings of the 9th European and Mediterranean 25-29 March 1996. Borrcll, P. (ed), (EUROTRAC, Cereal Rusts and Powdery' Mildews Conference. Garmisch-Partenkirchen, 1996) Poster 8-19. COST, European Commission IPO-DLO, Wage ­ Pilegaard K, Jensen N.O., Hummelshoj P. (1996) Ozone ningen: 58-60. fluxes over forested ecosystems and mechanisms of Mogelberg T., Sehested J., Nielsen O.J., Wallington removal. NOSA/NORSAC meeting, 15-17 Nov., T.J., Hurley M.D., Fracheboud J.M., Orlando J.J., Elsinore, Denmark., Riso-R-934(EN): 124-130. Tyndall G.S. (1996) Atmospheric chemistry of HFC- Pilegaard K, Jensen N O., Hummelshoj P.( 1996) Ozone 134a: Role of excited CF3CFHO radicals. Proceed ­ fluxes over forested ecosystems and mechanisms of ings of the NOSA/NORSAC Symposium, LO- removal. ISPRA 7th Symposium of Physico-Chemical Hojskolen, Elsinore, Denmark, 15-17 Nov, 1996. Behaviour of Atmospheric Pollutants, Venice. Italy. 2-4 Ed. by Poul Hummelshej. Rise-R-934(EN). Rise October 1996. Paper 0-39. National Laboratory, Roskilde, Denmark: 31-33. Platz J., Sehested J., Mogelberg T.F., Nielsen O.J., Nath B.N., Kunzendorf LI. Pluger W.L. Geochemical Wallington T.J. (1996) Atmospheric chemistry of signatures of extreme chemical weathering in SW 1,4-DI- and 1,3,5-trioxane. Proceedings of the coastal sediments of India. International symposium NOSA/NORSAC symposium, LO-Hojskolen. Elsi­ on geology and geophysics of the Indian Ocean - nore, Denmark, 15-17 Nov 1996. Ed. by P. 1996. Abstracts, GIO - 1996, Goa, India, 21-25 Oct Hummelshoj. Riso-R-934(EN). Rise National 1996. National Institute of Oceanography, Goa: 59. Laboratory, Denmark: 34. Nielsen T., Hertel O., Christensen C.S., Fgelov A.H., RoedJ., Andersson K.G., Togawa O. (1996) Weathering of Granby K, Hansen A.B., Skov H., Platz J. (1996) N- radionuclides deposited in inhabited areas. In: IRPA9. compounds and other air pollution components: 1996 International congress on radiation protection. Comparison of field measurements and model Proceedings. Vol. 3. 9 International congress on results. Center News. The Environmental Research radiation protection Association. Vienna (AT), 14-19 Programme. The Danish Centre of Air Pollution April. (IRPA, Seibersdorf): 167-169. Processes and Models 4 (2): 10-11.

122 Ris0-R-972(EN) Roed J., Goddard Lange ('., Byrne M.A. (1996) Sturup S. (1996). Application of HRICPMS for trace Mitigation of radiation dose through deposition to elemental analysis of biological samples. Kansas indoor surfaces. In: IRPA9. 1996 International congress City, USA 1 October. FACSS XXIII Final Program on radiation protection. Proceedings. Vol. 3. 9th and Abstract Book: 147. International congress on radiation protection Associa ­ Sturup S., Bibak A., Behrens A., Gundersen T. (1996). tion. Vienna (AT). 14-19 April. (IRPA. Seibersdorf): Multielemental determinations using HR-ICPMS. 47-49. 1996 Winter Conference on Plasma Spectro ­ RoedJ., Togawa O. (1996) A method for the determination chemistry, Florida. USA.. 8-13 Jan 1996: 294. of vertical distribution profiles of radioactive conta­ Wang R.R.-C., Bothrner R.. von, Dvorak J., Fedak G., mination in soils. In: IRPA9. 1996 International Linde-Laursen /., Muramatsu M (19%) Genome congress on radiation protection. Proceedings. Vol. 2. symbols in the Triticeae (Poaceac). Proceedings of the 9th International congress on radiation protection 2nd International Triticeae Symposium, Logan. Utah, Association. Vienna (AT), 14-19 April. (IRPA, USA, 20-24 June 1994: 29-34. Seibersdorf): 666-668 . Wessel Larsen N., Sejner Andersen T., Sehested J. Roed J, Andersson K.G., Sobotovitch E., Garger , (1996) Ozone formation with argon as bath gas. Matveenko Id. (1996) Distribution of radionuclides in Proceedings of the NOSA/NORSAC symposium urban areas and their removal. In: International 1996. NOSA/NORSAC symposium. LO-Hojskolen. conference of the European Commission and the Elsinore, Denmark. 15-17 Nov, 1996. Ed. bv P Belarus, Russian and Ukrainian Ministries on Hummclshoj. Riso-R-934(EN). Riso National Chernobyl Affairs, Emergency Situations and Health, Laboratory. Denmark: 114-116. Minsk (BY), 18-22 March. Karaoglou. A., Dcsmet, G„ Ostergard II, Pons J. (1996) Epidemic development of Kelly, G.N.. Menzel, H.G. (eds.) European a barley powdery mildew population. In: G.H.J. Commission, Brussels) (EUR-16544): 49-58. Kema, RE. Niks and R.A. Daamcn (eds). RoedJ., Andersson K.G., Togawa (). (1996) Weathering of Proceedings of the 9th European and Mediterranean radionuclides deposited in inhabited areas, hr: IRPA9. Cereal Rusts and Powdery Mildews Conference. 1996 International congress on Radiation Protection. COST, European Commission 1PO-DLO. Wagc- Proceedings. Vol. 3. 9th International congress on ningen: 279-283. Radiation Protection Association. Viemia (AT). 14-19 April. (IRPA, Seibersdorf): 167-169. Sehested J., Mogelberg T.E., Platz J., Nielsen O.J., 9.4 Research Reports Wallington T.J., Kaiser EAV. (1996) Dimethyl ether oxidation: Kinetics and Mechanism of the CH3OCH2 + 02 Reaction at 296 K and 0.38-940 Torr Total Andersson K.G., Roed J., Jacob /*., Meckbach R. (1996) Pressure. Proc. of the NOSA/NORSAC Symposium, Weathering of Cs-137 on various surfaces in inhabited LO-Hojskolen, Elsinore, Denmark. 15-17 Nov 1996. areas and calculated location factors, hi: Deposition of Ed. by Poul Hummclshoj. Riso-R-934(EN). Riso Radionuclides, their Subsequent Relocation in the National Laboratory, Roskilde. Denmark: 35-37. Environment and Resulting Implications. Final report. Siigur K, Nielsen 7., Gron C., Helweg C., Ilansen P.E., EUR-16604: 47-57. Jorgensen ()., Kirso U. (1996) Partitioning of poly ­ Andersson K.G., Roed J., Paretzke H.G., Tschhiersch J. cyclic aromatic compounds between soot particles (1996) Modelling of the radiological impact of a and water studied by applying aromatic macro - deposit of artificial radionuclides in inhabited areas. In: molecules as model substrates for soot particles. Deposition of Radionuclides, their Subsequent Proceedings of The NOSA/NORSAC Symposium, Relocation in the Environment and Resulting LO-Hojskolen, Elsinore, Denmark, Nov 15-17, Implications. Final report. EUR-16604: 83-94. 1996. Ed. by Poul Hummclshoj. Riso-R-934(EN). Bjerre A.B., Ploger A., Simonsen T., Woidemann A., Riso National Laboratory, Roskilde. Denmark: 128- Schmidt A S. (1996) Quantification of solubilized 1.30. hemicellulose from pretreated lignocellulose by acid Skov H, Christensen C.S., Granby K., Egelov Adi., hydrolysis and high-performance liquid chroma ­ Hansen A.B., Nielsen T., Platz J. (1996) tography. Riso-R-855(EN) 38 p. Tropospheric ozone and other photochemical Brown J., Jones A. J., Byrne M.A., Goddard A.J.II., Jacob products measured at the TOR station, LI. Valby, P., Tschiersch J., Lange C., RoedJ., Andersson K.G., Denmark. The oxidizing capacity of the Zombori Feher I. (1996) Review of current external troposphere. Abstracts of the 7th European Symp. dose models and recent experimental research on the on Physico-Chemical Behaviour of Atmospheric deposition and subsequent relocation of artificial Pollutants. European Commission: 0-37. radionuclides. In: Deposition of Radionuclides, their Sorensen P., Jensen E.S. (1996) 1 N-labelling of animal Subsequent Relocation in the Environment and manure: problems, solutions and use. 9th Nitrogen Resulting Implications. Final report: EUR-16604: 95- Workshop, Braunschweig, Germany, 9-12 Sep 1996.: 113. 67-70.

Ris0-R-972(EN) 123 Byrne MA., Goddard A.J.H., Lange, Roed, J. (1996) Glavas. Contract no. EV5V-CT93-0309 European Aerosol deposition on skin, hair and clothing. In: Commission. Deposition of Radionuclides, their Subsequent Roed J., Lange ('., Andersson K.G., Prip IL, Olsen S.: Relocation in the Environment and Resulting Implica­ Ramazaev V.P., Ponamarjov A. 1., Barskovskv A.N., tions. Final report: EUR-16604: 25-34. Mishine A.S., Vorobiev B.Ic, Chesnokov A.V., Potapov French .S'., Finck R., Hamalamen R.P., Naadland E., Roed V.N., Shcherbak SB. (1996) Decontamination in a Sato A., Sinkko K. (1996) Nordic decision Russian settlement. Riso report: RIS0-R-87O(EN). conference: An exercise on clean-up actions in an urban Stenholm M., Jensen P.A., Ilald P. (1996). Biomasses environment after a nuclear accident. Arlanda Hotel, Bramdsels- og Fyringskarakteristika. Fvringsforsog. Sweden. 30-31 Aug. 1995. Report of the NKS EKO 4 EFP-93, Journal 1323/93-0015. 75 pp. (in Danish). programme. STUK-A-132. 59 p. Zellner R., Herrmann IL., Hiding L.,Mirabel P., Buxton Jacob /'., Meckbach R, Hillmann (I, Andersson K.G., G., Salmon A., Sehested K., Rolle IF. (1996) Roed ./, Palagvi .S’, Afdro A. (1996) Radionuclide Removal and interconversions of oxidants in the distributions in undisturbed soil and related gamma atmospheric aqueous phase. Final report for dc EC dose rates in air. In: Deposition of Radionuclides, their R&D project R1NOXA 1.4.1994-31.3.1996. Riso- Subsequent Relocation in the Environment mid Dok-498, 177 pp. Resulting Implications. Final report. EUR-16604: 35- Z.etzsch C\, Dev’older I’., Pagsberg P., Barnes /.., 46. Glavas S. (1996) Cycling of OH mid HO: by NOx in Jacobsen /-., Ilolanan J., Sehested K. (1996) Oxidation the tropospheric oxidation of aromatics of Transition Metal Ions by Ozone and the Role of (CYTROXA) February 1994 - July 1996. Final Resulting Intermediates. Iron and Manganese. Final report. Riso-Dok-482. report of the EC R&D project. R1NOXA Removal and Interconvcrsion of Oxidants in the Atmospheric Aqueous Phase. Contract EF5V-CT93-0317. European Commission: 113-147. 9.5 Other Publications Jensen A., Gissel Nielsen G., Gundersen V, Nielsen O.J., Bjergbakke F., Ostergdrd //., Aarkrog A. (1996) Alhorzfar Af., Johnsson G., Fillumsen A, Grnn (’. Environmental Science and Technology Department (1996) Behandling af "brunt" vand ved membran- Annual Report 1995. R1S0-R-9O6(EN) 136 p. filtrcring. Vandteknik no. 6: 311-316 (in Dmiish). Jorgensen K.H., Larsen Li., Jensen P.A. (1996) Tar Andersen K., Bennelzen S., Thorsen l'., Hansen N., cracking at low temperature in gasifiers for biomass. Ploger A., (iron C. (1996). Vurdcring af analysc- Fasc 1. Riso Report. (ENS journal no. 1383/95- metodcr til bcstcmmclse af udvalgtc potcntiell sund- 0003). Riso-R-957(DA). hedsfarlige stoffer i plantcmaterialc. Fasc 1. Lange (l, Roed J., Byrne M.A., Goddard A.J.H. (1996) Vandkvalitetsinstituttet. ATV til Miljostyrclscn. Indoor deposition measurements and implications for Riso-Dok-447 (in Danish). indoor inhalation dose. In: Deposition of Radionuclides, Bjerre A.B. (1996) Biotcknologi forvandlcr plantcrcstcr their Subsequent Relocation in the Environment mid til flydcnde bnendstof. RisoNyt no. 1: 14-15 Resulting Implications. Final report. EUR-16604: 13- Lkngvild K (1996) Sidstc ar mod "Rode Rodgnmcr"/ 23. Skovcn 28(8): 356-357. (in Dmiish) Lange Roed J., Zambori P., Feher /., Paretzke I LG., Hansen //. (1996) Den radioaktive sky fra Tjcmobyl - dosis Tschiersch J., Rybdcek K„ Tomasek M, Navarcik /., og virkning. Kvanl 7 (1): 21-26, 28 (in Danish). Jansta V., Datelinka /., Cipdkovd A. (1996) Hansen If (1996) Tjcmobyl - dengang og nu. Ingcniorcn Rcsusspension of deposited material. In. Deposition of 22 (16). 12-13 (in Danish). Radionuclides, their Subsequent Relocation in the Hansen II.. (1996) Risiko i kullurcl belysning. Ingcniorcn Environment and Resulting Implications. Final report. 22 (1). 11 (in Danish). EUR-16604: 59-82. Jensen J. (1996) Coordinator's report: Chromosome 5. Lauridsen B., Damkjar A., Nielsen Nelsen S.P., Nonbol Barley Genetics Newsletter 25: 96-99. Thykier-Nielsen S. (1996) Vurdcring af konse- Jorgensen J.H. (1996) Coordinator's report: Disease and kvenser pa dansk omrade af et alvorligt havari pa pest resistance genes. Barley Genetics Newsletter kcmckraftv mrket Ingalina i Litaucn. (in Danish) R1S0- 25: 106-107. R-892(DA) 36 p. Jorgensen ()., Fgsgaard //., Larsen F. (1996) Synthesis Nielsen S.P. (1996) An Intercomparison Exercise on of deuteraled compounds for the F0TEK project. Radionuclides in Sediment Samples. R1S0-R-914(EN) Dansk Kemi. 77 (6/7). 12-14 (in Danish). 41 p. Jorgensen R.B. (1996) Miljommssig risikovurdcring af Pagsberg /’. (1996) Cycling of OH and HO: by No, in genetisk modificercdc planter. Tidsskrift for the Tropospheric Oxidation of Aromatics. In: CEC Landokonomi 1/96: 15-18 (in Danish). R&D Programme in the Field of Env ironment 1994- Nielsen S.P. (1996) Ringe risiko fra radioaktivt alia Id 1996. Final Report on CYTROXA. Ed. by C. dumpet i del Arktiske hav. RisoNyt No 2: 4-5 (in Zet/.sch. P. Dcvoldcr. P. Pagsberg, I. Barnes and S. Dtinish).

124 Ris0-R-972(EN) Nielsen T. (1996) Life on Mars - fup or facts. Brouwer A.M, Wiering P.G., Zwier J.A1., Langkilde Ingenioren no. 36, 13 (in Danish). F.W., Wilbrandt R. Localization versus Pagsberg P. (1996) Kortlzegning af kvcelstofoxiders delocalization in diamine radical cations. 14tli rolle i luftforurening. RisoNyt No 2: 10-11 (in International Conference on Radical Ions, Uppsala, Danish). Sweden, 1-5 Jul 1996. Sehested J., Mogelberg T. Nielsen O.J. (1996) Christensen C.S., Skov H., Granby K., Egelov A.If., Trifiuoroacetic acid - a danger to environment? Nielsen T., Lohse C. Urban and semiural Naturens Verden nr. 3: 100-103 (in Danish). measurements of volatile organic compounds in Sturup S., Larsen E., Kristensen LA'. (1996) 1CP-MS Denmark. 7 th European Symp. on Physico- for determination of residual concentrations in food ­ Chemical Behaviour of Atmospheric Pollutants stuffs. Dansk Kemi, 77 (8 ): 25-28. (in Danish) "The Oxidizing Capacity of the Troposphere", Venice, Italy, 2-4 October 1996. Christensen L.K., Bilde M, Mogelberg T.E., Platz J., 9.6 Presentations (Oral and Posters) Sehested J., Waliington T.J., Nielsen O.J. Atmospheric Chemistry of acetic anhydride. 14th International Symposium on Gas Kinetics, Aarkrog A. Radioactive contamination of the marine University of Leeds. England, 7-12 Sep 1996. environment. CIBA Foundation Symposium No. 203, Christiansen C., Kunzendorf If, Emeis XI-61. Endler on Health Impacts of Large releases of Radionuclides. R., Struck LI, Benesch /)., Neumann 71, Sivkov 11 St. Petersburg, Russia, 27-29 Feb 1996. Sedimentation rate variabilities in the Gotland Basin Ahring B.K., Licht D., Nielsen P., Schmidt A S., Bjerre (GOBEX). Baltic marine science conference. A.B. (1996), Conversion of wheat straw to ethanol. Ronnc, Denmark, 22-26 Oct 1996. Abstract Wet oxidation pretreatment followed by anaerobic available. thermoplilic fermentation of the hemicellulose Christiansen S. Transformation of the obligate parasite fraction. 9th European Bioenergy Conference, Ervsiphe graminis f.sp. hordei. University of Copenhagen 24-27 June 1996. Copenhagen, Denmark, 10 Oct 1996. Andersson K.G. Modelling external radiation doses in Egsgaard If, Larsen E., Sehested J., Nielsen O.J. contaminated urban areas: Implications for develop ­ Intermediates in dimethyl ether flames. Annual ment of decontamination strategies. 9th International meeting of the Chemical Danish Society . Odense, congress on Radiation Protection Association, Vienna, Denmark, 11 Jun 1996. Austria, 14-19 Apr 1996, (IRPA, Seibersdorf). Gissel-Nielsen G. Hvor gar det N og P hen, som vi lilforer Behrens, A. Bestemmelse af sporelementer i kartofler. 4. marken. Dansk Landbrugs Grovvareselskab. Copen­ Danske Symposium i Analytis Kemi. The Royal hagen, Denmark, 27 Feb 1996 (in Danish). Veterinary' and Agricultural University, Copen ­ Gron 61, Wassenaar L., Krog A/. Strukturanalyse af hagen, Denmark. 20-21 Aug 1996 (in Danish). humus fra gnmdvand. Grundvandskomitccns vintcr- Behrens, A. 1CPMS og Partial Least Squares Regression: mode ora Gnmdvandsforurening, Denmark. 4-5 Mar Standardiseringsmetodcr. Rise National Laboratory, 1996 (in Danish). Roskilde. Denmark, Jan 1996 (in Danish) Gron 61, Wassenaar 71., Krog M. Strukturanalyse af Bi/de M, Sehested J., Nielsen O.J., Waliington T.J. humus fra grundvand. 4. Danske Symposium i Weakly bound adducts. Annual meeting of the Analytisk Kemi. The Royal Veterinary and Chemical Danish Society. Odense, Denmark, 11 Jun Agricultural University, Copenhagen. Denmark. 20- 1996. (Poster). 21 Aug 1996 (in Danish). Bilile A/., Sehested J., Nielsen O.J., Waliington T.J. Hass H., Nielsen O.J. Fuel Additives for the Future. Weakly bound adducts formed from the gas phase Workshop on the photochemical ozone problem and reaction of F atoms w ith brominated methanes. 14th its control, Berlin, 24-27 Sep 1996. Int. Svmp. on Gas Kinetics, University of Leeds, Hauser T. Fitness of hybrids between oilseed rape U.K., 7-12 Sep 1996. (Poster E 30). (Brassica napus) and weedy B. campestris : Bilde A/, Sehested J., Nielsen O.J., Waliington T.J. Gas Possibilities for introgression of transgenes to the phase adducts between halogen atoms and weed. University of Minnesota, Si. Paul, USA, 6 halogenated methanes. Symbion, Department of Sep 1996. Chemistry , University of Copenhagen, 13 Dec 1996. Holcman J Ferry! Ion and Manganese (III) Reactions Bobrowski K., Holcman J., Poznanski J., Wierzchowski with Simple Compounds. RINOXA Meeting, K.L. Long range electron transfer (LRET) University of Essen, Germany, 22-23 Feb 1996. accompany ing Trp. -> TyrO. intramolecular radical Holcman J. Kinetics and modelling work. Meeting in transformation in hen egg-white lysozyme (HEWL). SMP Centre for Air Pollution Processes and Models. Workshop on Genes and their products in basic Copenhagen 29 Nov 1996. research and biotechnology, Warsaw, Poland, 28-30 Nov 1996.

Ris0-R-972(EN) 125 Ilolcman J. Oxidation of Transition Metal Ions by Jorgensen E. V, Jensen EX.. Effects of dung on N: fixation Ozone and Atmospheric Relevant Chemistry of in grass/clover pastures. IFOAM, Copenhagen. Resulting Intermediates. Institute of Nuclear Denmark, 11-15 Aug 1996. Chemistry and Technology. Warsaw. Poland. 3 Apr Keszthelyi T. , Wilbrandt R., Bally T. (1996) Resonance 1996. Raman spectra and quantum chemical analysis of Jacobsen E A mechanistic study of transition metal ion radical cations of short polyenes. Int. Meeting on ozonation. University of Roskilde. Denmark. 13 Dec Inlra- and Intermolecular Photo processes of Con ­ 1996. jugated Molecules, 14-18 July, Riccione, Italy. Jacobsen E. Effects of manganese on the oxidation Printed abstract. capacity of the troposphere. University of Roskilde, Keszthelyi T. Wilbrandt R., Bally T. Resonance Raman Denmark. 13 Dec 1996. and quantum chemical studies of short polyene Jakobsen I. Competition for soil P between roots and radical cations. 23rd European Congress on hyphac of arbuscular mycorrhizas. 1COM 1. University Molecular Spectroscopy, 24-30 August, Bcla- of California. Berkeley . USA. 4-9 Aug 1996. tonfiired, Hungary . Printed abstract. Jakobsen L Introduktion til projcktel "Pesticide Effects on Lange C ’. Size distribution of neutron activatable elements Agricultural Soil Ecosystems". Ekcnas Herrgard, in atmospheric aerosols sampled w ith a l()-stagc BLPI. Sweden. 20 May 1996. European Aerosol Conference. Delft University of Jakobsen J Pcsticiders indflydelse pa biologiskc inter- Technology, The Netherlands. 9-11 Sep 1996.. aktioner og proccsscr i rodzonen. Workshop for den Larsen J. Interactions between arbuscular mycorrhizal tvUTininistcricllc pesticide forskningsningsindsats. fungi and collembola. Microbial Ecology , Univ ersity of Kollekollc, Denmark. 8 Feb 1996. Lund, Sw eden, 13 Mar 1996. Jensen EX. Biological nitrogen fixation in orghanic Larsen J. Interactions between external mycelium of farming systems. RALA Institute (Agricultural arbuscular mycorrhizal fungi and collembola. 1st Research Institute of Iceland). Reykjavik, 23-26 Sep International Conference on Mycorrtuza (ICOM 2), 1996. University of California. Berkeley, USA, 4-9 Aug Jensen EX. On the interaction between crop residue 1996. decomposition on cover cropping. Workshop in EU- Larsen J. Interactions between external mycelium of Conccrtcd Action: Long-term reduction o f nitrate arbuscular mycorrhizal fungi and collembola. 9th leaching by cover crops. Gumpenstein, Austria. 4-7 International Colloqvium on Apterygola. Dublin, Nov 1996. Ireland, 28-31 Jul 1996. Jensen EX. The role of grain legume cultivation in the Mordzinski A. Kownacki K, Wilbrandt R„ Les A., nitrogen cycling of temperate soils. 9th Nitrogen Adamowicz L. (1996) On the structure of Workshop. Braunschweig,, Germany. 9-12 Sep 1996. photochromic schiff bases. Spectroscopy and ab Jensen EX., llogh-Jensen If. Jorgensen E.V., Schjorring initio calculations. Int. Meeting on Intra- and J.K., l inther EE. Management of biological nitrogen Intermolecular Photo processes of Conjugated fixation in pastures. IFOAM-Congress, Symposia SI 1, Molecules, 14-18 July, Riccione, Italy. Abstract Copenhagen. Denmark. 11-15 Aug 1996. available. Jensen PA.. Larsen Jorgensen K.ll. Tar reduction Mogelberg 71, Sehested J., Nielsen O.J., Tyndall (L, by partial oxidation. 9th European Bioenergy Orlando J.J., Eracheboud J.M., Wallington T. The Conference. Copenhagen, Denmark, 24-27 June NO effect. Annual meeting of the Chemical Danish 1996. Society, Odense, Denmark, 11 June, 1996. Johansen. .1. Possible mechanisms regulating the Mogelberg T.E., Sehested J., Nielsen O.J., Wallington transport of N by external hyphac of arbuscular T.J. Decomposition of the CF,CFHO radical. 14th mycorrhizal fungi. VAM-Workshop Regulation in Int. Symp. Gas Kinetics, University of Leeds. U.K., Fungi. Phenomena and mechanisms, Forschungs- 7-12 Sep, 1996. (Poster) zentrum Jiilich. 21-22 Nov 1996. Mich, Germany : Mogelberg T.E., Sehested J., Nielsen O.J., Wallington 10 (abstract) T.J., Eracheboud J.-M., Tyndall G.S., Orlando J. Johansen, A. Excidcd hyphac of AM fungi - techniques The NO effect. Symbion, Department of Chemistry , and N metabolism. COST-Action 8.21 Workshop on Univ ersity of Copenhagen. 13 Dec 1996. Transport and Metabolism in Arbuscular Nielsen O.J., Bilde M, Mogelberg T.E., Sehested J., My corrhizas, Tunc Landboskolc, Denmark, 28-30 Wallington T.J. Atmospheric Chemistry of 1,2- Nov 1996. dichloroelhanc. 14th Int. Symp. Gas Kinetics, Justesen A.E. Molecular studies of the powdery mildew- Leeds, U.K.. 7-12 Sep, 1996. (Poster) fungus Erysiphe graminis f.sp. hordei. Plant Nielsen O.J., Nielsen T. Studies of ozone-related Scientist Meeting, Royal Veterinary and Agricul ­ chemical processes in the laboratory and in the field. tural University. Copenhagen, Denmark. 14 Mar Ozone-meeting. Danish Society of Environmental 1996 Chemistry, University of Copenhagen. Denmark, 12 Nov 1996.

126 Ris0-R-972(EN) Nielsen O.J. Formation of CF3COOH during the Rasmussen S.K. Plant serpins. Dipartimcnto di Gcnctica Atmospheric Degradation of HFC-134a. World e Microbiologia, University of Pavia, Italy, 10 Dec Chlorine Council Workshop on Chloroacetic Acids 1996. in the Atmospheric Environment - Anthropogenic or Rasmussen S.K. Targeting of strcs-rclatcd barley Natural ? Kongresshaus, Garmisch-Partenkirchen, peroxidase Prx7 in barley and transgenic tobacco. 29-30 Mar 1996. Vienna, Austria, 5-10 Jul 1996. Nielsen O.J., Wallington T.J., Sehested J. Atmospheric Ravnskov S. Influence of arbuscular mycorrhizal fungi on Chemistry of HCFCs and HFCs - Laboratory Fseudomonas fluoresans (DF57) in soils with and Studies. Abstracts Book. 89th Annual Meeting & without roots. 1st International Conference on Exhibition, Air & Waste Management Association, Mycorrhiza (1COM 2), University of. California. Nashville, Tennessee, USA, 23-28 Jun 1996. Paper Berkeley, USA, 4-9 Aug 1996. RP130 A.03. RoedJ., Andersson K.G., Lange C. The Fate and Impact of Nielsen O.J., Sehested J., Magelherg T.K., Wallington Radio-contaminants in Urban Areas. CIBA Foundation T.J., Kaiser E. W. (1996) Dimethyl ether oxidation: Symposium No. 203 on Health Impacts of Large Kinetics and mechanism of the CH3OCH2 + 02 Releases of Radionuclides, St. Petersburg. Russia, 27- reaction. 7th European Symposium on Physico- 29 Feb 1996. Chemical Behaviour of Atmospheric Pollutants, Roed J., Andersson K.G., Togawa O. Weathering of Venice, Italy, 2-4 Oct 1996. (European Commission, radionuclides deposited in inhabited areas. 9th Brussels, 1996). Paper R-35. International congress on Radiation Protection Nielsen O.J., Wallington T.J., Hurley M.D., Sehested, Association. Vienna, Austria 14-19 Apr 1996. J., Magelherg T.E., Erachehoud, J.M., Orlando J.J., RoedJ., Goddard A.J.H., Lange Byrne M.A. Mitigation Tyndall G.S. Excited Alkoxy Radicals in the of radiation dose through deposition to indoor surfaces. Atmospheric Degradation of Halogcnated Com ­ 9th International congress on Radiation Protection pounds. First SPARC General Assembly of the Association, Vienna, Austria 14-19 Apr 1996 (IRPA. WCRP Project on Stratospheric Processes and Their Seibcrsdorf). Role in Climate, Melbourne, Australia, 2-6 Dec Roed J., Togawa O. A method for the determination of 1996. vertical distribution profiles of radioactive contamina­ Nielsen S.P. Radioaktivitet i Ostcrsocn: Model og tion in soils. 9th International congress on Radiation virkelighed. 9. Danske Havforskermode, Technical Protection Association, Vienna Austria, 14-19 Apr University of Denmark, 16-18 Jan 1996 (in Danish). 1996. (IRPA, Seibersdorf) Filegaard K. Deposition of O, and NOx . Workshop on Sandstrom B., Solgaard, F. Copper absorption from Atmospheric Deposition and Surface Exchange. high fibers diets. Elman, Germany 26-27 Sep 1996. Roskilde, Denmark, 11-12 Apr 1996. Schmidt A.S. Pretreatment of lignoccllulosc for Filegaard K. Hummelshoj F., Jensen NO. Nitric oxide conversion of hemicellulose to biocthanol. Nordic emission from forest soil. In: Transport and Energy Research Programme - Bioenergv processes/ transformation of pollutants in the troposphere. ethanol - seminar, VTT. Finland 12 Mar 1996. Abstracts of lectures and posters (Borcll, P„ ed ). Schmidt A S., Ahring B.K., Bjerre A.B. Recent studies EUROTRAC symposium 96, Garmisch-Partenkirchen, on Cenversion of Wheat Straw to ethanol Germany, 25-29 Mar 1996. (Poster 7-19). Workshop on Bioproducts from Lignocellulosics - Flatz J., Sehested J., Sehested K, Nielsen O.J. Progress in Developing Feasible Bio/chcmical Reactions of Dimethylether at 296-666 K. 14th Pathways, Sounion, Greece, 25-27 Sep 1996. International Symposium on Gas Kinetics, Schmidt, A.S., Bjerre, A.B. (1996). Pretreatment of University of Leeds, U.K.. 7-12 Sep, 1996. (Poster) agricultural crop residues for conversion to high- C41. value products. International Conference in Cereals: Flatz J., Sehested J., Nielsen O.J. Atmospheric Novel Uses and Processes. Manchester, U.K., 5-6 chemistry of 1,4-dioxane. University of Odense, Jun 1996. Denmark, 11 Jun 1996. Schmidt A.S., FulsJ, Bjerre A.B. (1996). Comparison of Flatz J., Magelherg T.E., Nielsen O.J., Sehested J., wet oxidation and steaming for solubilization of the Wallington T.J., Kaiser E.W. Experimental study of hemicellulose fraction in wheat straw and birch dimethyl ether in the gas phase. NOSA/NORSAC wood. 9th European Bioenergv Conference, symposium, LO-Hojskolcn, Elsinore, Denmark, 15- Copenhagen 24-27 June 1996. 17 Nov 1996. Sehested K. Oxidation of DMSO. DMS02, Floger A., Bjerre A ll., Egsgaard H., Nielsen T. Methansulfonic acid, Methansulflnic acid and Analytical chemical aspects of the oxidative Hydroxy MethanSulfinic acid by the OH radical. decomposition of tar compounds. 4th Danish RINOXA meeting, Zitell, Venice, Italy, 1-4 Oct symposium on Analytical Chemistry. The Royal 1996. Veterinary' and Agricultural University, Copen ­ Sehested J. Kinetics and spectroscopy of ozone hagen, Denmark, 20-21 Aug 1996 (abstract formation. University of Odense. Denmark. 11 Jun available). 1996.

Ris0-R-972(EN) 127 SeheslcclMogelberg T. F.. Fagerstrom K.. Lund A. The reactions of HO: radicals with FO:, CF,CFHO: and CF,0: radicals. 14th International Symposium on Gas Kinetics. University of Leeds. England, 7-12 Sep 1996. Sehested J.. Mogelberg Platz J., Nielsen O.J., Fgsgaard If., Wallington T.J., Kaiser F.W. Dimethyl ether oxidation: Kinetics and Mechanism of the CH3OCFL + 0; Reaction at 296K and 0.38- 0.40 Torr Total Pressure. 14th Int. Symp. on Gas. Kinct.. University of Leeds, U.K., 8 Sep 1996 C5. Skov II.. Fllerman T., Hansen A.B., Palmgren Nielsen 7., Platz ,/. Automatic gas chromatography in atmospheric monitoring. Workshop on Air Pollution Monitoring. Riso National Laboratory. Roskildc. Denmark, 11-12 Mar 1996. Stenhahn A/, Jensen P.A., Hald P. (1996). Deposition investigation in Straw Fired Boilers. 9th European Bioenergy Conference. Copenhagen, Denmark 24- 27 Jun 1996. Stitrup S. Grundstofbcstcmmelser ved ICPMS. FL1K - Forcningcn af lcvnedsmiddelingcniorcr og -kandida- tcr. Vanlose, Denmark, 26 Feb 1996. Sttirup S. (1996). Elemental trace analysis in biological samples by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR1CPMS). The Nordic Atomic Spectroscopy and Trace Element Conference. The Danish Society of Analytical Chemistry . Ebcltoft. Denmark, 2-6 Jun 1996. Ihingslnip /., Johansen A. P uptake by excised hyphae of AM fungi. - COST-Action 8.21 Workshop on Transport and Metabolism in Arbuscular Mycorrhizas, Time L;mdboskolc. Roskildc, Denmark. 28-30 Nov 1996. Ihingslnip /., Schweiger I\. Jakohsen I. Measurement of potential phosphorus transport by field populations of iirbuscular mycorrhizal fungi. 1st International Con­ ference on Mycorrhiza (ICOM 2), University of California. Berkeley. USA. 4-9 Aug 1996. 1 'inther F.P., Jensen E.S. Biological nitrogen fixation in an organic farming system. 11th LFOAM Conference, Copenhagen. Denmark, 11-15 Aug 1996. (Poster). Wallington T.J., Hurley AID.. Fracheboud J.M., Orlando J.J., Tyndall G.S., Sehested J., Mogelberg I F.. Nielsen O.J. (1996) Role of excited CF,CHFO radicals in the atmospheric chemistry of FTFC-134a. Book of abstracts. 14th International Sy mposium on Gas Kinetics. University of Leeds, England, 7-12 Sep 1996. Paper C4. Wilbrandt R. Time-resolved resonance Raman Studies and quantum chemical analysis of organic inter­ mediates. Annual meeting of the Danish Optical Society . Riso, Denmark, 21-22 Nov 1996. Oberg G., Gran C. Organically bound halogens in spruce forest throughfall. XIX Nordic Hydrological Conference, NHK 96. Akureyri, Iceland. 13-15 Aug 1996. 10 Papers accepted for Publication

Aarkrog A. Radioecologv. development and trends. Proc. monitored by the brown algae, Fucus vesiculosus. Proc. 7th Nordic Radiological Seminar. 26-29 August. Int. Symp. “Radionuclides in the Ocean" (RADOC 96- Reykjavik. Iceland. 97) Cherbourg-Oclevillc, France. 7-11 October. Aarkrog A., ('hen Q., Dahlgaard II, Nielsen S.P. Damgaard C, Ostergard H. Density dependent growth Evidence of "Tc in Ural river sediments. Journal of and life history evolution of polycyclic leaf Env ironmental Radioactiv ity; pathogens: A continuous time growth model. J. Ambus P. Spatial and Seasonal N20 and CH4 Fluxes in Phytopath. Danish Forest-. Grassland-, and Agroecosystems. Feilberg A., Holcman J., Lohse ('., Nielsen T., Sehested Journal of Environmental Quality . K. Reactions of N-heteroaromatics with HO in tire Ambus P. Patterns of N2Q and CH., flaxes along topo- aqueous phase in: Proceedings of EUROTRAC gnmdients in danish ecosy sytems. Journal of Environ­ Symposium '96. Ed by P.M. Borrcll, P. Borrell. T. mental Quality. Cvitas. K. Kelly, W. Seiler (eds). Computational Behrens A. Multivariate calibration standardization used Mechanics Publications, Southampton. for the reduction of standards needed for multivariate Granby A'., Egelov AIL, Nielsen '/., Lohse C. calibration in inductively coupled plasma mass spectro­ Carboxylic acids: Seasonal v ariation and relation to metry; Spectrochim. Acta B. photochemical products. J. Atmos. Chem. Bibak A. Competitive sorption of copper, nickel, and Gulin SB., Aarkrog A., Polikarpov G.G., Nielsen S.P., zinc by an oxisol. Commun. Soil Sci. Plant Anal. Egorov V.N Chronological study of 13 Cs input to tire Bibak A. Copper retention by Danish spodosols in rela ­ Black Sea deep and shelf sediments. Proc. hit. Symp. tion to contents of organic mater and aluminum, “Radionuclides in the Ocean" (RADOC 96-97) iron, and manganese oxides. Commun. Soil Sci. Cherbourg-Octeville. France, 7-11 October. Plant Anal. Hauser T.P., Jorgensen R.B., Ostergard 11. Preferential Bobrowski A... Hotcman J., Poznanski J., Wierzchowski exclusion of hybrids in mixed pollinations between K.L. Long range electron transfer between proline- oilseed rape (Brassica napus) and weedy B. bridged aromatic amino acids. In: Advances in campestris. Anver. J. Bot. Chemistry Scries (Photochemistry and Radiation He/weg ('., Nielsen T., Hansen P.F. QSPRs compared Chemistry ), cd. J.F. Wishart and D. Nocera. with HPLC experiments for the estimation of Kow Bobrowski K, Holcman J., Poznanski ./., Wierzchowski for N-PAC’s. Proc. of the 7th Int. Workshop on K.L. Pulse radiolvsis studies of intramolecular QSARs in Environmental Sciences. Ed. bv G. electron transfer in model peptides and proteins: 7. Schuurmann, Society of Environmental Toxicology Trp.-> TyrO. Radical transformation in hen egg- and Chemistry. white lysozy me. Effects of Ph, temperature. Trp-62 Helweg C., Nielsen '/., Hansen P.E. Determination of oxidation and inhibitor binding. Biophys. Chem. oclanol-vvatcr partition coefficients of polar poly ­ Brouwer A.M., Wiering P.G., Zwier J.M., Langkilde cyclic aromatic compounds (N-PAC) by high F.W., Wilbrandt R. Localization versus delocaliza ­ performance liquid chromatography . Chcmosphcrc. tion in diamine radical cations. Acta Chem. Scand. Hovmoller A I, Ostergard //., Munk L. Modelling Chen B Y., Cheng B.F., Jorgensen R.B., Heneen W.K. virulence dynamics of airborne plant pathogens in Production and cytogenetics of Brassica campestris- relation to selection by host resistance in alboglabra chromosome addition lines. Theor. Appl. agricultural crops. In: The gene-for-genc Relation ­ Genet. ship in Plant-Parasite Interactions. Crute I R , Holub Christensen C.S., Skov H., Granby A., Egelov A H., E.B.. Burdon J.J. (eds). Wallinglon. UK: CAB Nielsen 7 ’., Lohse C. Urban and semiural measure ­ International. ments of volatile organic compounds in Denmark. Jacobsen F, Holcman J., Sehested K. Activation Para ­ The oxidizing capacity of the troposphere. Proc. of meters of Ferry'1 Ion Reactions in Aqueous Acid the 7th European Svmp. on Physico-Chemical Solutions. Int. J. Chem. Kinet. Behaviour of Atmospheric Pollutants. European Jacobsen F, Holcman J., Sehested K. Manganese(II)- Commission. Superoxide Complex in Aqueous Solution J. Phys. Dahl S.W., Rasmussen S.K., llejgaard J. (1996) Chem. Heterologous expression of three inhibitory plant Jacobsen F., Holcman J., Sehested K. Effects of serpins. Proceedings. International Symposium on Manganese on the Oxidation Capacity of the the Chemistry and Biology of Serpins. Church. Troposphere. Proceedings of the 7th European Cunningham. Ginsburg. Hoffman. Tollefsen & Symposium on Physico-Chemical Behaviour of Stone, Plenum Publishing Corp. Atmospheric Pollutants: " The Oxidizing Capacity Dahlgaard II. Bergan T.D.S., Christensen G. Technetium- of the Troposphere, October 2-4. Zitcllc. Venice. 99 and 13 Cs time series at the Norwegian Coast Italy, B. Larsen (cd).

Ris0-R-972(EN) 129 Jensen ES. Nitrogen dynamics during initial decomposi ­ Nielsen S.P., Ohlenschheger A/, Karlberg K. The tion of 1'N-labelled pea and barley residues. Biology radiological exposure of man from ingestion of Cs-137 mid Fertility of Soils. and SR-90 in seafood from the Baltic Sea. International Jorgensen El'., Jensen KS. Do dung patches influence Symposium on Ionising Radiation: Protection of the N: fixation in grass/clover mixtures in the short term? Natural Environment. 20-24 May. Stockholm, Sweden. Proceedings. Legumes in sustainable farming systems. Nielsen 71, Granby K., Egelov A.II., Hummelshoj P . Occasional Symposium No. 30. 1996. British Grassland Skov II. Atmospheric nitrogen compounds, photo ­ Society. chemical oxidants and products. Tropospheric Jorgensen K. 11 Klovergnesmarkens N: fikscring: Nyt fra Ozone Research. D. Kiev (ed) Springer, Heidelberg. el dansk forskningsprojekt. Forskningsnytl om Nielsen 71, Hertel ()., ('kristensen C.S., Egelov A.II., ockologisk landbruk i Norden (in Danish). Granby K., Hansen A.B., Platz J., Skov II. Evalua ­ Jorgensen R ll , Hauser T., Landbo /,., Mikkelsen T.R., tion of a trajectory model to simulate formation of Ostergdrd II. Brassica napus and B. campeslris'. tropospheric ozone, other photochemical oxidants Spontaneous hybridisation, backcrossing and fitness and atmospheric reaction products. The oxidizing components of offspring plants. Environmental capacity of the troposphere. Proc. of the 7th Impact of Genetically-Modified Crops. Institute of Europemi Symp. on Physico-Chemical Behaviour of Terrestrial Ecology. Monks Wood. UK. 28-30 Nov. Atmospheric Pollutants. European Commission. 1995 Nielsen 71, Siigur K., Ilelweg Jorgensen ()., Hansen Kjtersgdrd J 1 1 II, Jespersen II.M, Rasmussen S.K., P.K., Kirso V. Sorption of polycyclic aromatic Welinder K.G. Sequence and RT-PCR expressioin compounds to humic acid as studied by high analysis of two peroxidases from Arabidopsis performance liquid chromatography. Environ. Sci. thaliana belonging to a novel evolutionary branch of Technol. plant peroxidases. Plant Mol. Biol. Nielsen 71, Hertel ()., Granby K., Hansen A.B., Platz J., Kristensen B.K., Brandi , J., Bojsen, A.., Thordal- Skov II, Stenholt C. Egelov A.II Comparison of Christensen, II., Kerhy. K.B., Collinge, D.B., measurements and modeling of ozone, other photo ­ Mikkelsen. J I)., Rasmussen, S.K.: Expression of a chemical oxidants, precursors and atmospheric defence-related intercellular barley peroxidase in reaction products during a summer high pressure transgenic tobacco. Plant Science. episode in Denmark. In: Proceedings of Landbo I... Andersen /» , Jorgensen R.B. Natural hybri­ EUROTRAC Symposium ‘96. Ed. by P.M. Borrell, dization between oilseed rape and a wild relative: P. Borrell. T. Cvitas. K. Kelly and W. Seiler. Com ­ hybrids among seeds from weedy B. campeslris. putational Mechanics Publications, Southampton. Hcrcditas 125 Roed J. Lange C, Andersson K. Decontamination in a Larsen J. Interactions between external mycelium of Russian settlement. 11th Meeting for Nordic Society arbuscular mycorrhizal fungi mid collcmbola. for Radiation Protection. 29-30 August, Reykjavik. Pcdobiologia. Iceland.. I.yngkjar M.F., Carver I.LAV., /even R.J. Suppression Scheel ILK, Ancellet G., Areskoug //., Beck,/., de A liter of resistance to Krvsiphe graminis f.sp. hordei I)., Egelov A IL, Esser P., Etienne A., Eerenczi Z. conferred by the mlo5 barley powdery mildew Geis II, Grab be G., Granby K., Gomiscek B., resistance gene. Physiol. Mol. Plant Pathol. Ilaspra /.., Kesele N., Klasinc /.., Laurila 71, Mogelberg T.K., Sehested J., Wallington T.J., Nielsen Lindskog A., Mowrer J., Nielsen 71, Roemer A7., O.J. Atmospheric Chemistry of HFC-134a: Kinetics Schmitt R., Simmonds P., Sladkovic R . Solberg S., of the Decomposition of the Alkoxy Radical Varotsos ('., de Waal L. TOR task group 1 report. CF3CFHO. hit. J. Chcm. Kinel. Tropospheric Ozone Research. Ed. bv D, Kiev, Nielsen O.J., Wallington I'.J. Formation of peroxy Springer, Heidelberg. radicals in the gas phase, ed. Zeev Alfassi, John Sehested J., Sehested K., Platz J., Nielsen O.J. Wiley and Sons Ltd. Oxidation of dimethy l ether: Absolute rale constants Nielsen O.J., Wallington T.J. UV absorption spectra of for the self reaction of CH3OCH2 radicals, the peroxy radical in the gas phase, ed. Zeev Alfassi, reaction of CH3OCH2 radicals with 02 , and the John Wiley and Sons Ltd. thermal decomposition of CH3OCH2 radicals. Int. J. Nielsen S.P. A comparison between predicted and Chem. Kinet. observed levels of 13 Cs mid ^'Sr in the Baltic Sea. Proc. Skov II., Egelov A. II, Granby K., Nielsen 71 Int. Symp. "Radionuclides in the Ocean ” (RADOC 96- Relationships between ozone and other photo ­ 97) Cherbourg-Octcvillc, France. 7-1 1 October chemical products at LI. Valby, Denmark. Atmos. Nielsen S.P. A sensitivity analysis of a radiological Environ. assessment model for Arctic waters. 7th Nordic Sorensen, P. Short-term anaerobic storage of 1 N-labcllcd Radiological Seminar, 26-29 August. Reykjavik, sheep urine does not influence the mineralization of Iceland. nitrogen in soil. Plant and Soil. Nielsen S.P. An Intercomparison Exercise on Radio ­ nuclides in Sediment Samples. 7th Nordic Radiological Seminar. 26-29 August. Rey kjavik, Iceland.

130 Ris0-R-972(EN) 11 Education

11.1 Titles of Honour FN02 + M studeret ved hjaelp af plus radiolyse. Riso National Laboratory and Aarkrog, A. N.V. Timofeeff-Ressovsky Memo­ University of Roskilde. rial Medal from Russia. Hansen, T., 1996: Cellulaer lokalisering af byg Nielsen, O.J. Henry Ford Technology Award in peroxidase Prx7 i transgen tobak. Riso Natio­ Detroit, USA. nal Laboratory and The Royal Veterinary and Agricultural University, Copenhagen. 11.2 Ph.D. Theses Jensen, P. Westphal, 1996: Transformation af tobak (Nicotiana benthamiana) med cDNA Dahl, S. Weis, 1996. Engineering and hetero ­ kodende for den stressrelaterede bygperoxi- dase Prx7. Riso National Laboratory and The logous expression of three protein Z-type plant Royal serpins. Identification and characterization og Veterinary and Agricultural University, Copenhagen. inhibitory activities. Riso National Laboratory Klausen, ./., 1996: 5’ RACE kloning af byg and Technical University of Denmark. L'rederiksen, J.M., 1996: The photophysics of (Hordeum vilgare L:) serpin protein Z7. Riso National truxene and a hexasubstituted derivative. Riso Laboratory and The Royal Veterinary National Laboratory and University of Copen ­ and Agricultural University, Copenhagen. hagen. Kloch, Kranip, H., 1996: Fjemelse af Ru fra Keszthelyi, 'lamas, 1996. The geometrical and lawandsprover med henblik pa kvantitativ vibrational structures of short polyene radical bestemmelse af Tc ved ICPMS. Riso National cations investigated by resonance raman Laboratory and Technical University of Denmark. spectroscopy and molecular orbital calcula­ tions. Riso National Laboratory and Univer ­ Kristensen, P. Kyvsgaard , 1996: The miniraliza- sity of Copenhagen. tion of faeces nitrogen in relation to the feed Lyngkjcer, M., 1996: Infection of barley by E. and faeces composition. Riso National graminis. Riso National Laboratory and The Laboratory and The Royal Veterinary and Royal Veterinary and Agricultural University, Agricultural University, Copenhagen. Copenhagen. Larsen, K. Block, 1996: Gen-for-gen systemets Mikkelsen, 7'., 1996: Flow of engineered genes anvendelse i byg/meldug systemet. Riso National Laboratory and The Royal Veterinary from crops to wild or weedy relatives - is there an environmental risk? A case study on and Agricultural University, Copenhagen. oilseed rape (Brassica napus) and weedy Siigur, K., 1996: Sorption of quinoline to organic Brassica campestris. Riso National Labora ­ matter. Riso National Laboratory and tory and University of Copenhagen University of Tartu, Estonia. Wetche, T.P., 1996: Adsorption af humus til mineraler. Riso National Laboratory and The Royal Veterinary and Agricultural University, 11.3 M. Sc. Theses Copenhagen. 109 p.

Andresen, T., 1996: Localization and effect of repetitive elements in the barley powdery 11.4 External Examiners mildew fungus, Erysiphe graminis f.sp. hordei. Riso National Laboratory and University of Odense. Engvild , K.C. Censor in plant physiology at the University of Aarhus. Feilberg, A., 1996: Spektrokimetiske undersogel- Giese, H. Censor in molecular biology and plant ser af HFC-236cb og HFC-227ca. Reaktioner biochemistry, The Royal Veterinary and Agri­ og UV-spektre af CF3CF2CFH02 og cultural University, Copenhagen. CF3CF2CF2O2 samt reaktionen F + N02+M -»

Riso-R-972(EN) 131 Censor in molecular biology and plant physio ­ Sept., Helsinki. Strategies and Methodologies for logy at University of Aarhus. Applied Marine Radioactivity Studies. Gran, C. Member of the panel of external Dahlgaard, H. (1996) Water transport, scavenging examines at the Danish Engineering Educa­ processes, water sediment interaction - part I: tions. radioactive tracers and coastal oceanography. Opponent at Ph D. thesis, Technical University of International Atomic Energy Agency, Interna ­ Denmark. tional Seminar on the use of Isotope Techniques Gundersen, V. Censor in chemical analysis at in Marine Environmental Studies, Athens, Technical University of Denmark. Greece: 11-22 November. Censor in mechanical engineering at Aalborg Dahlgaard, H. (1996) Water transport, scavenging University. processes, water sediment interaction - part II: Jensen, A. Censor in biology at all Danish radioactive tracers and bio-geo-chemistry. universities. International Atomic Energy Agency, Inter ­ Jensen, K.S. Censor in plant nutrition and crop national Seminar on the use of Isotope physiology, The Royal Veterinary- and Agri­ Techniques in Marine Environmental Studies, cultural University, Copenhagen. Athens, Greece: 11-22 November. Jorgensen, J.H. Censor in plant breeding and Dahlgaard, H. (1996). Introduction to Baltic Sea plant pathology at the Royal Veterinary and Studies. International Atomic Energy Agency, Agricultural University, Copenhagen. International Seminar on the use of Isotope Jorgensen, IIB. Censor in molecular biology and Techniques in Marine Environmental Studies, plant physiology at University of Aarhus. Athens, Greece: 11 -22 November. Nielsen, O.J. Censor in chemistry at University of Gron, (' Lectures on Natural Production of Chloro- Copenhagen and University of Odense. organic Compounds in Soil and Groundwater (in Nielsen, S.P. (1996) Opponent at Klas Rosen ’s Danish), 14 March, 1996, and Forest Soil Dr.science degree in Radioecology. Swedish Processes and Natural Products in Groundwater Agricultural University, Uppsala. 31 May, 1996 (in Danish), 3 December, 1996, at The Royal Rasmussen, S.K. Censor in molecular biology and Veterinary and Agricultural University, Copen­ plant biochemistry, The Royal Veterinary and hagen. Agricultural University, Copenhagen and Uni ­ Jakobsen, L Mykorrhiza. Lecture in Plant Biology 1 versity of Aarhus. at Institute of Plant Biology, The Royal Rosendahl, L. Censor in Biology at University of Veterinary and Agricultural University, Aarhus. Copenhagen 14 May 1996. Wilbrandt, R. Censor at University of Odense Jorgensen, J. Helms. Four lectures in Disease Gslcrgdrd, H. Censor in biology at University of Resistance Breeding at The Royal Veterinary Aarhus and University of Copenhagen. and Agricultural University, Copenhagen. Mogelherg T.E. Lecture in Stratospheric Ozone at University of Roskilde, 26 Nov 1996. 11.5 External Teaching and Lectures Nielsen, S.P. (1996) Ecological and Public Health Implications from Flasks lost at Sea. Paper Dahlgaard, H. (1996) Biological indicators: presented at a special consultative meeting biokinetics of radionuclides in mussels and (SCM) of entities involved in the Marine seaweeds. Regional Training Course, 9-20 Sept., Transport of Nuclear Materials covered by the Helsinki. Strategies and Methodologies for INF Code. International Maritime Organization, Applied Marine Radioactivity Studies. London, UK, 4-6 March. Dahlgaard, H. (1996) Radioactive tracers as a tool Nielsen. O.J. Course on Atmospheric Chemistry at in coastal oceanography. International Atomic the University of Copenhagen. Energy Agency, Regional Training Course, 9-20 Rasmussen, S.K. Lecture in Plant Biochemistry at The Royal Veterinary and Agricultural University, Copenhagen.

132 Ris0-R-972(EN) 12 Exchange of Scientists

Ambus, P., leave of absence as Post doc. at McLaughlin, W.L., National Institute of Michigan State University, U S A. (1 year). Standards and Technology, USA (2 weeks). Bell, A"., Imperial College of Science, London, Meunier, H.H., Assumption College, Worchester, U.K. (1 week) USA. (6 months) Bobrowski, K., Institute of Nuclear Chemistry Mogelberg, T. visiting Ph D. student at National and Technology, Warsaw, Poland (1 week) Center for Atmospheric Research, Boulder, Brouwer, A.M., University of Amsterdam, Colorado , USA (2 months). Holland (1 week) Mogelberg, T. visiting Ph D. student at Ford Byrne, M., Imperial College of Science, London, Motor Company, Dearborn, Michigan, USA U K (1 week) (1 month). Caffier, V, INRA, Grignon, France (3 weeks) Nath, B. Nagender, National Institute of Oceano ­ Chen, Ban-Yuan , Swedish Agricultural Univer ­ graphy, Goa, India (2 weeks) sity, Svaldv, Sweden (2 weeks) Nieolaisen, M, Technical University of Denmark Cheng, Bi-Bang , Swedish Agricultural Univer ­ (7 months) sity, Svaldv, Sweden (2 weeks) Nielsen, O.J. leave of absence as Managing Chevre, A M., INRA, Dijon, France (3 weeks) Director at Ford Research Center, Aachen, Draganic, I.G., Institute Vinca, Belgrade, Serbia, Germany (7 months). F.R. Yugoslavia (2 weeks) Olsen, S. Feltexpedition in Russia (3 weeks). Erchov, /./., Obnisk Institute of Nuclear En ­ Pedersen, C. visiting scientist at Department of gineering, Russia (1 week) Plant Science, University of Adelaide, Austra­ B’agerstrdm, K., Studsvik Material AB, lia (10 months) Nykoping, Sweden (1 week) Platz, J. visiting Ph D. student at Ford Motor Getoff, N., Institut fur Strahlenchemie, Vienna, Company, Dearborn, Michigan, USA (2 Austria (3 weeks) weeks). Grosell, M. University of Uppsala, Sweden (1 Ploger, A. visiting scientist at The Macaulay week) and McMaster University, Hamilton, Land Use Research Institute, Aberdeen, Ontario, Canada (2 weeks). Scotland (2 months). Hagen, A. Oslo University, Norway (6 weeks). Pogocki, D., Institute of Nuclear Chemistry and Hickel, B., CEA CE, Saclay, France (1 week). Technology, Warsaw, Poland (1 week) .Inner, E., Agricultural University of Norway (1 Pongsakul, P., Soil Science Division, Bangkok, week) Thailand (1 week) Jorgensen, B’. visiting PhD. student at Pons, J., University of Giessen, Germany (4 Agresearch, Ruakura Agricultural Research weeks) Centre, Hamilton, New Zealand (3 months). Rieseberg, /,., Department of Biology, Indiana Kahilunln, //., Agricultural Research Centre of University, U S A. (1 week) Finland (ARCF), Partala Research Station for Sato, K. Research Institute for Bioresources, Ecological Agriculture, Juva, Finland (1 week). Okayama University, Japan (2 months) Knllisl, //., University of Tartu, Estonia (8 Salnikov, N.L., Obnisk Institute of Nuclear months). Engineering, Russia (1 week) Knstyuk, ()., Institute of Cell Biology and Genetic Sehested, J. visiting scientist at National Center Engineering, Kiev, Ukraine (3 months) for Atmospheric Research, Boulder, Colorado, Lange, C. leave of absence as Post doc. at GSF- USA (2 months). Forschungszentrum fur Umwelt und Gesund- Sehested, J. visiting scientist at Ford Motor heit, Neuherberg, Germany (4 months) Company, Dearborn, Michigan, USA (1 Leinnva, .S', Laboratory of Belorussian State month). University, Ukraine (2 months) Siigur, K., Institute of Chemical Physics, Uni ­ Bower, C, Justus Liebig University, Giessen, versity of Tartu, Estonia (8 months). Germany (6 weeks)

Riso-R-972(EN) 133 Solar, S., Institut fur Strahlenchemie, Vienna, Wallington, 1'., Ford Motor Co., Dearborn, USA Austria (3 weeks) (2 weeks). Strzelczak-Burlinska, G., Institute of Nuclear Vest berg, M, Laukaa Research and Elite Plant Chemistry and Technology, Warsaw, Poland University, Agricultural Research Centre of (1 week) Finland (1 week). Szefer, P., Medical Academy, Gdansk, Poland (1 Zagorski, Z, Institute of Nuclear Chemistry and week)- Technology, Warsaw, Poland (1 week) IJnal. Z, Ankara Nuclear Research and Training Zwier, J., University of Amsterdam, Holland (4 Center, Turkey (4 weeks) weeks)

134 Ris0-R-972(EN) 13 Guest Lectures

Bendixen, C, Research Centre Foulum, Denmark: Hauge Madsen, K., The Royal Veterinary and Agri­ "The two hybrid system: advantages and applica ­ cultural University, Copenhagen, Denmark: tions". "Modeller til fomdsigelse af herbicidforbruget Bobrowski, K., Institute of Nuclear Chemistry ved dyrkning af herbicidtolerante afgroder" (in and Technology, Warsaw, Poland: “Mecha ­ Danish). nism of aqueous phase oxidation of sulphur in Jaenicke, R., University of Mainz, Germany: thioethers ”. “Atmospheric research at the University of Boren, H., Linkoping University, Sweden: “AOX Mainz ”. and Chlorinated Acetic Acids in Precipitation ” Japar, S., Ford Motor Company, Dearborn, Break, 7'., The Consumer Council, Copenhagen, Michigan, USA: “Fine Airborne Particulate Denmark: "Forbrugemes holdning til gentek- Matter: Can we accurately define the role of nologiske fodevarer" (in Danish). motor vehicles? ” Cqffier, V, INRA, Thiverval-Grignon, France: Knudsen, J.M., University of Copenhagen, Den ­ "Genetic structure of barley powdery mildew mark: “Exploration of life outside the Earth ” population in France". (in Danish). Dahlgard, ,/., University of Aarhus, Denmark: Palmgren, M, University of Copenhagen, "Varmeresistens hos Drosophila, akklimation og Denmark: "Iongradienter og v$kst". (in Danish) evolution" . (in Danish) Rieseberg, L, Department of Biology, Indiana Draganic. /., Institute Vinca, Belgrade, Serbia, University, Bloomington, USA: "The role of F R. Yogoslavia: “Radiationchemical ap ­ hybridization in plant evolution". proaches to comets and the early Earth ”. Sibbesen, £., Research Centre Foulum, Denmark: Getoff, N., Institut fur Strahlenchemie, Vienna, "Fosforomscetning i landbruget og fosfortab til Austria: “Pulse radiolysis in radiation vandmiljoet". (in Danish) biology ”. Weiner, J., The Royal Veterinary and Agricultural Gjessing, /{., Norwegian Institute for Water University, Copenhagen, Denmark: "When do Research, Norway: “Natural Organic Matter in large plants suppress the growth of their smaller Water; Ecological and Hygienic Effects”. neighbours?" Gregersen, /', The Royal Veterinary and Agri­ cultural University, Copenhagen, Denmark: "Ekspression af forsvarsgener i byg over for angreb af bygmeldugsvampen". (in Danish)

Ris0-R-972(EN) 135 14 Committee Membership

14.1 National Jorgensen, R.B. Member of Environmental Appeal Roard. Aarkrog, A. Danish National Council for I.arsen, E. Danish Society of Mass Spectrometry. Oceanology. Associated as censor in chemistry to the Danish AMAP steering group and AMAP Danish universities. evaluation groups on national terrestrial assess­ The Danish National Committee for Che ­ ment, marine assessment and analysis. mistry. Giese. H. OECD Committee for Biotechnology. Miller, A. Danish Medical Device Association Board of Danish Polar Center. (DMDA). Sterilization committee. Danish Society for the Conservation of Danish Standards Association. Committee S Nature, Committee for Environmental Issues. 259 on Sterilization of Medical Equipment. Gissel-Niclscn, G. Danish Academy of Technical (Chairman). Sciences. Nielsen, O.J. The National Committee for The Governing board of Danish Institute of Plant International Geosphere-Biosphere Program­ and Soil Science. me (IGBP). Danish Society for the Conservation of Pilegaard, K. Danish Centre for Atmospheric Nature, Committee for Environmental Issues. Research (DCAR) Working Group on effects. Gron. ('. Board of the Soil and Ground Water (Chairman). Contamination Commitee of the Danish National Strategy for Danish Agricultural Research Council of Technical Sciences. Working Group on Forest and Landscape. Danish Standardization Association. Commit­ Committee on the national strategy for public tee on Environmental Analyses. agricultural research. Subgroup on forest and .Jensen. A. Member of the board of directors for landscape. Pajbjerg Foundation, Denmark. Rasmussen, S.K. Society of Danish Engineers, DCAR, Danish Center of Atmospheric Board of Chemistry Section. Research. Board of Danish Cereal Society. DaFoLa, Danish Center for Forest and Land ­ Rosendahl, L. Member of Committee of Censors scape Ecosystem Research. in biology at all Danish universities. Invited member of The Rockwool Foundation Schmidt, A.S. Working committee in Biotechno ­ Research Unit discussion group "how to value logy, Chemical Engineering Group, The En­ "nature" in environmental policy". gineering Society of Denmark (IDA). Chairman of the evaluation panel conseming 0stergdrd, H. Programme Committee for Bio­ aquatic and terrestrial ecotoxicology in The technology Programme "Risk of introducing Danish Environmental Research Programme. genetically modified plants into the environ ­ Chairman of the steering committee for the ment" (head of programme). international evaluation of the Danish univer ­ Ph.D. Committee for Education, The Royal sity educations in biology. Veterinary and Agricultural University, Member of The Danish Academy of Technical Copenhagen. Sciences. Coordination group for resistance and vi­ Jensen, KS. Member of The Danish Agricultural rulence in cereals and cereal pathogens in and Veterinary Research Council. Denmark. Member of the scientific board for the Danish Committee Member for denining professor ­ Pesticide Research Programme. ship in population genetics at The Royal Jorgensen. J H. Danish Gene Bank Committee. Veterinary and Agricultural University, Coordination group for resistance and vi­ Copenhagen. rulence in cereals and cereal pathogens in Denmark.

136 Ris0-R-972(EN) 14.2 International Jorgensen, J.H. Co-editor of 'Euphytica'. International coordinator for disease and pest Aarkrog, A. Reference group NKS-ECO Pro­ resistance genes in barley. gramme. Working group cereal, Nordic Gene Bank AMAP drafting group on Radioactivity (Chairman). Assessment. Programme Committee for Jorgensen, R.B. Member of Panel of Experts, International Conference on "Environmental International Biosafety Forum, Third World Radioactivity in the Arctic". Tromso, Norway, Network. June 1997. Kunzendorf, H. Editorial Board of 'Marine Member of the evaluation group on the Georesources and Geotechnology'. Norwegian Resarch Programme on Radioactive iMrsen, E. European Commission, Access to Fallout 1992-1995. Editorial Board of Journal of Large-Scale Facilities, Technical Audit of the Environmental Radioactivity. HCM-LSF contracts. Member of IAEA Task Group A for Studies of Miller, A. CEN TC 204 WG2 Sterilization by the Radiological Situation at the Atolls of ionizing radiation. Member. Mururoa and Fangataufa. ISO TC 198 Sterilization of Health Care pro ­ Member of IAEA expert group on Research on ducts. Member. Worldwide Marine Radioactivity. Int. Symposium on Nuclear and Related Member of IAEA drafting group on background Techniques in Agriculture, Industry, Health paper to the International Conference “One and the Environment, 1997. Member of Decade after Chernobyl ”. Scientific Advisory Committee. Member of IASAP Advisory group, IAEA 13th Radiochemical Conference, Czech Re­ President for Nordic Society for Radiation public 1997. Member of Int. Advisory Board. Protection. Editor-in-Chief Radiation Physics and Bjerre, A.B. IEA - Bioenergy agreement Task Chemistry. XLU. Bioconversion activity. Alternative 10th International Meeting on Radiation member of committee. Processing 1997. Member of programme Giese, H. Editorial board, Hereditas and Plant committee. Pathology on line. International Electrotechnical Commission OECD sommittee for Biotechnology. (IEC), Subcommittee 15B, Working Group 2 Gran, C. Board member of the Nordic branch of on Endurance Tests. Radiation. Corresponding the International Humic Substances Society. member. Jakobsen, I Management committee for COST Organisation Internationale de Metrologie Action 8.21: Arbuscular mycorrhizas in Legale (OIML). TC-15. Measuring measure­ sustainable soil-plant systems. ments for ionizing radiation. Member. Jensen, A. Member of an EEC expert panel for Nielsen, O.J. EUROTRAC International Execu ­ the collaboration on higher educations tive Committee. between European countries and USA. COST-611 Working Party 2 Steering Com­ Jensen, KS. Editorial Board of'Plant and Soil'. mittee, Atmospheric and Photochemical Sectionboard for Soils and Fertilizers, Scan ­ Processes. dinavian Association of Agricultural Scien ­ Steering Committee for NORSAC (Nordic tists. Symposia on Atmospheric Chemistry). National delegate in OECD co-operative Nielsen, S.P. International Committee on Radio­ research programme. Biological Resource nuclide Metrology. Management for Sustainable Agricultural Articles 35 and 36 of the Euratom Treaty Systems. (Environmental Monitoring). Jensen, J. International coordinator, barley chro ­ Group of experts on Monitoring of Radioac­ mosome 5; International Barley Nomenclature tive substances in the Baltic Sea (MORS) Committee. (Project Leader). Baltic Marine Environment Protection Com­ mission, Helsinki Commission (RELCOM)

Riso-R-972(EN) 137 Nielsen , 7'. The board of Nordic Society of Organic Pollutants. Roed, J . The group on in-situ measurements (ICRU). IAEA and EEC VAMP Project Urban Group. (Chairman). The Fuel Cycle Safety Group. (OECD). Wilbrandt, R. International. Conference on Time- resolved Vibrational Spectroscopy. Member of Organizing Committee. 0stergard, H. COST, chairman of Management Committee "Population studies of airborne pathogens on cereals as a means to improve strategies for disease control". Editorial board of Agronomic.

138 Ris0-R-972(EN) 15 Seminars and Courses Organized

The Second Danish ICPMS User Meeting, Riso National Laboratory, Denmark, 31 January 1996 (S. Stump). SUCON/INTAS meeting at Riso National Labora­ tory, 26-29 March, 1996 (A. Aarkrog) EKO-1 meeting at Rise National Laboratory, 19-20 March, 1996 (S.P. Nielsen). ATV Meeting on Soil and Groundwater Conta ­ mination, Organic Screening Analysis, Gentofte, Denmark, 18 April, 1996 (C. Gren). Training course on Validation and Process Con ­ trol for Electron Beam Sterilization, Riso National Laboratory, 10-14 June, 1996 (A. Milter). Mini-seminar (MJKA-project) at Rise National Laboratory, 16-20 October, 1996 (./ Roed). ATV Meeting on Soil and Groundwater Contani- nation, Research Strategies, Evaluation and Applications, Copenhagen, Denmark, 29 October, 1996 (C. (iron). MARINA-BALT, meeting at Rise National Laboratory, 29-30 October, 1996. (S.P. Nielsen). COST-Action 8.21 Workshop “Transport and Metabolism in Arbuscular Mycorrhizas ”, Roskilde, Denmark, 28-30 November 1996 (I. Jakobsen). EKO-2 meeting at Rise National Laboratory, 2-3 December, 1996 ( S.P. Nielsen and Bergan, T.) Documentation Requirements for Radiation Steri­ lization, Riso National Laboratory, 6 Decem­ ber 1996 (A. Miller). EKO-1 meeting at Jyllinge (Hotel Sofryd), Denmark, 18-19 December (S.P. Nielsen and Palsson, S.E.). Ph D. course in Environmental Photochemistry, Riso National Laboratory, January-December 1996 ( T Nielsen) Ph D. course in Fate of Organic Contaminants in the Terrestrial Environment, Riso National Labora­ tory, January-December 1996 (T. Nielsen)

Riso-R-972(EN) 139 16 Scientific Results and Finances

16.1 Scientific Results

Results Results Results Results Results Results Results 1990 1991 1992 1993 1994 1995 1996 Basic research and development (man-month) 779 734 636 545 590 484 Research programmes (man-month) 74 273 549 599 659 655 848 Commercial contracts (man-month) 19 89 95 140 132 119 68 Technical support for research (man-month) 28 28 190 62 53 69 Management (man-month) 96 96 152 169 189 59 Total 93 1265 1502 1717 1567 1606 1528 Dissemination of results Papers in international journals and books 70 71 82 76 111 102 1 1 1 Papers in Danish journals and books 12 12 8 4 9 5 9 Riso reports, R , M and I 12 14 13 11 11 16 13 Danish books and reports 9 8 13 6 8 10 International books and reports 9 1 5 2 3 6 Other publications 11 4 10 26 21 10 Papers in conference proceedings 16 56 29 39 70 37 42 Other international conference contributions 52 46 72 65 70 108 93 Other Danish conference contributions 12 26 37 42 64 38 32 Patent proposals 1 1 1 Apprentices and trainees 6 12 15 7 16 14 Teaching courses (hours) 8 5 4 46 18 40 Lectures (hours) 17 27 11 61 80 External examiner (hours) 10 15 17 79 191 Conferences (arranged) 4 5 6 13 13 Networking and collaboration Ph D students (number) 20 22 20 25 24 23 Ph D. degrees (number) 3 6 3 5 4 5 5 Riso leave of absence (man-month) 21 15 26 11 8 47 46 Visiting scientists at Riso (man-month) 24 57 58 69 54 63 37 Scientific papers reviewed (number) 53 79 99 100 157 52 192 Committees for Ph.D.thesis. promotion of scientists, senior scientists, professors (number) 12 11 14 13 23 7 20 Committee memberships (number) 61 60 62 73 46 68 77 Contracts with companies 14 6 16 Collaboration with companies (man-month) 116 40 81 Contracts with research institutes (number) 51 60 62 Collaboration with research institutes (man-month) 595 652 571 Collaboration with public authorities (man-month) 7 7 Masters students (man-month) 8 18 15 76 96 52 Post docs (man-month) 1 5 22 1 12 150 Visiting scientists at Riso (numbers) 51 42 43 40 Post docs (numbers) 5 5 1 1 17 Masters students (number) 19 23 22 21 Doctors of science 1

140 Ris0-R-972(EN) 16.2 Finances

The activities of the Department are supported by a combination of basic government funding, Project funds from national and international research programmes and fully commercial industrial contracts. The numbers given in the tables are in units of 1000 Danish Kroner (DKK). One US$ is equivalent to 6.3 DKK.

1990 1991 1992 1993 1994 1995 1996

Income

Basic funding* 26,306 25,065 23,272 22,140 21,174 22,474 20,493

Pogrammes and contracts 14,132 15,312 18,554 17,921 24,307 28,737 32,000

Euuinment grants 0 0 1.039 1.372 983 4.179 2.315

Total 40,438 40,377 42,865 41,433 46,464 55,390 54,838

Expenditure

Salaries 31,861 31,083 31,848 31,233 32,616 35,168 38,151

Operating expenses 8,577 9,294 9,978 8,828 12,865 16,043 14,372

Euuinment 0 0 1.039 1.372 983 4.179 2.315

Total 40,438 40,377 42,865 41,433 46,464 55,390 54,838

* Basic funding is total funding minus overhead to Riso.

Additional funding has been obtained for Ph.D. grants, Post. doc. fellowships, apprenticies and trainees.

Approximately 80 per cent of the external funding comes from reseach councils, private foundations, national and international research programmes, and approximately 20 per cent comes from commercial contracts.

Ris0-R-972(EN) 141 17 Personnel

The Department includes approximately 145 scientific and technical staff members. The research activities are organized into 5 research programmes and supported by 3 special facility units.

Dr Ame Jensen is Head of Department

Research Programmes Atmospheric Chemistry and Air Pollution. Head: Dr Ole John Nielsen. Gene Technology and Population Biology. Head: Dr. Hanne Ostergard. Plant Nutrition and Nutrient Cycling Head: Dr. Gunnar Gissel Nielsen. Trace Analysis and Reduction of Pollution in the (ieosphere. Head: Dr. Vagn Gundersen. Ecology\ Head: Dr. Asker Aarkrog.

Special Facility Units Growth chambers. Riso Environmental Risk Assessment Facility (RERAP). greenhouses and the experimental farm, Dyskcergaard. Head: Dr. Vagner Haahr. Riso High Dose Reference Laboratory. Head: Dr Arne Miller Riso Integrated Environmental Facility (R1MI). Head: Dr. Kim Pilegaard

The list also includes short term employees

17.1 Scientific staff Engvild, Kjeld C Fenger, Jorgen Aarkrog, Asker Gavito, Mayra Ambus, Per Giese, Henriette Andersson, Kasper Gissel Nielsen, Gunnar Asmar, Mohammad Farouq (until 30.09.96) Gran, Christian Beier, Claus Gundersen, Vagn Bibak, Allan Hald, Pia (until 3 1.07.96) Bjergbakke, Erling Hansen, Heinz H Bjerre, Anne Belinda Hauser, Thure Bogo, Annette Heiberg, Ann-Charlotte Bork, Ulla (until 31.01.96) Holcman, Jerzy Chen, Qiangjiang Haahr, Vagner Christiansen, Solveig Krogh Jakobsen, Iver Dahlgaard, Henning Jensen, Erik Steen Damgaard, Christian (until 31.05.96) Jensen, Jens Egsgaard, Helge Johansen, Anders (until 29.02.96)

142 Ris0-R-972(EN) Justesen, Annemarie (until 31.03.96) Brandstrup, Oda Jorgensen, Bjame Brandt, Lis Jurgensen, Jurgen Helms Brink-Jensen, Merete Jurgensen, Rikke Bagger Baade-Pedersen, Pearl Keszthelyi, Tam as Christensen, Tove Kilen, Hans Henrik Clausen, Jytte Kunzendorf, Helmar Corfitzen, Hanne Lange, Christian Djurdjevic, Stanko Larsen, Elfinn Femqvist, Tomas Larsen, John Fosskov Jensen, Jette Larsen, Lotte Kjasr (until 30.04.96) Frederiksen, Trine (until 30.04.96) Lynggard, Bent Gade, Poul Lyngkjaer, Michael Gudiksen, Peter Mikkelsen, Teis Norgaard Hansen, Carl (until 11.06.96) Mikkelsen, Thomas (until 30.04.96) Hansen, Dorte Miller, Arne Hansen, Elly Mouritzen, Peter Hansen, Ina Nielsen, Ole John Hansen, Ingunn Innerdahl (until 15.08.96) Nielsen, Peter S. (until 31.03.96) Hasselbalch, Finn Nielsen, Sven P. Helbask, Kirsten (until 30.06.96) Nielsen, Torben Henriksen, Ebbe Nilsson, Karen Hougaard Pedersen, Henrik Offersgaard, Jesper Ibsen, Elly O’Hara, Robert Jacobsen, Michael (until 30.06.96) Pagsberg, Palle Jensen, Birgit Pedersen, Carsten Jensen, Ellen Muller Pilegaard, Kim Johansen, Torben Ploger, Annette Jorgensen, Vibeke Rasmussen, Lennart Karl sen, A age Rasmussen, Suren Kjaersgard Kjulhede, Alice Roed, Jum Koutras, Charlotte Rosendahl, Lis Kristiansen, Bo Schmidt, Anette Skammelsen Larsen, Erik Engholm Schweiger, Peter Larsen, Fritz Sehested, Jens Larsen, Ingelis Sehested, Knud Larsen, Inge Merete Sillesen, Alfred Heegaard Lilholt, Ulla Simonsen, Tina (until 30.04.96) Madsen, Michael Birch Stirrup, Stefan Mandrup Jensen, Karen Surensen, Peter (until 31.03.96) Meltofte, Liselotte Thingstrup, Ida Muller, Anette Wilbrandt, Robert Nielsen, Carina (until 07.11.96) Woidemann, Anders Nielsen, Jette Bruun Ostergard, Hanne Nielsen, Lars Muller Nielsen, Vagn A age Olsen, Anette 17.2 Technical Staff Olsen, Anne Olsen, Inge Andersen, Bente Olsen, Svend K. Andersen, Margit Elm Prip, Henrik Bitsch, Gunnar Sillesen, Anerikke

Risu-R-972(EN) 143 Sorensen, Poul 17.5 M.Sc. Students Tung, Tran Due Tuan Thygesen, Maria (until 15.12.96) Andersen, Thomas Sejner Vestesen, Hans Andersen, Trine Solberg Vinther Kristensen, Lis Bach, Louise Wojtaszewski, Hanne Buciek, Troels Christiansen, Lars Eriksen, Rasmus 17.3 Office Staff Forsting, Peter Ghasemi, Freshte Andersen, Annie Senten (until 31.08.96) Hansen, Henriette Seiler Bay, Kirsten Hansen, Rikke Stubbe Borring Sorensen, Marit Hansen, Steen Roed Brekmark, Anni Hansen, Tine Frandsen, Anette Jensen, Mads Toustrup Jakobsen, Inger Klausen, Janne Jensen, Hanne Kloch,Jorgen Komerup, Berit Krarup, Helene Kristensen, Ingrid Kristensen, Peter Kyvsgaard Krogh, Helle Larsen, Karina Block Petersen, Lis Monrad, Annette Scharff, Anne Marie Schneider, Kenn 17.4 Ph.D. Students Wetche, Torsten Ployart Wittenhorst, Vincent Behrens, Annette Bilde, Merete Brinch-Pedersen, Henrik 17.6 B.Sc. Students Butterworth, Lisa A Christensen, Lene Krogh Hamid, Shahida Dahl, Soren Weis Kipler, Yogini, Fauser, Patrik Larsen, Rene Vogt Feilberg Jorgensen, Anders Moradi, Manaz Grage, Mette M -L Nguyen, Quynh Guldmann, Lise-Lotte Helweg, Christian Hautorp-Grosell, Martin 17.7 Apprentices Holst, Pia Bachmann Jacobsen, Frank Christiansen, Charlotte Gronlund Johansen, Katja Salomon Demir, Gulhan Jorgensen, Finn Easton, Tine B. Kristensen, Brian K. Goldberg, Kristina Maria Landbo, Lars Hansen, Soren Loje Mogelberg, Trine Jensen, Linette Munksgaard Platz, Jesper Jensen, Mads Sabroe Ravnskov, Sabine Kendix, Elsebeth Simonsen, Anna Carina Wiborg Nielsen, Carina Thomsen, Heidi Thygesen, Maria

144 Riso-R-972(EN) 18 Acronyms

AAR AMAP Assessment Report ACDEP Atmospheric Chemistry and Deposition ACDEP Atmospheric Chemistry and Deposition model AM Arbuscular Mycorrhiza AMAP Arctic Monitoring and Assessment Programme ASTM American Society for Testing and Materials BW Brackish Water CAT Center for Advanced Technology CEC Commission of European Communities CEN European Standard Organization CF Concentration factor (e.g. Bq kg "1 fish : Bq l’1 water) CFC Compound: Chlorine, Fluorine, Carbon CLIMEX Climate Change Experiment COST European Cooperation in the Field of Scicntic and Technical Research CYTROXA Cy cling of OH and H02 by NOx in the Tropospheric Oxidation of Aromatics DANAK Danish Accreditation Scheme DEPH di-ethy 1-hexyl-phthalate DH Chromosome-doubled Haploid D1PSS The Danish Institute of Plant and Soil Science DMDA Danish Medical Device Association DMU National Environmental Research Institute DNA Dioxyribonuclcicacid DOAS differential optical absorption spectroscopy DS Danish Standards Association DTU Danish Technical University EPR Electron Paramagnetic Resonance EUROTRAC European Experiment on Transport and Transformation of Environmental Relevant Trace Conslitutents of Anthropogenic and Natural Origin EUROVOC Control Strategies for European Air Quality Based on the Tropospheric Oxidation Characteristics of Volatile Organic Compounds FABMS Fast Atom Bombardment Mass Spectrometry FAO United Nations Food and Agriculture Organization FORMONA Formation and Occurrence of Nitrous Acid in the Atmosphere FTIR Fourier Transform Infrared FW Fresh Water FWT Far West Technology F0TEK Fodcx areteknologiskc forsknings- og udviklingsprogram GC/MS Gas Chromatography/Mass Spectrometry GDC Gamma Dating Center GEUS Danmarks og Gronlands Geologiske Undersogelser GOBEX Gotland Basin Experiment GPS Global Positioning System GUS fl-glucuronidase GWP Global Warming Potentials HCFC Compound: Hydrogen, chlorine, fluorine, carbon HCM Human Capital and Mobility HDRL Rise High Dose Reference Laboratory FTF'C Hydrogen, fluorine, carbon HOMSEA Hy droxomcthansulfinic acid HPLC High Pressure Liquid Chromatography HR-ICPMS High Resolution Inductively Coupled Plasma Mass Spectrometry IAEA International Atomic Energy Agency IASAP International Arctic Seas Assessment Project IBSS Institute of Biology of the Southern Seas (Sevastopol, Ukraine) ICPMS Inductively Coupled Plasma Mass Spectrometry INRA Institute National dc la Recherche Agronomique

Ris0-R-972(EN) 145 INTAS International Association for the promotion of cooperation with scientists from the independent states of the former Soviet Union IPAE Institute of Plant and Animal Ecology (Ekaterinburg, Russia) ISO International Standards Organization LAS linear alkyl benzene sulfonates LIMS Laboratory Information Managing System LINE Long Interspersed Elements LSF Large-Scale Facilities MAST Marine Science and Technology Programme (under CEC) MIKA Miljokatastrofefonden - urban contamination MMC Mytomycine-C MORS Group of Experts Monitoring of Radioactive Substances in the Baltic Sea. MSE Mean Square Error N-PAC Nitrogen containing Polycyclic Aromatic Compounds NKS Nordic Nuclear Safety Project (RAD) NMR Nordisk Ministerrad NOM Natural Occurring Organic Matter NPL National Physical Laboratory NRPB National Radiological Protection Board, United Kingdom OC Organic Carbon ODS Octadccylsilylsilica OECD Organisation for Economic Cooperation and Development PAH Polycyclic Aromatic Hydrocarbons PCR Polymerase Chain Reaction PNA Poly Nuclear Aromatics POM Polycyclic Organic Matter PON Particulate Organic Nitrates QTL Quantitative Trait Loci RAPD Random Amplified Polymorphic DNA Technique RERAF Rise Ecological Risk Assessment Facility RFLP Restriction Fragment Length Polymorphism RIMI Rise Intergratcd Environmental Project RP-HPLC Reverse phase high performance liquid chromatography RVAU The Royal Veterinary and Agricultural University SBR Styrene butadiene rubber SCAR Sequence characterized amplified reaction SEP Standard Error of Prediction SINE Short Interspersed Elements SLU Swedish Agricultural University SMP Strategisk Mi 1 joforskningsprogram SMT Standards Measurement and Testing SNCR Selective Noil-Catalytic Reduction SOM Soil Organic Matter SUCON South Ural Contamination Studies svv Sea Water TF Transfer Factor (e.g. Bq m '/PBq yr"1) TGA Thermal Gravimetric Analysis TIMS Thermal Ionization Mass Spectrometry TMI Transition Metal Ions TNO Ncdcrlands Organization for Applied Scientific Research TSP Total Suspended Particulate Matter UMIST Univ ersity of Manchester, Institute of Science and Technology, UK VAM Vesicular-Arbuscular Mycorrhiza WO Wet Oxidation VOC Volatile Organic Compounds VTT Valtion Teknillinen Tutkimuskcskus, Finland YAC Ycast Artificial Chromosome

146 Ris0-R-972(EN) Bibliographic Data Sheet Riso-R-972(EN)

Title and authors(s) Environmental Science and Technology Department Annual Report 1996 Edited by A. Jensen, G. Gissel Nielsen, V. Gundersen, O.J. Nielsen, H. 0sterg&rd and A. Aarkrog

ISBN ISSN 87-550-2284-7 0106-2840 0906-8090

Dept, or group Date Environmental Science and Technology Department February 1997

Groups own reg. number(s) Projeel/contract no. (s)

Pages Tables Illustrations References 146 15 63 207

Abstract (Max. 2000 characters) The Environmental Science and Technology Department aspires to develop new ideas and methods for industrial and agricultural production through basic and applied research thus exerting less stress and strain on the environment. The Department endeavours to develop a competent scientific basis for future production technology and management methods in industrial and agricultural production. The research approach in the Department is mainly experimental. Selected departmental research activities during 1996 are introduced and reviewed in seven chapters: 1. Introduction, 2. Atmospheric Chemistry and Air Pollution, 3. Gene Technology and Population Biology, 4. Plant Nutrition and Nutrient Cycling, 5. Trace Analysis and Reduction of Pollution in the Geosphere, 6. Ecology, and 7. Other Acti­ vities. The Department ’s contribution to national and international collaborative research programmes are presented together with information about the use of its large experimental facilities. Information about the Department ’s contribution to education and training are included in the report along with lists of publications, publications in press, lectures and poster presentations at international meetings. The names of the scientific and technical staff members, visiting scientists, Postdoctoral fellows, Ph.D students and M.Sc. students are also listed.

Descriptors INIS/EDB AIR POLLUTION; ATMOSPHERIC CHEMISTRY; CHEMICAL ANALYSIS; CONTAMINATION; DISEASE RESISTANCE; ECOLOGY; ECOSYSTEMS; GENETICS; GEOCHEMISTRY; NUTRIENTS; NUTRITION; PLANTS; PROGRESS REPORT; RADIOECOLOGY; RADIONUCLIDE MIGRATION, RESEARCH PROGRAMS; RISOE NATIONAL LABORATORY

Available on request from Information Service Department, Riso National Laboratory (Afdelingenfor Informationsservice.ForskningscenterRisa), P.O. Box 49, DK-4000Roskilde, Denmark Telephone (+45) 46 77 46 77, ext. 4004/4005 Telex 43 116 • Telefax ( + 45) 46 75 56 27