ESNA

EUROPEAN SOCIETY FOR NEW METHODS IN AGRICULTURAL RESEARCH

XXVIIIth ANNUAL MEETING

and o International Union of Radioecology (IUR) i Working Group Soil-to-Plant Transfer JS Annual Meeting IN?

th th 26 August - 29 August 1998 o i NI CD ; Brno, Czech Republic fg \ oo! O! O i EUROPEAN SOCIETY FOR NEW METHODS IN CZ9928607 AGRICULTURAL RESEARCH ESNA Committee XXVIIIth ANNUAL MEETING 26th- 29th August 1998, Brno, Czech Republic President Prof. Dennis A. Baker - United Kingdom ORGANIZED BY

Mendel University of Agriculture and Forestry, Brno Vice-president Prof. Stanislav Prochazka - Czech Republic Society for Experimental Biology, Czech Academy of Sciences, Prague

Czech Academy of Agricultural Sciences, Prague Members Prof. M. Cristaldi - Italy Prof. W. Fiszer - Poland ORGANIZING COMMITTEE Dr. M. Gerzabek - Austria Prof. S. Ignatowicz - Poland Mendel University of Agriculture and Forestry Dr. N. G. Mitchell - UK Zemedelska 1, 61300 Brno, Czech Republic Dr. J. Schmidt - Austria Prof. E. Smedile - Italy Meeting Chairman Prof. A. S. Szabo - Hungary Prof. Stanislav Prochazka Prof. T. H. Thomas - UK Department of Botany and Plant Physiology Phone: +420/5 4513 6070 Fax: +420/5 4521 1128 E-mail: [email protected] ESNA Secretary Dr. Michael Poschl - Czech Republic Local Secretary Dr. Michael Poschl Department of Nuclear Methods Phone: +420/5 4513 3298 Fax: +420/5 4521 1128 E-mail: [email protected] GENERAL PROGRAMME 25th August 14:00-20:00 Registration

th 26th August 9:00-11:00 Registration 28 August 9:00-10:30 WG Nr. 1,3,4,6 sessions 11:00 Opening Ceremony 10:30-11.00 Coffee break 11:30-13:00 Plenary session - Invited papers 11:00-12:00 WG Nr. 1,3,4,6 sessions M. Marek: Global changes and natural ecosystems 13:00-14:00 Lunch M. Strnad: Cytokinins: Cell cycle regulators and 14:00-15:30 WGNr. 1,3,4 sessions potential herbicides 15:30-16:00 Coffee break P. Dvorak, A. Hampl: Recent advances in embryonic 16:00-17:30 WG Nr. 1,3,4 sessions induction - A potential role of growth factors 19:00 Farewell dinner 13:00-14:00 Lunch 14:00-15:30 WG Nr. 2,3,4,5 sessions 15:30-16:00 Coffee break 16:00-17:30 WG Nr. 2,3,4,5 sessions 18:00 ESNA Comittee meeting 19:00 Get-together party

27th August 9:00-10:30 WG Nr. 2,3,4,5 sessions 10:30-11:00 Coffee break 11:00-12:00 WG Nr. 2,3,4,5 sessions 13:00-14:00 Lunch 14:00- Excursion (Southern Moravia)

28th August 9:00-10:30 WG Nr. 1,3,4,5 sessions 10:30-11:00 Coffee break 11:00-12:00 WGNr. 1,3,4,5 sessions 13:00-14:00 Lunch 14:00-15:30 WGNr. 1,3,4,6 sessions 15:30-16:00 Coffee break 16:00-17:30 WG Nr. 1,3,4,6 sessions 19:00 Organ concert WORKING GROUPS & CHAIRMEN /SCIENTIFIC TOPICS

WG 1 RADIATION TECHNOLOGY WG 5 QUALITY OF AGRO-ECOSYSTEM Prof. A. S. Szabo /Prof. W. Fiszer Prof. S. Grego /Prof. F. Megusar /Prof. E. Smedile Food and feed irradiation, identification of irradiated food, pest control in Disturbances of nutrient cycling, technological aspects of agroecosystem agricultural crops, legal aspects, waste irradiation. sustainability, technological and ecological aspects of waste recycling, Prof. A. S. Szabo technical aspects related to spatial variability, bioremediation. University of Horticulture and Food Industry, Department of Food Prof. S. Grego Chemistry and Nutrition, Somloi ut.18, H-1118 Budapest, Hungary D.ABAC-Universita della Tuscia.Via S.Camillo de Lellis, 01100 Viterbo, Fax: +36 1 1666220 E-mail: [email protected] Italy Fax: +39 761 357242 E-mail: [email protected] WG 2 ADVANCED METHODS IN ANIMAL SCIENCES Prof. M. Cristaldi /Prof. B. Todorov WG 6 PEST MANAGEMENT Animal physiology, endocrinology, reproduction, metabolism, pathology, Prof. S. Ignatowicz /Dr. Marec genetics, radiobiology, technology. Physical, biological and genetical methods of control and related Prof. M. Cristaldi aspects. Dip. di Biologia Animale e dell'Uomo.Via A.Borelli, 50, 1-00161 Roma, Prof. S. Ignatowicz Italy Agricultural University of Warsaw, ul.Nowoursynowska 166, 02-766 Fax: +39 6 4457516 E-mail: [email protected] Warsaw, Poland Fax: +48/22 434 942 E-mail: [email protected] WG 3 SOIL-PLANT RELATIONSHIPS Dr. M. Gerzabek /Dr. N. G. Mitchell Soil science, plant nutrition (incl. microbial aspects), application of stable and radioactive isotope techniques, plant physiology, behaviour of pollutants in soil-plant system. Dr. M. Gerzabek Austrian Research Centre Seibersdorf, A-2444 Seibersdorf Fax: +43 2254 7803653 E-mail: [email protected]

WG 4 PLANT GENETICS, BREEDING AND PHYSIOLOGY Prof. T. H. Thomas /Dr. J. Schmidt Plant biotechnology, incl. advanced breeding, genotyping and propagation methods, plant biochemistry, physiology and molecular biology. Prof. T. H. Thomas lACR-Long Ashton Research Station, University of Bristol, Long Ashton, Bristol BS 18 9AF, United Kingdom Fax: +44 1275 394007 E-mail: [email protected] CONTENT

ABSTRACTS PRESENTED IN WORKING GROUP NO. 1 7 CHAIRMEN: PROF. A. S. SZABO AND PROF. W. FISZER

ABSTRACTS PRESENTED IN WORKING GROUP NO. 2 23 CHAIRMEN: PROF. M. CRISTALDI AND PROF. B. TODOROV

ABSTRACTS PRESENTED IN WORKING GROUP NO. 3 41 CHAIRMEN: DR. M. GERZABEK AND DR. N. G. MITCHELL

ABSTRACTS PRESENTED IN WORKING GROUP NO. 4 97 CHAIRMEN: PROF. T. H. THOMAS AND DR. J. SCHMIDT

ABSTRACTS PRESENTED IN WORKING GROUP NO. 5 167 CHAIRMEN: PROF. S. GREGO AND PROF. F. MEGUSAR AND PROF. E. SMEDILE

ABSTRACTS PRESENTED IN WORKING GROUP NO. 6 191 CHAIRMEN: PROF. S. IGNATOWICZ AND DR. MAREC Abstracts presented in working group No. 1

Chairmen: Prof. A. S. Szabo and Prof. W. Fiszer SUPPLEMENTATION OF FOOD FOR MILKING COWS WITH ORGANIC AND INORGANIC FORM OF IODINE 9 BOBEK, S., SECHMAN, A., BRZ6SKA, F.

COMPARATIVE STUDY ON THE THERMAL AND IRRADIATION STERILIZATION METHODS OF THE COLOURING ADDITIVE FROM MONASCUS PURPUREUS 10 FERDES, M., FERDES, O. S., MINEA, R.

THE COMBINED EFFECTS OF E-BEAM IRRADIATION AND MICROWAVES ON STARCH, FLOUR AND INGREDIENTS 11 FERDES, O. S., MARTIN, D., MINEA, R., TIRLEA, A., BADEA, M.

RHEOVISCOMETRIC MEASUREMENTS ON STARCH, FLOUR AND BLACK PEPPER TREATED BY ELECTRON-BEAM AND MICROWAVES 12 FERDES, O. S., CASANDROIU, T., OPRITA, N., TIRLEA, A., BADEA, M.

OCCURRENCE AND REMOVAL OF SOME GENOTOXIC COMPOUNDS IN GROUNDWATER 13 GEHRINGER, P.

FEED IRRADIATION AND DIET COMPOSITION IN DAIRY COWS 14 GRUBIC, G., VITOROVIC, G., VITOROVIC, D., DONIC, S.

ELECTRON-BEAM AND MICROWAVE TREATMENT OF SOME MICROBIAL STRAINS 15 MARTIN, D., FERDES, O. S., PLAMADEALA, S., FERDES, M., MINEA, R., TiRLEA, A., BADEA, M.

DISINFECTION OF RED DRIED HUNGARIAN SPICE PAPRIKA BY SYNERGOLUX TREATMENT REGARDING ALSO QUALITY ASPECTS16 SIMON, J., TOLNAY, P., SZAB6, A. S., JUHASZ-ROMAN, M.

EVALUATION OF DATA OF QUESTIONNAIRE NUTRITION AND HEALTH (1997-1998) CONCERNING THE TOPIC WHAT CONSUMERS THINK ABOUT FOOD IRRADIATION? 17 SZABO.A. S., TOLNAY, P.

RHEOLOGICAL PROPERTIES OF RIPENED, HARD CHEESES SUBJECTED TO HIGH HYDROSTATIC PRESSURE 18 SZCZAWINSKA, M. E., LENART, A., SZCZAWINSKI, J. COMPARISON OF THE EFFECT OF X-RAY AND ELECTRON BEAM IRRADIATION ON THE THIAMINE CONTENT IN CHICKEN DRUMSTICKS 19 VAN CALENBERG, S., VAN CLEEMPUT, O., MONDELAERS, W., HUYGHEBAERT, A.

137 IN VITRO AND IN VIVO EFFICIENCY OF CLINOPTILOLITE IN CS BINDING 20 VITOROVIC, D., VITOROVIC, G., MLADENOVIC, V., VUKICEVIC, O.

RADIATION INDUCED MAILLARD REACTIONS (THE KINETIC OF COLOUR FORMATION DURING HEATING) 21 TEGOTA, A., BACHMAN, S. Working group No.1 NOTES SUPPLEMENTATION OF FOOD FOR MILKING COWS WITH ORGANIC AND INORGANIC FORM OF IODINE BOBEK, S.\ SECHMAN, A.1, BRZOSKA, F.2 department of Animal Physiology, University of Agriculture, 30-059 Krakow, Poland 2National Research Institute of Animal Production, 32-083 Balice, Poland

In previous presentation we described the effect of iodination of salt-licks with potassium iodide (Kl) - an inorganic form of iodine, or ethylenediamine dihydroiodide (EDDI) - an organic form of iodine. The results have shown a low transfer of iodine from the fodder to milk which was 1.2 and 2.5 per cent for Kl and EDDI, respectively. Low transfer of iodine into milk leads to the necessity of increment the daily dose of iodine to 58 and 128 mg I/day in the form of EDDI and 121 and 300 mg I/day in the form of Kl to achieve the planned level from 30 to 150 or 300 u,g I/I of milk. Therefore the other experiment was performed, in which both iodine compounds were inserted in wafer capsules and given together with concentrated food. The following levels of iodine were given: 30, 60 and 100 mg I/day/cow, each for 6 days. The iodine compounds were prepared one week before onset of the experiment. The control iodine concentrations in milk were taken at first day of the experiment and at seventh day after offset of the iodine supplementation.

Results: 1. the iodine concentration in milk increased proportionally to its consumption by the cow; 2. the quantities of iodine transferred from the digestive tract to milk fluctuated between 26.1 to 29.3% for EDDI and 20.6 to 29.5% for Kl, respectively (correlation coefficients: r=0.995 to 0.997); 3. applying regression analysis the levels of 150 or 300 p.g I/I of milk were obtained when cows consumed 7 or 22 mg I/day/cow in the form of EDDI and 10 or 25 mg I/day/cow in the form of Kl, respectively; 4. at 18th day of the experiment passage of iodine originated from Kl into milk was by 17% lower (P.<0.01) in comparison to iodine from EDDI. This may indicate that already 4 weeks after preparation of Kl, it undergoes degradation (via sublimation). Since EDDI transiently suppresses milk yield and is less readily ingested by the cows in comparison with Kl, less stable Kl should be used. However, to avoid sublimation, ,,a stable form,, of Kl should be elaborated, otherwise food enriched with Kl should be consumed up to one month after supplementation. Working group No.1 NOTES COMPARATIVE STUDY ON THE THERMAL AND IRRADIATION STERILIZATION METHODS OF THE COLOURING ADDITIVE FROM MONASCUS PURPUREUS FERDES, M.2, FERDES, O. S.\ MINEA, R.1 National Institute of Laser, Plasma and Radiation Physics; 1 Atomistilor St.; P.O.Box MG-36; Ro-76900 Bucharest; Romania; E-mail: [email protected] institute of Food Chemistry, 1 Girlei St., Ro-71576, Bucharest, Romania

The paper presents and compares the experimental results obtained on the sterilisation of the food-grade red pigment from Monascus purpureus fungus, by two methods: a classical one, consisting of thermal sterilisation and a non-thermal sterilisation by irradiation. The Monascus purpureus red dye was obtained by biosynthesis of this fungus on natural culture media consisting of ground rice, for 30 days to 30 °C. It is assumed that the sterilisation of pigment powder obtained from Monascus purpureus biosynthesis could be performed either by thermal treatment or by gamma-ray or electron-beam irradiation, without pigment distruction. The thermal treatment has carried out to different temperatures ranging between 80 °C and 180 °C and for different duration. There have analysed the TGC/g (or CFU/g) for the product and the extinction of the colouring components in alcoholic extract, by spectrophotometry. It should point out that for applying a temperature between 100 -110 °C for 60 - 90 minutes the fungal spores are destroyed and the pigment preserved its properties, in the same time. Irradiation treatment, either by gamma-rays or by electron-beam has carried out in the dose range of 1 - 30 kGy. The irradiation assures a ,,complete" sterilisation for irradiation dose higher than 7.5, for the microbiological load presenting an initial value of 106 spores/g for the control sample. For irradiation treatment there have performed the same microbiological and spectrophotometrical analysis. Using the thermal treatment the colour loses could reach 20 % depending of the treatment duration, for a temperature between 100 -110 °C. Comparatively, the irradiation does not significantly affect the product colour. Concluding, both from hygienic (microbiological) and technological (colouring power) point of view the irradiation treatment could be considered more effective than the classical, thermal one and it could be apply for sterilising the red dye product obtained by Monascus purpureus biosynthesis. Working group No. 1 CZ9928608 THE COMBINED EFFECTS OF E-BEAM IRRADIATION AND MICROWAVES ON STARCH, FLOUR AND INGREDIENTS FERDES, O. S., MARTIN, D., MINEA, R., TIRLEA, A., BADEA, M. National Institute of Laser, Plasma and Radiation Physics; 1 Atomistilor St.; P.O.Box MG-36; Ro-76900 Bucharest; Romania; E-mail: [email protected]

The influences of both microwave field and electron-beam irradiation, separately and combined, mainly on physical parameters of corn starch, wheat flour and black pepper were studied. These treatments have been used to achieve the hygienic and microbiological quality requirements of these materials and for their dehydration. The electron-beam irradiation has been carried out by using a linear accelerator, ALIN-7 designed and produced in NILPRP Bucharest, Electron Accelerator Laboratory that presents the following parameters: electron mean energy ~ 6 MeV; mean bean current of 10 JJA; pulse period of 3.5 \is and a repetition frequency of 100 Hz. The main experimental parameters were: irradiation dose; dose rate and dose uniformity. For microwave experiments it was used a special designed microwave applicator consisting of a special cavity, a power controlled generator with a 2.45 GHz standard frequency CW magnetron of 850 W maximum output power, and the appropriate electronics which allow the control of the microwave power; the current intensity, and the exposure time. The experiments have been carried out in 5 variants: only microwave treatment; only electron- beam irradiation; combined microwave then electron beam treatment; combined electron-beam irradiation then microwave treatment and, simultaneously microwave and electron-beam. The samples have been treated by microwaves to 4 different power values, among 250 W to 550 W and for 5 different exposure time. The electron-beam irradiation took place into the dose range of 1 - 10 kGy, at the same dose rate of approximately 2 kGy/min. There were analysed the influence of these two physical fields on some common properties (r.h., pH), spectrophotometric (UV-VIS spectra), viscometric (rheograms) and microbiological (CFU/g) properties of the food materials. The physical investigations have been performed using common standardized methods. From microbiological point of view there were investigated the usual microbial parameters. It was pointed out a direct relationship between microwave power and exposure time with the physical effects, as well as a same relationship between irradiation dose and these effects. The microwave effects are mainly thermal effects, but it has observed a non-thermal effect, too. The main microbicidal action is based on the electron-beam effect, but the microwave treatment presents sometimes a significant influence both on the microbial population and on its sensitivity to irradiation. To the combined treatment seem to be present a synergistic effect of microwaves and electron-beams, this effect arising on non-thermal basis. It should be considered that combined treatments lead to reducing irradiation dose without losing the microbicidal effects, so it is possible to use this treatment in some cases of starch-based food processing.

li Working group No. 1 RHEOVISCOMETRIC MEASUREMENTS ON STARCH, FLOUR AND BLACK PEPPER TREATED BY CZ9928609 ELECTRON-BEAM AND MICROWAVES FERDES, O. S.\ CASANDROIU, T.2, OPRITA, N2., TIRLEA, A.1, BADEA, M.1 1National Institute of Laser, Plasma and Radiation Physics; 1 Atomistilor St.; P.O.Box MG-36; Ro-76900 Bucharest; Romania; E-mail: [email protected] 2Politehnica" University of Bucharest, Faculty of Biotechnical Engineering Systems; Laboratory of Agro-Food Physics; 313 Splaiul Independentei; Bucharest; Romania; E-mail: [email protected]

The paper presents the results on the investigation of the rheoviscometric behaviour of geiiffied suspensions of corn starch, wheat flour and black pepper, which were treated by microwaves and electron-beam irradiation. Irradiations have performed to ALIN-7 linac, in the dose range of 0 to 16 kGy with a dose rate of approximately 1.5 kGy/min. The samples were disposed in a cavity that is used for microwave treatment, too. The microwave irradiation has carried out inside a special designed system using a 2.45 GHz CW magnetron of 850 W maximum output power, and the samples were treated to different power values and for irradiation period varying among 10 to 120 s. There have carried out combined treatments, that means by microwave and electron-beam irradiations, too. The rheoviscometric measurements have carried out on geiiffied suspensions of starch, flour and black pepper, into equivalent starch concentration conditions. For the black pepper suspensions the pH value has adjusted to more than 12 units by adding 33 % NaOH solution. A rotationally viscometer Rheotest RV-2-type with different cylinder coupling and to 12 different share rate values, it has used in measurements. The rheological parameters that were considered to describe the electromagnetic field and irradiation effects on starch, flour and pepper are: the apparent viscosity, r\a (mPa-s) and the share stress, x (mPa). The experimental results were analysed in terms of rheograms, as share stress (T.) or apparent viscosity (r\a) dependency by irradiation conditions. In all the cases, the experimental results lead to the remark that the apparent viscosity and share stress dependency by the nature and intensity of the treatment could be considered as a result of physical fields effects on starch, because the flour and the pepper have a significant starch content (~ 80 % w/w for flour, respectively ~ 30 % w/w for pepper). All the investigated samples present similarities in their rheoviscometric behaviour, as it is show in rheograms. It should be assumed this similar viscometric behaviour of corn starch, wheat flour and black pepper could be attributed to the starch degradation under microwave and electron-beam irradiation. It has also pointed out a more significant effect of e-beam irradiation on the rheologic properties of the considered materials. In the same time it could be make some consideration on the thermal and non-thermal effects of microwaves and on the different combined (e-beam and microwave) treatments of starch, flour and pepper. Working group No. 1 NOTES OCCURRENCE AND REMOVAL OF SOME GENOTOXIC COMPOUNDS IN GROUNDWATER GEHRINGER, P. Austrian Research Centers Seibersdorf, A-2444 Seibersdorf, Austria

With regard to hydroxy free radical generation the ozone/electron beam irradiation treatment process is most efficient among the ozone based Advanced Oxidation Processes (AOPs). This process is able to mineralize trace amounts of perchloroethylene in groundwater in a single stage process without formation of any by-product to be disposed of. Moreover, experiments performed just recently have demonstrated that the ozone/electron beam process is also apt for total removal of genotoxic compounds detected in groundwater contaminated with perchlorothylene. The design of the world wide first ozone/electron beam full scale plant to purify groundwater contaminated with traces of perchlorothylene and additionally also with genotoxic compounds of unknown origin and structure is presented. The capacity of the plant which will be located in Bad Fischau-Brunn - a small community south of Vienna - will be 108 m3/h; the treated water will be supplied into the tap water distribution system of the community. In the present case the genotoxic components cannot detected by chemical methods. The Tradescantia micro nucleus assay used for the proof of genotoxicity needs about one month to yield the result. Under these conditions activated carbon filtration alone is not apt for remediation of such a contaminated water.

13 CZ9928610 Working group No.1 FEED IRRADIATION AND DIET COMPOSITION IN DAIRY COWS GRUBIC, G.1, VITOROVIC, G.2, VITOROVIC, D.1, DONIC, S.3 institute of Animal Production, Faculty of Agriculture, Zemun. 2Faculty of Veterinary Medicine, Belgrade, Yugoslavia. Veterinary Station "Napredak", Gevgelija, Macedonia.

In this paper the prospects of 137Cs activity reduction by ration formulation in dairy cows are presented. To be more realistic, the actual foodstuffs and their 137Cs activity were used according to measurements done during the Chernobyl accident. It was found that there is a significant possibility for substantial reduction of radioactivity in dairy cow rations during accidental situations. Using linear programming software for diet formulating by setting the criterion for lowest 137Cs activity, and choosing conserved foodstuffs (like silage) prepared before the accident, it is possible to formulate rations significantly less radioactive. At the same time, all the nutrient needs even for high yielding cows are met successfully, so that their production is not reduced. This simple method could be very successful in domestic animal protection during possible accidents, in case that there are sufficient quantities of conserved feeds on the farm. Working group No.1 ELECTRON-BEAM AND MICROWAVE TREATMENT OF SOME MICROBIAL STRAINS CZ9928611 MARTIN, D.1, FERDES, O. S.\ PLAMADEALA, S.2, FERDES, M.3, MINEA, R.1, TIRLEA, A.1, BADEA, M.1 National Institute of Laser, Plasma and Radiation Physics; 1 Atomistilor St.; P.O.Box MG-36; Ro-76900 Bucharest; Romania; E-mail: [email protected] 2The Ministry of Agriculture and Food, 24 Carol I Blvd.; Bucharest, Romania institute of Food Chemistry, 1 Girlei St., Ro-71576, Bucharest, Romania

There are presented the main experimental results obtained on the combined effects of microwaves and accelerated electron beams on the different microbial strains such as E. coli, Salmonella sp. and Monascus purpureus, which are important for food quality requirements. For the microwave treatment it is used a special designed microwave applicator with a 2.45 GHz frequency CW magnetron of 850 maximum output power and with associate electronics that allow to control the microwave power; the current intensity, and the exposure time, which are the main parameters taken into consideration. Meantime, the electron-beam irradiation have been performed to different irradiation doses and at a dose rate of 1.5 - 2.0 kGy/min by using a linac (ALIN-7 at NILPRP Bucharest, Electron Accelerator Laboratory) that presents the following main parameters: electron mean energy ~ 6 MeV; mean bean current of 10 |xA; pulse period of 3.5 us and a repetition frequency of 100 Hz. The experiments have been carried out in 5 variants: microwave treatment; electron-beam irradiation; combined microwave then electron-beam treatment; combined electron-beam then microwave treatment and, simultaneously microwave and electron-beam processing. The experimental results are presented and analysed in terms of TGC/ml, CFU/ml or the specific microbial load. The comparative results obtained pointed out that the microbicidai effect is enhanced by additional use of microwave energy to electron beam irradiation. Enhancement of inactivation rate is remarkable by microwave treatment or by simultaneous electron beam and microwave irradiation only at temperature over the critical value on which microorganisms begin to perish by heat only. Due to this fact, microwave parameters were controlled such as to keep the sample final temperature around this critical value. A clear and continuous decrease of microbial load by exposure time has been observed, but there is no clear evidence either the intensity or the power is directly related to the microbicidai effect of microwave. Simultaneous irradiation with electron beam and microwaves results in the reduction of temperature and time as well as in the decrease of the upper limit of required electron beam absorbed dose for an assumed microbiological quality parameter. The obtained results pointed out the presence of synergistic effect -on non-thermal basis- of these two physical fields. Considering these results it could be concluded the combined microwave-electron beam treatment could be considered as an effective method to reduce the microbial load.

15 Working group No.1 NOTES DISINFECTION OF RED DRIED HUNGARIAN SPICE PAPRIKA BY SYNERGOLUX TREATMENT REGARDING ALSO QUALITY ASPECTS SIMON, J.1, TOLNAY, P.1, SZAB6, A. S.2, JUHASZ-ROMAN, M.2 University of Horticulture and Food Industry, 'Department of Food Chemistry and Nutrition, department of Microbiology and Biotechnology, Budapest, Somloi str. 14-16, Hungary

In the lecture data are given about the effect of different doses of SYNERGOLUX treatment carried out in red dried Hungarian spice paprika samples. As a microbiological quality parameter, the total number count was used. Attention was paid to colour changes as well. SYNERGOLUX treatment means a combination of UV radiation and ozone gas treatment at the same time. Working group No.1 EVALUATION OF DATA OF QUESTIONNAIRE NUTRITION AND HEALTH (1997-1998) CZ9928612 CONCERNING THE TOPIC WHAT CONSUMERS THINK ABOUT FOOD IRRADIATION? SZABO, A. S., TOLNAY, P. University of Horticulture and Food Industry, Department of Food Chemistry and Nutrition, Somloi str. 14-16, Budapest, H 1118 Hungary

In the lecture detailed information is given about the evaluation of data of the questionnaire. Data are based on 2000 questionnaires distributed and collected in 1997 and 1998. Evaluation is carried out taking into account the following parameters: age, sex, education level, living place and financial income. As a conclusion we can establish that people have a rather limited information about food irradiation.

17 Working group No. 1 NOTES RHEOLOGICAL PROPERTIES OF RIPENED, HARD CHEESES SUBJECTED TO HIGH HYDROSTATIC PRESSURE SZCZAWINSKA, M. E.1, LENART, A.2, SZCZAWINSKI, J.1 1Warsaw Agricultural University, Faculty of Veterinary Medicine, Department of Food Hygiene, Nowoursynowska 166, 02-787 Warszawa, Poland, E-mail:[email protected] 2Warsaw Agricultural University, Faculty of Food Technology, Department of Food Engineering, Nowoursynowska 166, 02-787 Warszawa, Poland

Three different kinds of ripened, hard, sliced cheeses were placed in PA/PE pouches and sealed under vacuum. The samples were exposed to the increasing values of hydrostatic pressure (0-500 MPa) at ambient temperature. Slices of cheeses were compressed in a Universal Testing Machine (Zwick, modell 1445) to evaluate Theological properties of the experimental samples. Changes of the compressing force (F) during time were recorded. Analysis of the curves illustrating a change of the compressing force (F) in the function of time (t) as well as the maximum value (Fmax) of the compressing force recorded within the test time indicated that high pressure treatment did not affect rheologicai properties of the evaluated cheeses. The results confirm our earlier observations that sensory properties (colour, smell, taste and texture) of ripened, hard, vacuum-packed cheeses were not affected by high pressure treatment. Working Group No. 1

COMPARISON OF THE EFFECT OF X-RAY AND ELECTRON BEAM IRRADIATION ON THE CZ9928613 THIAMINE CONTENT IN CHICKEN DRUMSTICKS VAN CALENBERG, S.1, VAN CLEEMPUT, O.1, MONDELAERS, W.2, HUYGHEBAERT, A.1 1 University of Gent, Faculty of Agricultural and Applied Biological Sciences, Coupure links 653, B-9000 Gent, Belgium 2University of Gent, Faculty of Sciences, Proeftuinstraat 86, B-9000 Gent, Belgium

The effect of irradiation with X-rays and electron beams on the thiamine content of chicken drumsticks in function of the storage time at refrigerator temperature was examined. A measurement of the total colony count was used as an indication of the preservation time. The irradiation dose applied ranged from 0 to 7 kGy at dose rates of 0.05 and 5 kGy/min for X-rays and electron beams respectively. It was concluded that differences in the thiamine breakdown between the two types of irradiation increased as the dose increased. In the worst case, at the X-ray treatment of 7 kGy, the mean loss in thiamine was 46.06%.

19 Working group No.1 IN VITRO AND IN VIVO EFFICIENCY OF CLINOPTILOLITE IN 137CS BINDING CZ9928614 VITOROVIC, D1, VITOROVIC, G.2, MLADENOVIC, V.3, VUKICEVIC, O.4 1 2 3 4 Faculty of Agriculture, Belgrade, Faculty of Veterinary Medicine, Belgrade, VTI Institute, Belgrade, Institute for Technology of Nuclear and Other Raw Materials, Belgrade, Yugoslavia

In this are paper results from the investigation of micro amounts of 137Cs sorption on the powder of natural zeolite clinoptilolite in the electrolyte similar to stomach content of animals. Experiments were performed with the natural form of clinoptilolite from the Zlatopek mine (Vranjska Banja) and with sodium form, on the temperature of 37°C. Simultaneously, the investigation of the efficiency of the natural form of clinoptilolite in radioprotection of chicks alimentary contaminated with 137Cs. Clinoptilolite showed a great efficiency in 137Cs sorption from the electrolyte solution, where 96-98% of initial activity was sorbed. Protection effect in chicks alimentary contaminated with 137Cs, measured 3 and 7 hours after the contamination, was approximately 75% for meat and edible inner organs of chicks.

20 Working group Nr. 1 RADIATION INDUCED MAILLARD REACTIONS (THE KINETIC OF COLOUR FORMATION DURING CZ9928615 HEATING) TEGOTA, A, BACHMAN, S. Institute of Applied Radiation Chemistry, Laboratory of Food Irradiation, Technical University of todz, Poland

The reactions of non enzymatic browning called Maillard reactions include a series of chemical reactions taking place in food preservation, preparation and processing at higher temperatures. The final effect of proceeding Maillard reactions is mainly a change in the flavour and colour of food products. The products of these reactions have an essential effect on food organoleptic quality and their acceptance by consumers. They are characterized by strong antioxidant properties. Preliminary studies were mainly conducted on model system containing of selected compounds, simulating food, to allow the interpretation of the results in terms of the food (reducing sugars and amino acids). The products of these reactions have a critical influence on the quality of food products and their acceptance by consumers. There are prospects that in near future the radiation method will be accepted by joint FAO/WHO/IAEA (UNO- Organizations) and introduced on a commercial scale into international practice). In this communication the results are presented the investigations of the effect of ionizing radiation from 60Co on the acceleration of Maillard reactions in model system containing an aqueous solution of fructose F-0.03 mol/dm3 and alanine Ala 0.01 mol/dm3. Solutions F/Ala irradiated in the range of doses 5 to 30 kGy at a dose rate 1.4 Gy/s, were then heated for some hours at different temperatures: 40°, 60°, 80°, and 100°C. To asses the colour intensity of the solutions under investigation, the value of absorbance at 450 nm in 10 mm cuvettes was measured. These spectrophotometric studies showed that the colour intensity of irradiated solutions was dependent on the dose of radiation, the temperature and the heating time. The reaction constants estimated for the system studied increased with the increase of radiation dose and temperature. The activation energy of colour development determined for the range of temperatures from 60°C to 100°C decreased with dose from 70.6 kJ/mole for 5 kGy to 60.7 kJ/mole for 30 kGy. The results of spectrophotometric and chromatographic studies confirmed the formation of carbonyl products from fructose radiolysis and their participation in the acceleration of non-enzymatic browning reactions. The aldehyde products formed from the amino acids as a result of the Strecker degradation are responsible for the formation of odour characteristic for Maillard during heating. The changes in the concentration of fructose and alanine during irradiation of the solutions under study were proportional to the dose of radiation. The radiation yield of fructose decomposition was equal to G = 2,6 and that for alanine G = 0.22. In the irradiated solutions of F/Ala, serine has been found, which so far has not been mentioned as a product of alanine radiolysis. In conclusion it may be stated that the studies performed demonstrate the influence of radiation and acceleration of Maillard reaction during subsequent heating at 40°C up to 100°C of systems containing reducing sugars and amino acids. It should be taken under consideration in the studies on introducing radiation technology of food products preservation connected with further thermal treatment.

21 Abstracts presented in working group No. 2

Chairmen: Prof. M. Cristaldi and Prof. B. Todorov MEASUREMENT OF 137Cs ACTIVITY IN LIVING CHICKEN 25 BALAS, J., POSCHL, M.

BIOELECTRIC IMPEDANCE: SIMPLER APPROACH TO PREDICTION OF THE MEAT YIELD IN THE PIG HALVES 26 BEGO, U., TONKOVIC S., BIKIC M.

TRANSPORT OF 14C-METHIONINE, 14C-LYSINE AND THE ACTIVITY OF SOME ENZYME OF ISOLATED ENTEROCYTES AND MICROVILLOUS MEMBRANES OF BROILER CHICKS FED ON MIXES CONTAINING IONOPHORE ANTIBIOTICS. 27 CHOTINSKI, D., PROFIROV, Z., ALEXANDROV, S.

CHANGES IN THE CONCENTRATION OF THE THYROID HORMONES IN BLOOD PLASMA IN THE ONTOGENESIS OF BROILER CHICKS. 28 CHOTINSKI, D.

ROLE OF CYCLOOXYGENASE INHIBITION FOR THE DEVELOPMENT OF ERYTHROBLASTIC ISLANDS IN HYPERTRANSFUSED RATS29 KALAIDJIEVA, V.

INTRODUCTORY RESULTS OF SNAIL (HELIX ASPERSA MAXIMA) FARMING IN THE NATIONAL RESEARCH INSTITUTE OF ANIMAL PRODUCTION (CRACOW, POLAND) 30 LYSAK, A., LIGASZEWSKI, M., PRZYBY, A., SUROWKA, K.

HEAVY METALS LEVEL IN THE TISSUES IN SOME FARM-REARED FRESHWATER FISH SPECIES 31 LYSAK, A., MACH-PALUSZKIEWICZ, Z.

ACTIVITY CONCENTRATIONS OF 137Cs IN MEAT OF BROILER CHICKEN AFTER SINGLE AND CONTINUOUS APPLICATION. 32 POSCHL, M., BALAS, J.

IMPEDANCE PATTERNS OF VAGINA AFTER WEANING IN CONCEIVED AND NON-CONCEIVED SOWS 33 REZAC, P., OLIC, I.

IMPEDANCE PATTERNS OF VESTIBULAR MUCOSA DURING THE PERIPARTURIENT PERIOD IN MARES 34 REZAC, P., POSCHL, M.

23 CHANGES IN THE VAGINAL IMPEDANCE IN EWES AFTER OESTRADIOL TREATMENT DURING THE LUTEAL STAGE OF THE OESTROUS CYCLE 35 REZAC, P., POSCHL, M.

CHANGES OF CALCIUM AND CHLORIDE CONCENTRATIONS AFTER OESTRADIOL TREATMENT IN LAYING HENS 36 REZAC, P., CiZKOVA, J.

RELATIONSHIP BETWEEN PLASMA THYROID HORMONES AND OVARIAN STEROIDS DURING SEXUAL MATURATION OF THE DOMESTIC HEN (GALLUS DOMESTICUS) 37 SECHMAN, A., RZA.SA, J., MIKA, M., PACZOSKA-ELIASIEWICZ, H.

MORPHOLOGICAL AND FUNCTIONAL STUDIES OF POLYMORPHONUCLEAR LEUKOCYTES BY IMAGE ANALYSIS TECHNIQUE 38 SLADEK, Z., RYSANEK, D.

RESPONSE OF SYMPATHO-ADRENAL AXIS TO STRESS IN PREGNANT SHEEP 39 WIECZOREK E., NIEZGODA J., BOBEK S. Working group No. 2 MEASUREMENT OF 137Cs ACTIVITY IN LIVING CHICKEN CZ9928616 BALAS, J., POSCHL, M. Mendel University of Agriculture and Forestry, Department of Nuclear Methods, Zemedelska 1, 61300 Brno, Czech Republic, E-mail: [email protected]

The possibility of measuring the 137Cs activity in broiler chickens in vivo was examined. The experiment was conducted with 7-22 day-old broiler chickens (White Leghorn hybrid, race ISA VEDETTE). Three oral doses of 137Cs (total activity 5 kBq/chicken) were administered using a special applicator in the course of one day (at 8:00, 12:00 and 16:00). The radioactivity was determined using a Gamma Spectroscopy system with a stabilised Nal/TI detector (3x3 ins, i.e. 75x75 mm, in a lead box. Measurement was carried out 1d, 2d, 3d, 4d, 7d, 8d, 14d and 15d after the 37Cs dosage in live chickens (in vivo) fixed on special board. The detector was put closely to the body in the area of breast muscles. Radiocaesium activity concentrations in daily fresh excreta were also determined. A rapid uptake of the orally administered 137Cs (within a few hours) and also rapid loss of radiocaesium were observed. Dynamics and accuracy of the in vivo measurement of 137Cs activity are comparable with recently published results (Poschl et al., 1997) of the in vitro measurement of radiocaesium activity. Only initial 137Cs activity (during 2 days after 137Cs application) was higher with regard to the content of contaminated feed mixture in the gastrointestinal tract.

The Grant Agency of the Czech Republic under Grant No. 525/96/0640 supported this research.

Poschl M, Borkovec V, Zelenka J (1997) Dynamics and distribution of radiocaesium in broiler chicken. Radiat Environ Biophys 36:169-174

25 Working group No.2 NOTES BIOELECTRIC IMPEDANCE: SIMPLER APPROACH TO PREDICTION OF THE MEAT YIELD IN THE PIG HALVES BEGO, U.1, TONKOVlC S.1, BIKIC M.2 1Faculty of Electrical Engineering and Computing Zagreb, University of Zagreb, Unska 3,10000 Zagreb, Croatia. 2Faculty of Agriculture University of Zagreb, Svetosimunska c. 25, 10000 Zagreb, Croatia, E-mail: [email protected].

Development of the phase-sensitive electric devices enabled routine two-electrode measurement of electric impedance for the determination of PSE pork meat (Pfutzner et al., 1981, Feldhussen et al., 1987, etc.) and four- electrode measurement of impedance for the prediction of fat-free mass in the various species of domestic animals and their carcasses (Swantek et al., 1992, Berg and Marchello, 1994, etc.) Our long-lasting two-electrode researches have shown successfulness in the determination of PSE meat and composition of some muscles in the different animal species and their carcasses (Bego et al., 1995-1998). In this paper we investigated the possibility of usage of the method of bioelectric impedance in the prediction of percentage and mass of muscle tissue in the halves of Hypor lines fattened pigs (n=40) with the average carcass weight 76.79 kg, length 97.8 cm, back fat thickness 13 mm, muscle thickness 61.86 mm (measured using the ,,Two Point" method) and pH45 value 6.01. Measurement of the impedance module were done using HPRLC-Meter-4284A impedance analyzer at the frequencies of 15, 50, 100 and 200 kHz. Two electrodes were pointed into m. gracilis 8 cm ventral of pelvic symphisis. Multifrequency analyses showed that the data obtained at 200 kHz frequency are the most reliable. That frequency is the most suitable for the measurement of meat yield in the pig halves when using recalculated values of impedance, weight and length of the halves. Linear regression analysis shown exceptional accordance between the meat yield data obtained by the ,,Two Point" method and recalculated impedance measurements in m. gracilis and the weight (R2= 0.943, RSD= 0.91) and length (R2= 0.951, RSD= 0.83) of pig halves. Based on this research, we can conclude that two-electrode measurement of the impedance, done on m. gracilis at the frequency of 200 kHz with the recalculated values of weight and length of the halves, gives the reliable picture of the meat yield in the pig halves.

26 Working group No. 2 NOTES TRANSPORT OF 14C-METHIONINE, 14C-LYSINE AND THE ACTIVITY OF SOME ENZYME OF ISOLATED ENTEROCYTES AND MICROVILLOUS MEMBRANES OF BROILER CHICKS FED ON MIXES CONTAINING lONOPHORE ANTIBIOTICS. CHOTINSKI, D.1, PROFIROV, Z.2, ALEXANDROV, S.2 Voultry Research Institute, Department of Nutrition, Physiology and Biochemistry, Bulgaria 2Animal Breeding Institute, 2230-Kostinbrod, Bulgaria

Abstract was not delivered

27 Working group No. 2 NOTES CHANGES IN THE CONCENTRATION OF THE THYROID HORMONES IN BLOOD PLASMA IN THE ONTOGENESIS OF BROILER CHICKS. CHOTINSKI, D. Poultry Research Institute, Department of Nutrition, Physiology and Biochemistry, 2230-Kostinbrod, Bulgaria

Abstract was not delivered Working group No. 2 NOTES ROLE OF CYCLOOXYGENASE INHIBITION FOR THE DEVELOPMENT OF ERYTHROBLASTIC ISLANDS IN HYPERTRANSFUSED RATS KALAIDJIEVA, V. Dept. of Physiology, Faculty of Medicine, Thracian University, Armeiska str. 11, Stara Zagora 6000, Bulgaria E-mail: [email protected]

Although prostaglandins are well known as modulators of erythropoiesis, their precise action is still a subject of controversy. In order to study the role of prostaglandin's inhibition on hematopoietic recovery after suppression, we examined erythropoiesis in bone marrow erythroblastic islands in hypertransfused rats, injected with cyclooxygenase inhibitor indomethacin (4mg/kg/5days s.c). Suppression of erythropoiesis was performed by injecting rats intravenously with 2.5ml/100g of a 70% suspension of washed, packed isogenous red cells on two consecutive days. We considered the development of erythroblastic islands (El) as a reliable criterion for examining the terminal stage of erythropoiesis and the prostaglandin's impact on it. The study included estimation of plasma erythropoietin level (by RIA), peripheral blood and morphological analysis of the erythroblastic islands - number of El from I to V classes/femur, related indices: rate of CFU-E differentiation into erythroblasts, rate of repeated participation of macrophages in new El reconstruction, myelogramme, during a 10 da/s period. We observed a depressed El development after hypertransfusion which was potentiated after indomethacin treatment - an increased number of circulating reticulocytes, of El/femur, an enhanced reconstruction in involuted El and a reduced number of maturing El (p<0.01). Signs of activation of central El' macrophages were expressed after prostaglandin's inhibition - rate of their repeated participation in El reconstruction, rate of CFU-E differentiation into erythroblasts in El were found elevated on days 3-5 (p<0.01). Our study favours the concept about the stimulating effect of cyclooxygenase inhibition on erythropoiesis in bone marrow erythroblastic islands via activation of central macrophages within these cellular associations.

29 Work ng group No 2 NOTES INTRODUCTORY RESULTS OF SNAIL (HELIX,ASPERSA MAXIMA) FARMING IN THE NATIONAL RESEARCH INSTITUTE OF ANIMAL PRODUCTION (CRACOW, POLAND) LYSAK, A.1, LIGASZEWSKI, M1, PRZYBY, A.3, SUR6WKA, K.2 1 National Research Institute of Animal Production in Cracow, Department of Animal Production Technology and Ecology, 31-047 Krakow, ul. Sarego 2, Poland Agricultural Academy in Poznan, Chair of Fishery, Poland Agricultural Academy in Cracow, Institute of Refrigeration and Food Industry, Poland

Terrestrial snails from the genus Helix, native to the Mediterranean zone (Helix aspersa) and large areas of almost entire Europe (Helix pomatia), are desirable food iti France. This led to the overexploitation of natural snail populations. To prevent this situation, intensive farming of Helix aspersa, including its subspecies Helix aspersa maxima, has recently been promoted and developed n Europe. In Poland, an experimental farm belonging to the National Research Institute of Animal Production in Cracow has been actively involved in developing this new branch of animal production. Early-spring reproduct on started after 4 months lasting hibernation. Rearing of incubated snails is carried out indoors, while commeicial fattening and rearing of reproductors are carried out in both closed (hothouse) and open systems (s.c."park;s") in a total area of 200 sq m. In the short and cold 1997 season, which did not favour snail farming, 2.0 kg of snail biomass, including 60% snails whose commercial dimensions exceeded 14 g/head, was achieved per 1 sq m. The nutrient coefficient for the feed mixtures used was 1.4, compared to 2.0 for large production farms cooperating with the Institute. At the same time snails were farmed on plastic trays in laboratory conditions to test the efficiency of snail yield and feed conversion. The table below shows some of the results of this three-month experirnent, which used 4 experimental isoprotein and isoenergetic feeds (Agricultural Academy in Poznaii) compared to r.>rivately produced fodder. The initial weight of the hatch used in the experiment was 0,1 g/snail. The coefficient of VJariation for final body weight ranged from 61.0 to 73.1%, with body weight ranging from 0.2 to 24.2 g/snail. A statistically significant difference was found in the experimental feeds between the effect of feeding fodders from groups PI-PIII and PIV on the biomass and quality of snails produced. The PV feed led to the worst results, despite its high protein content. Fodder Increase proportion of biomass loses Food FODDER for snail over 14 g/snail (snail %) coeff. (%) g/m" g/snail Dry protein Ash Fat weight (g/kg) (g/kg) (g/kg) (g/kg) Pll 2071 9,3 51,3 24,7 0,75 948 152 331 40 Pill 2054 8,3 41,6 15,3 0,73 954 157 272 35 PI 1961 9,5 35,4 29,3 0,76 953 152 322 33 PIV 559 3,2 0,0 38,0 1,08 948 148 288 37 PV 454 2,1 0,0 20,0 1,56 946 211 139 23

30 Working group No 2 NOTES HEAVY METALS LEVEL IN THE TISSUES IN SOME FARM-REARED FRESHWATER FISH SPECIES tYSAK, A., MACH-PALUSZKIEWICZ, Z. National Research Institute of Animal Production, Department of Animal Production Technology and Ecology, 31-047 Krakow, ul. Sarego 2, Poland

The aim of the studies was to determine the Zn, Cu, Cd and Pb content of muscles and liver of brown trout (Salmo trutta morpha fario, L), brook trout (Salvelinus fontinalis, Mitch.) and carp (Cyprinus carpio, L). These species of fish, are the subject of an intensive farm rearing in Polish ponds. Two-year-old trout and three-year-old carp came from the the farms in southern Poland. The trouts were fed with high-protein pellets for salmonids, while the carps in particular ponds were given either cereal fodders or were keept on natural food. Their tissues were analysed using inverse voltammetry and atomic absorption spectrometry (AAS) after previous mineralization in the mixture of nitric and perchloric acids. The results are compared in the table below in the form of mean values from 3 years of research.

Species muscles (ppm of dry weight) liver (ppm of dry weight) Zn Cu Cd Pb Zn Cu Cd Pb brown trout 59,4 6,5 <0,1 1,2 164,0 143,0 0,1-0,4 3,4 brook trout 37,7 4,9 <0,1 1,2 105,1 114,3 LP,1-0,4 2,0 Carp: Natural feed 51,0 3,0 <0,1 1,1 366,5 29,4 <0,1 2,3 Cereal feed 40,6 2,9 <0,1 1,1 222,5 18,3 <0,1 2,1

The content of analysed metals was found to vary depending on the type of tissue and species of fish. The level of these metals, especially Zn and Cu, was many times lower in muscles then in liver of all the species. The concentration of Zn was 2 and of Cu 3-9 times lower in the trout than in the carp. The Cd and Pb content of liver and muscles was similar in both species. Statistically significant differences occurred in the concentration of Zn and Cu in the liver of brown and brook trouts. The level of Zn and Cu was significantly (PO.05) higher in the brown trout than in the brook trout, despite the fact that they were bred on the same farms. Similar, although non-significant differences were found in the level of Zn and Cu in trout muscles. The carp farmed in different conditions were found to vary as to the Zn and Cu content of their hepatopancreas. The higher amounts of these metals were found in the fish which were kept exclusively on natural food. None of the muscles of analysed fish was found to have metal levels in excess of current Polish standards.

31 Working group No.2 ACTIVITY CONCENTRATIONS OF 137Cs IN MEAT OF BROILER CHICKEN AFTER SINGLE AND CONTINUOUS APPLICATION. CZ9928617 POSCHL, M., BALAS, J. Mendel University of Agriculture and Forestry, Department of Nuclear Methods, Zemedelska 1, 61300 Brno, Czech Republic, E-mail: [email protected]

After the Chernobyl accident much interest was focused on the transfer of radiocaesium (137Cs) and its suppression in the soil-plant-animal food chain because the fallout resulted in high levels of the radionuclide in the environment of Europe. Although a lower contamination of broiler chicken can be expected after the accident, the potential hazard of 137Cs transfer by poultry products in the human diet should not be neglected. In literature the information about transfer in chicken is incomplete. In previous presentations (ESNA meetings in 1993-97) we described the transfer, distribution and half-live of radiocaesium in broiler chicken after application of artificially contaminated feed mixture or after feeding contaminated wheat from Chernobyl. Our results (Poschl et al., 1997) indicated the different dynamics of radiocaesium in breast meat compared to leg meat in the chicken after short-time application (3 oral applications in 1 day). The aim of the present study was to find if the results are similar also after single and repeated (long-time) applications of an artificially contaminated feed mixture. Two experiments were carried out with broiler chickens (White Leghorn hybrid, race ISA VEDETTE). In experiment 1, one artificially contaminated oral dose of 5160 Bq of 137Cs (activity concentration 1664 Bq.g"1) was administered to 18-day-old chickens. In experiment 2, artificially contaminated oral doses of 500 Bq Cs (activity concentration 161.3 Bq.g'1) were administered to 14-day-old chickens twice a day (at 8:00 and 20:00h.) for 10 days. In experiment 1 and 2, four chickens were slaughtered for activity determination in meat (breast and leg muscles) 6, 12, 24, 48 and 96 hours and 2, 4, 8, 10 days, respectively, after the 1st application of 137Cs. The uptake of single oral 137Cs was rapid and the maximal 137Cs activity concentrations were recorded in breast meat (0.783 Bq.g ) 24 hours and in leg meat (1.005 Bq.g"1) 6 hours after 137Cs application. From the 24th hour of the experiment, radiocaesium activity concentrations in breast and leg meat decreased with the biological half-life (Ti/2b) of 84 and 66 hours, respectively. During a 10-day application of continuous doses of 137Cs, the Cs activity concentrations increased and were 3.988 Bq. g'1 in breast meat and 5.610 Bq. g"1 in leg meat on the 2nd day, and 7.427 Bq.g"1and 7.698 Bq.g"1, respectively, on the 1001 day. Immediately after administration of radiocaesium 137 stopped, the Cs activity concentrations rapidly decreased with the biological half-life (T1/2b) of 4.5 and 3.8 days, in breast and leg meat, respectively. The results demonstrate that there is a faster uptake and higher 137Cs activity concentration into leg meat than into breast meat and faster release of radiocaesium after single and repeated (long-time) applications of artificially contaminated feed mixture with 137Cs in concentrations as mentioned above. This research was supported by the Grant Agency of the Czech Republic under Grant No. 525/96/0640. Pdschl M, Borkovec V, Zelenka J (1997) Dynamics and distribution of radiocaesium in broiler chicken. Radiat Environ Biophys 36:169-174 Working group No.2 NOTES IMPEDANCE PATTERNS OF VAGINA AFTER WEANING IN CONCEIVED AND NON-CONCEIVED SOWS REZAC, P., OLIC, I. Mendel University of Agriculture and Forestry, Department of Morphology, Physiology and Veterinary Medicine, Zemedelska 1, 61300 Brno, Czech Republic

The important problem that pig breeders are facing is to determine the suitable time for artificial insemination of the sows after weaning. A number of diagnostic methods are available for monitoring events occurring during the periovulatory period. The aim of the present investigation was to determine whether there are any differences in vaginal impedance near the cervix uteri between conceived and non-conceived sows after weaning. The standing reflex was detected in the presence of a sexually mature boar twice a day. Insemination was carried out twelve hours after oestrus onset and re-insemination after another twelve hours. The impedance was measured by a four-electrode method on the vagina near the cervix uteri. Impedance measurements were carried out every day from weaning to the 8th day after oestrus onset. Pregnancy was confirmed by the date of farrowing. After weaning, the vaginal impedance in sows which subsequently conceived (n=72) was found to fall (P < 0.01). The lowest values of vaginal impedance were observed 1-2 days before oestrus. During oestrus a marked increase in vaginal impedance followed (P < 0.01). In the course of early pregnancy no changes in the vaginal impedance were recorded. After weaning, the vaginal impedance in sows which subsequently non-conceived (n=14) was found to fall (P < 0.05) with a consecutive increasing and decreasing of its values during pro-oestrus. During oestrus a marked increase in vaginal impedance followed (P < 0.05). In the course of dioestrus no changes in the vaginal impedance were observed. The decrease of vaginal impedance during pro-oestrus in the pregnant sows was significantly greater in comparison with the non-pregnant sows (P < 0.05).

33 Working group No.2 NOTES IMPEDANCE PATTERNS OF VESTIBULAR MUCOSA DURING THE PERIPARTURIE-INT PERIOD IN MARES REZAC, P.1, POSCHL, M.2 Mendel University of Agriculture and Forestry, Department of Morphology, Physiology and Veterinary Medicine1, Department of Nuclear Methods2, Zemedelska 1, 61300 Brno, Czech Republic, E-mail: [email protected]

Parturition is a critical step in reproduction. Serious associated problems in mares are dystocia and fetal death. A number of diagnostic tool are available for monitoring events occurring prior to delivery. The aim of this study was to determine whether marked histological and histochemical changes in the genitalia are followed by changes in the vestibular impedance before and after foaling. The impedance was measured by a two-electrode method on the vaginal vestibule 5 cm from the vulva. Impedance measurements were carried out daily from 10 days before the expected parturition to 4 days after foaling. The impedance of the vaginal vestibule in mares (n=11) decreased significantly 8 days before parturition (P < 0.05). Afterwards no marked changes in the vestibular impedance were observed until parturition. After foaling the vestibular impedance increased significantly (P < 0.05).

34 Working group No.2 NOTES CHANGES IN THE VAGINAL IMPEDANCE IN EWES AFTER OESTRADIOL TREATMENT DURING THE LUTEAL STAGE OF THE OESTROUS CYCLE REZA6, P.1, POSCHL, M.2 Mendel University of Agriculture and Forestry, Department of Morphology, Physiology and Veterinary Medicine1, Department of Nuclear Methods2, Zemedelska 1, 61300 Brno, Czech Republic, E-mail: [email protected]

Cyclic changes in vaginal impedance during the oestrous cycle were observed in cattle, pig, sheep, horse, mouse, dog, rat, guinea pig, monkey, and fox. It was found that a decrease in vaginal impedance during the perioestrous period was caused by oestradiol produced by the growing preovuiatory follicles. The objective of the experiment reported below was to determine whether exogenous oestradiol given during the luteal phase of the oestrus cycle is able to induce a decrease in the vaginal impedance in ewes. Synchronisation of oestrus was carried out by cloprostenol given twice in an interval of 10 days. Oestrus was detected by a teaser twice a day. The ewes (n=12) received i.m. 3.5 mg oestradiol twelve days after oestrus onset. The impedance was measured by a four-electrode method on the vagina mucosa 10 cm from the vulva. Impedance measurements were carried out 4 and 2 hours before and 1, 2, 4, 6, 8, 10, 12, 14, 20 and 22 hours after oestradiol injection. A mild decrease in vaginal impedance was recorded during the first 14 hours after oestradiol treatment. A significant fall in vaginal impedance was found 20 hours after oestradioi injection. A marked decrease of vaginal impedance in all ewes was observed 20 hours after oestradiol treatment (P < 0.01).

35 Working group No.2 NOTES CHANGES OF CALCIUM AND CHLORIDE CONCENTRATIONS AFTER OESTRADIOL TREATMENT IN LAYING HENS REZAC, P., CiZKOVA, J. Mendel University of Agriculture and Forestry, Department of Morphology, Physiology and Veterinary Medicine, Zemedelska 1, 61300 Brno, Czech Republic

There is a critical control within a very narrow range of circulating calcium in the laying hens. No less than four endocrine systems (parathyroid hormone, calcitonin, vitamin D3, and oestradiol-17p) control these processes. The objective of this study was to determine the response of the plasma levels of calcium and chloride in laying hens to injection of oestradiol. Laying hens were given an i.m. injection of 0.6 mg (n=8) and 0.06 mg (n=8) oestradiol dipropionate and the control received an i.m. injection of sterile saline (n=8). Treatment was conducted between 3 and 6 h after oviposition. Blood samples were taken from the vena basilica. Each animal was sampled immediately before and 1 h, 2 h, 3 h, 24 h and 96 h after treatment. The ion concentrations were determined by atomic absorption spectrophotometry. Plasma concentrations of calcium decreased gradually and the minimum level was achieved 3 h after saline treatment (P < 0.01) and 24 h after oestradiol treatment (P < 0.01). The calcium concentration returned to the pre- treatment level 24 h after saline injection and 96 h after oestradiol injection. The concentration of calcium 24 h after treatment was significantly lower in oestradiol treated groups in comparison with the saline treated groups (P< 0.01). Concentrations of chloride in saline and oestradiol treated groups decreased one hour after treatment and then increased markedly to 24 h after treatment (P < 0.01). A fall in chloride levels was recorded 96 h in all three groups of laying hens (P < 0.01).

36 Working group No. 2 NOTES RELATIONSHIP BETWEEN PLASMA THYROID HORMONES AND OVARIAN STEROIDS DURING SEXUAL MATURATION OF THE DOMESTIC HEN {GALLUS DOMESTICUS) SECHMAN, A., RZASA, J., MIKA, M., PACZOSKA-ELIASIEWICZ, H. Department of Animal Physiology, University of Agriculture, Al. Mickiewicza 24/28, 30-059 Krakow, Poland, E-mail: [email protected]

In birds thyroid hormones, thyroxine (T4) and triiodothyronine (T3), are essential for the control of many physiological processes such as development, differentiation, growth, metabolism and termoregulation.(McNabb, 1992). There are many evidences that thyroid hormones play an important role in the regulating of the onset of puberty and reproductive function in birds (Sharp and Klandorf, 1985; Kirby at al., 1996), however, the exact nature of hormonal interactions between ovarian and thyroid axes remains obscure. The aim of the present study, therefore, was to investigate the relationship between thyroid and ovarian function in the domestic hen during the last phase of the sexual maturation which is associated with the elevated activity of the hypothalamo-ovarian axis. The study was carried out on 24 Astra S pullets between 15 and 22 week of age. Birds were kept in 14L10D lighting regime and provided with water and food ad libitum. Blood samples were collected from the wing vein at 16, 18 and 20 week of age (i.e. during the time preceding the onset of laying), and at 22 week of age (i.e. 1.5 week after the onset of laying). Thyroid hormones (T4 and T3) and ovarian steroids (progesterone - P4, estradiol - E2 and testosterone - T) were measured in blood plasma by means of RIA methods. Results: A gradual decrease of T4 and T3 concentration in blood plasma of hens was observed. In comparison with the starting T4 level (52.94 ± 5.71 ng/ml at 16 week of age), 1.5- and 2.4-fold lower levels at 20 and 22 week of age were found (PO.01); the appropriate T3 declines (initial level: 2.44 ± 0.35 ng/ml) were 1.9- and 5.1-fold (PO.01). At first E2 concentration (0.208 ± 0.009 ng/ml at 16th week) increased 1.5-fold at 18 week then decreased 2.6-fold at 22 week of age (PO.01). The level of P4 gradually increased from 16th week (0.31 + 0.008 ng/ml) till 22 week of age; the appropriate concentrations at 20 and 22 week of age were 8.1- and 10.5-fold higher (P<0.01). The level of T (0.41 ± 0.02 ng/ml) increased 1.9-fold at 20 week of age and then decreased 3.4-fold at 22 week (PO.01). The following significant correlation coefficients were found between: T4 and P4 -> r = -0.76; T3 and P4 -» r = -0.70; T4 and E2 -> r = 0.53. There was no significant correlation between T3 and E2 as well as between thyroid hormones and testosterone. The data obtained suggest that during the last phase of sexual maturation of the domestic hen there is a negative relationship between hypothalamo-thyroid and hypothaiamo-ovarian axis.

37 Working group No. 2 NOTES MORPHOLOGICAL AND FUNCTIONAL STUDIES OF POLYMORPHONUCLEAR LEUKOCYTES BY IMAGE ANALYSIS TECHNIQUE SLADEK, Z.1, RYSANEK, D.2 1Mendel university of Agriculture and Forestry, Brno, Czech Republic Veterinary Research Institute, Brno, Czech Republic

Differential counts and morphological and functional characteristics of somatic cells collected from secretions of intact or infected mammary glands of barren heifers were studied using the image analysis technique (LUCIA G, LABORATORY IMAGING, Prague, Czech Republic). Samples were collected by lavage with sterile buffered saline (PBS). The collected material was centrifuged, the supernatant decanted, the sediment resuspended in PBS and one drop of the suspension was used to prepare a smear. The smears were dried and stained according to Pappenheim. Image analysis allows recording, filing, correction and processing of microscopic pictures. Macrophages dominated in differential somatic cell counts in samples collected from intact or infected mammary glands. Typical polymorphonuclear leukocytes at various stages of degradation were observed. Macrophages were represented by monocytary and vacuolized cell types. The mammary glands were infected by both minor and environmental pathogenic agents. Findings of bacterial agents engulfed in polymorphonuclear leukocytes allowed the determination of the ratio of phagocytizing and non-phagocytizing cells and the calculation of the phagocytary index. Working group No. 2 NOTES RESPONSE OF SYMPATHO-ADRENAL AXIS TO STRESS IN PREGNANT SHEEP WIECZOREK E., NIEZGODA J., BOBEK S. Department of Animal Physiology, University of Agriculture, 30-059 Krakow

Isolation of sheep from the fiock produce an emotional stress characterised by increased activation of sympatho- adrenal system (SAS). The dimension of response to such a stress seems to depend on physiological state of the animal. In the present experiment we wanted to compare the response of SAS in non-pregnant and pregnant ewes to emotional stress. The experiment was carried out on 30 ewes divided into 3 groups: (1) non-pregnant ewes, (2) ewes in the early stage of pregnancy, (3) ewes in the late stage of pregnancy. Sixty minutes of isolation from the flock served as the emotional stressor. Blood was taken from the jugular vein at 60 minute before the isolation and then at 5, 15, 30 and 60 minute of stress and at 60 minute after cessation of the isolation. Adrenaline (A) and noradrenaline (NA) were determined by means of radioenzymatic procedure using Catechola 3H kit - Immunotech. The obtained data have shown that basal levels of A and NA (1.49±0.13 and 2.61±0.1 nmol/l, respectively) in non- pregnant and pregnant ewes did not differ. However, significant decreased response of the SAS to stress in pregnant ewes was noted. The mean integrated response of A in non-pregnant ewes was 169.67±64.71 nmol/l/180 minutes and was 5 times higher than in pregnant ewes. The integrated response of NA in non-pregnant ewes was 164.66+80.07 nmol/l/180 minutes and was 2 times higher than in pregnant animals. The obtained results clearly point to the diminished response of SAS of pregnant sheep to emotional stress.

39 Working group No.3 NOTES THE ECONOMIC ANALYSIS OF FERTILISER EFFECT ON THE YIELD OF SOME IMPORTANT CROPS GROWN IN TOKAT PROVINCE ESENGON, K., AKAY, M. Agricultural Economics Dept., Agricultural Faculty of Gaziosmanpasa University, 60100 Tokat, Turkey

Agricultural sector has an important role in the supplying of food needs of the people. And also population growth forces people to grow much more agricultural products. Increasing agricultural production is possible by means of increasing production area or having more yield per area. By the way, as an input agricultural lands of countries are limited so only one way to get more production is increasing yield per area. One of the most important method of increasing productivity in agricultural land is to provide increasing yield by effecting the nutrition characteristics of soil. This needs fertiliser application. The aim of this study is to determine the efficiency of farm fertiliser application on yield by economic analysis in Artova District of Tokat Province, Turkey. Data were collected by means of Personal Interview Method by the researchers. Wheat in dry conditions, wheat in irrigated conditions, barley and sugarbeet covered 83.50 % of total sowing area grown among field crops of the region. These crops were investigated by functional analysis. In the first step, Quadratic Function y = a+bx^cxi2, which explains the relation between yield and fertiliser was found for 4 crops separately. Being suitable for the Law of Diminishing Returns and having an advantage for reaching the Technical Optimum in relation to input-output are the reasons for preferring Quadratic Function. In the second step, Economic Optimum was determined by the functions formed. For this process the first derivation of function according to Xi was taken and the level of fertiliser use equaling the ratio of product input prices was determined. That is to say: Economic Optimum Level was found from equation: dY/dX| = PX/PY. In this study, the price and quantity of fertliser were taken as 1 kg nutrient. The value of yield and price belong to the 1996 production period. In the final step, the levels of fertiliser use in Economic and Technical Optimum were compared with farm application and then the results were interpreted. It was determined that farmers have not come close to the Economic and Technical Optimum. It can be said that doing enough soil analyses in fertiliser application is the reason of this situation and as a result the nutrient status of soil was explained. And also that the parity of product-fertiliser is in favour of fertiliser is thought as a second factor. As a result it was found that farmers did not have sufficient sources for fertlisers. It was proposed that soil analysis should be compulsory and legal arrangements to increase the level of fertliser subsidies should be done.

40 Abstracts presented in working group No. 3

Chairmen: Dr. M. Gerzabek and Dr. N. G. Mitchell COMPARASON OF THE POLICIES OF FERTILISER SUBSIDY AND PRICE SUPPORT FOR SELF-SUFFICIENCY IN SUNFLOWER: A CASE STUDY OF TURKEY 46 AKCA, H., GURLER, A. Z., SAYILI, M.

EFFECT OF NP FERTILIZATION ON THE YIELD, YIELD COMPONENTS AND NPK CONTENT OF WHEAT CROP GROWN UNDER TOKAT ECOLOGICAL CONDITIONS 47 BROHI, A. R., AKTAS, A. SAVASLI, E.

EFFECT OF NP FERTILIZATION ON THE YIELD COMPONENTS AND NPK CONTENT OF MAIZE CROP GROWN UNDER TOKAT ECOLOGICAL CONDITIONS 48 BROHI, A. R., AKTAS, A. SAVASLI, E.

EFFECT OF DIFFERENT PHOSPHORUS FERTILIZERS ON YIELD AND NPK CONTENT OF WHEAT CROP GROWN UNDER ECOLOGICAL CONDITIONS OF TOKAT/TURKEY 49 BROHI, A. R., TOPBAS, M. T., SAVASLI, E., AKTAS, A.

RESEARCHES - WITH IEFERT MODEL - CONCERNING THE VARIATION OF PHYSICAL AND ECONOMICAL EFFICIENCY OF FERTILIZERS USE 50 BUDOI, G. H.

MODELLING THE CROP RESIDUES AND ORGANIC FERTILIZERS MINERALIZATION IN SOIL 51 BUDOI, G. H., DANUSO, F., GIOVANARDI, R., DAVIDESCU, V., MADJAR, R.

FRUITS CONTAMINATION FOLLOWING AERIAL DEPOSITION AND REMAINING ACTIVITY AFTER WASHING OR PEELING 52 CARINI, F., ANGUISSOLA SCOTTI, I., D'ALESSANDRO P. G.

137CS SOIL - WHEAT PLANT TRANSFER IN GREENHOUSE CONDITIONS 53 COJOCARIU, T., CUTRUBINIS, M., TURTUREANU, A.

TRANSFER OF RADIOCESIUM IN AQUATIC AND FOREST ECOSYSTEMS OF SOUTH-WEST GERMANY 54 DRISSNER, J., KLEMT, E., MILLER, R., ZIBOLD, G.

FERTILIZER AND POLLUTANT POWER OF A SEWAGE SLUDGE: ESSAY IN COLUMNS WITH SANDY SOIL 55 ESTEBAN-MOZO, J., GOMEZ-LUCAS, I., MATAIX, J.

41 A PRELIMINARY CLASSIFICATION OF SOIL ECOSYSTEM AS A MEANS TO ESTIMATE EXPECTED SOIL TO PLANT TRANSFER OF RADIONUCLIDES 56 FRISSEL, M. J.

QUALITY ASSURANCE OF DATABASES FOR RADIONUCLIDE TRANSFER; PROTOCOLS FOR SOIL TO PLANT UPTAKE EXPERIMENTS 57 FRISSEL, M. J.

USE OF STABLE ISOTOPE TECHNIQUES IN SOIL ORGANIC MATTER STUDIES 58 GERZABEK, M.H.

INFLUENCE OF ELEVATED RADIONUCLIDE CONTAMINATION ON NATURAL PLANT POLESSKY STATE RADIOECOLOGICAL RESERVE. 59 GONCHAROVA, N., MATSKO, V., ZHEBRAKOVA, I., MONTIK, T.

ACCUMULATION OF Cs, Sr INTO LEAVES AND GRAIN OF WINTER WHEAT UNDER ACT OF N, Zn, Li, Na 60 GRODZINSKY, D., TKATCHUK, K., ZHMURKO, N., BOGDAN, T., GURALCHUK, ZH.

USE OF STABLE ISOTOPES FOR ESTIMATING WATER AND NITROGEN TRANSPORT IN PLANTS 61 GRYGORYUK I. P., PETRENKO N. I., SHVEDOVA O. YU., TKACHEV V. I., YAROSHENKO O. A.

FERTILISER INDUSTRY IN TURKEY, THE FOLLOWED POLICIES AND THE ECONOMETRIC ANALYSIS WHICH AFFECT THE FACTORS OF PRODUCTION VALUE IN THIS INDUSTRY 62 GURLER, A. Z., KIZILASLAN, H., GULSE, S., UZUNOZ, M.

THE UPTAKE OF NITROGEN BY SPRING BARLEY AND THE BALANCE OF 15N AS AFFECTED BY SOIL pH LEVEL 63 HEJNAK, V., LIPPOLD, H.

APPROACHES TO THE PREDICTION OF RADIOCESIUM UPTAKE BY PLANTS FROM CONTAMINATED SOILS 64 HERREN, T., RIESEN, T.

RELATIONSHIP OF Be IN SOIL TO ITS CONTENT IN SELECTED VEGETABLE PLANTS 65 HLUSEK, J., RICHTER, R. SOIL-TO-PLANT TRANSFER FACTORS: LIMITATIONS OF A SIMPLE CONCEPT 66 KIRCHNER, G., EHLKEN, S.

LONG-TERM INFLUENCE OF LIMING ON BIOLOGICAL AVAILABILITY OF RADIOCAESIUM IN FOREST SOILS 67 KONOPLEV A. V., DRISSNER J., KLEMT E., KONOPLEVA I. V., MILLER R., ZIBOLD G.

COMPARATIVE STUDY OF THE RADIONUCLIDE UPTAKE AND DISTRIBUTION WITHIN PLANTS FOR BARLEY AND MAIZE VARIETIES68 KOSTYUK O.

CENOTIC AND PHYSIOLOGICAL CONTROL OF THE RADIONUCLIDES MIGRATION INTO SYSTEM SOIL-PLANT 69 KRAVETS, A. P.

SOIL AND PLANT FACTORS INFLUENCING THE ACCUMULATION OF RADIONUCLIDES BY THE HIGHER PLANTS. CONCEPTUAL AND MATHEMATICAL MODEL 70 KRAVETS, A. P.

STRESS MODIFICATION OF THE KINETIC OF 137CS ABSORPTION AND PRIMARY PATTERN OF INTERCOMPARTMENTS DISTRIBUTION 71 KRAVETS, A. P.

METHODS AND MEANS OF MODIDFICATION OF ACCUMULATION RADIONUCLIDES BY PLANTS IN CONNECTION OF PHYTODECONTAMINATION OF SOILS. 72 KUTLAKHMEDOV, Y., MIHEEV, A., ZEZINA, N., PAVLENKO, N.

PROBLEM OF A RADIOCAPACITY IN A SYSTEM SOIL - PLANT FOR BOG ECOSYSTEM 73 KUTLAKHMEDOVA-VYSHNYAKOVA V.

THE UPTAKE OF HEAVY METALS (Pb, Cu, Ni, Cr) BY GRAPEVINE 74 LICINA, V., JAKOVLJEVIC, M., ANTIC-MLADENOVIC, S.

OBSERVATIONS OF Cs MIGRATION IN COLUMN EXPERIMENTS: II. FROM SURFACE DEPOSITION 75 MARCHANT, J. K., BUTLER, A. P., SHAW, G., TOMPKINS, J. A., WADEY, P., WHEATER, H. A.

43 EFFECT OF LONG-TERM FERTILIZATION WITH DIFFERENT NORMS AND RATES OF NUTRIENTS ON SLIGHTLY LEACHED CHERNOZEM FERTILITY 76 NANKOVA, M., KIRCHEV, H.

GENETIC VARIATION FOR DRY MATTER AND NITROGEN ACCUMULATION OF DOUBLE HAPLOID WHEAT LINES 77 NANKOVA, M., MILKOVA, V., IVANOV, P.

REDUCTION OF Cs-137 LEVELS IN PLANTS AND FUNGI AFTER POTASSIUM FERTILIZATION IN A SWEDISH FOREST 78 NIKOLOVA, I.

134CS UPTAKE BY PLANTS FROM SOIL APPLYING DIFFERENT ABSORBENTS 79 ONCSIK, M.

TRANSFER OF 226RA TO PLANTS FROM TWO TYPES OF SOIL 80 ROSIAK, L, PIETRZAK-FLIS, Z.

PREDICTING RADIOCAESIUM AND POTASSIUM ROOT UPTAKE USING THE BARBER-CUSHMAN MODEL. 81 ROCA JOVE, M. C, VALLEJO CALZADA V. R.

137Cs AND 90Sr ROOT UPTAKE BY BEANS IN SOILS WITH CONTRASTING PROPERTIES 82 SAURAS YERA, T., VALLEJO, R., WAEGENEERS, N., MADOZ-ESCANDE, CH.

NUTRIENT MOBILITY IN SOIL-PLANT SYSTEM ACCORDING TO SOIL MAINTENANCE SYSTEMS AND NATURE OF FERTILIZERS 83 SERDINESCU, A.

SOIL TO PLANT TRANSFER OF 134CS FOR OLIVE AND ORANGE TREES AFTER FOUR YEARS' EXPERIMENTATION 84 SKARLOU, V., NOBELI, C, ANOUSSIS, J., HAIDOUTI, C.

EFFECT OF NPK FERTILISATION ON 3 APPLE DISEASE RESISTANT VARIETIES GROWN IN CONTAINER 85 STANICA, F.

TRANSFER OF 137CS FROM SOIL TO PLANTS IN DIFFERENT TYPES OF SOILS 86 TODOROVIC, D., POPOVIC, D., RADENKOVIC, M., DJURIC, G. NEW SCALABLE TRANSFER FACTORS: A STUDY OF THE KEY PROCESSES AFFECTING SOIL-TO-PLANT TRANSFER FACTORS AND THE DEVELOPMENT OF A NEW SCALABLE APPROACH 87 TOMPK1NS, J. A., WADEY, P., BELL, J. N. B., BUTLER, A. P., SHAW, G., WHEATER, H. S.

EFFECT OF SPECIFIC AND VARIETAL FEATURES OF CROPS ON CAESIUM-137 ACCUMULATION 88 TSYGANOV A. R., CHERNUKHA G. A., KRUGLENYA V. P.

THE EFFECT OF PUTRIFIED PLANT RESIDUES ON THE GERMINATION SPEED AND POWER OF WHEAT (GEREK 79) 89 UYANOZ, R., ACAR, R., ER, F., BROHI, A. R.

OBSERVATIONS OF Cs MIGRATION IN COLUMN EXPERIMENTS: I. FROM A CONTAMINATED WATER TABLE 90 WADEY, P., BELL, J. N. B., BUTLER, A. P., MARCHANT, J. K., SHAW, G., TOMPKINS, J. A., WHEATER, H. S.

UPTAKE OF RADIOCAESIUM BY LETTUCE CROPS: THE EFFECT OF K IN SOIL SOLUTION 91 WAEGENEERS, N., CAMPS I VILA, M., SMOLDERS, E., MERCKX, R., SAURAS, T. MADOZ-ESCANDE, C.

USING SOIL-TO-PLANT TRANSFER TO IDENTIFY AREAS VULNERABLE TO RADIOCAESIUM DEPOSITION IN WESTERN EUROPE 92 WRIGHT, S., CREAMER, R., SANCHEZ, A., HOWARD. B.

PERSPECTIVES AND POSSIBILITIES OF ECOMODEL IN ESTIMATION AND PROGNOSIS TRANSFER OF RADIONUCLIDES IN SYSTEM SOIL-PLANT 93 YASKOVETZ, I., GIRIY, V., ZAITOV, V.

CHANGES IN SULPHUR BALANCE IN PLANT PRODUCTION OF THE CZECH REPUBLIC 94 ZELENY, F., ZELENA, E., CERMAK, M.

THE ECONOMIC ANALYSIS OF FERTILISER EFFECT ON THE YIELD OF SOME IMPORTANT CROPS GROWN IN TOKAT PROVINCE 40 ESENGUN, K., AKAY, M.

45 Working group No. 3 NOTES COMPARASON OF THE POLICIES OF FERTILISER SUBSIDY AND PRICE SUPPORT FOR SELF- SUFFICIENCY IN SUNFLOWER: A CASE STUDY OF TURKEY AKCA, H., GURLER, A. Z., SAYILI, M. University of Gaziosmanpasa, Faculty of Agriculture, Department of Agricultural Economics, Tokat, Turkey

Today, common goal of Turkey and the world is to provide enough food for the need of the increased population. The role of fertilisation in increasing the crop production is 58 % hence fertilisation is the essential input in the production of Turkish agriculture. This is why Turkish government has been paying subsidy for enhancing the fertiliser use. Sunflower is one of the agricultural products in which Turkey is less sufficient. For example, the self-sufficiency ratio for this commodity was 62% in 1995. In order to achieve self-sufficiency in sunflower, domestic production must rise to the some level as consumption. This can be done either by raising the producer price or by lowering the fertiliser price. The State has given subsidy premium for fertiliser. The subsidy premium was determined as 20% in September 1994 but this rate later was increased to 30 % on 15 October 1994 and 50% in 1995. The fertiliser consumption has not been at the desired level although the increasing rate of subsidy has increased fertiliser consumption. In this study, the amount of domestic production and consumption of sunflower, the amount of fertiliser consumption, world prices of sunflower and fertiliser, yield elasticity with respect to urea use and urea price, and demand elasticity for urea have been used as data. In this paper, policy analysis was used to determine which of these instruments (price support policy) could attain the goal at lower cost to the Turkish government and with minimum losses or maximum gains in social welfare. Working group No.3 NOTES EFFECT OF NP FERTILIZATION ON THE YIELD, YIELD COMPONENTS AND NPK CONTENT OF WHEAT CROP GROWN UNDER TOKAT ECOLOGICAL CONDITIONS BROHI, A. R., AKTAS, A. SAVASLI, E. Soil Science Department, Agricultural Faculty of Gaziosmanpasa University, 60250 Tokat, Turkey

A study was carried out to see the effect of different rates of NP fertilization on some agronomic parameters, yield and NPK content of wheat crop. N rates significantly increased the single head weight from 1.01 cm (control) to 1.61 cm (360 kg N/ha), earhead length from 6.29 cm (control) to 9.94 cm (240 kg N/ha), spikelet number from 13.31 (control) to 19.01 (240 kg N/ha), number of kernels in spike from 24.70 (control) to 35.99 (360 kg N/ha), plant height from 61.22 to 94.60 cm (360 kg N/ha), 1000 kernel weight from 44.60 gram (control) to 48.50 gram (240 kg N/ha) and hectolitre weight from 80.63 kg (control) to 82.69 kg (120 kg N/ha), respectively. P rates have no significant effect on these parameters, except hectolitre weight. P rates significantly increased the harvest index from 13.86 to 17.15% in control and at 160 kg P2O5/ha rate, resp. N rates significantly increased grain yield from 896 kg to 3073 kg/ha in control and at 240 kg N/ha, whereas P rates increased grain yield from 2023 kg (control) to a maximum of 2618 kg/ha at 160 kg P2Oij/ha rate of application. N and K content of grain increased with increasing rates of NP fertilization. P content of grain decreased with N rates and increased with P rates. Increasing rates of P increased the P content of soil after harvest of crop. A P content of 8.24, 8.24, 12.20 and 13.53 was found at 0, 50, 100 and 150 kg P2Os/ha rates, respectively.

47 Working group No.3 NOTES EFFECT OF NP FERTILIZATION ON THE YIELD COMPONENTS AND NPK CONTENT OF MAIZE CROP GROWN UNDER TOKAT ECOLOGICAL CONDITIONS BROHI, A. R., AKTAS, A. SAVASLI, E. Soil Science Department, Agricultural Faculty of Gaziosmanpasa Universiuty, 60250 Tokat, Turkey

A study was carried out to see the effect of different rates of NP fertilization on some agronomic parameters, yield and NPK content of maize crops. N rates significantly increased the cob diameter from 4.58 to 4.74 cm, cob length from 21.00 to 22.19 cm in control and at highest N rate of 360 kg/ha, respectively. P rates have no effect on these parameters. N rates significantly increased the maize grain yield from 10305 kg/ha (control) to 13896 kg/ha (240 kg N/ha) and P rates increased the grain yield from 9399 kg (control) to 15824 kg/ha (240 kg P2O{5/ha). The N rates increased the content of grain N from 0.94 to 1.17% and P from 0.24 (control) to 0.29% (highest rate). After the harvest of the crop the soil analysis results showed that N rates increased the NO3N content of soil (at 0-20 cm depth) from 9.09 to 56.00 ppm, from 8.76 to 30.68 ppm NO3N (at 20-40 cm depth) and from 5.94 to 20.1 ppm NO3N (at 40-60 cm depth) for control and 240 kg N/ha, respectively. The P rates increased the NO3N content at 0.20 cm depth and decreased it at 20-40 cm depth, and again increased it at 40-60 cm depth when compared to control treatment. However, after the harvest NP fertilization has no significant effect on available P content of soil. However, an average of 10.89, 5.88 and 3.23 ppm P was found at 0-20, 20-40 and 40-60 cm depth, respectively.

48 Working group No.3 NOTES EFFECT OF DIFFERENT PHOSPHORUS FERTILIZERS ON YIELD AND NPK CONTENT OF WHEAT CROP GROWN UNDER ECOLOGICAL CONDITIONS OF TOKAT/TURKEY BROHI, A. R., TOPBAS, M. T., SAVASLI, E., AKTAS, A. Soil Science Dept., Agricultural Faculty of Gaziosmanpasa University, 60250 Tokat, Turkey

The main object of this study is to compare the effect of different phosphorus fertilizers on yield, yield components and NPK of wheat crop grown under ecological conditions of Tokat, Turkey. Increasing rates of P increased the grain yield from 2050 kg/ha (control) to 2696 kg/ha (100 kg P2O5) with no effect among phosphorus fertilizers. P rates significantly increased spike length from 8.13 (control) to 9.06 cm (150 kg P2C>5/ha), number of kernels in spike from 32.97 (control) to 37.50 (150 kg P2O5/ha), weight of 1000 grains from 47.05 g (control) to 49.22 g (150 kg P2C>5/ha), harvest index from 15.63% (control) to 17.93% (50 kg P2C>5/ha). P rates have no effect on number of spikelets, single head weight and length of ear head. No significant effect for these parameters was observed among P fertilizers. P rates significantly increased the N content of grain from 1.23 to 1.63% and the P content from 0.45 to 0.51% and decreased the K content from 0.44 to 0.38% in control and 150 kg P2O5/ha rates, respectively. Similarly grain protein content increased from 7.11 to 9.38% at the above rates. Triple superphosphate fertilizer gave the highest protein content of 8.980 as compared to 8.37% of DAP and 8.26% of composite (20:20:0) fertilizer. After harvest of the crop the soil P content was found to be 0.44, 10.59 and 12.84 ppm with DAP, TSP and 20:20:0 P fertilizer, respectively.

49 Working group No. 3 NOTES RESEARCHES - WITH IEFERT MODEL - CONCERNING THE VARIATION OF PHYSICAL AND ECONOMICAL EFFICIENCY OF FERTILIZERS USE BUDOI, G. H. University of Agricultural Sciences and Veterinary Medicine, Department of Agrochemistry, Bd. M3ra§ti 59 sect. 1, 71331 Bucharest, Romania

The lEFERT model allows to do researches concerning the variation of physical and economical efficiency indexes of fertilizers use, in a crop series (winter wheat, corn, sun-flower, soya-been, potato, etc), related to the applied fertilizer rate, the cost of fertilization and soil nutrient supply, using the Mitscherlich-Baule function (in ICPA modification), MB-ICPA, as response function and when is used the relationship derived from this function to calculate the optimum economic fertilizers rates - DOE; the paper presents the results for winter wheat. lEFERT use the ICPA system to establish the action coefficients and soil nutrient supplies as a function of relevant agrochemical indexes, based on long-term field experiences. The yield increase rise toward a maximal value, at the same time with rate of fertilizer increase, according to the law of diminishing returns, while the average specific yield increase, SSp (kg/kg N, P2O5 or K2O), and the marginal yield, PMg, decrease continuously at the same time with the rate increase. The technical coefficient of the average specific yield increase, CSp (kg N, P2O5 or K2O/1 kg average yield increase), as weil as the technical coefficient of the marginal yield increase, increases simultaneously with the rate. The elasticity of the yield increase, Ep, is continuously decreasing toward a minimum level. The fertilization costs/kg N, P2O5 or K2O, decreases continuously with the rate, while the costs/kg yield increase decreases rapidly first, becomes minimal for the rate corresponding to maximal return per dollar invested, DMPS, and then increases continuously; this index can be used as criteria to establish DMPS. The total profit/ha obtained by fertilization increases simultaneously with the rate with diminishing returns, riches the climax at DOE, and then decreases gradually. The K rates for which the profit comes to have positive values are greater then for P, and the P rates are greater then N rates. The profit per dollar invested, PSp, varies both with the fertilizer rate and the fixed costs for fertilizer application, CF. For high CF values and small rates, the PSp is negative, then increases rapidly with the rate, reaches the maximum at DMPS, then decreases continuously slower and slower. CF highly influences PSp; a high dispersion of PSp values takes place as a function of CF with small rates; the differences decrease with the rate increase toward the DOE. PSp decreases simultaneously with the CF increase. The higher negative influence of CF on PSp occurs for small fertilizer rates. As long as CF is higher as much PSp reaches the peak for a higher percentage of DOE. A maximum PSp on a medium N, P and K-supplied soil is, in N, for rates between 10 and 25 % from DOE-N, in P for rates between 18 and 43 % from DOE-P, and in K for rates between 50 and 100 % from DOE-K. In comparable conditions, DMPS-K > DMPS-P > DMPS-N. The marginal profit per dollar invested, PrMg, is negative for small rates, increases rapidly to a maximum, then decreases continuously with a rate smaller and smaller, becomes zero for DOE, then becomes negative. The profit/ha and the profit/dollar invested, as well as the PrMg are, for comparable rates, higher in N then for P and in P then for K. Working group No. 3 NOTES MODELLING THE CROP RESIDUES AND ORGANIC FERTILIZERS MINERALIZATION IN SOIL BUDOI, G. H.\ DANUSO, F.2, GIOVANARDI, R.2, DAVIDESCU, V.1, MADJAR, R1. 1University of Agricultural Sciences and Veterinary Medicine, Department of Agrochemistry, Bd. MaYa§ti 59 sect. 1, 71331 Bucharest, Romania 2Udine University, Department of Vegetal Production and Agricultural Technologies, Via Delle Scienze 33111, Udine, Italy

In the Nitrogen CROp Simulation model - NICROS, the soil organic matter is considered as being constituted by the following fractions: humus, crop residues and applied organic fertilizers; as concerning the crop residues two fractions are taken into account: the easily decomposable fraction - 77 % of the total dry matter - and the resistant decomposable fraction - 23 %. Each fraction of soil organic matter has specific mineralization rates. Knowing that organic fertilizers have effect for many years, the yearly amount decomposed from the total applied is computed as a function of number of years from application; by mineralization of medium fermented manure, in the first year is mineralized about 33 % of the total nitrogen, during the second year around 20 %, and in the third year 15%. The mathematical model used for the soil organic matter mineralization is practically a quadruple exponential model, whose precision is higher then of the simple or double exponential models used in literature. In order to estimate the nitrogen mineralized in time, the model computes the amounts of organic matter mineralized from each fraction based on the specific mineralization rates, dynamic rates which change as related of the main abiotic factors: soil temperature, moisture, clay content. Then, starting from the specific nitrogen content of the fractions, it compute the total amount of nitrogen mineralized from each of them. This is what essentially distinguishes the NICROS model from others models. This paper presents the differential equations used to compute the daily amounts of residues and organic fertilizers mineralized and the equations used to compute the amounts mineralized in a given period of time; the computing equations for the corresponding amounts of nitrogen mineralizations are also showed. It deals with the influence of abiotic factor - temperature, soil moisture, day content - on the specific mineralization rates and present the original relationships used by the NICROS model to describe these influences. Finally, presents simulations done for real conditions of the reddish-brown soil from Moara Domneasca\

51 Working Group No. 3 NOTES FRUITS CONTAMINATION FOLLOWING AERIAL DEPOSITION AND REMAINING ACTIVITY AFTER WASHING OR PEELING CARINI, F., ANGUISSOLA SCOTTI, I., D'ALESSANDRO P. G. Universita Cattolica de! Sacra Cuore, Institute of Agricultural and Environmental Chemistry, Faculty of Agricultural Sciences, Via Emilia Parmense, 84,1-29100 Piacenza, Italy, E-mail: [email protected]

The contamination of fruits by aerial pathway can result from direct deposition of radionuciides on the fruit surface or from deposition on other above-ground parts of the plant. Directly deposited radionuciides can be absorbed by the fruit skin and transported to the interior. The surface contamination of fruits can also be transferred to internal tissues during peeling. Radionuciides deposited on the other aerial parts of the plant can be absorbed by them, mainly by leaves, and translocated to fruits. The concentration of radionuciides in fruits can, in its turn, be affected by domestic or industrial processes, removing certain parts of the raw food. Fruits can be eaten fresh as picked, after washing or peeling, natural dried or squeezed, or after processing by the food industry. Neglecting the losses during food processing can lead to overestimation of the calculated dose. The knowledge of the distribution of radionuciides in pulp and skin is useful to develop methods dealing with reduction of radionuciides in food in case of accident. The aim of the present research was to assess: the rate of translocation of radionuciides from leaves to fruits, the loss of direct deposited radioactivity on fruit in time, the loss from fruits after washing, peeling or squeezing. Apples, pears, and grape vines were grown in large pots kept in open field and protected by an antihail net. Plants were contaminated via leaves and fruits with 134Cs and 86Sr, both in form of chloride salts in aqueous solution. The foliar and fruit contamination was carried out on 20 July, when plants were bearing well-developed green fruits. Pears were contaminated only via leaves, because plants bearing fruits were not sufficient for replicates. The contamination was carried out by putting microdrops of the radioactive solution on 10 leaves of 3 branches or on the upper half of 3 fruits. The activity administered/plant was 110.1 kBq of 134Cs and 221.5 kBq of 85Sr. Fruits and leaves were picked at ripening on 10 September 1996, separating treated and non-treated fruits and leaves. Fruits were peeled or squeezed or washed. Fruits, their fractions and leaves were analysed separately by gamma spectrometry. Translocation from leaves to fruits is from one to two orders of magnitude higher for 134Cs than for 85Sr. There is also some evidence of translocation of both radionuciides from fruits to leaves. The loss of 134Cs and 85Sr from leaves after direct deposition is of about 96-97% in apples and pears, but is higher in vines. Generally speaking the residual activity in fruits after direct contamination is lower than 1% of that applied both for 134Cs and for 85Sr. Data suggest that Sr, like 134Cs, tends to concentrate most in peel, even if 134Cs seems to be partially translocated into the pulp. Washing of spiked fruits removes more 134Cs than 85Sr from peels. Working group No.3 137CS SOIL - WHEAT PLANT TRANSFER IN GREENHOUSE CONDITIONS CZ9928618 COJOCARIU, T., CUTRUBINIS, M., TURTUREANU, A. Institute of Food Research, Food Biophysics Laboratory, Garlei 1, 71576 Bucharest, Romania

The paper presents the results of 137Cs soil-wheat plant transfer in greenhouse conditions. Soil to plant transfer factors of Cs were determined in pot experiments, using a loamy soil with known properties. The variants of work were: A. control soil: (40K:603,02+49,45 Bq/kg dry weight; 137Cs: 17,13+2,57 Bq/kg dry weight) B. 137Cs contaminated soil:(40K:657,64+64,45 Bq/kg dry weight; ™Cs: 1248,11 +74,89 Bq/kg dry weight) 137 40 137 C. Cs contaminated soil and then KNO3 treated ( K:745,37+60,00 Bq/kg dry weight; Cs:961,09+57,67 Bq/kg dry weight) The wheat was sown three months after soil contamination. The 137Cs and 40K concentrations were measured by y spectrometry in 30 days old wheat. The experiments were repeated for three growing periods. The soil to wheat plant transfer factor of 137Cs for the A variant was 0,170 and for the B variant was 0,075. Adding potassium to a soil contaminated with radiocaesium (C variant), the radiocaesium transfer factor is reduced at about 0,019.

53 Working group No. 3 TRANSFER OF RADIOCESIUM IN AQUATIC AND FOREST ECOSYSTEMS OF SOUTH-WEST GERMANY CZ9928619 DRISSNER, J., KLEMT, E., MILLER, R., ZIBOLD, G. FH Ravensburg-Weingarten, University of Applied Sciences, Center of Radiometry, P.O. Box 1261, D-88241 Weingarten, Germany, E-mail: [email protected]

The contamination of the natural environment with radionuclides from the Chernobyl fallout in the south-west of Germany is among the highest in central Europe. Radioecological transfer processes of cesium radionuclides in aquatic and forest ecosystems have been investigated since 1987 and will be presented. Different limnological properties of lakes determine the time-dependency of the contamination of water and of fish. In Lake Constance, a large and deep mesotrophic hardwater lake, cesium radionuclides were rapidly removed from the water column. For whitefish an effective half-time of the specific 137Cs activity of about 4 months was observed and carnivorous fish never exceeded the maximum contamination of herbivorous fish. In Vorsee, a small and shallow eutrophic lake, even in 1997 the contamination of the water was in the order of 100 Bq/m3, a factor of 500 higher than in Lake Constance. As a consequence, the general contamination of fish is higher than in Lake Constance, and e.g. the contamination of pike decreases with an effective half-time of about 3 years. The surveillance of the roe deer contamination shot within an area of 40 x 40 km2 was performed by analyzing more than 6300 meat samples. The seasonal structure of the roe deer contamination shows a general decline which is superimposed by periodic maxima in autumn correlated with the mushroom season. Neglecting the autumn maxima, the geometric means of the aggregated 137Cs transfer factors soil - roe deer of a selected region show an ecological half-time of about 3.5 years. Similar values are found for different grazing plants. The binding of radiocesium at the organic matter of the top forest soil horizons is considered to be decisive for the transfer to plants and finally to roe deer. As a result of a forest fertilization in 1984 reduced transfer factors soil - plant were still observed in 1997. Working group No. 3 NOTES FERTILIZER AND POLLUTANT POWER OF A SEWAGE SLUDGE: ESSAY IN COLUMNS WITH SANDY SOIL

ESTEBAN-MOZO, J.f GOMEZ-LUCAS, I., MATAIX, J. University of Alicante, Department of Agricultural Chemistry and Biochemistry. Ap. Correos 99, E-03080 Alicante, Spain.

The potential for contaminants present in sewage sludge to leach and cause groundwater contamination as well as nutrient status of a sandy soil following sludge application to agricultural land have been assessed. Three treatments were carried out in columns which represents three rain simulations: A (annual mean), B (two times annual mean) and C (ten times annual mean). A comprehensive analysis was performed: -.Soil (texture, pH, electric conductivity, OOC, OOM, polychlorinated dibenzo-p-dioxins and dibenzofurans, Cd, Cr, Pb, Zn, Cu, Ni, Mn, + 2 Fe, Co, K, Na, Ca, Mg, NH4 , NO3", NO2", SO4 ", Cl", HCO3> Leached (pH, electric conductivity, Cd, Cr, Pb, Zn, Cu, + 2 Ni, Mn, Fe, Co, K, Na, Ca, Mg, NH4 , NO3", NO2", SO4 ~, CI", HCO3"). However, the experiment indicated that under routine operational practice with typical sludge application rates, and the usual range of compound concentration detected in sludge, contaminants concentration are unlikely to be hazardous. Furthermore, although nutrient status of the soil was improved (OM, N, P, Mg), sewage sludge could not be used as a only fertilizer.

55 Working group No. 3 A PRELIMINARY CLASSIFICATION OF SOIL ECOSYSTEM AS A MEANS TO ESTIMATE EXPECTED CZ9928620 SOIL TO PLANT TRANSFER OF RADIONUCLIDES FRISSEL, M. J. Private consultant, Torenlaan 3, NL-6866-BS Heelsum, Netherlands

There exists a wealth of soil to-plant transfer values (TF's) which describe the uptake of radionuclides by various crops. Existing files of these data are not structured and therefore application of these data to estimate a transfer for a particular crop at a particular soil of for a scenario in the near of far future is difficult. To facilitate the estimation of expected soil to plant transfer factors a soil ecosystem classification system has been developed Estimates of transfers can be based on these classes. The classification is based on reference TF- values. For Cs, cereals appeared to be a suitable reference crop, for Sr green vegetables were used. To derive a classification it should be convenient if of each ecosystem a reference value was available, e.g. the TF of transfer flux of cereals. In fact of many soils cereals can not be grown, from other locations data of cereals are missing. Therefore a concept of conversion ratios has been developed which allows the calculation of the TF value of a particular crop group from data of other crop groups. The term ecosystem is used in a very narrow sense. A coil ecosystem is considered here as a set of coil and ecological conditions which define a particular uptake regime. Such an ecosystem may have been used to determine TF values of one or more crop groups. If a soil is fertilised the uptake conditions change, and therefore a fertilised soil is considered as another ecosystem than the same soil but not fertilised. Conversion of the TF value of one crop group is only possible if all TF's are expressed as the same units. The TF may be expressed as (Bq/kg crop)/(Bq/m ) or as (Bq/kg crop)/(Bq/kg soil). In the latter case the definition of the IUR is useful. It states that the amount of radionuclides which is present in the actual rooting zone is assumed to be present in the top 20 cm layer of soil. For grass the top 10 cm layer is used. A TF based on the concentration in the actual rooting zone is not useful, in this case TF's can not be compared to each other and conversion ratios can not be calculated. It is customary to distinguish between equilibrium transfer data and time dependent transfer data. In fact the availability of both Cs and Sr decreases with time during 20 to 30 years (The longest time series available). Therefore it might be more appropriate to use transfer parameters for ..Steady state releases" and ..Accidental releases". Steady state release values can be used for the assessment of releases for normal operation of a nuclear power plant or for releases from a waste depository. The classification, presented in detail in the full paper, is based on environmental threshold values, rather than on regression analyses. Results are preliminary and need further consideration.

56 Working group No. 3 QUALITY ASSURANCE OF DATABASES FOR RADIONUCLIDE TRANSFER; PROTOCOLS FOR SOIL TO PLANT UPTAKE EXPERIMENTS CZ9928621 FRISSEL, M. J. Consultant radioecology, Torenlaan 3, NL-6866-BS Heelsum, Netherlands

1. Introduction Radioecologists are faced with some specified problems on quality assurance. Sometimes it is clear that experimental or interpretation errors occur. E.g. if someone reports a difference of the behaviour of P-31 and P-32, or a difference between Sr-89 or Sr-90 or deviating behaviour of one of the Cs nuclides. All such effects appeared to be artefacts, and radioecologist do not have to spent time on a possible explanation. But what to do if someone reports a transfer value which is outside the expected range? Even if the experiment is repeated and the strange effect disappears one is never sure that the unexpected result was not caused by some special ecological condition which was overlooked. An obvious recommendation is to carry out all experiments according to a protocol so that artefacts are avoided and all important ecological conditions are collected. One might think that phyhicist have shown radioecologists the way how to develop protocols. A detailed instruction how to carry out an expenment is coupled to standardised experiments. All participating scientists carry out the same experiment applying the same experimental conditions. The result is that physicist reach accuracy's of which radioecologists can dream only. One might think that uptake experiments, carried out ion identical growing chambers by a number of radioecologists, may show the way on how to do reliable experiments. This it not as simple as it looks. Even if, with the same soil, within the same growth chamber e.g. 6 repetitions of a simple growth experiment are being repeated it is difficult to obtain the same yields, in particular if products as grain and potatoes are cultivated. One might argue at the uptake of radionuclides, if expressed per gram product, will vary less than the yield. This is probably true and probably it is possible to develop, provided the type of soil and crop are carefully selected, a standard uptake experiment which can be used as a test to verify someone's ability to do uptake studies in a growth chamber. Yet this does not solve the problem. For assessment calculations one need locally determined parameters, which reflect the local farming practice. Soils, locally preferred crop varieties, types of management, they ail differ from location to location. A radioecological protocol will therefore have to differ very much from a physical protocol. In a next section a protocol is described which is based on a recent IAEA-IUR project [1].

2. Protocol for radioecological soil to plant uptake experiments This protocol is based on an earlier protocol by the IUR Working Group Soil to Plant Transfer, on recommendations of IAEA and, mainly, on the IAEA-IUR CRP on the determination of transfer parameters in tropical environments. At a workshop of IUR on the production of a database of radionuclide fluxes some modifications were made to meet requirements for the determination of time dependent parameters.

57 Working group No. 3 USE OF STABLE ISOTOPE TECHNIQUES IN SOIL ORGANIC MATTER STUDIES CZ9928622 GERZABEK, M.H. Austrian Research Centers, A-2444 Seibersdorf, Austria

Due to the rapid development of mass spectrometry and its applications, the use of stable isotope techniques in environmental studies, plant nutrition and soil science has increased considerably during the last decades. Both, applying stable isotopes as tracers or using the natural abundance, especially of C and 15N, opens the chance of quantifying even single processes within the complicated soil system. Plants differ distinctly in their C-isotopic composition. The largest difference occur between plant species with different photosynthetic pathways. C3- and C4-plants are differentiated by approximately 14%o on the 5-scale (app. - 27%o13C versus -13%o13C). Modern elemental analyser - mass spectrometer combinations reach acurracies of at 13 least 0.1 %o 5 C. Therefore, the difference between C3 and C4 plants is sufficient to be used for tracer studies. Several investigations of SOM turnover under field conditions were undertaken using the fact that the vegetation cover changed between C3 and C4 plants (Baldesdent and Mariotti, 1996). Cerri et al. (1991) described the discrimination between SOM originating from indigenous vegetation (forest, C3) and sugar cane (C4) after 50 years of cropping introducing two SOM compartments of different stability. Another example is the change from prairie vegetation (C4) to different C3-crops and the evaluation of the carbon origin at or near equilibrium (Wagner, 1991). 15 13 More recent studies use N-labelled C4-plant residues or C-labelled C3-plants (Hopkins et al., 1997) to elucidate the fate of carbon and nitrogen in soils developed under C3-vegetation. Both in situ experiments (Aita et al., 1997) and laboratory incubations (Stemmer et al., 1998) were used to evaluate carbon and nitrogen fluxes from crop residues. Physical fractionation of bulk soil into particle sizes proved to be of advantage to follow short and long- term dynamics of crop residues within SOM. Changes in the natural abundance of 13C and 15N within soil profiles can elucidate leaching or mineralization of humic substances (Gerzabek et al., 1989). Changes in the natural abundance of stable isotopes are also possible due to the application of organic manures, quantification, however is not easy because of the small isotopic differences between soil and manure carbon and nitrogen (Gerzabek et al., 1997). 15N labelling of soil nitrogen has been widely used in the last two decades to quantify biological nitrogen fixation. Considerable progress has been made due to the isotope dilution method in estimating absolute rates of mineralisation, immobilisation, nitrification and nitrate reduction in soil, which cannot be directly deduced from non- tracer experiments. A very interesting topic seems to be the measurement of nitrogen released from organic residues, like N-fixing plants (Elwaraky and Haunold, 1991), sewage sludge (Gerzabek et al., 1997) or other manures. Further potential of stable isotope techniques in SOM dynamics studies can be envisaged in the field of quantifying the pools contributing to N-mineralization, the role of soil microbial biomass in this context, the sulphur cycle ( S), climate change and the impact of nitrogen oxide pollution on SOM dynamics, especially in semi-natural environments. Working group No.3 INFLUENCE OF ELEVATED RADIONUCLIDE CONTAMINATION ON NATURAL PLANT POLESSKY CZ9928623 STATE RADIOECOLOG1CAL RESERVE. GONCHAROVA, N., MATSKO, V., ZHEBRAKOVA, I., MONTIK, T. Institute of Radiobiology of the Academy of Sciences of Belarus, Plant Radiobiology Laboratory, Zhodinskaya str 2, 2200141 Minsk, Belarus, E-mail: [email protected]

Our investigations confirm that plant complexes as a whale are relatively resistant to radiation exposure. The majority of the species on the absence of visible violations on population and ecosystematic levels in plants communities it is impossible to rule out the possibility of changes in phytocenosis with predominance of the most radioresistant species. The group of meadow dominants was selected-representatives of families of Gramineae, Compositae, Primulaceae, Rosaceae in which the137 Cs migration from soil to overgrown phytomass relates closely with the sum of atmospheric precipitation's. It is necessary to take into account that in the conditions of chronic irradiation the vegetation of majority of meadow dominants (representatives of families of Gramineae and Leguminosae ) is completed by the formation of valued seed posterity able to produce a new generation. The radiology situation in meadow grassland was evaluated in the territory of Polessky State Radioecological Reserve. In a 9-year population monitoring experiment was observed that the radiosensitivity of different plants species is different . This fact determined by the different specificity of genetic systems and bioecological peculiarities of species. The plants species with narrow ecological amplitude, high ploidy, apomictic breeding are the most radiosensitive, ones as well as the plants which grow in Southern Belarus as a limit of there natural dissemination. Decrease in number was noted for the majority of such species, or elimination from plants communities. The anthropogenic load removal from the evacuation territories followed by the radical phytocenoses reconstruction is of important ecological significance as the ionising radiation effect. It may be inferred that long - time chronic action of radionuclides on plants in the fall out zone will depend on specific features of their accumulation by some plant species, the age related radiosensitivity and some other factors, associated with their growing conditions such as soil types, forms of radionuclide fall out, chemical and physical effect.

59 Working group No.3

ACCUMULATION OF Cs, Sr INTO LEAVES AND GRAIN OF WINTER WHEAT UNDER ACT OF N, CZ9928624 Zn, Li, Na GRODZINSKY, D., TKATCHUK, K., ZHMURKO, N., BOGDAN, T., GURALCHUK, ZH. Institute of Plant Physiology and Genetics, National Academy of Sciences of Ukraine, 31/17 Vasylkivska St., Kyiv, 252022, Ukraine; E-mail: [email protected]

The aim of our research is to study the effect of nitrogen and microelements Zn, Li, Na on Cs, Sr accumulation into leaves and grain of winter wheat plants. The experiments have been carried out on cv Lutencens 7 winter wheat grown on grey forest soil. In order to study the influence of nitrogen on Cs and Sr accumulation a background of P60 K60 added in autumn different doses of nitrogen (30,60,120 kg/ha) were applied in spring. The influence of micronutrients on Cs and Sr accumulation was studied by addition 3 kg/ha Zn and 2 kg/ha Li into soil under ploughing on background of N60 P60 K60 Besides the foliar application with 0,05% Na2SO4 was carried out. Cation content (Cs, Sn, Zn, Li, Na) in soil and plant organs was determined by atom absorption spectrophotometer AAS-30. It has been found that Cs, Sr content in control plant leaves made up 15,0 and 21,0 mg per 1g of dry matter at the early stages of plant development. As plants aged, the content of those elements into leaves decreased strongly (3- 4 times). At early stages of plant development, nitrogen caused the increase in Cs content into leaves by 8,9-11%. At the stages of heading to grain filling the increase of Cs content was obserwed only at high nitrogen dose, while low nitrogen doses had no effected on Cs accumulation in leaves. In should be noted that nitrogen (N60 and N120) decreased Cs content in grain by 32-33%. As for Sr content into grain one must note that its amount was 3 to 4-fold less than that of Cs. It is important to note that nitrogen had no effected on Sr content in grain. Zn and Li addition into soil as well as foliar nutrition with Na had different effect on Cs and Sr content in winter wheat leaves and grain. The addition of Li decreased Cs and Sr content into old leaves by 13% and 25% accordingly. The addition of Zn and Na decreased Sr content into old leaves but not effected on Cs content in them. Zn, Na and Li reduced Sr content in grain also, by 16,11 and 7% accordingly. So our research has been shown possibility of Cs and Sr accumulation regulation into above - ground organs of winter wheat plants. Working group No.3 USE OF STABLE ISOTOPES FOR ESTIMATING WATER AND NITROGEN TRANSPORT IN PLANTS CZ9928625 GRYGORYUK I. P., PETRENKO N. I., SHVEDOVA O. YU., TKACHEV V. I., YAROSHENKO O. A. Institute of Plant Physiology and Genetics of the National Academy of Sciences, Vasilkovskaya Str, 252022 Kiev, Ukraine

Reculiarities in the response of different wheat cultivars and maize hybrids to water deficiency were studied in laboratory and vegetation experiments. Their resistance to external environmental factors was estimated by changes in nitrogen (^N) and water (HDO) accumulation, transport and distribution in plant organs. The water supply was maintained at 60% FWC (control) and was reduced to 30% FWC (experiment) in the absence of plant watering during different stages or with use of polyethylenglycol. Decrease in water potential of medium from -O.05 (control) to -0.5, -0.9 and -1.6 MPa resulted in inhibition of water absorption, transport and distribution in spring wheat organs. After 24-hour stress, root absorption of water of drough-resistant varieties as compared to non- drought resistant ones was more sensitive, during 5, 10 and 15 min intervals after HDO introduction in nutrition medium. Strong depression of water-exchange was observed at weaker stress in non-resistant variety. HDO absorption of the low part of the stem at short exposure resembled that of roots. The 24-hour stress revealed the tendency to sharper inhibition of absorption of labelled water in leaves of resistant variety. At a more durable stress the intensity of leaf water-exchange resistant variety was stabilized, while in non-resistant variety it was reduced considerably. The intensity of HDO and 15N exchange under stress conditions depended on the lability of regulator mechanism of water transport. Genotypic specificality of N use by wheat and maize plants depending on water supply and inclusion of ^$N in total and protein N was found. The 15N content in total N in spring wheat cultivars under optimum water supply and under drought made up 3.65 to 6.20; 1.69 to 3.47, respectively. The ^N content in protein N under the above conditions was 3.03 to 5.96 and 2.36 to 2.93, resp. At water stress the main mass of labelled N in plant roots and stems was localized, while its intake into reproductive organs was inhibited. The presented results may be used for clearing up of mechanism of water and nitrogen transport under conditions of insufficient water supply and for the maximum realization of genetic potential of the cultivar and hybrids in their adaptation to drought.

61 Working group No.3 NOTES FERTILISER INDUSTRY IN TURKEY, THE FOLLOWED POLICIES AND THE ECONOMETRIC ANALYSIS WHICH AFFECT THE FACTORS OF PRODUCTION VALUE IN THIS INDUSTRY GORLER, A. Z., KIZILASLAN, H., GULSE, S., UZUNOZ, M. University of Gaziosmanpasa, Faculty of Agriculture, Tokat, Turkey

Fertilizer industry has become encouraging industry, as it is one of the most important input of agriculture in the planned term. With its particular potential and technological level this industry has the power of competition. It faces some problems as for the production, investment, trade, marketing and other activities. In addition to the kinds of producing final goods; differences in raw material and semi-manufactured goods result in a complex sector in the production process. Importing fertilizer which is the need of agriculture sector and importing semi-manufactured goods and sum of raw material which is used in fertilizer production has an important place imported goods. In this study, in the years between 1986-1996, production, demand, raw material, foreign trade, number of firms and employment have been investigated together with the problems and development of the fertilizer industry which has a complex structure and is dependent on foreign trade trend equation has been calculated to define the position of the sector in future. In addition to this a model was created to determine production and investment with the factors concerning the sector, which affect the production values. For this model it was made use of Cobb-Douglas Production Function. As a results, using the important factors that affect the production value of the sector, suggestions were brought an investment and production policies.

62 Working group No. 3 NOTES THE UPTAKE OF NITROGEN BY SPRING BARLEY AND THE BALANCE OF 15N AS AFFECTED BY SOIL pH LEVEL HEJNAK, V.1, LIPPOLD, H.2 1Czech University of Agriculture, Departament of Botany and Plant Physiology, 165 21 Praha, Czech Republic 2Saxon Provincial Institute for Agriculture, Gustav - Kuhn - Str. 8, 04159 Leipzig, Germany

In the years 1996 and 1997 was studied the effect of soil pH level ( pH > 6,5 and pH < 4,5) and nitrogen fertilization (rates of 0,85,170 and 255 mg N per pot, i.e. 0, 30, 60 and 90 kg N.ha"1) on the uptake of nitrogen and the balance of 15N in system soil - plants. Spring barley, Jubilant variety, was grown in small - plot fields trials in pots (area 1/35 m2) without bottom recessed in soil profile at the experimental station. It was by 15 plants per pot. Ammonium sulphate was used as the N fertilizer, enriched 25% by stable isotope 15N. The total nitrogen uptake by spring barley was much more higher in soil with a better fertility. For instance, the spring barley intaked on the average yearly on the acid soil from 591 to 714 mg N per pot and on the neutral soil from 813 mg (at control variation) to 1.157 - 1.188 mg N per pot (with growing rates of nitrogen fertilizer). The nitrogen fertilization in soil with neutral reaction has a direct influence on higher intake of soil - contained nitrogen by plants. This so - called ..priming effect reached 236 -338 mg N per pot. ..Priming effect didn't manifest or little only (to 56 mg N per pot) in the soil with low pH level. The share of soil nitrogen in the total uptake by the harvest of spring barley was a very high and it was between 97 and 91 % on the soil with high pH level and between 97 and 89% on the soil with low pH level. With growing rates of nitrogen fertilization the share of soil nitrogen decreased. The uptake nitrogen from fertilizer is dependent on soil pH level and rates of nitrogen fertilization. The spring barley intaked on the average yearly on the soil with neutral reaction 37-67-108 mg N, but on the soil with acid reaction only 20 - 47 - 64 mg N from fertilizer at the application ammonium sulphate containing gradated rates of nitrogen - 85,170 and 255 mg N per pot. The balance of fertilizer nitrogen (average for two years in % of supplied nitrogen) show that the use of nitrogen from fertilizer by spring barley is higher in soils with higher pH level (39 - 45 %) compared with the soil of low pH level (24 - 27 %). At higher pH level rested in soil much more nitrogen of fertilizer (27 - 37 %) than at acid soil (15 %). That is why in soil with low pH level was loss of nitrogen fertilizer 58 - 61 %, but in neutral soil 24-31 % only.

63 Working group No. 3 APPROACHES TO THE PREDICTION OF RADIOCESIUM UPTAKE BY PLANTS FROM CZ9928626 CONTAMINATED SOILS HERREN, T., RIESEN, T. Paul Scherrer Institut, Dept. of Radiation Protection and Waste Management, Radioecology Group, CH-5232 Villigen-PSI, Switzerland. E-mai!: [email protected]

Due to the accident at the Chernobyl Nuclear Power Plant in 1986 radiocesium was deposited over large areas of the European continent. Meanwhile it was shown that in most soils radiocesium was almost completely retained in the uppermost soil layers. An extensive uptake by crop plants could result in an accumulation of this element in the human food chain. Due to its chemical similarity to the macronutrient potassium, plants may take up radiocesium. Nevertheless, radiocesium contents detected in crop plants are much lower than expected from total soil contents. The ratio between dissolved and sorbed radiocesium in the soil is expressed by the Ko-value which is usually used for modeling of radiocesium soil-to-plant transfer. However, the correlation between dissolved radiocesium in the soil solution and the radiocesium content of plants is still very weak. It is generally accepted that the strong retention of radiocesium is mainly due to its binding to specific sites of clay minerals. Although a lot of work was done on this topic, little is known under which conditions cesium may be released from these sites and may therefore also be available for plant uptake. Additionally, the interactions between soil properties and element uptake into plant roots as well as the uptake mechanisms themselves are not known in detail. Additional work in this field is necessary to improve the prediction of plant uptake in case of an acci- dental release of radiocesium and to provide the background for new decontamination techniques such as phytoremediation. Our approaches are mainly focused in two directions: (1) investigation in alternative methods for the determination of the KD or of other parameters characterizing the ratio between bound and solved radiocesium. The fraction of available radiocesium determined by the different methods is correlated with contents of plants grown on another sample of the same soil. (2) Search for a plant that combines high concentrations of cesium in the shoot with considerable biomass accumulation. Such a plant could be used for remediation of a contaminated site if it gets possible to increase the desorb radiocesium from the clay minerals. Such a plant may also be used as a bioindicator of the effectively available radiocesium concentration of a contaminated soil.

64 Working group No.3 NOTES RELATIONSHIP OF BE IN SOIL TO ITS CONTENT IN SELECTED VEGETABLE PLANTS HLUSEK, J., RICHTER, R. Mendel University of Agriculture and Forestry, Department of Agrochemistry and Plant Nutrition, Zemedelska 1, 613 00 Brno, Czech Republic

In the past years the occurrence of beryllium in the environment has become a very topical issue. In plants it occurs in very small amounts, under natural conditions 0.001 - 0.4 mg.kg"1 of dry matter. Higher concentrations of Be are very toxical for plants,, the aim of the present paper was to study the level of beryllium in lettuce and kohlrabi plants based on increasing Be levels in soil (control; 2, 7, 14, 28 mg Be.kg'1 of soil). The results of a pot experiment carried out in 1997 are given in the table. Weight of Weight of kohlrabi Level of Be in Level of Be in lettuce (g/plant) (g/plant) lettuce (mg.kg*1) kohlrabi (mg.kg"1) Variant of trial fresh dry fresh dry dry fresh dry fresh matter matter matter matter matter matter matter matter 1. Control 87.6 9.19 54.3 7.20 0.035 0.0037 0.020 0.0026 2.2 mg Be.kg"1 79.8 6.49 67.5 8.50 0.085 0.0069 0.025 0.0031 3. 7 mg Be.kg"1 73.2 5.91 55.5 7.56 0.390 0.0315 0.040 0.0054 4.14 mg Be.kg"1 76.6 6.62 60.6 8.09 1.210 0.1046 0.140 0.0187 5. 28 mg Be.kg"1 75.0 5.84 26.3 3.57 2.035 0.1584 0.380 0.0516 Control = natural level of Be - 0.622 mg.kg of soil Increasing levels of Be reduced the weight of lettuce by 9-16% on average, as well as dry matter production which decreased by 28-37%. Differences between values in lettuce variants with Be are insignificant. The average weight of kohlrabi and dry matter production in tubers increased in variants 2, 3, 4. The highest yields were obtained with a 1 level of 2 mg Be.kg" in the soil; fresh and dry matter increased by 24% and 18%, respectively. The highest Be concentrations (28 mg.kg"1) had a strongly depressive effect on kohlrabi. The weight reduction of the tubers and content of dry matter in this variant were statistically significant; both values are approximately half of the values found in soil with a natural level of Be. The levels of Be in plants showed that the accumulation of Be was higher in lettuce (in soil with a natural level of Be 1.42 times higher, with 7 mg Be.kg"1 in soil 5.83 times; the other values were within this range). The level of Be in lettuce ranged between 0.0037 and 0.1584 mg.kg"1 of fresh matter and increased with increasing level of Be in the soil. The dependence was similar in kohlrabi at levels of 0.0026 - 0.0516 mg Be.kg"1 of fresh matter. The relation between Be in soil and its level in lettuce plants and kohlrabi tubers was statistically significant.

65 Working group No. 3 SOIL-TO-PLANT TRANSFER FACTORS: LIMITATIONS OF A SIMPLE CONCEPT CZ9928627 KIRCHNER, G.\ EHLKEN, S.2 University of Bremen, Dept. of Physics / FB 1, Postfach 330440, D-28334 Bremen, Germany E-mail: [email protected] 2ZKH St-Juergen-Strasse, Abt. Oekologie, D-28205 Bremen, Germany

The use of soil-to-plant transfer factors is common practice in radioecology. Their mathematical formulation is based on the fundamental assumption that the concentration of a radionuclide in a plant relates linearly on - and only on - the average concentration of the radionuclide in the rooting zone of the soil. However, the large range of transfer factors reported in the literature shows that the concentration of a radionuclide in a soil has an only minor impact on its uptake by a plant, whereas other processes not included in the definition of the soil-to-plant transfer factor are of greater importance. This paper gives a review of processes, which influence root uptake of trace substances by plants. It is concluded that soil-to-plant transfer factors are a valuable tool for radiological screening models which are used for rapid impact assessment after accidents, but due to their simplistic definition do not assist in understanding root uptake processes of radionuclides by plants.

66 Working group No. 3 LONG-TERM INFLUENCE OF LIMING ON BIOLOGICAL AVAILABILITY OF RADIOCAESIUM IN FOREST SOILS CZ9928628 KONOPLEV A. V.1, DRiSSNER J.2, KLEMT E.2, KONOPLEVA I. V.3, MILLER R.2, ZIBOLD G.2 1SPA «Typhoon», 82 Lenin av., 249020 Obninsk, Russia 2FH Ravensburg-Weingarten, University of Applied Sciences, P.O. Box 1261, D-88241 Weingarten, Germany 3AII-Russia research institute of agricultural radiology and agroecology, 249020 Obninsk, Russia

Presented are the results of research of 137Cs transfer from soil to plants on a plot of a spruce forest (Picea abies) in Baden-Wuerttemberg (Germany) in 13 years after fertilization (83% CaCO3, 8% MgO, 6% K2O, 3% P2O5, 2.5 t/ha, 1984). It is shown that liming results in long-term decrease of 137Cs uptake by plants by a factor of 8-20 in comparison with a control plot. The results of liming are the morphological changes of litter and characteristics of rooty soil layer: the parameters of caesium selective sorption and composition of a soil solution determining biological availability of 137Cs. On the basis of model of radiocaesium transfer from soil to plants the prediction of 137Cs accumulation factors for forest plants has been made. Theoretical assessments are compared with experimental data.

67 Working group No.3 COMPARATIVE STUDY OF THE RADIONUCLIDE UPTAKE AND DISTRIBUTION WITHIN PLANTS CZ9928629 FOR BARLEY AND MAIZE VARIETIES KOSTYUK O. Environmental Science and Technology Department, Riso National Laboratory, DK 4000 Roskilde, Denmark (On leave from the Radiobiology and Biophysics Department, Institute of Cell Biology and Genetic Engineering, Ukr. Acad. of Sci., 148 Zabolotnogo st., Kyiv, 252143, Ukraine. E-mail: [email protected])

Differences in the Cs-134 and Sr-85 uptake by three barley and two maize varieties were investigated in a water culture experiment. The maximum differences were about 30% (for radiocaesium) and 50% (for radiostrontium) for barley varieties. The differences between maize varieties were negligible. The maximum difference between varieties of these two species of crops was approximately 30% for radiocaesium and 170% for radiostrontium with higher radionuclide uptake by maize. All barley varieties accumulate radiocaesium nearly 3.5 times more effectively than radiostrontium, whereas for maize varieties radiocaesium was accumulated about 2 times more effectively. There is a large difference in radionuclide distribution within the plants: the amount of radiocaesium in the green part of plants of both species was approximately 30% of the total, while for radiostrontium it was about 80%. As a result approximately the same amount of these radionuclides were present in the green part of plants, despite the large difference in the uptake of these ra dionuclides by whole plants. It is concluded that crop selection, as a measure to reduce radionuclide contamination of the food chain should not occur without taking into consideration the different radionuclide distributions within the plants.

68 Working group No.3

CENOTIC AND PHYSIOLOGICAL CONTROL OF THE RADIONUCLIDES MIGRATION INTO SYSTEM CZ9928630 SOIL-PLANT KRAVETS, A. P. Institute Cell Biology and Genetic Engineering NAS, Department of Radiobiology Vasilcovsky st. 17/31, Kiev - 252022.GSP-22, Ukraine, E-mail: [email protected]

Some biological- cenotic and physiological, - factors which determine of the availability of the radionuclides for a plant and general capacity for the pollutants accumulation have been investigated and analysed. Metabolites of soil microorganisms and especially root excretion of higher plants increase the rate of destruction of solid forms of pollution and enhance leaching of radionuclides from solid matrix. It have been demonstrated in condition of contamination heterogeneity of the Chernobyl fallout that: - During the period of the vegetation the plants of different species of f. Poacea increase of chemical mobility and biological availability of radionuclides in 1.5-2.7 times; - Additional increase of the concentration of soil micro-organisms (micromycetes) lead to enhance contents of mobile form pollutant in soil and level of radionuclides accumulation by higher plants; - Increase of density of sowing (and competition, respectively) of the plants different species lead to enhance of the radionuclides availability and level of radionuclides accumulation by plants in 1.7-2.4 times, too. Other aspect of the formation of the level of plant pollution consists on the peculiarities of the radionuclides absorption and accumulation by plant biomass namely. Effect of the high density of sowing, high level of the watering and gamma irradiation to alterations of the level of the radionuclides accumulation and, at the same time, to the cation exchange capacity (CEC) of the plant biomass have been investigated in laboratory and green house experiment. In parallel, increasing CEC and radionuclides accumulation by 1.5-2.7 times has been demonstrated. These facts suggest, that biological factors are power lever of the control of the pollutants availability and accumulation and may be take into account under development of the modern agricultural technology for clear products formation. Investigation of the different aspects of the biological control of the radionuclies migration into soil -plant system have been proposed as necessary steps in the development of the modern management methods for the soils restoration.

69 Working group No.3 SOIL AND PLANT FACTORS INFLUENCING THE ACCUMULATION OF RADIONUCLIDES BY THE HIGHER PLANTS. CONCEPTUAL AND MATHEMATICAL MODEL CZ9928631 KRAVETS, A. P. Institute of Cellular Biology and Genetic Engineering NAS, Vasilkovsky st 31/17, Kiev-252022, GSP-22, Ukraine E-mail: [email protected]

The successive development, adaptation and parameterization dynamic model of the radionuclides migration into systems soil-plant has been proposed. Conceptual part of description taking as a basis main principles, such as: 1. Plant absorb dissolved part of mineral pollution only. Absorption and accumulation of the mineral pollution is indissoluble connected with plant growth and biomass production. 2. Plants have ability to control of biological availability of any mineral elements (radionuclides and heavy metals, including) This reaction is non-linear, have constitutive and inducible forms and its level has been determined genotype and functional state of plants. 3. Soil radionuclides solution is weak from chemical point of view. Functional dependence of concentration of absorbed radionuclides from soil solution concentration is linear. Coefficient of this linear function depends on how well the plant is provided with functional significant mineral elements. 4. Main part of mineral elements migrates to root system with diffusion and moisture flow by means of convection current due to plants transpiration. It is known, transpiration is connected with biomass production. The mathematical model describing of the pollutants migration into system soil-plant and accumulation of radionuclides by one-year plant during vegetative period has three equations and take into account main soil and plant characteristics.

7f) Working group No.3

137 STRESS MODIFICATION OF THE KINETIC OF CS ABSORPTION AND PRIMARY PATTERN OF CZ9928632 INTERCOMPARTMENTS DISTRIBUTION KRAVETS, A. P. Institute of Cellular Biology and Genetic Engineering NAS, Vasilkovsky st 31/17, Kiev-252022, GSP-22, Ukraine E-mail: [email protected]

Kinetic of 137Cs absorption by seedling of pea and possibility of alteration of the qualitative and quantitative characteristics of this process has been investigated. It is demonstrated, that gamma-irradiation of dry seeds of pea in doses 25-50 Gy lead to following global alteration of kinetic and concentration curves of the active and passive 137Cs uptake by plant roots. First and second phases of kinetic curve, which are reflection of passive stage of absorption and type of concentration curve in genera!, have been alternated. Level of radionuclide accumulation on this passive stage and retention for passage apopiast-symplast increase significantly. Alterations of Michaelis-Menten parameters of the concentration curves have been observed, too. These changes are reflection of alteration of radionuclide distribution into system symplast-apoplast. Time of the 137Cs retention for transition root-stem increase from 0.5 min. to 1 min. This investigation suggest that cation-exchange capacity of the parts and structures of a plant is one of the main factors, which determinate of primary intercompartment distribution of 137Cs as nonbiogenous mineral elements

71 Working group No. 3 METHODS AND MEANS OF MODIDFICATION OF ACCUMULATION RADIONUCLIDES BY PLANTS CZ9928633 IN CONNECTION OF PHYTODECONTAMINATION OF SOILS. KUTLAKHMEDOV, Y.\ MIHEEV, A.1, ZEZINA, N.1, PAVLENKO, N.2 institute of Cell Biology and Genetic Engineering of NASU, Kiev, Ukraine institute of Bioorganic and Oil Chemistry of NASU, Kiev, Ukraine

In an outcome of long-term field and laboratory investigations developed by us and system of "levels" of management is tested by efficiency of Phytodecontamination (PD), including means of influence on form of radionuclides being in soil, on a crop of biomass of plants and on Tf transfer factor of radionuclides. Field experiment has allowed us to reach Cd (Coefficient of decontamination) in 4-5 units in 5 years of experiments with different cultures (crop rotation). The optimum scheme of application PD in conditions of Ukrainian Polessye actuates: - Means of increase a lot of the bioavailable forms radionuclides in soil (microorganisms, irrigation etc.); - Means of increase of a plants crop (optimum crop rotation, fertilizers, irrigation) Means of increase Tf (increase of biomass and surface of root system, at the expense of an operation of the physical factors, selection optimum irrigation, density sowing of plants, optimum scheme of the precursors, special PD crop rotation); - Agrotechnical means (mode of cleaning of crop, frequency hay-crops, mode irrigation, mode of cleaning, selection of the best sorts plants, oriented on problems PD). Offered system of means and methods will derivative real model of the future process engineering Phytodecontamination of radionuclides c contaminated soils of Ukrainian Polessye and may be converted on other conditions in different countries. Working Group No. 3 PROBLEM OF A RADIOCAPACITY IN A SYSTEM SOIL - PLANT FOR BOG ECOSYSTEM CZ9928634 KUTLAKHMEDOVA-VYSHNYAKOVA V. Institute of Radioecology, Ukrainian Academy of Agrarian Sciences, Tolstoy str. 14, 252033, Kiev -33, Ukraine

It was carried out the evaluations of the factors of the various components of pasture bog ecosystem on an example of Volynsk area. It was shown that soil and water introduces the noticeable contribution to accumulation of radionuclides in plants on bog ecosystem. The evaluation of integral distribution of radionuclides (Cs-137) and factors of a radiocapacity on components of bog ecosystem has made Fsoil=0,5; Fwater=0,1; Fplants = 0,25; Froot = 0,15 (soil, water, plant and rooted system). The factor of a radiocapacity determines a part of radionuclides from a general reserve concentrated in a particular component of the ecosystem. It is necessary to underline the higher values transfer factors of accumulation for plants in bog ecosystem on a comparison with terrestrial ecosystem (Tf = 1,5-18). Thus the contribution of soil to formation Tf makes from 60 % up to 80 %, and remaining pathway in plants from a water phase. It can be connected to a high radiocapacity of soil of bog ecosystem and as a corollary the rather small concentration of radionuclides in a water on a comparison with soil.

73 Working group No.3 NOTES THE UPTAKE OF HEAVY METALS (Pb, Cu, Ni, Cr) BY GRAPEVINE LICINA, V., JAKOVLJEVIC, M., ANTIC-MLADENOVIC, S. University of Belgrade, Faculty of Agiriculture, Departement of Plant Physiology and Agrochemistry, Yugoslavia E-mail: vlicina @afrodita.rcub.bg.ac.yu

In the glasshouse experiment it the uptake of heavy metals by grapevine was investigated. The grafts with estimated initial content of investigated elements (Pb, Cu, Ni, Cr) where planted in 3dm3 pots supplied with soil and inert substrate (perlite : send 1:1). Fertilizers were added to the soil as a dose of 100 kgN/ha, 50 kg P/ha and 100 kg K/ha, while the inert substrate were supplied with Ried-York nutrient solution. Used brown-forest soil was wight very low heavy metal content, having 4,6 ng/g of available Pb (28 pg/g was a total Pb), 1.10 ng/g of available Ni (58 ng/g was a total Ni), Cr has 0.3 (ag/g of Cr as a available fraction where 47 |xg/g was its total content, and soil, also, has 2.2 |*g/g of available Cu of its 22.6 ng/g total content. Available fraction of investigated elements were estimated by using DTPA extraction. Before planting, in pots with soil were added a dose of 100 pg/g of Pb, Cu, Ni and 20 n g/g of Cr in the form of C4H6O4 Pb x 3H2O, CuSO4, NiSO4 x 7H2O and K2Cr204 respectively. The same salts were added into the nutrient solution, but in lower concentrations (20 \xglg of Pb, Cu, Ni and 5 ng/g of Cr). Each variant have five repetitions, with the duration of experiment from March to September. At the end of the experiment, after separation of organs (root, stem, shoots, leaves), the plants were analyzed. The results were submitted to the statistical analyzes, and the two following factor were discussed: the concentration of heavy metals in organs and the their uptake by their biomass.

74 Working group No. 3 OBSERVATIONS OF Cs MIGRATION IN COLUMN EXPERIMENTS: II. FROM SURFACE CZ9928635 DEPOSITION MARCHANT, J. K., BUTLER, A. P., SHAW, G., TOMPKINS, J. A., WADEY, P., WHEATER, H. A. Imperial College of Science, Technology and Medicine, TH Huxley School of Environment, Earth Sciences and Engineering, Environmental Analysis Section, Silwood Park, Ascot, Berkshire, SL5 7TE, U.K. E-mail: [email protected]

This paper describes experimental and modelling work carried out to investigate the unsaturated zone movement and bioaccumulation of radiocaesium, as part of the EU PEACE project on nuclear safety. Results are presented of the migration of surface deposited Cs, in response to artificial rainfall events, in a set of undisturbed soil columns containing two soil types both cropped with perennial ryegrass (Lolium perenne var. Profit). The experiment was carried out under controlled conditions and climatic, hydrological and biotic variables were monitored in detail throughout the experiment. Destructive sampling of replicate columns enabled time-dependent radiochemical information to be obtained. The results provide interesting information on the effects of the interaction between the mechanisms of sorption and root uptake, and hydrological fluxes. Cs has been observed to move a significant distance down the soil profile and the effects of macropore flow and time dependent sorption have been investigated using physics-based models of the system. The experiment has been run in parallel with a similar column experiment looking at upward migration of Cs from a contaminated water table funded as part of the Nirex safety assessment programme.

75 Working group No.3 NOTES EFFECT OF LONG-TERM FERTILIZATION WITH DIFFERENT NORMS AND RATES OF NUTRIENTS ON SLIGHTLY LEACHED CHERNOZEM FERTILITY NANKOVA, M., KIRCHEV, H. Institute for Wheat and Sunflower "Dobroudja", General Toshevo 9520, Bulgaria

In order to determine the effect of long-term fertilization with macroelements on the changes in soil fertility of slightly leached chernozem as well as on yields and quality of wheat, in 1967 a field trial was started with different norms and ratios of N, P and K fertilization. The trial includes testing of 4 nitrogen and phosphorus norms - 0, 6.12 and 18 kg/dka N and P2O5 and 3 potassium norms - 0, 6 and 12 kg/dka K2O, in full scale (4 x 4 x 3 = 48 variants). The experiment is conducted according to the method of net square in 4 replications, the size of the trial plot being 20 m2. In order to characterize the changes in soil fertility, in 1997 after wheat harvest, soil samples were taken from ail tested variants at depth 2 m, every 20 cm. The pH, forms of mineral N and mineralization ability of soil (mg/1000 g soil), as well as available P2Os and K2O (mg/100 g soil) were determined. The introduction of N, P and K fertilizers at different norms and rates for 30 years lead to considerable changes in nutrition regime of the slightly leached chernozem. The obtained results reveal an increased content of mineral nitrogen forms when increasing nitrogen fertilization norms as a result of increased rate of mineral reserves in soil. Considerable changes were established in values of mineralization ability of soil along the investigated profile as compared to the non-fertilized control variant (NoPoKo). A certain decrease of the values for surface soil acidity was also established as a consequence of long-term fertilization at high N norms. Available P content significantly differs from control variant. Both independent and combined N and P fertilization contribute to the increased amount of available phosphates in soil. In independent application of P and P-K fertilizers as well as in combination with N, the content of available phosphates in soil is affected mainly by the norm of P fertilization. A tendency was established for reduction of the values for this index with a presence of N, and N and K in the combinations at different norms of P fertilization. This is most probably due to the more intensive usage of nutrient elements in variants with combined fertilization resulting from the creation of considerably higher vegetation biomass.

76 Working group No.3 NOTES GENETIC VARIATION FOR DRY MATTER AND NITROGEN ACCUMULATION OF DOUBLE HAPLOID WHEAT LINES NANKOVA, M., MILKOVA, V., IVANOV, P. IWS "Dobroudja", near General Toshevo 9520, Bulgaria

In the present study, 10 DH lines of wheat from the crosses 7-P211 x H-81/32-4 (41-519; 41-181; 41-191) and 7- P211 x H-81/32-24 (41°-306; 41-344; 41°-355; 41°-470; 41°-426; 41°-413 and 41°-234) were used. The investigation included the cv. Siavaynka-196 as a quality standard. The lines were obtained by using a combination of embryo and anther cultures. Immature embryos were obtained after hybridization and were plated on a MacKinnon et al. (1987) medium for further development. Five months later F-| plants were used as donors for anther culture. Double haploid lines were produced as a result. Biochemical and technological analyses of DH lines were made. Results showed that DH lines had better quality characteristics than initial parents. Glu-1 score of high molecular glutenines is 10 in some of the new lines (41-181; 41-191; 41°-413). The study gives basic information on the accumulation of dry matter and N in the DH lines included in the experiment. The aim of the study was also to develop data necessary to understand the physiological and genetic relationships between N movement and grain yield and protein concentration. Results from the field and vegetative experiment, where the genotype reaction to low and high N nutrition is examined, show statistically proved differences between the lines and their parental forms. The greatest quantity of dry matter and grain yields were established in lines 41-191 and 41°-344. Increased accumulation of N between anthesis and maturity suggested continued uptake of N from soil. The same lines stand out with the highest values of GHI - 49.22 and 46.46, respectively, and NHI - 79.26 and 81.80, respectively. Considerable genotype differences were established in the nitrogen amount from the vegetative mass per unit of mature grain. Line 41 °-344 used 24.82 mg N/g mature grain. Comparing the genotypes by their nitrogen expense for forming 100 kg of grain together with the respective straw produced, it was established that the same line is the most economical one - 2.03 for grain and 0.39 kg for straw. Grain protein content increases considerably as a result of the increasing nitrogen dressing norms, especially evident at fertilization with 400 mg N/1000 g soil. Most of the new lines exceeded, according to these values, not only their parental forms, but the quality standard Slavaynka-196.

77 Working group No.3 REDUCTION OF Cs-137 LEVELS IN PLANTS AND FUNGI AFTER POTASSIUM FERTILIZATION IN A SWEDISH FOREST CZ9928636 NIKOLOVA,!. Pouskharov Institute of Soil Science and Agroecology, 7 Shosse Bankya. 1080 Sofia, Bulgaria and Karl J. Johanson Department of Radioecology, Swedish University of Agricultural Sciences, P.O. Box 7031, S-750 07 Uppsala, Sweden

The uptake of Cs-137 in plants in forest ecosystems are much higher than in agricultural ecosystems. One reason could be that the concentrations of mineral nutrients usually are at much lower levels in forest soils compared to soil from arable land. On the other hand there are often rather weak correlation between the concentrations of exchangeable potassium in forest soils and the levels of Cs-137 in for example dwarf-shrubs. The variation in potassium levels in the forest soils are however rather small. By fertilization with potassium it is possible to increase this variation. The aim of the present study was to investigate the effects of potassium fertilization on the uptake of Cs-137 on a rather nutrient poor forest ecosystem - a rocky area with a rather shallow soil layer with high organic content. The potassium was spread in May 1992 by normal agricultural equipment trying to obtain 200 kg of potassiumklorid per hectare. On the fertilized area 3 plots, each about 200 m2, were selected and used for sampling of bilberry, lingonberry and heather. One sampling were performed before the spreading and then at least once a year up to 1997, During the mushroom season, fruitbodies of the most common species of fungi were collected if they growed within the 3 plots. A closely located rocky area was selected as a control area. The Cs-137 levels in bilberry and lingonberry showed only a minor decrease during the 1992 vegetation period. In contrary, heather show a well marked decrease of about 50 % already the first year. In mushrooms - Lactarius rufus and Rozites caperatus the decrease were even more pronounced. In 1997, 5 vegetation periods after the fertilzation, the Cs-137 levels in bilberry, lingonberry and heather were 2767, 1741 and 13,213 Bq kg"1 respectively in control area. In the fertilized area the corresponding levels were 633, 926 and 3,224 Bq kg"1 respectively corresponding to 23, 53 and 24 % of control levels. Even fruitbodies of Suillus variegatus, Cortinarius semisanguineus, Lactaries rufus and Rozites caperatus showed Cs-137 levels around 30 to 50 % of that in the control area. Potassium fertilization seems thus to be effective as a counter measure at least on the type of forest used in this study. The reduction of 137Cs levels in plants and fungi persist for quite a long period and therefore it could be a usable counter measure in some specific situations.

78 Working group No.3 134 CS UPTAKE BY PLANTS FROM SOIL APPLYING DIFFERENT ABSORBENTS CZ9928637 ONCSIK, M. Irrigation research institute, 5540. Szarvas, Hungary; E-mail: [email protected]

A kind of utilisation of the contaminated soil radionuclides can be amelioration. It means not only liming but also using different soil-additives. Among of this additives are widespread the different zeolite minerals. In their chemical compound the natural zeolites form incoherent spatial structure of SiO4 ion groups. Selective, sorption, cation changing and specific surface properties of the zeolites connected with their nature can be explained with this structure. In Hungary we can find territories suitable for zeolite mining in the Tokaj-mountains. We have started to study the isotope uptake by plants using different zeolite clay minerals (montmorillonite, mordenite, clinoptilolite). Final aim of the experiment was to reduce the rate of isotope uptake by plants applying the additives given to the soil. According to the experiment's results made in vegetation pots the Cs134 activity of green peas were reduced averaging by 16% comparing with the control in the soils enriched with zeolite (mordenite type). Under field conditions we studied the radioactivity of millet and carrot yields in the soils ameliorated with montmorillonit clay mineral. It was found that the radioactive contamination of yield in soil treated with zeoilite reduced by 10% averaging in case of millet, while it was 24% for the carrot roots comparing with the control.

79 Working group No. 3 22b6 TRANSFER OF " RA TO PLANTS FROM TWO TYPES OF SOIL CZ9928638 ROSIAK, L, PIETRZAK-FLIS, Z. Central Laboratory for Radiological Protection, Department of Radiation Hygiene, Konwaliowa 7, 03-194 Warsaw, Poland, E-mail: [email protected].

The transfer of 226Ra to plants used as food (carrot, parsley, wheat) and as fodder (grass, maize) has been studied. Plants were grown on two types of soil: sandy soil (I) and sandy loam soil (II) in an open field (exposure to dry and wet deposition, resuspension and soil adhesion) and in polyethylene tent with an underground irrigation system (isolation from wet deposition and from water splash on soil). The plants were grown simultaneously on the open and sheltered fields. The average concentration of total 226Ra in Soil I was equal to 8.48±0.50 Bq kg"1dw and concentration of exchangeable 226Ra was 0.62±0.07 Bq kg"1^, while in Soil II they were equal to 12.2±0.56 Bq kgV and 0.66±0.05 1 226 Bq kg" dw, respectively. Ra was determined in above-ground parts of plants after washing in distilled water, in the rinse obtained from washing, in insoluble residue separated from the rinse, and in roots. The data obtained allowed to determine the incorporated radionuclide into the plants and on their surface. The statistical analysis of data indicates a lack of differences in the incorporated 226Ra for plants growing on the open field and in the tent. This indicates that 226Ra enter the plants mainly via root system, whereas incorporation through leaves and stems is negligible. Table 1 gives some results. Table! Average concentrations of 22SRa incorporated into plants and washed out from the plants' surface mBq kg"1^ Soil I Soil II Incorporated Washed out Incorporated Washed out 1 1 1 1 Plant mBq kg" dw mBq kg' dw mBq kg' dw mBq kg' dw Grass, cut I 536±49.8 29.6±16.6 240±46.4 87.3±69.4 Grass, cut II 635±141 101±55.1 463±20.6 78.5±38.3 Parsley root 1044±55.9 1009±110 leaves & stems 1977±91.4 223±40.1 2559±420 1022±131 Wheat straw 918±65.4 87.8±54.0 554±11.8 grain 96.5±7.97 18.0±3.32 90.7±4.96 In majority of the cases the concentration of Ra was in the plants growing on Soil I higher than on Soil II although the concentration of totali 226,Ra was in Soil I about 1.4 times lower than in Soil II. The concentration of exchangeable "22™6 was in both soils almost the same. The total external contamination of the plants was at average about 30% of the incorporated Ra and about one third of the external contamination was soluble. Transfer Factors related to exchangeable and total Ra in soil were calculated. Working group No.3

PREDICTING RADIOCAESIUM AND POTASSIUM ROOT UPTAKE USING THE BARBER-CUSHMAN CZ9928639 MODEL. ROCA JOV£, M. C, VALLEJO CALZADA, V. R. Departament de Bioiogia Vegetal, Facultat de Biologia, Universitat de Barcelona, Avg. Diagonal, 645, 08028, Barcelona, Spain

Mechanistic models can be applied in the study of radionuclide soil-plant transfer for the obtention of predictive estimates of radionudide plant contamination. The present study measured Kand 134Cs transfer to pea plants growing in pots within two different soils (Calcic Luvisol and Fluvisol) and compared the results obtained with the prediction of Barber-Cushman model. Good correlation among simulated and measured values was observed for both ions. For K, model predictions were valid for ail development stages sampled, but for 134Cs the quality of the prediction depended on the plant stage. Estimates of model parameters varied depending on plant age and soil type. The model was also run for 134Cs using the Michaelis-Menten parameters obtained for K. In this case, the predicted values were about three times higher than those observed and regression was still significative (1^=0.553, n=11, P<1.6%). These results show a positive plant discrimination of K versus 134Cs for plant absorption and for the soils studied. However, as regression was significant, the use of K absorption parameters to estimate the 134Cs absorption appears as a good option in radiocaesium plant uptake estimation.

81 Working group No.3 137Cs AND 90Sr ROOT UPTAKE BY BEANS IN SOILS WITH CONTRASTING PROPERTIES CZ9928640 SAURAS YERA, T., VALLEJO, R., WAEGENEERS, N., MADOZ-ESCANDE, CH.

Radionuclide soil to crop transfer was analysed in large undisturbed soil monoliths installed in lysimeters under controlled climatic conditions. The selected soils are representative of large agricultural areas in the European Union: Loamy-sand Orthic podzol from Belgium (B), Loamy Calcic Luvisol from Spain (E), Silty-loam Orthic Luvisol from Germany (DNW), and Sandy-loam Eutric Fluvisol from the UK (UK). The soil to plant transfer of radionuciides is a multifactorial process dependent on the radionuclide availability in the soil solution, or the solid/liquid de distribution coefficient KD (KD = [RN] in soil (Bq kg"1)/[RN] in soil solution (Bq I"1) and on the Concentration Factor (CF = Bq kg"1 plant/Bq I"1 soil solution) which is the root uptake ability of the crop. Both KD and CF parameters are affected by the concentration of K, Ca and Mg in soil solution. 137Cs concentration in soil solution varied from 400 Bq I'1 in the B soil to about 9 Bq I"1 in the E soil while 137Cs Solid /liquid distribution coefficient KD ranged from about 480 in the loamy-sand B soil to about 19000 I kg"1 in the Loamy E soil. On the other hand, K concentration in soil solution (m«) varied from about 25 mmol I"1 in the B soil to 0.6 mmol I"1 in the E soii. UK and D soils presented intermediate values for the three parameters. A negative linear relationship has been found between Kg and K status in soil solution at a logarithm scale, high level of K resulted in a low KD and thus in a promotion of Cs in soil solution. On the contrary, low K concentration in soil solution 137 produced high KD and low Cs concentration in soil solution. The plant concentration factor was also affected by the K concentration as according as in the soil solution. High K content in soil solution resulted on low CF and on the opposite side, low K resulted on high CF. The relationship between CF and K was fitted to the following equation: log CF = -b0 log mk + b^ 2 For bean plants the regression parameters were b0 = 0.81 and bi = 2.06 (r = 0.69, n = 16, p < 0.001). For bean pods the parameters were b0 = 0.73 and bi = 1.83, and the regression model explained 70% of the variance in log CF. The 137Cs root uptake crop reflected the specific scenario of each soil, the Loamy-sand B soil showed significantly higher Concentration Ratio (CR = Bq kg"1 plant / Bq kg"1 soil) followed by the Sandy-loam UK soil and the DNW Silty-loam. The lowest root uptake was found in the Loamy E soil. ^Sr concentration in soil solution varied from 211 in the DNW soil to 3890 Bq 1-1 in the B soil. Accordingly, the KD H n a varied in the opposite way and was negatively related to the Ca+Mg concentration in soil solution (m(ca+Mg))- '9 ^ and Mg concentration in soil solution resulted in lower KD. With respect to the CF, as for 137Cs, the best fitting was a linear regression of the logarithms of CF and m(Ca+Mg): Log CF = -b0 log m(Ca+Mg) + bi. The proposed model explains 89% and 93% (n = 10 p<0.001) for bean plant and pod respectively. The fitted parameters are b0 = 1.06 and bi = 2.56 for bean plants and bo = 1.15 and bi = 2.17 for bean pods. According to the obtained results, K, Ca and Mg in soil solution play an important role on the 137Cs and 9QSr root uptake, in addition soil type and specific growing conditions are important factors governing the transfer.

82 Working group No.3 NOTES NUTRIENT MOBILITY IN SOIL-PLANT SYSTEM ACCORDING TO SOIL MAINTENANCE SYSTEMS AND NATURE OF FERTILIZERS SERDINESCU, A. Research Institute for Viticulture and Enology, Valea Calugareasca, Romania

Utilization of certain alternative systems of soil maintenance in vineyards can significantly influence the hydric and trophic regime of the soil. Therefore, our experimenta aimed at estimating nutrient mobility in soil-piant system under the influence of basic systems of soil maintenance - bare fallow, application of herbicides, covering with annual or perennial crops, associated or not with the application of organic and chemical fertilizers. Four systems of soil maintenance were studied: bare fallow, total herbicide application (Simanex 8 kg/ha + Gramoxone 6 I/ha), green fertilizers (pea) and guided covering with perennial crops (Lolium sp. 50% + legumes 50%) for 5 years. Each system included 3 fertilization treatments: unfertilized (control), organic fertilization (40 t/ha manure for 4 years), and chemical fertilization (N-|60p160K160 *or 4 years). Green mass annually incorporated in soil in case of the green fertilizer (21.5 t/ha/year) or periodically in case of covering with perennial crops (18 t/ha/year) determined soil enrichment in organic matter and nutrients due to mineralization. Application of manure as well as of chemical fertilizers contributed to an increase in green mass by 20-30%. Annual share of biological N brought by annual (pea) or perennial legumes (alfalfa) oscillated between 125-139 kg N/ha. The hydric regime of soil was at its best in case of herbicide treatment (19.3%) and deficit in case of covering with perennial crops (13.4%). Total application of herbicides led to the best maintenance of water in soil even in the superficial layer (O- 10 cm). As a consequence of these modifications on the hydric and trophic regime of the soil, grapevine nutrition was more intense in case of green fertilizer for soil maintenance systems (5.74%, with nutritional equilibrium of 63:15:22) and in case of chemical fertilization for the treatments (5.27%, with nutritional equilibrium of 62:19:19). The different systems of soil maintenance also influenced the percentage distribution of the elaborated substances, nutrient carrier, in vegetative and perennial organs of grapevine. So, green fertilizer determined the increase of N and K in vine roots and stem, while the application of herbicides and covering with perennial crops determined the increase of P in these organs. In the vegetative organs (shoots and leaves) the percentage distribution of NPK was differentiated, being higher for N in case of green fertilizer and for P and K in case of covering with perennial crops. One can say that the different systems of soil maintenance which are rendered more efficient by fertilization treatments materialized new relations of reciprocity between growth and fertility, the synergic effect being more obvious in case of the green fertilizer system which gave the highest yields.

83 Working group No.3 SOIL TO PLANT TRANSFER OF 134CS FOR OLIVE AND ORANGE TREES AFTER FOUR YEARS1 EXPERIMENTATION CZ9928641 SKARLOU, V., NOBELI, C, ANOUSSIS, J., HAIDOUTI, C.1 N.C.S.R. "Demokritos" Agia Paraskevi 15310 Athens, Greece Agricultural University of Athens, lera Odos 75, 11855 Athens, Greece

The objective of this research was mainly to calculate values of transfer parameters of134Cs from soil to tree crops (olive and orange trees) in a long term glasshouse pot experiment, started in 1994. The influence of the soil characteristics on '!34Cs uptake was also studied with the selection of two soils differing in their physical and chemical properties. Both evergreen trees behaved similarly in the studied soils showing that a higher or a lower uptake is not crop specific. The capacity of the studied trees to absorb ^34Cs through the roots seems to be significantly influenced by the soil type. Transfer factors were very low in the calcareous heavy soil and much higher in the acid light soil (up to 10 times for olives and 40 for the edible part of oranges). The difference in TFs is higher between the two soils than between the two tree species. ^3^Cs concentration kept increasing in the orange trees up to the 4th year of growth, while it seems to reach an equilibrium, with no further increase in the olive trees. Although the behavior of these two tree species are similar the difference in the final processed product is extreme. A significant amount of 134Cs was measured in olives grown on light-acid soil but this was not transferred to the 1 olive oil. On the contrary the edible part of the oranges showed the highest 34QS from nearly all plant parts.

84 Working group No.3 NOTES EFFECT OF NPK FERTILISATION ON 3 APPLE DISEASE RESISTANT VARIETIES GROWN IN CONTAINER STANICA, F. University of Agricultural Sciences and Veterinary Medicine, Faculty of Horticulture, 59 Marasti Str., 71331 Bucharest, Romania, E-mail: [email protected]

In a 4-year factorial trial with 3 resistant apple varieties, GENEROS, PIONIER and FLORINA, grafted on MM. 106 rootstock and grown in container, the effects of major nutrients on soil characteristics, leaf content and productive shoot formation were studied. Nitrogen, phosphorus and potassium were applied in 7+1 combinations (NP, NPK, PK, P, N, NK, K and control). The moment of application and annual fertilisation doses were: -15 April -100 ppm N, 50 ppm P and 50 ppm K; -15 May -100 ppm N, 100 ppm P and 100 ppm K; -15 June - 100 ppm N, 50 ppm P and 50 ppm K. Containers with brown-reddish soil from Baneasa - Bucharest had a 20 liter capacity. N fertilisation has a positive influence on total shoot nitrogen content of the 3 resistant varieties studied. The Romanian apple variety "Generos" accumulated two times less total N than the other two varieties. At the same time, N fertilisers have a negative effect on shoot phosphorus and magnesium content. P application increased 3 times the content in shoots, respecting the control. The Romanian apple variety "Pionier" accumulated the highest quantity of potassium (1.48%). The same variety had the highest Ca content on the V3-PK variant. K application had no significant effect on the "Fiorina" variety. P and K application stimulated formation of flowering branches, while N had an opposite effect. The most precocious variety was "Pionier" but the most productive was "Fiorina".

85 Working group No.3

137 TRANSFER OF CS FROM SOIL TO PLANTS IN DIFFERENT TYPES OF SOILS CZ9928642 TODOROVIC, D.1, POPOVIC, D.z, RADENKOVIC, M.1, DJURIC, G.2 Environmental and Radiation Protection Laboratory, Institute for Nuclear Sciences »Vinca«, P.O.Box 522, 11001 Belgrade, Yugoslavia department of Physics and Department of Radiology and Radiation Hygiene, Faculty of Veterinary Medicine, University of Belgrade, Bul.JA 18,11000 Belgrade, Yugoslavia, E-mail: [email protected]

The results of the investigations of the transfer of 137Cs from soil to plants in different types of soils are presented. The investigations were carried out in two mountainous regions in the West and South region of the country (MtTara, 1280 m and MtSara, 2000 m). The investigations are part of the long term project on the distribution of natural and man made radionuclides in »soil - plants« ecosystems. The activity of 137Cs in the samples of soils and plants was determined by a HPGe detector (ORTEC, relative efficiency 20%) by standard gamma spectrometry. Three main types of soils were examined: shale, limestone and the mixed one and a certain number of plants: grass, meadow flora, pinewood, blueberries, an endemic species of Mt.Sara and the bioindicators: moss and lichten. The transfer factors defined as ratios od 137 Cs concentrations in plants and soil from the same locality were calculated. The factors are in the range of 0.1 - 2.0 due to the type of soils and plants (3.0 - 10.0 for bioindicator plants). The vertical distribution of137 Cs in the first 15 cm of soils indicate the slow migration of Chernobyl cesium through soils except on riversides due to the wash-out effect. Generally the concentration of 137,Cs in soils strongly depends on the configuration of the terrain. Working group No.3

NEW SCALABLE TRANSFER FACTORS: A STUDY OF THE KEY PROCESSES AFFECTING SOIL- CZ9928643 TO-PLANT TRANSFER FACTORS AND THE DEVELOPMENT OF A NEW SCALABLE APPROACH TOMPKINS, J. A., WADEY, P., BELL, J. N. B., BUTLER, A. P., SHAW, G., WHEATER, H. S. Imperial College of Science, Technology and Medicine, Department of Civil Engineering, London SW7 2BU, U.K. E-mail [email protected]

Transfer factors are widely used in radiological assessment modelling. They provide important information on the behaviour of radionuclides within the biosphere and give a measure of the overall efficiency of contaminant movement from donor to receptor. There are international databases of soil-to-plant transfer factors (IAEA, 1994, IUR 1989) containing over 8,000 values for different soil, plant and radionuclide combinations. However a study of these values shows that the factors can differ widely even for the same conditions and this has cast doubt upon their applicability. The major drawback of transfer factors is that they do not explicitly represent the physical, chemical and biological processes affecting soil-to-plant transfer of radionuclides, but contain aggregated information on a number of such processes. This paper aims to identify the key processes, affecting radionuclide transport, that are contained within the soil-to-plant transfer factor values for generic soil and crop types. It also presents the values obtained by applying various methods for transfer factor calculation and iooks at developing a new approach in which transfer factors are estimated by reference to the underlying mechanistic processes. This approach is applied to a range of soil and crop data from lysimeter and column experiments of radionuclide migration. This work was carried out as part of the Nirex safety assessment programme.

87 Working group No. 3

EFFECT OF SPECIFIC AND VARIETAL FEATURES OF CROPS ON CAESIUM-137 CZ9928644 ACCUMULATION TSYGANOV, A. R., CHERNUKHA, G. A., KRUGLENYA, V. P. Belarussian Agricultural Academy, Gorki, Mogilev region, Belarus

Our research was aimed at determining the level of caesium accumulation in crops (cereals, legumes, barley) considering their variety features. Experiments were conducted in zones radioactively contaminated with caesium- 137 at the level of 555-1480 MBq/m and above. The radiometric analysis was made with gamma-spectrometer. Hay was tested to determine caesium presence in cereals and leguminous crops while grain and straw were tested in experiments on barley. The research done gave reason to claim that caesium accumulation depends on the crop variety and variety features. So, the level of caesium accumulation in the hay of cereal grasses is 1,5-2 times as low as that in leguminous grasses. Red clover was marked for the maximum level of caesium content (397 Bq/kg) while reed fescue had the lowest level of caesium accumulation (145 Bq/ kg). Besides the tendency snowing the dependence of caesium accumulation on the kind of agricultural area was noticed. The general natural phenomenon is as follow: the content of caesium in hay from cultivated areas is lower than in that from natural ones. As a result of the research done different doses of caesium accumulation in parts of plants were determined. The content of caesium in barley straw was about twice as much as that in grain. Moreover, differences in radiocaesium accumulation depending on the barley varieties were revealed. Working group No.3 NOTES THE EFFECT OF PUTRIFIED PLANT RESIDUES ON THE GERMINATION SPEED AND POWER OF WHEAT (GEREK 79) UYANOZ, R.1, ACAR, R.1, ER, F.1, BROHI, A. R.2 1SeIcuk University, Agricultural Faculty, Dept. Soil Science, Konya, Turkey 2Gazi Osman Pasa University, Agricultural Faculty, Dept. of Soil Science, Tokat, Turkey

Effects of some plant residues putrified at different periods (10, 20, 30 days) on germination speed and power of wheat {Triticum aestivum L). The following plants were used: fenugreek (Trigonella foenum-graecum L), common vetch (Vicia sativa L), field pea (Pisum arvense L), Narbonne vetch (Vicia narbonensis L), Hungarian vetch (Vicia pannocia Crantz.), sugar beet (Seta vulgaris L.) and wheat (Triticum durum Desf.). The effect of putrified plants on germination speed was statistically significant (F = 2.49). The minimum germination (89.7%) of wheat was found with fenugreek and the maximum (95.3%) with wheat.

89 Working group No.3

OBSERVATIONS OF Cs MIGRATION IN COLUMN EXPERIMENTS: I. FROM A CONTAMINATED CZ9928645 WATER TABLE WADEY, P., BELL, J. N. B., BUTLER, A. P., MARCHANT, J. K., SHAW, G., TOMPKINS, J. A., WHEATER, H. S. Imperial College of Science, Technology and Medicine, TH Huxley School of Environment, Earth Sciences and Engineering, Environmental Analysis Section, Silwood Park, Ascot, Berkshire, SL5 7TE, U.K. E-mail [email protected]

The behaviour of radiocaesium within the unsaturated near surface environment and its uptake by plants is a topic of importance, in order to study its behaviour a series of column experiments are being undertaken. Results are presented from a set of undisturbed soil column experiments investigating Cs migration in three different soil types, all cropped with perennial ryegrass (Lolium perenne var. Profit). This experiment investigates the upward movement of Cs from a contaminated water table, as part of the Nirex safety assessment programme. The experiment was carried out under controlled conditions and climatic, hydrological and biotic variables were monitored in detail throughout the experiment. Time dependent information on crop uptake of Cs was established by destructive sampling of replicate columns. The results provide interesting information on the effects of the interaction between the mechanisms of sorption, root uptake and hydrological fluxes. The results have shown significant upward movement of Cs in a number of the columns and modelling has indicated that this mobility could be related to time dependent sorption effects. The experiment has been run in parallel with a similar column experiment looking at the leaching of surface deposited Cs in response to rainfall, funded as part of the EU PEACE project on nuclear safety. Working group No. 3 UPTAKE OF RADIOCAESIUM BY LETTUCE CROPS: THE EFFECT OF K IN SOIL SOLUTION CZ9928646 WAEGENEERS, N.\ CAMPS I VILA. M.2, SMOLDERS, E.3, MERCKX, R.3, SAURAS, T.4, MADOZ- ESCANDE, C.5 1Labo Radio-ecologie, DepartementStralingsbescherming, SCK-CEN, Boeretang 200, B-2400 Mol, Belgium. 2Departament de Quimica Analitica, Universitat de Barcelona, Diagonal 647, E - 08028 Barcelona, Spain. 3Labo voor Bodemvruchtbaarheid en Bodembiologie, Katholieke Universiteit Leuven, K. Mercierlaan 92, B-3001 Heverlee, Belgium. 4Departament de Biologia Vegetal, Universitat de Barcelona, Diagonal 645, E - 08028 Barcelona, Spain. 5SERE-IPSN, CEA-Cadarache, 13108 St. Paul lez Durance, France

The effect of varying K supply on 137Cs uptake by lettuce (Lactuca sativa L, cv. Batavia, Gloire du Dauphine) was studied in solution culture, in a potted soil experiment and in a greenhouse lysimeter experiment under close-to-real conditions. Lettuce was grown for 13 days in nutrient solution spiked with 137Cs. Treatments were four concentrations of potassium (K) in solution (25 uM, 50 uM, 250 uM and 1000 uM). Yields were marginally affected by K supply. The Cs concentration factor (CF, ml/g) decreased 66-fold in the shoot and 432-fold in the roots over the whole K concentration range. The decrease was most pronounced between 25 uM K and 250 uM K. In a subsequent experiment, lettuce was grown for 20 days under the same climatic conditions in two sandy-loam soils (A, B) contaminated with 134Cs. Both had similar characteristics but differed widely in K supply. Soil solution K concentrations were 100 uM (A) or 3000 uM (B). The radiocaesium soil-to-plant Transfer Factor (TF, g plant dry weight/g soil) was 0.320 in soil A and 0.016 in soil B. The higher 137Cs availability at the lower K supply (soil A) was contrasted by lower 137Cs concentrations in soil solution of soil A than of soil B. Radiocaesium transfer to lettuce grown to maturity was analysed on 5 different lysimeter soils under controlled climatic conditions. The soils were artificially contaminated with 137Cs in 1994. The TF's varied between 0.032 and 0.191, and were not related to K concentrations in soil solution. The CF decreased about 100-fold with K concentrations increasing from 0.3 mM to 18 mM. Predictions of soil-to-plant transfer factors based on soil solution composition and nutrient solution results were qualitatively correct but underestimated the observed values.

91 Working group No. 3 (Abstract for UIR)

USING SOIL-TO-PLANT TRANSFER TO IDENTIFY AREAS VULNERABLE TO RADIOCAESIUM CZ9928647 DEPOSITION IN WESTERN EUROPE WRIGHT, S., CREAMER, R., SANCHEZ, A., HOWARD. B. Institute of Terrestrial Ecology, Merlewood Research Station, Windermere Road, Grange-over-Sands, Cumbria, LA11 6JU, United Kingdom, E-mail: [email protected]

Following the deposition of radionuclides from a nuclear accident some geographical areas may be more vulnerable to radioactive contamination than others. Differences in soil type and agricultural practice across Europe will influence the importance of such areas as sources of contaminated foodstuffs. This poster will introduce the concept of applying the critical load methodology within radiation protection for the rapid identification of areas, which may be vulnerable to the deposition of 137Cs following an accident. This vulnerability can be defined in terms of elevated levels in food products or elevated fluxes. It has frequently been reported that the rates of uptake of radiocaesium from soils with a high clay content is lower than from soils with a coarser textural composition and a high organic matter content. It is therefore important to be able to quantify the rates of transfer of radiocaesium from different types of soil. Tag values have therefore been collated from the literature into a database and allocated to one of four soil groups reflecting differences in radiocaesium soil-to-plant transfer. The collated Tag values have been used to estimate representative transfer values for each of the four soil groups. The critical load methodology is based upon the principle that if the sensitivity of an ecosystem to a pollutant is known, it is possible to determine the critical load - the maximum pollutant load that will not cause long-term damage. The spatial variation in critical load across an area can be compared to pollutant deposition data to identify areas where the deposition load exceeds the critical load. Adopting this approach within radiation protection could provide rapid assessment of the potential effects of deposition resulting from nuclear releases. The potential of such an approach is illustrated using the example of 137Cs contamination of cow milk in Western Europe following a hypothetical nuclear accident. A Geographical Information System (GIS) is used to integrate the spatial variation in deposition, transfer and production. Critical loads are developed in terms of the 137Cs activity concentration in cow milk and the 137Cs flux from an area. These highlight the relative simplicity of the methodology for the rapid identification of vulnerable areas, which could also be adopted as a useful modelling tool allowing a consideration of different contamination and response scenarios.

92 Working group No. 3 NOTES PERSPECTIVES AND POSSIBILITIES OF ECOMODEL IN ESTIMATION AND PROGNOSIS TRANSFER OF RADIONUCLIDES IN SYSTEM SOIL-PLANT YASKOVETZ, !., GIRIY, V., ZAITOV, V. Institute of radioecology UAAS, Kiev, Ukraine, E-mail: [email protected]

Abstract was not delivered.

93 Working group No.3 NOTES CHANGES IN SULPHUR BALANCE IN PLANT PRODUCTION OF THE CZECH REPUBLIC ZELENV, F., ZELENA, E., GERMAIN M. Research Institute of Crop Production, 161 06 Prague, Czech Republic

In the past, the sulphur were more than sufficient to meet the requirements of crops in the Czech Republic. The balance of sulphur as a plant nutrient was fairly active, there was annual surplus at least about 277 0001, which mean minimum about 39 kg S/ha (Silar, 1973). Therefore most concern was focused on the harmful effects of acidic deposition on field crops and natural ecosystems. Sulphur dioxide emission in the Czech Republic peaked in the late 1980s. Total SO2 emissions have decreased from 2,171 million tones in 1986 to 946 179 tones in 1996 and further large decreases are expected in the next years. That is positive from the ecological point of view, but on the other hand one of the main sources of sulphur for crop nutrition has been and will be removed. Currently about 1/3 of the Czech Republic land area (mainly south Bohemia and south Moravia regions) receives less than 15 kg S/ha from the atmosphere, which is smaller than the amount required by most grown crops, especially of winter rape which is grown on nearly 9% of the arable area. Use of fertilizers that mean also sulphur deeply decreased as a result of the important social changes and price liberalization and in 1996 only 61 kg N, 11,8 kg P2O5 and 8,0 kg K2O/ha of agriculture land were applied. If there is annual average 50 kg S/ha leaching, negative sulphur balance can be expected on at least 50% of the land. Therefore, it can be supposed that S fertilization will be a new and important part of future plant nutrition programs also in our country. Influence of sulphur and nitrogen application on plant growth, survival of seedlings, S status and on yields of winter oilseed rape will be discussed. Abstracts presented in working group No. 4

Chairmen: Prof. T. H. Thomas and Dr. J. Schmidt HISTOLOGICAL ANALYSIS OF DIFFERENT TYPES OF SYNGONIUM PODOPHYLLUM SCHOTT. CALLUS 103 ANTOFIE, M. M., ANGHELUTA, R.

BIOACTIVE SUBSTANCES TESTED AT IN VITRO EXPLANT CULTURE 104 ARDELEAN, A., CORNEANU, G., CORNEANU, M., MURESAN, N., BRONZETTI, G., FURUKAWA, H., JUNKO, E., BICA, D., MINEA, R., GABOR, L, FRANGOPOL, C.

"IN VITRO" REACTIVITY ON SOME RARE PLANTS OF SPONTANEOUS FLORA 105 AZEMA, T., CIUBOTARU, A., KIRTOACA, V., MALAHOVA, L

HORMOMAL REGULATION OF ASSIMILATE PARTITIONING 106 BAKER, D. v

INTERACTION OF CYTOKININS AND AUXIN IN PEA LATERAL BUDS AFTER BREAKING AND RENEWAL OF APICAL DOMINANCE 107 BALLA, J., BLA^KOVA, J., PROCHAZKA, S.

CYTOKININS IN TOBACCO AND WHEAT CHLOROPLASTS: OCCURRENCE AND CHANGES DUE TO LIGHT/DARK TREATMENT 108 BENKOVA, E., WITTERS, E., VAN DONGEN, W., KOLAR, J., BRZOBOHATY, B., VAN ONCKELEN, H., MOTYKA, V., MACHACKOVA, I.

KANAMYCIN UPTAKE BY CONTROL (SENSITIVE) AND TRANSGENIC (RESISTANT) POTATO IN THE PRESENCE OF INCREASING AMOUNTS OF EXTERNAL Ca2+ 109 BERENYI, M., KEPPEL, M., KOPPER, E., BURG, K., SCHMIDT, J.

POTATO TRANSFORMATION WITH CECROPIN AND ATTACIN GENES 110 BERENYI, M., KOPPER, E., KEPPEL, M., SCHMIDT, J.

EFFECT OF LOW pH ON THE STATE OF SHOOTS, ROOTS AND ENERGY TRANSFORMATION SYSTEMS IN CHLOROPLASTS OF PLANTS OF DIFFERENT GENOTYPES 111 BUDAGOVSKAYA, N. V.

STUDIES ON VEGETAL AND MICROBIAL UREASE OBTENANCE 112 CAMPEANU, G. H., EREMIA, M., DINU, L.-D., BURCEA, M.

STUDIES ABOUT CALLUS AND PLANTLET REGENERATION USING THE MATURE EMBRYOS IN WHEAT (T. AESTIVUM) 113 CASIAN, H., MORARU, I., STROE, E.

97 APOPTOSIS AND ORGANOGENESIS PROCESS IN CACTACEAE AT IN VITRO CULTURE 114 CORNEANU, G. C, CORNEANU, M., MOGOANTA, L, CRAClUN, C, CRACIUN, V.

IN VITRO DEVELOPMENT OF THE CELLS AND EXPLANTS IN SIMILAR CONDITIONS TO THOSE FROM EXTRATERRESTRIAL ENVIRONMENT 115 CORNEANU, M., CORNEANU, G. C, CRACIUN, C, MORARIU, V. V., ARDELEAN, A., MARINESCU, G,, BADEA, E., CRACIUN, V., BICA, D., MINEA, R., HANESCU, V.

RELATION BETWEEN ELECTRICAL CONDUCTIVITY AND pH IN CONTAINERISED CULTURE OF THREE ORNAMENTAL SHRUBS 116 COSTEA, G., NEATA, G., DAVIDESCU, V., MADJAR, R.

SUNFLOWER EXTRANUCLEAR MUTANT COLLECTION. PHOTOSYNTHETIC REVERTANTS: BACK MUTATIONS OR SUPPRESSION7117 DANILENKO, N.

IDENTIFICATION OF POTATO CYST NEMATODES (PVCN) USING MOLECULAR BIOLOGY 118 DOBRIN, I.

STUDIES REGARDING IMPROVEMENT OF THE TECHNOLOGICAL BARLEY QUALITY INDICES THAT AFFECT BEER QUALITY 119 DRAGHICI, E., BEGEA, M., STROIA, I.

,,IN VITRO" CULTURE ESTABLISHMENT OF COTINUS COGGYGRIA SHOOTS 120 DUMITRASCU, M.

POLYTENE CHROMOSOMES AND NODULIN GENE AMPLIFICATION IN ROOT - NODULES OF FABACEAE: AN ELECTRONOMICROSCOPIC AND GENOME ANALYSIS APPROACH. 121 GAVRILA, L, VLADIMIRESCU, A. L, REBEDEA, I.

THE MODIFICATION OF RADIATION ACTION IN PLANTS BY USING BIOLOGICALLY ACTIVE SUBSTANCES. 122 GONCHAROVA, N., ZHEBRAKOVA, I., MONTIK, T., SHAPCHITS V.

SOME FACTORS AFFECTING CALLUS MEDIATED (INDIRECT) AND DIRECT SOMATIC EMBRYOGENESIS IN PEA (PISUM SATIVUM L.) 123 GRIGA, M. SYSTEMIC ACQUIRED RESISTANCE IN SUNFLOWER (HELLANTHUS ANNUS) 124 GUSCHA, M. I., DYACHENCO, A. I., DMITRIEV, O. P.

SOME RELATIONSHIPS BETWEEN HELIANTHIN AND RESISTANCE TO PHOMOPSIS IN SUNFLOWER GENOTYPES 125 HAGIMA, I., RADUCARU, F.

USE OF RAPD MARKERS IN CHARACTERIZATION OF EARLY SOMATIC EMBRYO CULTURES OF NORWAY SPRUCE 126 HANACEK, P., TRUKSA, M., PROCHAZKA, S.( HAVEL, L.

CORRELATION BETWEEN PLANTING DISTANCE, LIGHT QUANTITY RECEIVED BY CROWN, LEAVES TOTAL CONTENT IN CHLOROPHYLL AND PHOTOSYNTHESIS AT PLUM TREE 127 HOZA, D.

CORRELATION BETWEEN TOMATO PLANT CHARACTERISTICS AND NUTRITIONAL SUBSTRATES 128 HOZA, G., POPESCU, V.

FLOW CYTOMETRY - A RELIABLE METHOD FOR QUICK AND EFFICIENT DETECTION OF SOMACLONAL VARIATION IN NORWAY SPRUCE 129 HRISTOFOROGLU, K., BURG, A., SCHMIDT, J.

THE BEHAVIOUR OF SOME DWARF PEACH AND NECTARIN SELECTIONS IN THE SOUTH AREA OF ROMANIA 130 CHIRA, L, CEPOIU, N., CHIRA, A., DELIAN, E.

HYBRIDIZATION BETWEEN TETRAPLOI HEUANTHUS SPECIES AND CULTIVATED SUNFLOWER H. ANNUUS L 131 CHRISTOV, M.

AGROBACTERIUM - MEDIATED TRANSFORMATION OF NORWAY SPRUCE POLYEMBRYONIC CULTURE 132 KARLOVSKA, L, TRUKSA, M., HAVEL, L.

THE UPTAKE AND METABOLISM OF 3H-BENYZLAMINOPURINE IN THE EXPLANTS OF TOBACCO (NICOTIANA TABACUM L.) AND CUCUMBER (CUCUMIS SATIVUS L.) 133 KLEMS, M., MACHAeKOVA, !., EDER, J., PROCHAZKA, S.

99 THE CHANGES OF ABA LEVEL IN THE ROOT-XYLEM SAP AND PLANTS OF WINTER BARLEY CV. LUNET TREATED BY LOW TEMPERATURE 134 KLEMS, M., FLORES, J., BALLA, J., KREJCJ, P., KADLECOVA, Z., PRASLL, I.

ORIGINAL METHOD FOR DETERMINATION OF SULFOXIDES 135 KOSYAN, A., TARAN, N.

DOES MELATONIN PLAY IN PLANTS A SIMILAR ROLE AS IN ANIMALS? 136 MACHACKOVA, I., KOLAR, J., EDER, J., PRINSEN, E., VAN DONGEN, W., VAN ONCKELEN, H.

THE EFFICIENCY OF ,,HAIRY ROOTS „ TRANSFORMATION TRANSFER SYSTEM IN NICOTIANA TABACUM L. AND VITIS VINIFERA L. ,,IN VITRO" CULTURED TISSUES 137 MAXIMILIAN, C, BREZEANU, A., ROSU, A., VATAFU, I.

CYTOLOGICAL ANALYSIS OF LEAF DISC-DERIVED REGENERANTS OF NICOTIANA PLUMBAGINIFOLIA 138 MAZIKTOKEI K, GYULAI, G., KISS, E., TOTH LOKOS, K., HESZKY L. E.

HYBRID PROGENIES FROM HORDEUM VULGARE L. X HORDEUM BULBOSUM L. INTERSPECIFIC CROSSES 139 MIHAILESCU, A., GIURA, A.

CROSSING ABILITY BETWEEN HELIANTHUS DECAPETALUS L. AND HELIANTHUS ANNUUS L. 140 NIKOLOVA, L, CHRISTOV, M., JAMBASOVA, T.

THE SULPHOLIPID CONTENT AND THE PLANT DROUGHT RESISTANCE 141 OKANENKO, A., TARAN, N.

AFLP ANALYSIS OF RECJPROCAL SUBSTITUTION LINES IN WHEAT (T. AESTIVUM L) 142 OVESNA, J., SULAKOVA, S., KUCERA, L, KOSNER, J.

THE IMPLICATION OF THE KILLER PHENOMENON ON YEASTS FROM WINE-MAKING 143 POMOHACI, N., BURCEA, M., CORNEA, P., RADOI, F., COSOI, C.

PULSED-FIELD GEL ELECTROPHORESIS - METHOD FOR THE TAXONOMIC IDENTIFICATION OF THE OENOLOGICAL YEAST STRAINS 144 POMOHACI, N., BURCEA, M., CORNEA, P., POMOHACI, D.

100 THE CHARACTERISATION FROM A CHROMATICAL POINT OF VIEW OF SOME ROMANIAN ROSE WINES 145 POMOHACI, N., RADOI, F., BURCEA, M.

IN VITRO ESTABLISHMENT AND CHARACTERIZATION OF HAIRY ROOT CULTURES INDUCED WITH AGROBACTERIUM RHIZOGENES STRAINS 146 ROSU, A., CORNEA, P..CAMPEANU, G. H..CEPOIU, N..BREZEANU, A..MAXIMJLIAN, C.

A COLD- AND DROUGHT-RESISTANCE OF THE SAMPLES FROM THE CAMBRIDGE PHASEOLUS GERMPLASM COLLECTION 147 RUSSKIKH, I., SHANGINA, N., DAVYDENKO, O.

GENETICS TRANSFORMATION AND PLANT IMPROVEMENT 148 SANGWAN; R. S., SANGWAN-NORREEL, B. S.

LIPID COMPONENTS OF CHLOROPLAST MEMBRANES IN STRESS CONDITIONS UNDER THE EFFECT OF IMMOBILIZED BENZYL- AMINOPURINE 149 SERGA, O.

THE FERTILIZER INFLUENCE ON SOME PHYSIOLOGICAL PARAMETERS OF ENSILAGE CORN THAT IS GROWN ON THE MEADOW CALCEROUS SOFT-LOAMY ON LOESS LOAM CHERNOZEM OF THE NORTH PART OF UKRAINIAN FOREST STEPPE 150 SMETANSKAYA, I. N.

ORGANOGENESIS VIA CALLUS IN KIWIFRUIT HYBRID PLANTS (ACTIMDIA DEUCIOSA CHEV. X ACTINIDIA ARGUTA SIEB. E ZUCCH.) 151 STANICAF.

DETERMINATION OF GENETICALLY DISTANCES IN ROMANIAN SOYBEAN (GLYCINE MAX) GENOTYPES 152 STROE, E., GRESSHOFF, P.

BARLEY AND HOP VARIETIES CULTIVATED IN ROMANIA FOR BREWING 153 STROIA, I., BEGEA, M., DRAGHICI, E.

APPLICATION OF SOME BIOTECHNOLOGICAL METHODS AS AN ALTERNATIVE APPROACH IN RESISTANCE BREEDING OF PEAS TO FUNGAL PATHOGENS 154 SVABOVA, L, GRiGA, M.

101 GALEGA ORIENTAL® LAM. - A NEW VALUABLE FORAGE PLANT 155 TELEUTA, A.

THE PROMOTIVE EFFECT OF COMBUSTION PRODUCTS FROM PLANT MATERIAL ON THE RELEASE OF SEEDS FROM DORMANCY.156 THOMAS, T. H., THORNTON, M. A., PETERS, N. C. B., VAN STADEN, J.

STUDIES ON THE BIOCHEMICAL TRANSFORMATIONS DETECTED IN POLYANTHES TUBEROSA L. PLANTS 157 TOMA, F. L, NEATA, G., DAVIDESCU, V., DIMA, I.

TESTING THE REGENERATIVE CAPACITY IN JN VITRO" CONDITIONS OF THE POLYANTHES TUBEROSA L. PLANTS 158 TOMA, F. L, ROSU, A.

SUNFLOWER EXTRANUCLEAR MUTANT COLLECTION. PHOTOSYNTHETIC REVERTANTS: BACK MUTATIONS OR SUPPRESSION?159 TRIBOUSH, S., ULITCHEVA, I., DANILENKO, N., DAVYDENKO, O.

CHROMOSOME ISOLATION FROM NORWAY SPRUCE (PICEA ABIES (L.)KARST.) ROOT TIPS. 160 GBERALL,!., HAVEL, L.

A SIMPLE METHOD FOR DNA ISOLATION FROM ALGAE USING CHELEX -100. 161 VLADIMIRESCU, AL, GAVRILA, L, GRECU, C.

IMAGE ANALYSIS IN CHARACTERIZATION OF NORWAY SPRUCE POLYEMBRYONIC CULTURES 162 VLASlNOVA, H., TRUKSA, M., HAVEL, L.

THE AUXIN AND CYTOKININ ECONOMY DURING THE GROWTH CYCLE OF VBI-0 TOBACCO CELL CULTURE 163 ZAZIMALOVA, E., PETRA$EK, J. BftEZINOVA, A.

THE EFFECT OF ETHREL ON CUCUMBER PLANT GROWTH AND FLOWERING 164 ZELENA, E., ZELENY, F., VECEROVA, D. Working group No.4 NOTES HISTOLOGICAL ANALYSIS OF DIFFERENT TYPES OF SYNGONIUM PODOPHYLLUM SCHOTT. CALLUS ANTOFIE, M. M., ANGHELUTA, R. Institute of Biology, Bucharest, Romania

This paper presents the main morphogenic events that appear in Syngonium podophyllum Schott callus for different types of media used in initiation and subcultivation. By cultivating syngonium on different types of media, containing MS 1962 and N69 minerals, different combination of hormones (2,4D 0.1 mg/l, NAA 0.1 mg/l, Kinetin 3 mg/l and BA 1 mg/l) vitamins MS 1962 and N69, and different quantities and qualities of carbon source (glucose 30 g/l and sucrose 30-60 g/l) we obtained distinct forms of non-friable callus. All types of callus were morphogenics and macroscopically it was difficult to establish the shoots origin because of their high density and very small size. As a general feature, for all MS media we obtained a yellowish type of callus and for all N69 media a green type callus. We also made different variants of macroelements for N69 media and obtained interesting results for 3/4, 1/2, 1/4 macroelement variants using as hormones 2,4D 0.1 mg/l, Kin 3 mg/l and BA 1 mg/l. We did not obtain macroscopical differences when using a high supply of nitrogen source. Using hematoxyline as dye we made histological investigations and we relived for calluses originating from MS media all stages of somatic embryogenesis and for N69 originating callus we only obtained organogenic events, very highly expressed.

103 Working group No. 4 NOTES BIOACTIVE SUBSTANCES TESTED AT IN VITRO EXPLANT CULTURE ARDELEAN, A.1, CORNEANU, G.2, CORNEANU, M.2, MURESAN, N.2, BRONZETTI, G.3, FURUKAWA, H.4, JUNKO, E.5, BICA, D.6, MINEA, R.e, GABOR, L.6, FRANGOPOL, C.7 University of West ,V. Goldis", Bvd. Revolutiei, 81,2900-Arad, Romania; University of Craiova, Genetics Section, A.I. Cuza 13,1100-Craiova; Romania* 3lstituto di Mutagenesi e Differenziamento, Via Svezia 10, 56124-Pisa, Italy; "Meijo University, Pharmacology Dept., 468-Nagoya, Japan; Osaka City University, Living Sciences Dept.,468-Osaka, Japan; Politehnica" University, 1900-Tlmi8oara, Romania; sc-*5r- Electroputere, 1100-Craiova, Romania

Some bioactive substances extracted from plants or obtained through synthesis, presented a stimulatory, antimutagen or anticarcinogen activity (in vivo or in vitro experiments). The magnetic fluids, used recently in biotechnology (at supplementation of the culture medium at in vitro cell and tissue culture), present a stimulatory activity of the organogenesis processes. Also the magnetic fluid acting as hormone substitute and for the recovery of the senescent and partial vitrified plants (experiments in Fragaria ananassa, Mammillaria duwei, a/o). Their effect is dependent on their features (composition, ratio between Fe2+: Fe3+; the presence of other elements, the dispersed phase, their concentration in magnetite particles a/o), genotype and the explant type, the analysed organogenesis process, a/o. The researches performed over some genotypes and explant type, point out that the magnetic fluids with petroleum or oleic acid as dispersed phase induced supper values for organogenesis and growth rhythm, while the magnetic fluid on water basis manifest a lower effect at in vitro explant culture over the same processes (Mammillaria duwei, Chrysanthemum hortorum, tpomoea batatas, a/o). Also, they manifest a protective effect against some stress factors. Thus, in the experiments performed in vivo in Nigella damascena, a magnetic fluid on petroleum basis or a complex magnetic fluid on oleic acid basis (Zn : Mn : Fe3+), in a concentration of 10%, presents in the time of seeds irradiation with X-rays (40 Gy), manifests a radioprotective effect (reduce the chromosomal aberration and initiate the recovery phenomena). In other experiments with the simulate action of some extraterrestrial factors (cvasi-null or double geomagnetic field, screened geoelectrical field, negative thermic shocks of-196° C, a/o), in vitro inoculated explants on a culture medium supplied with a magnetic fluid on oleic acid or petroleum basis, not presented alteration of the cell ultrastructure, indifferent of the values of the geomagnetic field, while a magnetic fluid on water basis, induced severe alteration of the cells from parenchyma, the alteration being more severely in a natural geomagnetic field. Some naphtoquinones derivatives were tested both in vivo (Nigella damascena) and in vitro culture (Mammillaria duwei, Drosera rotundifolia, Dendrobium sp., a/o.) They manifest an antimutagen effect through the reducing of the chromosomal aberrations value, as well as a stimulatory effect at in vitro culture. Also, were tested the action of some leaves total extracts or chiorophyllin. A total extract from Spinacia oleracea or Brassica rapa HV nKomatsuna" leaves were tested for their antimutagen or stimulatory effect at in vitro culture, in Chrysanthemum hortorum and tpomoea batatas (stem fragments with a node, inoculated on an MS medium supplied with 1.08 mg/l KIN and 1.0 mg/l IAA. The shooting process and shoot length are stimulated by Komatsuna 0.6 g/l leaves extract in /. batatas, while at Ch. hortorum this presence is little influenced by the two natural antimutagenic extracts. The antimutagen effect of the Spinacia leaf total extract (600 mg/l) or sodium-copper-chlorophyllin (20 mg/l), against X-rays (104 Gy) was tested at in vitro culture of the Dendrobium sp. somatic embryos. The chiorophyllin presence in the culture medium, enhanced the organogenesis processes, while the Spinacia oleracea leaves extract enhanced the embryogenesis processes, more evident in the irradiated variants, in comparison with the Control.

1/1/1 Working group No.4 NOTES "IN VITRO" REACTIVITY ON SOME RARE PLANTS OF SPONTANEOUS FLORA AZEMA, T., CIUBOTARU, A., KIRTOACA, V., MALAHOVA, L. Institute of Botany, Academy of Sciences, Republic of Moldova, P.O. Box 3466, 2060 Chisinau-60, Moldova

At the end of this century we run the risk of continuous loss of species from the spontaneous fauna and flora. These are species for particular decoration or those utilized by man in different fields: food, medicine, and cosmetics. The increasing demands of pharmaceutical industry in the secondary metabolites of spontaneous plants and its protected status in several European countries including Moldova has required further development of tissue cell culture techniques. Six species of plants of spontaneous flora were introduced "in vitro". The ability of different explants to regenerate was studied: leaf fragments, stem shoots with nodal buds, flower segments; fruit portions; seeds as well as bulbs scales. Positive results have been obtained only for 3 species out of 6. Cephalanthera damasonium (Mill.) Druce (Orchideaceae). In nature this plant regenerates from a single apical bud. The seeds contain a poorly developed embryo and endosperm, which is not sufficient for embryo nutrition, so the seeds have very low fertility (2-3%). Inoculated on Whait medium the seeds formed a small conglomerate of cells after 4-5 weeks of cultivation. Gibberellic acid and Kinetin ratio served as a basis for seed stratification - a stage undergone by them in nature. Long transfers on Murashige Skoog (MS) medium supplemented with 0,2 mg/l - Kin; 0,25 mg/l - IAA; and 0,25 mg/l - BAP; 0,05 mg/l - NAA and 0,05 mg/l Gk3 pH 5,8 resulted in the proliferation of some weakly developed callus. Paeonia peregrina (Mill.) (Paeoniaceae). A plant used in popular Bulgarian medicine. It contains the poisonous substance - pennon, as well as tannin, used as strong dyes. The plant also contains etheric oils, glutamine and arginine. It is very decorative which is at the basis of selecting peonies in horticulture. The chemical composition of the plant influenced the manipulation "in vitro" as the inocula provoked the formation of secondary metabolites, which inhibited callus proliferation. We managed to obtain the callus with 0,25 mg/l - BAP and 0,1 mg/l - NAA pH 5,6. Lilium martagon L. (Liliaceae) An especially decorative rare plant recommended for the embellishment of natural parks. The plant blossoms in the period with sufficient rainfalls but the young plant blossom every 4-5 years. It has been shown that only 5-7% of the plant blossom in the vegetal population. Young nodal segments of shoots, juvenile leave and bulbous segments have been inoculated on different media. Control of dormancy development during bulblets tissue culture is highly desirable. Their development does not start immediately but after a period of rest. At the final stage they form "in vitro" a lot of proliferation observed on the MS basal medium supplemented with 1,0 mg/l BAP and 0,25 mg/l NAA. Further multiplication of bulblets with a differentiated leaflet can be effected on 1/4 liquid MS medium free of hormones. These results are of great importance for: a) the repatriation of the plants to their proper area; b) "in vitro" conservation of their genotype; c) obtaining artificial seeds to spread the species; d) selecting decorative plants with a high potential to disease resistance, temperature variations, humidity and other climatic conditions.

105 Working group No.4 NOTES HORMOMAL REGULATION OF ASSIMILATE PARTITIONING BAKER, D. Department of Biological Sciences, Wye College, University of London, Ashford, Kent TN25 5AH, UK

The partitioning of assimilates between photosynthetic source organs and utilizing sink organs is widely accepted to be regulated by endogenous plant hormones. The key intermediate steps involved in assimilate transport, such as phloem loading and unloading, have been shown to be responsive to applied hormones, although the role of endogenous hormones in these processes remains essentially unresolved. Analyses of vascular saps supplying source and sink organs have demonstrated the presence of the major endogenous hormones and/or their precursors. lndo!-3yl-acetic acid, a number of gibberellins, cytokinins and abscisic acid, as well as the precursor for ethylene production have been found in these vascular saps, allowing the sites of hormonal synthesis and putative target tissues to be deduced. Exogenously applied hormones are also easily loaded into these vascular pathways and may be translocated over considerable distances from a point of application. Observations such as these indicate a possible coordination system between source and sink regulated by the synthesis and transport of endogenous hormones. Results will be presented on analyses of vascular saps from RICINUS COMMUNIS using a range of physico- chemical methods. These results will be compared and contrasted with those obtained by the application of exogenous hormones, or their precursors, and critically evaluated and interpreted in the light of current models of source : sink regulatory processes.

106 Working group No. 4 NOTES INTERACTION OF CYTOKININS AND AUXIN IN PEA LATERAL BUDS AFTER BREAKING AND RENEWAL OF APICAL DOMINANCE BALLA, J., BLAZKOVA, J., PROCHAZKA, S. Department of Botany and Plant Physiology, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00 Brno, Czech Republic, E-mail: [email protected]

Hormon involvement in apical dominance (AD), the phenomenon of control exerted by apical portions of the shoot over the outgrowth of lateral buds, has been studied over sixty years. The usual model for AD studies are decapitated plants where the focus is on the hormonal changes (CKs, IAA) in lateral buds immediately after breaking of AD. Less attention is paid to changes taking place during the longer interval covering the renewal of AD. The model used in our work were pea lateral buds of 2nd primary scales showing the highest growth potential. The influence of apex was eliminated by detaching the buds from 21 days-old mother plants followed by their transfer to half strength MS medium without any hormones. Development of buds under in vitro conditions was observed for 4 weeks. At the end of culture period 75% of regenerated buds developed visible flower buds. Stages of differentiation were characterized in week intervals by anatomical studies, analysis of endogenous level of CKs (Z, iP, DHZ, BA, mT, oT, their ribosides and 9-N-glucosides) and IAA. The main elevated CKs were iP, BA and mT, or their derivatives, where, after 1 week in culture, the levels increased in case of iPR 3-fold, BA 6-fold and mTR 4-fold. Uptake of 3H-BA, 3H-Z and 3H-1AA was observed in each stage of differentiation.

107 Working group No. 4 NOTES CYTOKININS IN TOBACCO AND WHEAT CHLOROPLASTS: OCCURRENCE AND CHANGES DUE TO LIGHT/DARK TREATMENT BENKOVA, E.1, WITTERS, E. 2, VAN DONGEN, W.2, KOLAR, J.3, BRZOBOHATY, B.1, VAN ONCKELEN, H.2, MOTYKA, V. 3, MACHACKOVA, I.4 institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135,61265 Brno, Czech Republic 2 Department of Biology, University of Antwerpen, B2610 Wilrijk-Antwerpen, Belgium 3 Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojova 135,16502 Praha 6, Czech Republic "Norman Borlaug Institute for Plant Science Research, De Montfort University and Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojova 135,16502 Praha 6, Czech Republic, E-mail: [email protected]

Many cytokinin effects are connected with chloroplasts: cytokinins are necessary for proper chloroplast development and functioning. But until now, it was not rigorously shown, if chloroplats contain cytokinins. Purified intact tobacco chloropiasts contain broad spectrum of cytokinin molecules as shown by liquid chromatography/tandem mass spectrometry: isopentenyladenine, isopentenyladenosine, zeatin riboside, isopentenyladenosine-5'-phosphate, zeatin riboside-5'-phosphate, dihydrozeatin riboside-5'-phosphate and N- glucosides of zeatin, dihydrozeatin and isopentenyladenine. No O-glucosides were detected. The levels of zeatin-N9-glucoside (ZN9G) and dihydrozeatin-N9-glucoside (DZN9G) were significantly increased and those of other N-glucosides decreased in comparison with the leaf tissue. Tobacco chloropiasts from leaves at the end of 12 h dark period have higher CK content (when expressed on chlorophyll) than those isolated at the end of the light period. The most striking difference was found in the levels of ZN9G and DZN9G. Interestingly, some O- glucosides were detected in chloropiasts isolated after 12 h of darkness. Similar, but less dramatic changes were found in chloroplasts isolated from wheat leaves. Tobacco chloroplasts also display cytokinin oxidase activity. Our results show that chloroplasts not only contain a broad spectrum of cytokinins, but are also probably able to metabolically interconvert individual forms and also degrade cytokinins by cytokinin oxidase.

ins Working group No.4 NOTES KANAMYCIN UPTAKE BY CONTROL (SENSITIVE) AND TRANSGENIC (RESISTANT) POTATO IN THE PRESENCE OF INCREASING AMOUNTS OF EXTERNAL Ca2+ BERENYI, M., KEPPEL, M., KOPPER, E., BURG, K., SCHMIDT, J. Austrian Research Centers, Seibersdorf, Dept. Biotechnology, A-2444 Seibersdorf

Aminoglycoside antibiotics (like kanamycin (Km)) are frequently used for the selection of transgenic plant cells. However, for a number of species Km selection is inefficient. It was reported that the phytotoxic effects of Km were strongly dependent on the Ca2+ concentration in the growth media. (M.Joersbo and F.T.Okkels Physiologia Plantarum 97: 245-250 1996.) The object of the present study was to analyse the effect of the external Ca2+ concentration on the shoot and root elongation of both kanamycin sensitive and resistant potato plants in the presence of 172uM Km (100mg/l). Therefore we studied the interaction of kanamycin uptake and Ca2+ concentration in both sensitive and resistant potato plants by measuring the free and bound kanamycin levels of the plants in the presence of increasing concentrations of Ca2+ in the medium. Using two kanamycin resistant transgenic, and four sensitive potato varieties, we measured the levels of the kanamycin in whole plants as well as in the root and in segments of the stem. The plants were grown in medium containing different calcium (3-15 mM) and standard kanamycin (172uM) concentrations. The Km levels were measured by bacterial growth inhibition test using Km sensitive E. coli. The bound Km was measured by releasing it by elevating the calcium concentration of the plant homogenate to 40-120 mM. Increasing concentrations of external Ca2+ reduced the toxic effect of the antibiotic on the root growth of both sensitive and resistant plants and facilitated the kanamycin uptake (sum of bound and free Km) of the sensitive ones. The highest free and bound Km levels were found at the stem base in case of both the resistant and the sensitive plants. The bound Km decreased dramatically while the free Km was not changed by the increasing Ca2+ concentration of the growth medium. In the stem the Km levels decreased with the height yielding no measurable Km at the top of the stem. In the stem base the bound Km was always higher in the sensitive plants compared to the resistant ones, whereas the level of the free Km concentration was about the same in both the resistant and sensitive plants. The difference between the levels of the bound Km of the sensitive and resistant plants could be explained by supposing that the NPT II enzyme (neomycin phosphotranspherase) which is present in the Km resistant transgene potato plants is membrane bound.

109 Working group No.4 NOTES POTATO TRANSFORMATION WITH CECROPIN AND ATTACIN GENES BERENYI, M., KOPPER, E., KEPPEL, M., SCHMIDT, J. Austrian Research Centers, Seibersdorf Dept. Biotechnology, A-2444 Seibersdorf

The cecropins are a family of low molecular weight (4kD) basic inducible peptides first found in the haemolymph of silkmoth pupae (Hyalophora cecropia) as a response to a bacterial infection. The attacins were also isolated from the giant silkmoth pupae, where they are acting synergistically with the cecropins. In the present study, we have transformed potato plants with synthetic cecropin B and attacin E genes either alone or in combination with each other in order to mimic the natural immune system of the insect, where the antibacterial peptides acts together. Two different versions of the modified cecropin B (C38 and C4) were applied in the transformation experiments. The C38 gene (synthetic cecropin without signal peptide) was combined with the enhanced CaMV double35S promoter while the C4 gene (synthetic cecropin gene with hordothionin signal) with the mas2-mas1 dual promoter. We have made various constructs combining the C4 and attacin genes with the mas2- mas1 promoter pair, where the mas1 promoter is a weak constitutive while the mas2 promoter is an inducible and stronger promoter. On this way, we made cecropin-cecropin and cecropin-attacin combinations. These constructs were introduced into the pMOG 402 vector, which contained the selective NPT II kanamycin (Km) resistance gene and Agrobacterium transformation system was used to obtain transgene potato plants. In the Km selected transformants both the transcription of the attacin and cecropin genes and the presence of the cecropin peptide in the root, stem, leave and tuber of the transgene potato plants were tested. We have also analysed some selected transgenic clones in in vitro stem base resistance test against Erwinia infection. With Southern analysis, we have found usually 1-3 insertions of the constructs. Northern analyses of both the cecropin and attacin expression showed strong transcription in the stem, leaf and root tissues with all promoter combinations, but in the tuber we have found transcription only in the plants where the cecropin gene was driven by the CaMV 35S promoter. In the case of the mas2-mas1 promoter driven C4 gene we have found transcription only in a single done, where the promoter pair contained only a single cecropin gene in the mas2 position. We have tested the cecropin content of the mas2C4-mas1C4, mas2C4-mas1- and CaMV 35S-C38 construct containing clones analysing the level of the peptide in the stem, leaf, root and tuber respectively. The sensitivity of the competition ELISA was about 5-10 ng/ml, the calibration curve was linear between 10 and 1000 ng/ml. We could detect cecropin from the stem and leaf tissues, however no or marginal amounts were detected from root and tuber. Control experiments showed that during the purification we lost more than 90% of the peptide, so we could not exclude low levels of cecropin expression also in the root and tuber. Usually we could detect cecropin between 1-20 ng/g fresh plant from the plant organs. The highest cecropin content obtained was 200-500 ng/g plant leaf with the CaMV 35S-C38 construct, but only in a single clone. Considering the 90% lost of cecropin during purification, we can expect 20-400 ng/ml and in a single clone up to 1mg/ml cecropin concentrations in the stem and leaf tissues. In vitro stem base resistance test with the Erwinia ssp causing blackleg yielded a better survival compared to control plants. Comparing the different combinations it is possible to conclude that the survival of the mas2C4-mas1 attacin and mas2C4-mas1C4 combinations were better than that of the mas2attacin-mas1C4.

110 Working group No. 4 NOTES EFFECT OF LOW pH ON THE STATE OF SHOOTS, ROOTS AND ENERGY TRANSFORMATION SYSTEMS IN CHLOROPLASTS OF PLANTS OF DIFFERENT GENOTYPES BUDAGOVSKAYA, N. V. Institute of Plant Physiology of the Russian Academy of Sciences, Botanicheskaya 35, Moscow 127276, Russia, E-mail: [email protected]

Acid soils occupy vast territories in many countries. Besides, sulfurous-acid anhydride and some other industrial gases cause "acid rains" when getting to the atmosphere. All these factors create serious problems for agriculture. Low pH provoke poor development and low productivity of plants. In this connection evaluation of the state of plant under acid pH conditions and selection of tolerant genotypes is an important task. Different in tolerance cultivars of buckwheat, wheat, pea and maize were grown at pH 3.0, pH 4.0 and also at pH 6.0 (control). Acidification of shoot and root tissues, slowing down in the growth of these organs and deviation of relationships of dimensions of shoots and roots and of pH of shoot and root tissues from normal values were observed for susceptible plant cultivars grown on acid media. The magnitude of this deviation was proportional to the degree of the observed functional disturbances in plants. Low pH caused a decrease in the activity of light- induced proton uptake and ATP formation in chloroplasts. Disturbances in energy transformation systems were expressed more in plants with a low content of SH-compounds in chloroplasts. For plants of tolerant cultivars, grown under low pH, the pH of shoot and root tissues and the growth rates of these organs were preserved at the level of control plants or alkaiinization of shoot and root tissues and increase in the plant growth rate were observed. Under low pH proportions of dimensions of shoots to roots and of tissue pH of shoots to roots for tolerant cultivars are equal to the control variant or "adaptive" proportion values are maintained which slightly differ from "normal" values. Proportions of the measured parameters are values characteristic of the cultivar. They are more stable indexes than initial parameters used for calculations (the measured parameters change with the age of plants). Structure-functional proportions of shoots and roots express interrelations of the main parts of plant - photoautotrophic and heterotrophic. They are more informative indexes for testing plants as unitized dynamic systems than local indexes of the state of different tissues and organs.

111 Working group No. 4 NOTES STUDIES ON VEGETAL AND MICROBIAL UREASE OBTENANCE CAMPEANU, G. H.\ EREMIA, M.2, DINU, L.-D.1, BURCEA, M.1 1University of Agronomical Sciences and Veterinary Medicine Bucharest, Faculty of Biotechnology, Marasti, 59, 71331, Bucharest, Romania, E-mail: dinue @ fx. Ro 2Chimico-Pharmaceutical National institute of Research and Development Bucharest

Urease (E.G. 3.5.1.5.) used in diagnostic kits for the determination of urea in physiological fluids as blood, can be obtained from vegetable sources {jack-bean, Canavalia ensiformis, Citrullus vulgaris) and microbial sources (Corynebacterium sp., Brevibacterium sp., Arthrobacter sp., Proteus vulgaris). Methods of enzyme isolation depend on source, urease from water melon (Citrullus vulgaris) seeds was isolated using organic solvent (diclormethan ) and for the bacterial enzymes, which is an intracellular enzymes, we used mechanical methods. For the purification of the vegetable urease we used ion-exchange chromatography (DEAE - Sephadex). In our research work we obtained purified urease from, water melon (Citrullus vulgaris) seeds and two bacterial strains urease producing Brevibacterium lactofermentum I.C.C.F. 7, Corynebacterium melassecola I.C.C.F. 16) as a result of screening program. Working group No. 4 NOTES STUDIES ABOUT CALLUS AND PLANTLET REGENERATION USING THE MATURE EMBRYOS IN WHEAT (T. AESTIVUM) CASIAN, H.\ MORARU, I.2, STROE, E.1 1University of Agronomical Sciences and Veterinary Medicine Bucarest, 59 Bd. Marasti, 71331 Bucarest, Romania 2Research Institut for Cereals and Industrial Crops, 8264, Fundulea, Romania

Our experiment was initiated to identify the genotypes with caliusation capacity frcm mature embryos and to verify regeneration ability of this type of callus. Additionally a number of medium variants were tested for that. Regarding caliusation capacity 8 genotypes from 14 tested genotypes were selected for "in vitro" comportment. A high frequency of callus production was observed using MS to cultivate embryos, comparatively with LS medium.

113 Working group No. 4 NOTES APOPTOSIS AND ORGANOGENESIS PROCESS IN CACTACEAE AT IN VITRO CULTURE CORNEANU, G. C1., CORNEANU, M1, MOGOANTA, L2., CRACIUN, C3., CRACIUN, V3. 1 Craiova University, Genetics Sect., 1100-Craiova, Romania, E-mail: [email protected].; Craiova University, Medicine Dept., 1100-Craiova, Romania; 3"Babes-Bolyai° University, Biology Dept., 3400-Cluj-Napoca, Romania.

Previously research point out the apoptosis importance in the embryogenesis process in different plant species (Picea abies, Cupressus sempervirens a/o, Havel et al., 1996, 1997). Present research point out the apoptosis implication in the organogenesis processes in Mammillaria duwei (Cactaceae; in vitro culture) at the leading system formation and of the thorns (or spines as leaves completely metamorphosed), characteristics for these plants. In vitro culture in M. duwei was initiated through using of the stem fragments, inoculated on an MS basal medium supplied with 1.8 mg/l IAA, 0.5 mg/l NAA and 0.011 mg/l KIN. At the basis of these explants, were developed a friable, unorganogen callus as well as a compact, organogen callus. Apoptosis and oraanogenesis process in M. duwei. The organogen callus present at outside an unistratified epidermis covered with a thin cuticle. In organogen callus, the organogenesis process begins with the leading system forming. In the middle of an organogen area, take place the tracheids forming (xylogenesis process), through folding and thickened of the cell longitudinal walls. These cells elongation is accompanied by an apoptosis process of the adjacent parenchymatic cells. Apoptosis and embrvoqenesis process in M. duwei. In Cactaceae, mammilla is an auxoblast offshoot having a short axis to end in a vegetative tip (by example in M. duwei), or in a vegetative cone. In M. duwei the mammilla length is among 2-8 mm and areolas present 1.5 - 2.5 mm in length and 8 mm breadth. The radial thorns number is 28-36. Can be present or not a central spine of 10 mm length. In M. duwei {in vitro) an apoptosis process was emphasised in the time of the organogenesis processes from mammilla, (a) The vascular system present origin in the intercellular spaces of the fundamental parenchyma under epidermis and is formed through an apoptosis process. These spaces result as following of apoptosis process of some parenchyma cells, from the subepidermal region of the stem. These intercellular spaces, are continued with the libero-ligneous vessels, (b) The spines (thorns) are formed also through an apoptosis process. In the tip of a young mammilla, is an organogenic zone, in which through a succession of modifications take place the spines formation. The cells from this region, disposed on 3-4 rows, are in an intense mitotic division. As result of the mitotic divisions, resulted two cells populations, disposed in two regions alternate one to other, covered with a unicellular layer. In one region, the cells are big, with pelicular cytoplasm and poor in cellular organites, similar to those from the aquiferrous parenchyma. In the other region, the cells are disposed on 3-4 rows, perpendicular on the organogenic zone surface. They are flattened, with the thickened walls. Subsequent, the walls of the flattened cells begin very strong sclerified and they are transformed in spines. The unicellular layer situated oven they are distort through apoptosis, and the cells from the intermediary regions are exfoliated. The formed spines grow in length through cells multiplication from the generative region, until they catch up to normal size, characteristic for the considered specie. This process present (probably) a genetic determinism, being a result of the cactus's adaptation at the special environment conditions. The evaluation of this process, permit an explanation for the leaves reduction in Cactaceae family, and their adaptation at other environment conditions, with a different degree of the leaf reduction, from Peireskia sp. with leaves and spines to globular forms as Mammillaria sp., as well as others related species. Working group No. 4 NOTES IN VITRO DEVELOPMENT OF THE CELLS AND EXPLANTS IN SIMILAR CONDITIONS TO THOSE FROM EXTRATERRESTRIAL ENVIRONMENT CORNEANU, M.\ CORNEANU, G. C.\ CRACIUN, C.2, MORARIU, V. V.3, ARDELEAN, A.4, MARINESCU, G,5, BADEA, E.5, CRACIUN, V.2, BICA, D.6, MINEA, R.7, HANESCU, V.1 1Craiova University, Genetics Sect., 1100-Craiova, Romania, E-mail: [email protected].; ^Babes-Bolyai" University, 3 Biology Dept, 3400 Cluj-Napoca, Romania; ITIM, P.O. Box 700, 3400-Cluj-Napoca, Romania; "University of West nV. Goldis", 2900-Arad, Romania; "Craiova University, Biochemistry Dept., 1100-Craiova, Romania; ^Politehnica" University, RCHCMF, 1900-Timisoara, Romania; ^Politehnica University", Chemistry Technological and Environment Engineering, 1900-Timisoara, Romania.

Different authors performed experiments regarding to the effect of different geomagnetic field values, geoelectrical field and other factors from extraterrestrial environment, about living systems, as well as their interaction with other internal or environmental factors. Previously research performed of ours collective in bacteria and plant cells and tissue, reveal the geomagnetic field effect (of different value) on the cell ultrastructural, as well as his biochemical and metabolic activity. In different bacteria genotypes maintained for different time (97 - 1800 hours) in a near-null geomagnetic field, were recorded differences in their ultrastructure and development, depending on genotype and time action of the geomagnetic field (Corneanu G.C. et al., 1996). Also, was point out the interaction between magnetic field and other environmental factors. Previously research performed in different plant species, evidentiated a stimulatory effect of magnetic fluid at in vitro or in vivo culture, dependent on the magnetic fluid features and its concentration, genotype and explant type, analysed biological process, a/o (Corneanu et al., 1993-1998). The interaction between the different values of the geomagnetic field (natural, cvasi-null), and the supplementation of the culture medium with different magnetic fluids type, affect the explant ultrastructure at in vitro culture in Mammillaria duwei (Comeanu G.C. et al., 1995, 1996), as well as the organogenesis process in Mammillaha duwei and Drosera rotundifolia (Corneanu M. et al., 1996). Recently, was studied the effect of an initial period of development in Dioscorea alata cells (266 hours), Dendrobium sp. somatic embryos and Krainzia longiftora callus (816 hours) at different values of geomagnetic field (natural, cvasi-null or double) and geoelectrical field (natural or screened). The effect is dependent on the genotype, the geomagnetic and geoelectrical field value, analysed features a/o. Thus, in Dioscorea alata cells, the best values for the synthesis activity and burgeonesis process recorded at variant developed in an initial phase (216 hours) in a cvasi-null geomagnetic field, with a screened geoelectrical field. In Dendrobium sp. somatic embryos, the screening of the geoelectrical field in an initial stage, in a natural geomagnetic field, enhanced the organogenesis activity. Between the explant in vitro development and organogenesis processes on one hand, and peroxidase isozymes activity on other hand was established a positive correlation. Also, was studied the effect of three factors (geomagnetic field, geoelectrical field, negative thermic shocks of -196° C) on the eukaryote cell in vivo or in vitro development in the presence or not of a magnetic fluid. The seeds of Nigella damascena, or Arachis hypogaea, previously maintained or not in a magnetic fluid (3 hours), were exposed (time of 1-60 min, respectively 20 min) at the single or combined action of the natural or cvasi-null geomagnetic field, screened or not geoelectrical field and a thermic shock of -196° C. The presence of a magnetic fluid on petroleum basis, at the concomitant action of a cvasi-null geomagnetic field with screened of a geoelectrical field at in a -196 C, enhanced the organism response capacity, manifest through enhanced the peroxidase isozyme activity (their number and activity). This research presents an importance in the spatial biology.

115 Working group No. 4 NOTES RELATION BETWEEN ELECTRICAL CONDUCTIVITY AND pH IN CONTAINERISED CULTURE OF THREE ORNAMENTAL SHRUBS COSTEA, G., NEATA, G., DAVIDESCU, V., MADJAR, R. University of Agronomical Sciences, Faculty of Horticulture, Bd. Mara§ti 59, 71331 Bucharest, Romania.

Three ornamental woody species of one year old: Euonymus japonicus Thunb., Pyracantha crenatoserrata (Hance) Rehd. and Ilex aquifolium L, were grown in three different container substrates, consisted in peat, leaves compost, fallow and sand in various proportions and were weekly fertilised with two nutritive solutions (the control was watered only with water) in a bifactorial experiment (2 x 3), with four replication. Samples of substrates for each species were monthly analysed for pH (H2O), electrical conductivity (EC) in ms/ cm and mineral nutrients content ( N, P, K ). The growth dynamics of the plants was pursued by measuring the cumulated length of the shoots for Euonymus and Pyracantha and by counting the leaves of Ilex. We found a close relation between pH and electrical conductivity values during the growing season. There were variations depending on the utilised nutritive solution and the moment of the culture. In June and July pH and EC values were constant, but in August the pH values decreased and the EC values increased for the same variants, in all species.The increase of soluble salts concentrations in the substrates was the result of nitrates and phosphates accumulation, which were less absorbed. However, the plants were not affected; Euonymus and Pyracantha presented a linear, continuous growth, while Ilex had a "S" shaped growth. Working group No.4 NOTES SUNFLOWER EXTRANUCLEAR MUTANT COLLECTION. PHOTOSYNTHETIC REVERTANTS: BACK MUTATIONS OR SUPPRESSION? DANILENKO, N. Institute of Genetics and Cytology, Kuprevich, 220141 Minsk, Belarus, E-mail: [email protected]

Abstract was not delivered

117 Working group No.4 NOTES IDENTIFICATION OF POTATO CYST NEMATODES (PVCN) USING MOLECULAR BIOLOGY DOBRIN, 1. University of Agricultural Sciences and Veterinary Medicine, Bucharest, Romania

Cyst nematodes are economically the most important group of plant parasitic nematodes in Romania and in temperate regions of the world. With their wide host range including many important crop species. The ability to accurately identify potato cyst nematodes (PCN) gene pools throughout the world are essential for their efficient management and control. The polymerase chain reaction (PCR) was used to amplify a region of PCN DNA coding for ribosomal RNA. Then, sequence differences between amplified products from different PCN isolates were examined by cutting the DNA with restriction enzymes. The PCN product from nine Globodera rostochiensis Wollenweber and Globodera pallida Stone (locality from Romania, Europe and South America) population was digested with eight restriction enzymes and the resulting fragments assessed efficiacy in distinguishing between PCN species and among PCN populations.

118 Working group no. 4 NOTES STUDIES REGARDING IMPROVEMENT OF THE TECHNOLOGICAL BARLEY QUALITY INDICES THAT AFFECT BEER QUALITY DRAGHICI, E., BEGEA, M., STROIA, I. University of Agronomic Sciences and Veterinary Medicine - Bucharest, Department of Vegetable Crops, Str. Marasti nr. 59, Sector 1, cod. 71331 Bucuresti - Romania.

The work is presenting a study made during a three years period on five autumn barley for beer hybrids and varieties. The interaction genotype - environmental factors and the acclimatisation to the Romanian locally climacteric conditions were studied. The biological material was studied from biochemical point of view, regarding the stability of the barley and resulting malt quality indices and their influence on the beer quality. The determinations were made within a micro malting resort as well as within a beer - processing laboratory. The analysed varieties and hybrids, have been compared to the internal and international standards, normally used within the beer industry. A statistical analysis regarding the evaluation of the stability parameters of every genotype from the experience and their confrontation was included.

119 Working group No. 4 NOTES JN VITRO" CULTURE ESTABLISHMENT OF COTINUS COGGYGRIA SHOOTS DUMITRASCU, M. University of Agricultural Sciences and Veterinary Medicine, 59, Mara§ti, 71331, Bucharest, Romania

The Jn vitro" shoot culture of Cotinus coggygria was established from both juvenile and adult plant material. Initial explants from epicormic shoots formed on the basal zone of the trunk had a higher capacity for "in vitro" establishment than explants from crown branches. Initial cultures produced a high quantity of polyphenols despite the shoots pre-treatment with a 50 mg/l ascorbic acid solution. The MS culture medium was supplemented with different combination of antioxidants: polyvinylpyrrolidone (PVP), citric acid + ascorbic acid, PVP + citric acid, charcoal. The best results were obtained when 150 mg/l PVP were added. Also, the detrimental effect of the polyphenols was reduced by frequent transfer to fresh medium. Shoot multiplication was obtained by culturing explants on Quoirin&Lepoivre modified medium supplemented with IBA 1mg/l, BAP 0,5 mg/l and GA3 0,5 mg/l. The multiplication coefficient increased from the first to fourth subculture. Genotype and the explant's age influenced the Jn vitro" performance of clones. Working group No. 4 NOTES POLYTENE CHROMOSOMES AND NODULIN GENE AMPLIFICATION IN ROOT - NODULES OF FABACEAE: AN ELECTRONOMICROSCOPIC AND GENOME ANALYSIS APPROACH. GAVRILA, L, VLADIMIRESCU, A. L, REBEDEA, I. Bucharest University, General Genetics & Evolution Department. Aleea Portocalilor 1 - 3, Code 77206, Bucharest, Romania.

Root - nodules of some Fabaceae species were throughly investigated in order to certify the existence of polytene structures inside the nuclei of host - cells, as it was claimed by Gavrila and Mihaescu (1992). There is now a solid body of evidence, certifying the polytenic organization of nuclei of certain types of cells inside the nodule. A gradient of polyteny levels from the surface toward the centre of nodule was detected, associated with a gradient in the size of the cells and nuclei. The fine structure of polytene nuclei and nucleoli is typical for the polytene nuclei of other cell types in the plants (e. g. embryo suspensor, root metaxylematic cells, stamen hairs etc.), with obvious extrachromosomal multiple nucleoli. An attempt to detect the amplification of other genome sectors was performed, using routinal genomic analysis methods.

121 Working group No.4

THE MODIFICATION OF RADIATION ACTION IN PLANTS BY USING BIOLOGICALLY ACTIVE CZ9928648 SUBSTANCES. GONCHAROVA, N., ZHEBRAKOVA, I., MONTIK, T., SHAPCHITS V. Institute of Radiobiology of the Academy of Sciences of Belarus, Plant Radiobiology Laboratory, Zhodinskaya str 2, 2200141 Minsk, Belarus, E-mail: [email protected]

Productivity of two varieties of pea, after acute gamma- irradiation with a dose of 8 Gy was investigated. Their radiosensitivity differed by a factor of 1,4-1,6. The presowing treatment of seeds with biologically active substances (BAS) influenced the main parameters of crop. The degree of the effect depended on BAS nature, their concentration and pea variety. However the modification of productivity in irradiated plants by using BAS did not result in significant effects and difference did not exceed 8 times. The modern technologies of plant breeding involve the measures oriented to increasing the farm crops resistance towards biotic factors, the measures stimulating growth, development and productivity both due to balanced mineral nutrition and through using regulators of growth - physiologically or biologically active substances. When applying plant growth stimulators, it is their ability to increase organism resistance to stress-factors. It should be noted that the effect of BAS of various classes on cytokinetic, biochemical and physiological reactions, as well as the processes of mineral nutrition elements transport, suggests their potential influence on the radionuclides transfer by the chain soil-plant-yield. In our experiments a great number of biologically active compounds have been tested. We were found that it increased and disease resistance of plants and simultaneously activated the physiological and biochemical processes. In conclusion, use of BAS in cropping technologies gives a principal possibility for regulating the intensity of radionuclides transfer to the yield.

122 Working group No. 4 NOTES SOME FACTORS AFFECTING CALLUS MEDIATED (INDIRECT) AND DIRECT SOMATIC EMBRYOGENESIS IN PEA (PISUM SATIVUM L.) GRIGA, M. Agritec Ltd., Zemedeiska 16, 787 01 Sumperk, Czech Republic, E-mail: [email protected]

Somatic plant cell contains entire genetic information for its morphogenesis into whole plant. This competence may be expressed in in vitro conditions as cfe novo organogenesis or somatic embryogenesis. Internal ontogenetic state of initial explant (biochemical, physiological and cytological status) which responds to external inductive signal influences basically the expression of morphogenetic developmental programme and the resulting way of regeneration. Even delicate changes in internal and/or external conditions can substantially change the final regeneration pathway, in the case of somatic embryogenesis the direct or indirect (callus mediated) formation of somatic embryos. In general, the ability to form direct embryos decreases continually from the ontogenetically younger to ontogenetically older tissues / organs, which situation is usually connected with different demand for external inductive signal (auxin, cytokinin). The regeneration pathway may affect the genetic status of regenerated plants; in general, the callus mediated regeneration (organogenesis, indirect somatic embryogenesis) is considered to increase the probability of production phenotypically / genetically altered plants. Direct somatic embryogenesis - on the other hand - should guarantee clonal fidelity and genetic stability with donor plant tissue. Both direct and indirect somatic embryogenesis was described in pea. Embryogenic caili were induced by various auxins (NAA, 4-CPA, 2,4-D, 2,4,5-T, Picloram) from immature zygotic embryos, shoot apices, lateral buds and nodal segments (1-8). NAA and Picloram induced direct somatic embryos from immature cotyledons and shoot apical meristems, respectively (9, 10). The serial initiation of direct somatic embryos preferentially in the leaf primordia area of shoot apical meristem supports the idea that meristem isolation together with the auxin signal terminates organogenic developmental programme and switches it on embryogenesis. There is an evidence that plants from pea embryogenic calli may exhibit some somaclonal variation on cytological (1) and whole plant level (11). Up to date no data are available about the genetical status of plants derived from pea direct embryogenesis. Strong genotype effect on pea embryogenic potential in vitro was reported by several authors (2, 3, 4, 6, 9,10).

(1) Kysely W, Myers JR, Lazzeri PA, Collins, GB, Jacobsen HJ (1987) Plant Cell Rep. 6, 305-308. (2) Lehminger- Mertens R, Jacobsen HJ (1989) Plant Cell Rep. 8, 379-382. (3) Kysely W, Jacobsen HJ (1990) Plant Cell Tissue Organ Cult. 20, 7-14. (4) Stejskal J, Griga M (1992) Biol. Plant. 34, 15-22. (5) Ozcan S, Barghchi M, Firek S, Draper J (1993) Plant Cell Tissue Organ Cult. 34, 271-277. (6) Van Doorne LE, Marshall G, Kirkwood RC (1995) Ann. Appl. Biol. 126, 169-179. (7) Loiseau J, Marche C, Le Deunff Y (1995) Plant Cell Tissue Organ Cult. 41, 267-275. (8) Nadolska-Orczyk A, Milkowska L, Orczyk W (1995) Acta Soc. Bot. Pol. 63, 153-157. (9) Tetu T, Sangwan RS, Sangwan-Noreel BS (1990) J. Plant Physiol. 137, 102-109. (10) Griga M (1998) Biol. Plant (in press). (11) Griga M, Stejskal J, Beber K (1995) Euphytica 85, 335-339. "Supported by grant No. 206/96/K188 and 522/96/1101 (GA CR) and No. IC15-CT96-096 (INCO- COPERNICUS)

123 Working Group No.4 NOTES SYSTEMIC ACQUIRED RESISTANCE IN SUNFLOWER {HELLANTHUS ANNUS) GUSCHA, M. I., DYACHENCO, A. I., DMITRIEV, O. P. Institute of Cell Biology, Dept. of Biophysics, 148 Zabolotnogo St., 252022, Kyiv, Ukraine E-mail: [email protected]

Plant resistance to different pathogens are realized by wide variety of mechanisms. Among them systemic acquired resistance (SAR) has become a subject of increasing inquiry. It is known that SAR can be induced by many pathogens and chemicals in different plant species. We investigated the possibility of induction systemic resistance to Phomopsis helianthi by salicilic acid (SA) in sunflower plans. For SAR induction three separate 10 mkl drops of 0,1% SA (pH 6,2) were applied to cotyledons of sunflower at 14-day old seedling stage and for the two succeding days.Eight days after last application of elicitor 20mkl drop of Phomopsis conidia suspension (10-6 conidia/mi) were applied to the first leaf of induced and control plants. Inoculated plants were incubated for 3 days in moist chamber and 7 days after challenge inoculation leaf area, covered by lesions were determined. It was shown that treatment of sunflower cotyledons with SA induced resistance of the first leaf to a challenge inoculation by Phomopsis helianthi. The decrease of disease development were detected in ten among twelve independent experiments and varied from 18 to 53%. When discs from systemically induced leaves were treated with mixture of cell wall- degrading enzymes, they releazed fewer protoplasts than discs from control leaves. It looks like, one of the possible mechanisms of acquired resistance in sunflower plants is reinforcement of leaf cell walls. Thus, the results obtained suggest that treatment of sunflower cotyledons with salicilic acid induced SAR to Phomopsis helianthi. Working group No.4 NOTES SOME RELATIONSHIPS BETWEEN HELIANTHIN AND RESISTANCE TO PHOMOPSIS IN SUNFLOWER GENOTYPES HAGIMA, I., RADUCARU, F. Research Institute for Cereals and Industrial Crops, Fundulea-Calarasi, Romania

Helianthinin, the main sunflower seed storage protein, is characterised by significant polymorphism. In SAS - PAGE electrophoresis, 10 genotypes (obtained "in vitro") were analysed using isolates and filtrates of Phomopsis in different experimental variants. The comparative analysis revealed three groups of components corresponding to basic poiypeptides and acidic ones. The infection determined a clear difference in number and intensity of bands. For each genotype we obtained a specific pattern. We mention also that the ratios of frequencies of different helianthinin banding patterns in a variety can serve as an index of its genotypic structure.

125 Working group No. 4 NOTES USE OF RAPD MARKERS IN CHARACTERIZATION OF EARLY SOMATIC EMBRYO CULTURES OF NORWAY SPRUCE HANACEK, P., TRUKSA, M., PROCHAZKA, S., HAVEL, L. Department of Botany and Plant Physiology, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00 Brno, Czech Republic, E-mail: [email protected]

Discovery of the polymerase chain reaction (PCR) lead to a great boom of the techniques utilizing the principle of PCR for the DNA studies such as DNA fingerprinting for phylogenetic, taxonomic purposes as well as the management of genetic resources and in vitro cultures. One of the techniques based on the PCR amplification of DNA is Random Amplified Polymorphic DNA known also as RAPD or AP-PCR. It is based on the amplification of random fragments of the genomic DNA using short primers of arbitrary sequence. Such fragments are with a certain probability spread through the plant genome and can serve as valuable genetic markers. One of the main advantages of RAPD is that no previous information on the genomic sequences is required and so, it can be applied in the species without a long history of genetic studies. The minute amount of DNA required for the assay makes possible non-destructive genetic analysis of individual plants or even seeds. We present our use of RAPD in characterization, collection management and monitoring of early somatic embryos of Norway spruce cultures. The RAPD markers proven to be useful in particular set of genotypes can be subctoned, sequenced and primers for specific PCR - a more robust method - can be prepared. RAPD can also serve as a source of markers for in situ labelling of chromosomes.

126 Working group No. 4 NOTES CORRELATION BETWEEN PLANTING DISTANCE, LIGHT QUANTITY RECEIVED BY CROWN, LEAVES TOTAL CONTENT IN CHLOROPHYLL AND PHOTOSYNTHESIS AT PLUM TREE HOZA, D. University of Agricultural Sciences and Veterinary Medicine, 71331 Bucharest, Bd. MarSsti 59, sector 1, Romania

The comparative study on three plum varieties and five planting distance made an evidence for their different behaviour. The plum planting at small distance (over 910 trees/ha) did not ensure sufficient light for the crown, the light quantity received by leaves being under 50% from the light. Generally, it was observed that leaves content in total chlorophyll and the photosynthesis intensity decrease at the same time with the growing of trees number/ha. At the same planting distance, the varieties had different values for the studied parameters, which showed that the phenotype influenced the development of the physiological process in leaves.

127 Working group No. 4 NOTES CORRELATION BETWEEN TOMATO PLANT CHARACTERISTICS AND NUTRITIONAL SUBSTRATES HOZA, G., POPESCU, V. University of Agricultural Sciences and Veterinary Medicine, 71331, Bucharest, Bd. Mara§ti 59, sector 1, Romania

Tomato plants were grown on several substrates (peat, admixture of peat with forest compost, perlite and sand) all of these fertigated with two nutrient solutions namely Coic-Lesant and Cesniakov. At the time of planting nursery tomatoes have been established correlation coefficients between plants and nutrient substrates making evident the positive influence of nitrogen on plant high and on photosynthesis intensity and also on leaf number and on stem diameter. Also, at the time of transplant, the correlation between plant characteristics and nutrient substrates on total chlorophyll content in leaves and photosynthesis were established. When checking the nursery tomato plants under production conditions it has been established a positive influence of photosynthesis on of early fruit production as well as on total yield of tomato fruits, while a negative influence of nursery plant age on tomato fruit yield and on average weight of fruit has also been found.

128 Working group No.4 NOTES FLOW CYTOMETRY - A RELIABLE METHOD FOR QUICK AND EFFICIENT DETECTION OF SONIACLONAL VARIATION IN NORWAY SPRUCE HRISTOFOROGLU, K., BURG, A., SCHMIDT, J. Austrian Research Centers, Biotechnology, A-2444 Seibersdorf, Austria

Production of plantlets via somatic embryogenesis as a mass propagation system needs a well developed, easy method to distinguish and sort out somaclonal variation. Two to 10 years old Norway spruce cell lines were screened for genetic stability and uniformity. DAPI-stained nuclei of embryogenic suspensor masses (ESM) and of needles and buds of regenerated plantlets were analysed with a Partec Ploidy Analyser (PA). Detected cell lines with a significantly higher DNA-content of > 4 % compared to the standard cell line PF6/96 (induced 1996, 2n=24) were selected for caryological studies to clarify the genetic variation. Flow cytometer analyses allow a quick and efficient screening for somaclonal variation. Additionally, separate analyses of all ESM explants of a cell line can avoid the production of plantlets from mutated cell lines.

129 Working group No. 4 NOTES THE BEHAVIOUR OF SOME DWARF PEACH AND NECTARIN SELECTIONS IN THE SOUTH AREA OF ROMANIA CHIRA, L, CEPOIU, N., CHIRA, A., DELIAN, E. University of Agricultural Science and Veterinary Medicine, MarSsti Blv., 59, Sector 1, Bucharest, Romania

The objective of these researches which were performed at the Faculty of Horticulture - Bucharest, in an experimental orchard (four years old), was to study the behaviour of some varieties and hybrids of dwarf peach and nectarin, with the aim to select and introduce in culture the most valuable of them. Among the advantages of the cultivating of dwarf varieties, we refer to : the intensivisation of the culture, the simplification of the work in the orchard, the high quality of fruits, the possibility to protect against the natural calamities adding to the possibility to growing them even in the baicony of the flats. The obtained results emphasized Fiorina (peach var. - 9,3 kg/ tree) and VT- M - R 10 P 11 (nectarin hybrid -12,1 kg/ tree), if we refere to the yeld. Refering to the fruits quality it was emphasized the peach var. Genetic dwarf, with average weight of fruit 145g soluble dry matter -11,6%; and for nectarin - the hybrid VT - 84 - N -08, with average weight of fruit 100 g and 11,5 % soluble dry matter. A remarkable ornamental feature have had the peach variety Fiorina and the nectarin variety Garden Beauty.

130 Working group No.4 NOTES HYBRIDIZATION BETWEEN TETRAPLOI HELIANTHUS SPECIES AND CULTIVATED SUNFLOWER H. ANNUUS L. CHRISTOV, M. Institute for Wheat and Sunflower "Dobroudja", near General Toshevo, Bulgaria

Hybridization was carried out between the tetraploid species of the genus Helianthus and cultivated sunflower Helianthus annuus L Hybrid material was obtained in all cross combinations. The 5-j plants developed altogether normally. The sterility in the first and second hybrid generations was overcome by using backcrossing with cultivated sunflower. New sunflower forms of generations 1 to 14 were obtained. Some of the material was used as initial material for sunflower breeding purposes.

131 Working group No. 4 NOTES AGROBACTERIUM - MEDIATED TRANSFORMATION OF NORWAY SPRUCE POLYEMBRYONIC CULTURE KARLOVSKA, L, TRUKSA, M., HAVEL, L. Department of Botany and Plant Physiology, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00 Brno, Czech Republic

Investigation of morphogenetic events may help to understand maturation, rooting and dormancy of conifers. Observing GFP (green fluorescent protein) could broaden the spectrum of experimental tools in this field, because this is the marker detectable in vivo. Transformation of polyembryonic culture was attempted to select the most suitable Agrobacterium tumefaciens strain to be used in further experiments. Three non-oncogenic strains of A. tumefaciens MP5, EHA105 and LBA4404 with binary plasmids pGKB5, pGT89 and pB!121, respectively, all containing p-glucuronidase (GUS) reporter gene within their T-DNA were co-cultured 1day with polyembryonic culture. Later performed GUS assay revealed only LBA4404 as the efficient strain. Polyembryonic cultures expressed GUS at a high level, but just some of them were able to grow. Expression of reporter gene was observed in all parts of embryos.

132 Working group No. 4 NOTES THE UPTAKE AND METABOLISM OF 3H-BENYZLAMINOPURINE IN THE EXPLANTS OF TOBACCO (NICOTIANA TABACUM L.) AND CUCUMBER {CUCUMIS SATIVUS L.) KLEMS, M. \ MACHACKOVA, I.2, EDER, J.2, PROCHAZKA, S.1 1 Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00 Brno 2 Inst. of Experimental Botany, Academy of Science of the Czech Republic, Prague

The uptake and metabolism of 3H-BAP were studied in the expiants of stem pith and leaves of tobacco and the hypocotyls and cotyledons of cucumber. The expiants were kept for short-term (2, 5, 8 and 20 hours) on MS medium with 0.8 rngJ"1 2,4-D, 0.5 mg.l'1 BAP and 13.2 mg.l"1 asparticacid (inductive medium). Uptake of 3H-BAP was the highest in tobacco leaves and cucumber cotyledons. More intensive metabolism was found in the cucumber expiants as the spectrum of 3H-activity of hypocotyls and cotyledons was more stuctured. The major metabolite was 3H-benzylaminopurine riboside (3H-BAPR). The highest content of this metabolite was found in cotyledons of cucumber after 20 hours of culture. The 3H-BAPR was formed with lowest intensity in tobacco stem pith. The intensive 7-glucosyiation 3H-BAP was described in the tobacco leaves expiants after 20 hours of culture. At this time the content of 7-glucoside-3H-BAP and free 3H-BAP was equal. The content of 3- g!ucoside-3H-BAP (3H-BAP3G) and 9-glucoside-H-BAP (3H-BAP9G) was not significant. For the study of morphogenic effect of growth regulators (BAP and 2,4-D), the expiants were subcultured after short-term induction to MS medium without growth regulators. The formation of the callus on basal part of cucumber hypocotyls and tobacco stem pith tissue was observed. Rhizogenesis was observed on cotyledons and formation of shoots on tobacco leaves. Long-term culture (3 weeks) of tobacco leaves on induction medium produced the cailus and globules. The microscopic analysis of globules indicated the presence of meristematic and tracheidal cells. This research was supported by the grant of Grant Agency of the Czech Republic (No. 206/96/K188).

133 Working group No. 4 NOTES THE CHANGES OF ABA LEVEL IN THE ROOT-XYLEM SAP AND PLANTS OF WINTER BARLEY CV. LUNET TREATED BY LOW TEMPERATURE KLEMS, M.1, FLORES, J. \ BALLA, J.\ KREJCJ, P.1, KADLECOVA, Z.2, PRASIL, I.2 1 Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00 Brno 2 Research Institute of Crop Production, Dmovska 507,161 06 Prague

Transport and changes of abscisic acid (ABA) levels were studied in plants of winter barley cv. Lunet treated by low temperature (+5 °C). Plants with two developed leaves, third leaf beginning to grow were used. Fiuridone (10 mg.l'1) was used in the experiment for carotenoids (precursors of ABA) synthesis inhibition. Root-xylem exudates were collected in silicone tubes using mild vacuum. The endogenous level of ABA was determined by the radio-immuno assay according to the method of QUARRIE et GALFRE (1988). The distribution and transport of ABA was studied using 14C-ABA taken up by roots. Endogenous level of ABA increased in aboveground parts already 3 hours after treatment by low temperature. The aboveground part of plants treated by fiuridone had higher level of ABA 3 hours after low temperature treatment and after 24 hours the ABA level decreased. The increasing of ABA level in roots was determined in plants treated by fiuridone (culture in +20 °C) or low temperature (non-treated by fiuridone). The combination of fiuridone and low temperature treatment resulted in lowering of ABA level in the roots. Fiuridone treatment did not reduce the ABA level in root-xylem exudate, low temperature treatment increased mildly the ABA level in root-xylem exudate. The uptake of 14C-ABA was lowest after low temperature treatment. Distribution of 14C-activity described poor transport 14C-ABA from the roots to the aboveground parts in low temperature treatment. The highest accumulation of 14C-activity in roots was in the plants cultured at +20 °C. Application of fiuridone at +20 °C resulted the change of 14C-activity distribution in plants, C-activity in aboveground part and roots was equal.

This research was supported by the grant of Grant Agency of the Czech Republic (No. 522/97/0842). Working group No.4 NOTES ORIGINAL METHOD FOR DETERMINATION OF SULFOXIDES KOSYAN, A., TARAN, N. National Taras Shevchenko University, Department of Plant Physiology and Ecology, Volodymyrska 64, 252033 Kyiv, Ukraine, E-mail:valentin @ sbird.kiev.ua

Original method for determination of sulfoxides based on the property of this substances to react with the product of triptophane's and paradimethilaminobenaldehide's condensation is proposed. An oxidize-deoxidize reaction results in formation of bis-2-triptophani!-4-dimethilarninophenilcarbonil - blue-violet dye with the absorption maximum at 575 nm. Sensibility of method is 5 (iM/g, relative mathematics error at three determinations is +3,3%. Approbation of the method proposed was performed while determination of the quantitative sulfoxide content in garlic organs at various vegetation stages. The proportional increase of the sulfoxide content during the whole period of vegetation observed, whereas in roots and unreal stem the phenomenon like this was noted only while growth at spring vegetation. Drastic decrease of the sulfoxide content in unreal stem (100% ) taking place during the period from May to July in connected to processes of bulb forming which lead to migration of sulfoxides from unreal stem tissues to bulb tissues. Dying out of roots unreal stems and leaves at the end of vegetation is accompanying by the sulfoxide content decrease down to row quantities in these organ issues. The method allows to determine the sulfoxid content in 50 samples daily and can be used for studying of features of the sulfoxide biosynthesis and accumulation in garlic tissues and for estimation of quality of bulbs as agricultural products and row material for pharmaceutics industry.

135 Workig group No.4 NOTES DOES MELATONIN PLAY IN PLANTS A SIMILAR ROLE AS IN ANIMALS? MACHACKOVA, I.1, KOLAR, J.2, EDER, J.2, PRINSEN, E.3, VAN DONGEN, W.3, VAN ONCKELEN, H.3 1Norman Borlaug Institute for Plant Science Research, De Montfort University and Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojova 135, 16502 Praha 6, Czech Republic, E-mail: [email protected] 2 Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojova 135, 16502 Praha 6, Czech Republic 3 Department of Biology, University of Antwerpen, B2610 Wilrijk-Antwerpen, Belgium

Melatonin (5-methoxy, N-acetyltryptamine) is an animal hormon involved in regulation of photoperiodic and rhythmic events. Its production is high during night and low during day. The actual level of melatonin is a signal of day-time and the broadness or position of the night maximum is the signal of the length of the night, i.e. of the season. Recently, two independent laboratories demonstrated the presence of melatonin (5-methoxy, N- acetyltryptamine) in plants. Under daily light/dark regime a rhythm in melatonin level in the above-ground parts of Chenopodium rubrum plants was demonstrated, with maximum in the hours 4-6 of the dark period and very low levels during light. Further, we found an endogenous rhythm in melatonin level in continuous darkness with maxima 4, 43 and 74 h. In regimes with dark periods shorter than 12 h the maximum of melatonin level was shifted to earlier hours of darkness. Thus, it seems that the position of the maximum is changing with the length of the photoperiod. Radioactive serotonin was converted in C. rubrum plants to melatonin: the highest conversion rate was observed in the 4th h of darkness. Measurable melatonin levels were found in both xylem and phloem exudates from C. rubrum plants. Applied melatonin is actively degraded, not only in the plant tissue, but also on the surface of the plants, which may be the cause of its very low effectiveness, when applied on the plants. Thus, melatonin system in plants may function in a similar way as in animals. Working group No. 4 NOTES THE EFFICIENCY OF ,,HAIRY ROOTS „ TRANSFORMATION TRANSFER SYSTEM IN NICOTIANA TABACUM L. AND VITIS VINIFERA L. ,,IN VITRO" CULTURED TISSUES MAXIMILIAN, C.1, BREZEANU, A.1, ROSU, A.2, VATAFU, I. laboratory of Genetic Engineering and Plant Morphogenesis, Biology Institute, 296 Splaiul Independentei, P.O. Box 56-53, Bucharest, Romania 2University Agronomic Sciences and Veterinary Medicine, Biotechnology Faculty, Bd. Marasti 59, Bucharest, Romania

The Ri plasmids of Agrobacterium rhizogenes transferred in host cells produce ,,hairy roots" at the infected sites. This experimental system is widely used in plant biotechnologies, like genetic transformation and secondary metabolites biosynthesis. In our paper the transformed capacity of some wild and genetic modified strains of Agrobacterium rhizogenes were analyzed for one cuitivar of Nicotiana tabacum (Xanthi) and three cultivars of Vitis vinifera {Coarna Rosie, Metiot, Grasa de Cotnari). In this respect, the following aspects are discussed in the paper: - evaluation in our experimental conditions of transformation ability of A. rhizogenes 8196, R 1601, 62, 113 and Ti piasmid of A. tumefaciens (ATCC 13333); - establishment of reactivity of the used biological systems in relation to plant genotype and explant types (leaves, shoots and petioles); - characterization of peroxidase isoenzymes content of tobacco and grape vine nhairy roots" induced ,,in vitro"; - quantitative analyse of the total protein content in ,,hairy roots". The results obtained allowed us to establish for each plant genotype a properly experimental system including the type of the explants, the highest virulent bacterial strain and optimal experimental conditions.

137 Working Group No.4 NOTES CYTOLOGICAL ANALYSIS OF LEAF DISC-DERIVED REGENERANTS OF NICOTIANA PLUMBAGINIFOUA MAZIKTOKEI K, GYULAI, G., KISS, E., TOTH LOKOS, K., HESZKY L E. University of Agricultural Sciences, Department of Genetics and Plant Breeding, 2103. Godollo, Hungary, E-mail: [email protected]

Plants regenerated from cultured protoplasts and plant tissues can show considerable somaclonal variation (Larkin, Scowcroft, 1981). Cytological studies provide the opportunity of establishing the degree this variation due to gross changes in the genome.The most frequent variation reported among somaclone involves changes in chromosome number and structure, i.e. polyploidy, aneuploidy, translocation, deletions, inversions and duplications. Change in chromosome numbers result in genomic instability. This report describes the results of studies in tobacco plants regenerated from leaf discs via adventitious shoot formation. Shoot cultures Shoots of haploid (n=1O) and diploid (2n=20) Nicotiana plumbaginifolia Vivani, Bench were maintained on a hormone-free agar medium containing MS elements , 2,0 % sucrose and 0,8 % agar at pH=5,8. Shoots were grown at 24 °C under a light intensity of 2000 lux with 16/8 h photoperiod. Leaf discs Discs of 0,8 cm in diameter were cut from the first fully developed leaves and placed with abaxial surface downwards on the medium containing cytokinin at the concentration of 10 mM BA. (Gyulai et al. 1995). Cytology Chromosome counts were made using shoots regenerated on the cytokinin treated leaf discs. The small leaves were pre-treated in 0,05% colchicine for 2,5 h at 20 °C, fixed in 3:1 95% ethanol:glacial acetic acid for 24 h at 20 °C, and then were stained in 4 % aceto-carmine for 24 h at 20 °C. Leaves were then squashed in 45 % acetic acid. 10 plants were used for the adventitious shoot regeneration, the variability of the chromosome number of these initial plants and leaf disc-derived regenerants were compared both in haploid and diploid N. plumbaginifolia. Mitotic chromosome numbers of initial hapioid plants ranged from 4 to 20, the average number of haploid cells was 51,85 %. The amount of the haploid cells among regenerants was 36,95 %. Chromosome numbers of initial diploid tobacco plants ranged between 16-23, the portion of the 20 chromosome cells was 75,73 %, while among regenerants ratio of the 20 chromosome cells was 51,89 % (with chromosome numbers 9-29). More wide range of chromosome numbers were established in the leaf disc derived regenerants then in the initial leaves both in haploid and in dipioid A/, plumbaginifolia. The regenerants of haploid tobacco seems to be more variable in chromosome number then regenerants of diploid tobacco. There were no correlation in chromosome numbers of initial leaves and those of regenerants of the same leaf discs. 1. Larkin, P.J., Scowcroft, W. 1981. Somaclonal variation - a novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet. 60:197-214. 2. Gyulai, G., Kiss, J., Jekkel, Zs., Kiss, E, Heszky, L.E. 1995. A selective auxin and cytokinin bioassay based on root and shoot formation in vitro. J. Piant Physiol. Vol. 145:379-382. Working group 4 NOTES HYBRID PROGENIES FROM HORDEUM VULGARE L. X HORDEUM BULBOSUM L. INTERSPECIFIC CROSSES MIHAILESCU, A., GIURA, A. Research Institute for Cereals and Industrial Crops, Fundulea 8264, Calarasi, Romania

The major use of the crosses between barley (H. vulgare L.) and perennial wild species Hordeum bulbosum L. was doubled haploid production. Bulbosum system enabled homozygosity and uniformity to be achieved rapidly from early generation of hybrid material or from final products of the selection, respectively breeding lines. H. bulbosum is closely related to cultivated barley and posseses useful traits such as: resistance to several fungal and viral pathogenes (powdery mildew, leaf rust, net bloch, scald and barley yellow/ wild mosaic virus), to Russian wheat aphid and winter hardiness. Another important aim of this interspecific hybridization was been to introgress desirable genes into cultivated barley from wild species H. bulbosum. In this case there were several bariers associated with successful gene transfer. These included partial interspecific incompatibility (locus Inc), hybrid chromosome instability (parental genome ratio and strong influence of temperature), low or absence of chromosome pairing (recombination) and hybrid infertility. !t was established parental genome (H. vulgare = W, H. bulbosum = BB), ratios which are likely to favour haploid or hybrid formation. For the latter, genome ratios of 1V: > 1B are required, whereas chromosome elimination of bulbosum genome occurs when these ratios are 1V: 1B or 2V: 1B. Temperature of < 17.57C during the first days after pollination are more suitable for obtaining hybrids from diploid crosses. This study presents preliminary results of hybridization between diploid H. vulgare L. (2n = 2x = 14) and diploid (2n = 2x = 14) and tetraploid (2n = 4x = 28) H. bulbosum L. for a programme of gene introgression from wild species to cultivated barley. In 1997 at RICIC - Fundulea, in field conditions 12 spring barley genotypes were pollinated with diploid and tetraploid H. bulbosum and 32 putative hybrids were regenerated after "in vitro" embryo culture. Part of hybrids was perennial, the following grouping could be made on morphological characters: Hybrid type 1: prostrate growth habit, leaves slightly curled and dark green, visible hairs on leaves, excessive tillering, completly male sterile. Hybrid type 2: upright growth, foliage slightly broader than type 1, hairs distinctly visible on leaves and stems, completly male sterile. Triploid type: tall plants, intermediar tillering, broader leaf shape, excessive (visual) hairs on leaves and shoots, bulbosum habitus and type of spike, with rare dehiscent anthers. One triploid plant were partial fertile. Several hybrid plants were characterised cytologically (mitotic chromosome number) and have compraised: diploid hybrids (VB) with chromosome numbers varied between 2n = 9 and 2n = > 14 (6 plants) and triploids (VBB) with 2n = 19, 20 and 21 chromosomes (9 plants). Diploid hybrid progenies will be cytogenetically analysed (meiozis and C - banding) for eventually presence of aneuploid and/ or chromosome rearangements. The fertility of these hybrids could be restored by colchicine treatments or by pollination with cultivated barly. Triploids will be backcrossed to barley and following backcrosses, chromosome substitution plants and recombinants will be identified cytologically.

139 Working group No.4 NOTES CROSSING ABILITY BETWEEN HEUANTHUS DECAPETALUS L. AND HEUANTHUS ANNUUS L. NIKOLOVA, L, CHRISTOV, M., JAMBASOVA, T. Institute for Wheat and Sunflower "Dobroudja", near General Toshevo, Bulgaria

Interspecific hybridization was carried out between the tetraploid species Helianthus decapetalus L and cultivated sunflower Helianthus annuus L Reciprocal crosses were performed. Hybrid material was obtained in Fi,F2lF3 and F4 generations. Cytological investigations showed abnormalities in meiotic behaviour and triplod chromosome number of the PMC. Morphological investigations were carried out on the hybrid material, showing a big diversity of forms in F2 and F3 generations.

140 Working group No.4 NOTES THE SULPHOLIPID CONTENT AND THE PLANT DROUGHT RESISTANCE OKANENKO, A., TARAN, N. Taras Shevchenko University, Plant Physiology and Ecology Department, Volodymyrska 64, UA 252033, Kyiv, Ukraine, E-mail:[email protected]

Drought factors can influence the type of acyl lipids in the memebranes of the plant tissue and their fatty acid composition especially. The study devoted to superoptimal temperature action shows the increase of monogaiactosyl diacylgiycerol [MG] (by 20%),digalactosyl diacylglycerol [DG] (by 30%) and sulpholipid [SL] (by 50- 200%) content [Pearcy, 1978]. The last, attracted our attention, sulphoquinovosyl diacylglycerol [SQDG], comprised between 8 and 24% of the four major chloropiast lipids and contained a substantial quantity of high melting point fatty acids (16:0 + 18:0), the range lying between 26 (T. aestivum) and 62% (Carica papaya) [Kenrick, Bishop 1986]. With the increase of growth temperature of Nerium oleander from 20 to 45°C the major changes in molecular species was an increase in the proportion of dipalmitoil SQDG from 12 to 20% and decrease in that of linoleil- palmitoil SQDG from 40 to 30%. Thus, SQDG could be the main lipid component involved in the phase transition in the thylakoid polar lipids. Besides, sulpholipid molecules as well as galactolipid and phospholipid molecules within the functioning thylakoid membrane are associated with proteins that are involved in the electron transport of PS I and II. SQDG is found in the RC PC I and PC fl and effective as stabilizing agent of the photophosphorilation coupling factor.Therefore we consider as well-grounded the decision to study the action of drought factors upon the winter wheat leaves and chloroplast lipid content paying special attention to the SL changes. Data obtain show the high temperature action induced the SL accumulation in leaves of drought resistant (DR) plants while in sensitive plants (DS) one could see the decrease of their content. The water deficit induced some SL accumulation in DR plants accompanied by the drastic decrease in sensitive plants. Results obtained in the field experiments in the drought region evidence the SL increase to be as at the stage of stooling, so at the stage of milk ripeness comparing with watering plots. The SL content changes in chloroplasts while heating at temperatures 40, 45 and 50°C was described by curve with one apex at 45°C with following drop at 50°C in all resistant plants. The drastic fall of this compound content from control to heating at 50°C in sensitive plants was observed. The water deficit action induce the reaction slightly resembled those under the heating in all varieties. Combine action of heat and water deficit induced the SL accumulation in the almost all resistant plant chloroplasts, while the SL content decreased in sensitive variety more drastically (twice), than in the cases of single action of heat or water deficit. Thus the shifts observed under the heat are similar to those under the water deficit action and their combine action induced the SL accumulation sometimes much more significant than single factor action. The phenomenon gives us good grounds for conclusion about special role of these compounds in adaptive reaction to extreme factors and possibility to predict the drought tolerance of new wheat varieties.

141 Working group No.4 NOTES AFLP ANALYSIS OF RECIPROCAL SUBSTITUTION LINES IN WHEAT {T. AESTIVUM L) OVESNA, J., SULAKOVA, S., KUCERA, L, KOSNER, J. Research Institute of Crop Production, Dmovska 507,161 06 Praha 6 Ruzyne, Czech Republic, E-mail: [email protected]

Molecular markers and DNA fingrprinting play an important role in plant genome studies. Several techniques have been developed including RFLP, RAPD, SSR and AFLP. The last one seems to be an usefull tool for such a purposes as it has been shown on several plant species. The possibility to use AFLP for wheat genome analysis was studied with the aim to prepare chromosome specific markers. Reciprocal substitution lines of winter wheat Mironovskaya and Bezostaya for chromozomes of fifth homeologous group were used. Different alleles of Vrn (vernalization) genes are present on these chromosomes in the cultivars. ( Mironovskaya - vrn1, Vrn2, vrn3, resp. Bezostaya - Vrn1, vrn2, Vrn 3 ). Such a plant material could be suitable for chromosome specific markers preparations, which could be latter on used for the mapping of the genes of interest. Combination of restriction sites EcoRI and Mse I were used for AFLP analysis. After pre-selectiove amplification selective ones were carried out. Altogether 132 combinations of selective primers were tested to find polymorphism between Mironovkaya and Bezostaya and to choose the best primers for substitution lines analysis . Perkin Elmer protocols were used to prepare fluorescent dye labelled amplification products. They were separated on Perkin- Elmer ABI Prism 310 Genetic Analyser and analyzed using Genescan and Genotyper software. Two basic types of polymorpfics signals were found in amplification patterns. (1) Qualitative - that means presence or absence of fragments of certain lenght, which were very reproducible (100%) (2) Qualitative - that means that peak height ratio was at least 1 : 2. Not all such detected differences were reproducible, so for substitution lines analysis qualitative types were used. Out of 10050 amplified products 222 - 2.2% (group 1 - quantitative) and 576 - 5.7% (group 2 - qualitative)), were polymorphic e.g. together 7.9% of fragments. We founded 1.7 polymorphic amplified fragments (of group 1) per reaction (ranging from 0-27). We were able to identify chromosome specific ones among polymorphic fragments but less amount than we expected from the number of substituted chromosomes including in the studies . The work was supported by a grant of National Agency for Agricultural Research of Czech Republic No. EP 7235

142 Working group No. 4 NOTES THE IMPLICATION OF THE KILLER PHENOMENON ON YEASTS FROM WINE-MAKING POMOHACI, N., BURCEA, M., CORNEA, P., RADOI, F., COSOI, C. University of Agronomical Sciences and Veterinary Medicine Bucharest, Faculty of Biotechnology, Marasti, 59, 71331, Bucharest, Romania, E-mail: flore @ usab.ro

Twenty-five yeast strains isolated from Panciu Wine Region ( Romania) were used in a screening program. After the chemical tests, organoleptycal and GC/MS analysis seven strains were selected and used in following experiments. One of the criteria in the screening program was the detection of the killer phenomenon of this strains as wel| as the sensitivy of the new isolates to known Killer strains. Two type of Killer strains were used: Ki type (Sacccharomyces cerevisiae 9744c, Sacccharomyces cerevisiae 1070/c and Sacccharomyces cerevisiae X 208) and K2 type (Sacccharomyces cerevisiae SMR4). The sensitive strain tested for detection of Killer character was a collection strain Sacccharomyces cerevisiae 17/17. All the new isolates from Panciu Region were resistant to the killer strain Sacccharomyces cerevisiae SMR4 but sensitive to K^ type strains. Moreover, with an exception, none of the new isolates were Killer (they were not able to inhibit the growth of Sacccharomyces cerevisiae 17/17). The strain proved to be Killer was also the most valuable strain from oenologica! point of view.

143 Working group No. 4 NOTES PULSED-FIELD GEL ELECTROPHORESIS - METHOD FOR THE TAXONOMIC IDENTIFICATION OF THE OENOLOGICAL YEAST STRAINS POMOHACI, N., BURCEA, M., CORNEA, P., POMOHACI, D. University of Agronomical Sciences and Veterinary Medicine Bucharest, Faculty of Biotechnology, Marasesti, 59, 71331, Bucharest, Romania

We have selected seven yeast strains from four wineyards of the Panciu area-Romania, of read and white grape juice. After the taxonomic identification by using classical morpho-physiological tests (Lodder, 1974 and the API-20 system) we have noticed that the strains belong to the same species: Saccharomyces cerevisiae and Sacoharomyces cerevisiae var. ellipsoideus. After oenological testing in identical tehnological conditions we have obtained wines wich differ from chemical (GC/MS) and organoleptycal point of view. A variant pulsed-field gel electrophoresis (TAFE- Transverse Alternating Field Electrophoresis) was used in order to identify the selected strains. All the seven strains were used were totally different as regards their cariotype proving a high intraspecific genetic polymorphism among the Saccharomyces cerevisiae strains. This method proves to be rapid, relative simple and conclusive, as well. Working group No. 4 NOTES THE CHARACTERISATION FROM A CHROMATICAL POINT OF VIEW OF SOME ROMANIAN ROSE WINES POMOHACI, N., RADOI, F., BURCEA, M. University of Agronomical Sciences and Veterinary Medicine Bucharest, Faculty of Biotechnology, Marasti, 59, 71331, Bucharest, Romania, E-mail: flore @ usab.ro

The colour of rose wines is given by the relation between different phenolyc compounds (flavones, anthocians, tannins) and the equilibre between yellow, red and violet compounds. The paper approaches the study of chromatically characters of rose wines obtained from Cabernet Sauvignon variety using modern techniques of analysis (which are based on spectrophotometry). It is analyzed in every detail each chromatic compounds (tannins, anthocians, total polyphenols value, shade, coloured intensity, modified coloured intensity, % 420, %520, %620). Finally we reached the conclusion that concerning the rose wines the most important is the shade of the wine (ratio between yellow compounds and red compounds is 0,5 - 2) and not the color intensity. Also, in order to obtain rose wines, it must be considered the percentage of violet compounds which gives the wine personality from a visual point of view.

145 Working group No. 4 NOTES IN VITRO ESTABLISHMENT AND CHARACTERIZATION OF HAIRY ROOT CULTURES INDUCED WITH AGROBACTERIUM RHIZOGENES STRAINS ROSU, A., CORNEA, P..CAMPEANU, G. H..CEPOIU, N..BREZEANU, A.1,MAXIMILIAN, C.1 University of Agricultural Sciences, Faculty of Biotechnology, Bd. Marasesti 59 , 71331 Bucharest, Romania. E-mail:arosu @ usub. ro institute of Biology Spl.lndependentei 296, 79651 Bucharest, Romania.

The "hairy root" phenotypes formed in response to infections by Agrobacterium rhizogenes and established as aseptic cultures in vitro, have an important biotechnological potential both for producing a large variety of phytochemicals and for regenerating transformed plants with phenotypic alterations of practical value. Fully expanded leaves, internode segments and roots from "in vitro" grown plants of Morus nigra , Vitis vinifera and Eustoma grandiflora were infected with two wilde -type strains of A. rhizogenes. Bacteria free root cultures were obtained after subculturing the transformed roots on modified L S medium supplemented with Cla-foran. Adventious shoots developed readily on the hairy roots in Eustoma grandiflorum and the regenerated plants are studied for distinctive morphological alterations. Following the infection of leaf and steam explants in Vitis vinifera, transformed roots developed, displaying an enhanced morphogenetic potential compared to the untransformed tissues, expressed by adventitious shoots from which whole plants were regenerated. The A. rhizogenes mediated transformation of leaf explants in Morus nigra resulted in the development of putative transformed roots that are under investigation for the production of desirable secondary compounds endowed with antiviral properties.

14fi Working group No. 4 NOTES A COLD-AND DROUGHT-RESISTANCE OF THE SAMPLES FROM THE CAMBRIDGE PHASEOLUS GERMPLASM COLLECTION RUSSKIKH, I., SHANGINA, N., DAVYDENKO, O. Soya-North Co., Ltd, Institute of genetics & cytology, National Academy of Sciences of Belarus, Kuprevicha str., 14., Minsk, 220141 Belarus

The industrial beans cultivating in Belarus is limited by the absence of available local varieties for growing in our climatic area. We have the experience of adaptive soybean breeding for Belarus, and we have began such breeding work for beans. In 1996 we had received 880 samples specially selected for Belarussian conditions from the Cambridge Phaseolus Germplasm Collection (CPGS) from Great Britain. And 700 samples from it proved to be available for growing in Belarus. These samples are used as source material for new varieties creation. The germplasm collection was sowed in the southern region of Belarus. This region is considered as the most perspective for the industrial beans cultivation, but part of this territory is polluted by radionuclides. However, beans are cultivated in small quantities everywhere for private use, up to north regions of Belarus. The main limiting factors for industrial beans cultivating were determined already during the first year of testings. They are: the low rainfall in the south area with light sandy loam and foam soils, and reduction of active temperature amounts while moving this culture to the north. In this connection it is considered perspective to study a cold- and drought-resistance of the beans samples, and the inheritance of these characters (the accessible references give not enough information on this case). The 20 most productive varieties were selected for determination of theirs cold- and drought-resistance. The drought-resistance on the early stages of development (germination) had been studied by seed sprouting in the sucrose solutions (with concentrations 86,6 and 108,1 gr./l, that corresponds to 7 and 9 atm.). The standard strategy by Vavilov Research Institute of Plant Industry (St.-Petersburg) was modified slightly. We defined not only the influence of sucrose concentrations on germination and dry mass of prosprouts, but also the intensity of plants development by measurements of length of prosprouts and number of additional roots. The cold-resistance was determined by the way of seed sprouting under the conditions with different temperatures on the same stage. The temperature 20 °C was taken for optimum conditions, and for extremal conditions-12 °C. As a result of the present studies we have calculated drought-resistance and cold-resistance factors. The factor of the seeds sprouting reduction (FSSR) changed from 1.67 (most cold-resistance variety 6919 (number CPGS)) to 9.29 (Turkish brown). This factor for drought-resistance changed from 0.94 (Snowbunting) to 1.71 (Bountiful (1823)) - for 7 atm. variant, and from 1.24 (6895) to 9.43 (4280) for 9 atm. variant. We have found out the absence of the connection between FSSR and other resistance factors. If this variability is genetically controlled, we can to try to combine these factors in one variety in the most perspective form. We can't be sure that the genotypic differences in resistances studied for the prosprouts correlate with the same characters in the grown-up plants. That's why we'll continue our investigations of these varieties in the field conditions.

147 Working group No.4 NOTES GENETICS TRANSFORMATION AND PLANT IMPROVEMENT SANGWAN, R. S., SANGWAN-NORREEL, B. S. Laboratoire Androgenhse et Biotechnologie, Universiti de Picardie Jules Verne, 33, rue Saint-Leu, Hot des Poulies, 80039 Amiens Cedex, France.

It is only during the last decade that genetic engineering of plants has been developed, only recently that it has become an important tool for crop improvement. Several molecular techniques are widely used to introduce recombinant DNA molecules into a variety of plant species; however, one of the most effective means of gene transfer into plants is the utilization of soil bacteria from genus Agrobacterium and essentially A. tumefaciens or A. rhizogenes, as vectors. The fundamental problem, in the production of fertile transgenic plants, especially in recalcitrant species, does not seem to lie so much in the delivery or perhaps intergration of the introducted DNA, but rather in the regeneration of transgenic plants from the transformed cells. In most transformation experiments, in vitro regeneration of plants has become the main limiting factor for success. At present the major source of highly regenerative cells has been immature zygotic embryo or meristematic tissues but a routine Agrobacterium- mediated transformation protocol for these tissues is lacking. Circumvention of the difficulties inherent in in vitro regeneration by transforming the easily regenerable explants such as zygotic embryos would therefore be highly advantageous. Although meristematic and embryogenic cells were thought to be insensitive to Agrobacterium infection, it has been demonstrated recently that they are susceptible to Agro-infection and under certain in vitro conditions e.g. with phytohormone treatment prior to infection by Agrobacterium, the embryonic cells become competent for infection. We have studied the feasibility of zygotic, somatic and pollen embryo transformation in higher plants. In 1998, as a first model system we choose to transform the zygotic embryos of Arabidopsis then the pollen embryos of Datura and Nicotiana and lastly we have exploited the direct somatic embryogenesis system from zygotic embryos in Datura. We have reported on 3 transformation systems : 1) routine zygotic embryo transformation and regeneration of fertile transgenic plants 2) transformation of cotyledonary stage pollen embryos 3) a successful combination of the technique of direct somatic embryogenesis with Agrobacterium-mediated gene transfer. This later method does not require wounding nor a callus phase. Using these methods, we have recently introduced several genes of interest e.g. herbicide, insect and virus resistances in both model and economically important plant species.ln this meeting.We would outlined the possible impact of transgenic technology on plant improvement. Working group No.4 NOTES LIPID COMPONENTS OF CHLOROPLAST MEMBRANES IN STRESS CONDITIONS UNDER THE EFFECT OF IMMOBILIZED BENZYL-AMINOPURINE SERGA, O. Taras Shevchenko University, Volodymyrska St.64, 252033 Kiev, Ukraine

Essential decrease of photosynthetic activity under water stress may depend on destructive changes of photosynthetic membrane lipid components. The effect of physiologically active compounds on plants provides regulation of photosynthesis and its drought resistence by influence on chloroplast membrane system. The protective ability of synthetic growth regulator immobilised 6-benzyIaminopurine (BAP) under water stress in relation to chloroplast lipid matrix of drought resistant winter wheat was investigated. Winter wheat (Triticum aestivum) plants were subject to water stress in vegetative experiments. Intact chloroplasts were isolated fron leaves and content of polar lipids was determined. It was shown that water stress results in degradation of phospholipids, especially in chloroplasts of plants of sensitive genotype and decrease of mono- and digalactolipid (MGDG:DGDG) ratio as well as linolenic acid content in polar lipids. Treatment of leaves by immobilized BAP prevented degradation of phospholipids and MGDG:DGDG ratio decrease in chloroplasts of resistant wheat genotype. Treatment with synthetic 6-BAP stimulated incorporation of unsaturated fatty acids in chloroplast polar lipids regardless of variety resistance. The results suggest that BAP in plant metabolism blocks destructive changes of membrane lipid components and optimises conditions of biochemical process.

149 Working group No.4 NOTES THE FERTILIZER INFLUENCE ON SOME PHYSIOLOGICAL PARAMETERS OF ENSILAGE CORN THAT IS GROWN ON THE MEADOW CALCEROUS SOFT-LOAMY ON LOESS LOAM CHERNOZEM OF THE NORTH PART OF UKRAINIAN FOREST STEPPE SMETANSKAYA, !. N. Dept. of Agrochemistry and Quality of Agricultural Production, National Agricultural Academy, Kiev, Ukraine

Modern ecological situation in Ukraine (industry, Chernobyl accident and high technologies demanding high quantity of fertilizers) demands to study the effect of high doses of chemical fertilizers on some anatomy-morphological and physiology-biochemical changes, directed on forming corresponding by size and work intensity photosynthetic apparatus and on using photosynthetical products for forming yield quantity and quality. Important for effective fertilizer se is balanced application taking into consideration soil-climatic conditions and plant biological peculiarities. The aim of our research was to make a model of heavy metal (HM) ranslocation in the system soil-plant (anatomy- morphological and physiology-biochemical parameters), yield-yield quantity; fertilizer influence on dynamics of HM mobility and the optimal fertilizer doses for high yields balanced with nutrient elements and characterized by high quality level. The object of our study is corn Odeska-80 grown on meadow calcerous soft-loamy on loess loam chernozem of the north part of Ukraine forest steppe (pH=7.1, in the soil horizon 0-25 cm humus 4.2%, total nitrogen 0.20%, hydrolysable nitrogen 6.5, ammonia nitrogen 4.1, nitratenitrogen 0.42 mg/100g soil; active solid phase phosphorus 3.4 and exchange potassium 11.2 mg/100g soil). Selected 7 plots in crop rotation are fertilized with different quantity of organic (pig manure) and inorganic (ammonium nitrate, phosphorus, potassium salt) fertilizers according to the scheme: 1. Absolute control 2.301 manure /ha background (bg) 3. Bg+Pgg, 4. Bg+PggKgn, 5. Bg+Ni20p90^90- *>• Bg+N-)80p135K135' 7- N120p90K90- Tne experiment was organised in crop rotation field, each variant with 3 replicates. We studied the dynamics of N, P, K, Ca and heavy metals (Zn, Cu, Co, Ni, Cd, Pb, Hg) in plant organs (leaves, stems, cobs, roots) and in soil (0-25 cm and 25-50 cm horizons). Fertilizers improve HM accessbility for plants and content in plant tissues in the period of milk-waxy maturity (MW) are for Cu 2.25mg/kg in control and 7.80mg/kg for Bg+Nigop135K135- Data °f dynamics of pigments show that chlorophyl a and b concentrations are 1037 and 445 mg/kg dry matter in control and 1760, 702mg/kg at Bg+N-]20p90^90'' carotines are 61mg/kg dry matter in control and 88 Bg+N-|20p90^90; carotinoides are 98 and 117 mg/kg at MW. Dynamics of enzyme activity increased by fertilizer action: peroxidase from 12.12 to 14.73 ml titr.H2C>2, catalase from 3.81 to 5.33 ml titr.H2O2 in corn leaves at MW. The square of leaf assimilation surface increased from 36.8 to 53.16 th m2/ha at Mv, the average pure photosynthetical productivity from 5.92 to 8.14 g/ m2/day. Total biomass at MW increased from 441 to 662 c/ha and dry matter from 112 to 182 c/ha. Protein amounts increased from 9.4 to 11.9% dry matter in leaves, from 4.2 to 6.6% in stems and from 8.9 to 11.2% in cobs. The obtained data could allow us to conclude that fertilizers can be applied in selected doses so as not make a harmful effect as sources of heavy metals. Working group No.4 NOTES ORGANOGENESIS VIA CALLUS IN KIWIFRUIT HYBRID PLANTS (ACTINIDIA DEUCIOSA CHEV. X ACTINIDIA ARGUTA SIEB. E ZUCCH.) STANICA, F. University of Agricultural Sciences and Veterinary Medicine, Faculty of Horticulture, 59 Marasti Str., 71331 Bucharest, Romania, E-mail: [email protected]

Using four different explants: root, intemode, petiole and leaf blade, excised from a kiwifruit hybrid (Actinidia deliciosa Chev. x Actinidia arguta Sieb. e Zucch.) cultured in vitro, the effects of culture medium, pH and subculture number on callus production were studied. The maintaining of callus proliferation, organogenetic capacity during seven subcultures and the callus regenerated shoot behavious in proliferation and rooting processes was also studied. Callus proliferation, its quality and the regenerated shoot number were different, depending on experimental factors.

151 Working group No. 4 NOTES DETERMINATION OF GENETICALLY DISTANCES IN ROMANIAN SOYBEAN {GLYCINE MAX) GENOTYPES STROE, E.\ GRESSHOFF, P.2 1 University of Agronomical Sciences and Veterinary Medicine, Genetics and Plant Breeding, 59 Bd. Maristi, 71331 Bucuresti, Romania University of Tennessee, Plant Molecular Genetics, 269 Ellington Bldg., Knoxville, TN 37901-1071, USA

Molecular polymorphism in eight soybean (Glycine max L.) genotypes were studied using arbitrary primer analysis (DAF) coupled with polyacryiamide gel electroforesis and DNA silver staining. Genetic distances consistent with pedigree data were established. Five arbitrary, linear primers (with 8 or 10 nucleotides) and four hairpin primers (possessing a fold-back loop at their 5' end) were utilized for DNA amplification using 55°C as a high stringency annealing temperature. The material was represented by F4 generation and parents used for an intraspecific hybridization. Both linear as well as hairpin primers produced good DAF profiles of about 40 scorable bands each. The DAF analysis revealed a high degree of polymorphism for all genotypes and showed the relations between the parents and F4 generation. This approach is of value for marker-assisted-selection as well as gene discovery in soybean." Working group No.4 NOTES BARLEY AND HOP VARIETIES CULTIVATED IN ROMANIA FOR BREWING STROIA, I.1, BEGEA, M.1, DRAGHICI, E.2 institute of Food Research - Bucharest 2University of Agronomical Sciences and Medicine Veterinary - Bucharest

The paper presents technical information regarding barley and hop for brewing cultivated in Romania. Main areas specific for malting barley and hop are presented. There are described soil and climate conditions specific to Romania and data regarding physical - chemical and technological quality of cultivated varieties. The results represent average values for three years. The presented results are long - term results and we estimated that we have European competitive varieties. The research work in the field of brewing barley has a 30 years tradition in Romania and in the field of hops a 20 years tradition.

153 Working group No.4 NOTES APPLICATION OF SOME BIOTECHNOLOGICAL METHODS AS AN ALTERNATIVE APPROACH IN RESISTANCE BREEDING OF PEAS TO FUNGAL PATHOGENS SVABOVA, L, GRIGA, M. AGRITEC Ltd., Zemddelska 16, 787 01 Sumperk, Czech Republic, E-mail: [email protected]

A set of experiments with Fusarium oxysporum and F. solani culture filtrates applied on several types of in vitro cultures was utilized in resistance improvement to pathogens of peas (Pisum sativum, L). Variable influence of filtrates on the different plant levels - intact plants in tube tests, multiple shoot culture and somatic embryos in vitro - was recorded. The sensitivity in the tube test was evaluated as the decrease of the infection degree in three variants - inoculation with fungus, addition of filtrates in 10% v/v concentration and control without any treatment. The effect of mycelial culture on growth reduction of test tube plantlets was much more evident as compared to the filtrate treatment. The results allowed to sort the studied pea cultivars in resistant, tolerant and sensitive groups. Multiple shoots and somatic embryos differed in the responsiveness considering the filtrate used. Multiple shoots were cultured on media with addition of filtrates and then transferred on rooting media. Viability was evaluated via the inhibition of rooting and number and length of roots. The most sensitive and evident character was the length of roots. Addition of filtrates to the induction media for somatic embryogenesis in relatively high concentrations did not suppress the initiation of somatic embryos completely. In some variants with lower filtrate concentration stimulating effects were noticed. Among the attributes under study some correlating results were recorded. Surviving plants obtained after in vitro selection were grown to maturity in the greenhouse and will be subjected to further testing in natural conditions. The filtrates were analysed for the content of some phytotoxins (T2-toxin, fusaric acid, deoxynivalenol) by TLC and HPLC. Supported by grant No. 6046 of Ministry of Agriculture CR and grant No. 521/96/K117 of Grant Agency CR .

154 Working group No.4 NOTES GALEGA ORIENTAL® LAM. - A NEW VALUABLE FORAGE PLANT TELEUTA, A. The Botanical Institute of Academy Science of R.Moldova, 14, Padurilor street., Chisinau, MD-2002 Moldova

As the result of the introduction, long cultivation, repeated reproductions of the specimens from the wild flora and of the populations from different ecological and geographical zones were elucidated the adaptationis the perennial species Galega orientalis Lam. in Moldovien's conditions. With the individual breeding's method from the local populations was created the variety Speranta, which is a regionized in R.Moldova and possesses: -high total plant weight (63-70 t/ha with humidity 75- 79%, or 160-170 q/ha dried substance) and rich in protein (22-23%); the 100 kg of green forages equate 21-22 fodder unit; - early harvest the green forages (more early with 20- 25 days that lucerne and with 10-15 days that perennial grass family; long utilization of the plantation (10-12 years); good frost-resistant (to -30 C), the immunites, resistance to pests; high foliage (70%), which not defoliate for removal hay; enriching the soil with nitrogen and calcium, ensure his protection against erosion; nectariferous plant; - absolutly absent the galegin (alcaloid) in green forage and hay and high eatability it of horned and small cattle; no expensiv spend for maintenance plantation. We have elaborated the ecological harmless tehnology of the cultivation for the new plant the purpose of obtaining seeds (5-6 q/ha) and green forages (70 t/ha).

155 Working group 4 NOTES THE PROMOTIVE EFFECT OF COMBUSTION PRODUCTS FROM PLANT MATERIAL ON THE RELEASE OF SEEDS FROM DORMANCY. THOMAS, T. H., THORNTON, M. A., PETERS, N. C. B., VAN STADEN, J. University of Bristol, IACR Long Ashton Research Station, Bristol, BS41 9AF, UK.

Seed germination responses to smoke are well documented, particularly for species found in regions where vegetation fires are prevalent. However, the effect of smoke does not seem to be dependent on seed size, shape or form; annual or perennial; sensitive or tolerant to fire. Indeed, previously we demonstrated that the dormancy of light-sensitive seeds, such as lettuce and celery, could be broken by treatment with various dilutions of saturated solutions of smoke. Seed germination can also be stimulated by aqueous extracts from charred wood. It appears that the active element in smoke is a volatile compound, or mixture of compounds, produced when the plant material is heated to 200C. To date it has not been characterised chemically Rapeseeds can be induced into dormancy with polyethylene glycoi and this dormancy can then be broken by treatment with GAs or GA-active compounds. Solutions made from disolving smoke from burnt material of various species in water, or the residues from the energy-generating gasification process of willow chips, also broke this PEG-induced dormancy. The highest biological activity was obtained from aqueous extracts of pyrolised willow, which reaches temperatures of 800-1500C during the gasification process. This suggests that the active compound/s are very heat-stable. Products of combustion from wheat, barley, rosemary and heather also stimulated the germination of lettuce seeds and rapeseeds.

156 Working group No. 4 NOTES STUDIES ON THE BIOCHEMICAL TRANSFORMATIONS DETECTED IN POLYANTHES TUBEROSA L. PLANTS TOMA, F. L, NEATA, G., DAVIDESCU, V., DIMA, I. University of Agronomical Sciences Bucharest, Bd. Marasti 59, sector 1, 71331 Bucharest, Romania.

We used an indigenous population in Bucharest area. Eight experimental variants has been studied with plants regenerated " in vitro" and in field obtained bulbs, storages at 8- 10 ° C and 20 -22 ° C in the rest period. We determined the dry substance (%), soluble sugar, nitrogen, phosphorus and potassium - total and soluble forms, in rhizome, bulbs and leaves. The results of the analyses showed a growing dynamical for the dry substances and soluble sugar, excepting the sprouting moment. Concerning the three principal macroelements, the analyses shown a strong subside on July and August, because in these months the plants metabolism was maximal.

157 Working group No. 4 NOTES TESTING THE REGENERATIVE CAPACITY IN ,,IN VITRO" CONDITIONS OF THE POLYANTHES TUBEROSA L. PLANTS TOMA, F. L, ROSU, A. University of Agronomical Sciences Bucharest, Bd. Marasti 59, sector 1, 71331 Bucharest, Romania.

The method of "in vitro" micropropagation represents a promising way to shorten the time necessary for the production of flower bulbs in Polyanthes tuberosa L. plants. By rescuing the bud explants of 1-2 mm and culturing them on Murashige & Skoog medium supplemented with different concentrations of NAA, BAP and KIN results in the stimulation of shots formation in various proportion. Stimulation of maximal number of shoots was observed on the medium with 0,2 mg/l NAA + 1,5 mg/l KIN + 2,0 mg/l BAP. The present work shows the possibility to regenerate a rich propagation material that can be more rapidly and efficiently transferred in the field for cut flowers production. Working group No.4 NOTES SUNFLOWER EXTRANUCLEAR MUTANT COLLECTION. PHOTOSYNTHETIC REVERTANTS: BACK MUTATIONS OR SUPPRESSION? TRIBOUSH, S., ULITCHEVA, I., DANILENKO, N., DAVYDENKO, O. Institute of Genetics and Cytology, National Academy of Sciences, Kuprevich 14, 220068 Minsk, Belarus

Previously we had studied a collection of 12 NMU-induced extranuclear photosynthetic mutants and ascertained that the mutation point was located in the chloroplast DNA. In the present study the same mutagen was used for the re-treatment of the en:chlorina-7 mutant and a range of revertants (complete or partial) was selected. The maternal nature of the inheritance of these reversions was proved by reciprocal crossing. One of the revertants fully restored the green colour of the leaves (complete revertant-2), while the others, although still partly deficient in chlorophyll content, were more productive than the mutants. According to our previous results, the cpDNA of en:chlorina-7 is definitely re-arranged. A new restriction site appears for the Hindi!! endonuclease; consequently there is no 6.7 kb fragment, while two new fragments with 3.2 kb and 3.5 kb appear. The detailed restriction analysis of revertants revealed two types of cpDNA/Hindlll patterns: a retained mutant DNA with two additional fragments and a DNA lacking one fragment as seen in en:chlorina-7 and the initial cpDNA as determined in the parent inbred line 3629. Restoration of the normal cpDNA pattern correlates with the complete reversion of leaf color (CR-2), while the mutant cpDNA is retained in the partial revertants. In view of these results, together with attempts to localise the mutation by comparing the restriction map of Helianthus annuus cpDNA with chloroplast physical maps of other plant species, we propose that the chlorina mutation is situated in the DNA region containing a set of photosystem (PS) genes. An explanation of the exact mechanism of the induced chloroplast mutation and reversion will not be possible until the fragments are sequenced, but we suppose that in the case that the CR-2 revertant true reversion has occurred. As far as we know this is the first case of induced reversion detected at the cpDNA level in higher plants.

159 Working group No. 4 NOTES CHROMOSOME ISOLATION FROM NORWAY SPRUCE (PICEA ABIES (L.) KARST.) ROOT TIPS. OBERALL, I., HAVEL, L. Mendel University of Agriculture and Forestry, Department of Botany and Plant Physiology, Zemedelska 1, 613 00 Brno, Czech Republic, [email protected]

The procedure of chromosome isolation consist of two fundamental steps: 1) accumulation of metaphase cells, 2) isolation of chromosomes from the cells. Accumulation of cells in metaphase implies the use of physical or chemical agent inhibiting mitotic spindle action. After chemical treatment chromosomes can be released by mechanical disruption of plasma membranes in hypotonic buffer containing agent weaking the membrane and stabilizing the released chromosomes. Root tips of Norway Spruce (Picea abies (L) Karst.) seedlings grown for 1 week on wet filter paper were used for chromosome preparation. The root tips were then treated by 0.05% colchicine for 24 h at 25 °C. Roots were separated from plantlets, washed in distilled water, fixed for 15 min at 4 °C (3% formaldehyde, 10 mM Tris, 10 mM Na2EDTA, 100 mM NaCI, 0.1% Triton X-100) and washed 3 times 20 min in Tris buffer (10 mM Tris, 10 mM Na2EDTA, 100 mM NaCI) again at 4 °C. Excised root tips were placed in LB01 buffer (15 mM Tris, 2 mM Na2EDTA, 0.5 mM spermine, 80 mM KCI, 20 mM NaCI, 0.1% Triton X-100) and chromosomes were released by homogenization with T8 Ultra Turrax dispergator. Obtained suspension was filtered through 48 jam nylon mesh and chromosomes were separated by centrifugation in sucrose gradient (20 - 40 - 60%) at 57g for 15 min. The highest number of intact chromosomes was found in the 40% sucrose layer. The chromosomes were washed in PRINS buffer (10 mM Tris, 50 mM KCI, 2mM MgCI2.6H2O), centrifuged (157 g, 15 min) and 40 (J of pellet was resuspended, stained with DAPI and observed by fluorescence microscopy.

160 Working group No. 4 NOTES A SIMPLE METHOD FOR DNA ISOLATION FROM ALGAE USING CHELEX - 100. VLADIMIRESCU, AL, GAVRILA, L, GRECU, C. Bucharest University, Genera! Genetics & Evolution Department., Aleea Portocalilor 1 - 3, Code 77206, Bucharest, Romania.

We present a simple and reliable method for DNA - template preparation after Berthold et al. (1993), by using CHELEX -100 (BIORAD) - a synthetic resin with chelative properties against endonudeases. The protocol was applied at various photosynthetic microorganisms: Synechocvstis PCC 6803, Spirulina platensis (cyanobacteria) and: Chlamydomonas reinhardtii 137c* and 3359, Dunaliella salina and Chlorella kessleri (green algae). After isolating DNA by CHELEX - 100 method (about 40 min.) this DNA was used as template in RAPD - PCR protocols. In this way, we have obtained reliable patterns for each alga, such specific pattern has been produced even between two different strains of Chlamydomonas reinhardtii. A modified pattern in transformed (using E. coli plasmid pBR322 and electroporation technique) and nontransformed Dunaliella salina has also been obtained.

161 Working group No.4 NOTES IMAGE ANALYSIS IN CHARACTERIZATION OF NORWAY SPRUCE POLYEMBRYONIC CULTURES VLASlNOVA, H., TRUKSA, M., HAVEL, L. Mendel University of Agriculture and Forestry, Department of Plant Physiology, Zemedelska 1, 613 00 Brno, Czech Republic, E mail: [email protected]

Primary goal of our experiments was to establish sustainable polyembryogenic cultures producing embryos converting to well developed plantlets of selected Norway spruce trees. The selected trees come from Beskydy mountains of Northern Moravia. Approximately 100 of polyembryogenic cultures were established from 4 trees that showed good environmental tolerance and disease resistance. High variability between cultures established from seeds of single mother tree as well as originating from different parts of a single zygotic embryo were found. These polyembryonic cultures differed in water content, growth rate, subculture interval, mitotic index and in their response to osmotic compounds and abscisic acid in maturation medium. Sorting of cultures to groups with the similar growth characteristics seems to be helpful to optimise the culture conditions (transfer interval, maturation medium, desiccation). Growth differences were detected by: 1) increase of fresh weight to 1 mg of starting weight of polyembryonic culture, 2) image analysis of area and density of cultures, 3) dry weight - water content measurement. The differences between mitotic indexes of cultures were also observed. The comparison of different approaches suggested that image analysis could be a helpful method for prompt basic characterisation of Norway spruce polyembryonic cultures. Acknowledgement: This study was supported by the Grant Agency of the Czech Republic (grant No. 522/96/K186).

162 Working group No.4 NOTES THE AUXIN AND CYTOKININ ECONOMY DURING THE GROWTH CYCLE OF VBI-0 TOBACCO CELL CULTURE ZAZiMALOVA, E., PETRASEK, J. BREZINOVA, A. Institute of Experimental Botany ASCR, Laboratory of Hormonal Regulations in Plants, Rozvojova 135, CZ 16502 Prague 6, Czechia, E-mail: [email protected]

The key role of phytohormones of both auxin- and cytokinin-type in the regulation of plant cell development was first reported many years ago. There is an extensive knowledge about the endogenous levels, uptake and efflux of both phytohormones and mechanisms of reception of both phytohormonal signals. However, the data are scattered to the various plant cells, tissues, organs, etc., analysed at different stages of their development. We have studied these parameters on the auxin-dependent, cytokinin-autonomous VBI-0 tobacco cell strain grown in a suspension culture. This model cell strain displays high friability and specific filamentous phenotype and the processes of cell division and polar cell growth are well time-separated. When cultivated under optimal conditions, the endogenous auxin (indole-3-acetic acid, IAA) level reached its maximum in the stationary growth phase almost in parallel with the maximum activity of the auxin efflux carrier. In contrast, the maxima of endogenous levels of individual cytokinins (N6-(A2-isopentenyl)adenine, zeatin and their ribosides) either preceded or coincided with the individual onsets of cell division within the exponential growth phase. Also the highest activity of membrane-associated cytokinin-binding site appeared in the exponential phase and coincided with the most intensive cell division. The activity of membrane-associated auxin-binding site increased in the late exponential and early stationary phase when the polar cell growth began. The results suggest the regulatory role of auxins in both cell division and elongation and the role of cytokinins in the control of cell division. The VBI-0 tobacco cells posses the mechanisms governing both the intracelluiar levels of auxins and cytokinins and the sensitivity of the cells to both these signals (including respective putative receptors) in relation to the individual stages of cell development. Acknowledgement: the research was supported by the Grant Agency of the Academy of Sciences of the Czech Republic (project No.: A6038706) and by the Grant Agency of the Czech Republic (project No.: 206/98/1510).

163 Working group No.4 NOTES THE EFFECT OF ETHREL ON CUCUMBER PLANT GROWTH AND FLOWERING ZELENA, E., ZELENt, F., VECEROVA, D. Research Institute of Crop Production, 161 06 Prague, Czech Republic

Ethylene is known to stimulate female flower formation in members of the Cucurbitaceae. We examined the effect of Ethrel, as a source of ethylene, on growth and flower formation on two varieties of picking cucumber differing in capability to form female flowers. The non-hybrid variety Melnicka on contrary to the hybrid Fatima grows faster and uniform, however it is known, there are problems with fruit formation in this variety. Cucumber Melnicka even did not form any female flowers in the cultivation conditions of growth chambers. Application of Ethrel in the form of spray to the seedlings in the stage of developing of first true leaf caused in the both varieties a similar reaction as far as growth of leaves and branches, however opposite reaction as far as female flowers formation. Ethrel caused a significant stimulation of female flowers on variety Melnicka, especially on plants grown on the higher level of nutrients and inhibition of female flowers formation on variety Fatima, especially when grown on the basal nutrients supply. Therefore experiments were conducted to examine the response of cucumber variety Melnicka to the timing of Ethrel application. The effect of Ethrel soaked to the seeds with that applied as a spray in the stage of cotyledonary leaves and, similarly as in the forgoing experiments, in the stage of developing of first true leaf was compared. Only in the last case the treatment stimulated female flower formation. In all cases Ethrel at concentrations that stimulated female flower formation inhibited the actual leaf growth. However growth of the new leaves was accelerated and the total weight of the shoots was not influenced significantly.

164 Abstracts presented in working group No. 5

Chairmen: Prof. S. Grego and Prof. F. Megusar and Prof. E. Smedile RESEARCH AND RESULTS REGARDING THE CORNICHON - TYPE CUCUMBER CULTURE ON RE-USED ORGANIC SUBSTRATE IN GREENHOUSES 169 ATANASIU, N.

TOMATO SOILLESS CULTURES WITH NUTRIENT SOLUTIONS ON RECYCLING ROOTING SUBSTRATE IN AN OPEN CIRCUIT 170 ATANASIU, N., LUCHIAN, V., CHIRA, A., POPESCU, V., DOBRESCU, A.

RESEARCHES ABOUT THE RECYCLE OF SOME ORGANIC WASTE IN GREENHOUSE VEGETABLE CULTURE 171 DAVIDESCU, V., NEATA, G., DIMA, I.

NEW TYPES OF NITRIC FERTILIZERS AND BIOLOGICALLY ACTIVE SUBSTANCES AS ELEMENTS OF ECOLOGICALLY PURE TECHNOLOGY OF GROWING WINTER WHEAT 172 DRAGA, M., SHUMIK, S., MUSIENKO, M.

DYNAMIC OF 1SN-UREA IN SILT LOAM AND CLAY SOILS DURING TWO YEARS ROTATION 173 GREGO, S., BADALUCCO, L, MOSCATELLl, M. C, MARiNARl, S. FALCHINI. L.

PROTECTION METHOD OF MOLLIC ALLUVIAL SOIL STRUCTURE IN VEGETABLE AGROECOSYSTEMS CULTIVATED WITH CARROT174 CHIVULETE, S., CIOFU, R., DITU, D., CHITANU, G., MATE!, M., MOTELICA, M.

A BIOMONITORING SYSTEM FOR URBAN TRAFFIC POLLUTION 175 IERADI, L.A., CRISTALDI, M., SABELLI, S., PALMI, i., GROSSI, R., CAMPANELLA, L.

TRANSGENIC CROPS AND ENVIRONMENTAL RISKS: WHAT TO MONITOR 176 JAVORNIK, B.

STATION NETWORK DATA: REPRESENTATIVE OF DATA AND THEIR SPATIAL INFORMATION 177 KAJFEZ-BOGATAJ, L, BERGANT, K.

COMPARISON OF THE PRIMARY AROMA COMPOUND MAPS OF MUSCAT OTTONEL MUST AND WINE BY GASCHROMATOGRAPHY/ MASS SPECTROMETRY 178 KOVACS, T., KALLAY, M., SZABO, S. A., KORANY, K.

PROCESSING OF ORGANIC SECONDARY AND RENEWED RAW MATERIAL FOR ALTERNATIVE POWER-CARRIERS MANUFACTURE179 LAGNO, YU., ISHKOV, V., SOBOLEV, W.

167 IMPROVEMENT POSSIBILITIES OF SEVERAL PHYSICAL CHARACTERISTICS OF NUTRIENT POTS AND THEIR EFFECT ON THE QUALITY OF CABBAGE TRANSPLANTS 180 LUCHIAN, V., CIOFU, R., CHIVULETE, S., MOTELICA, M.

ENVIRONMENTAL IMPACT ASSESSMENT FOR A BIOMASS FUELLED POWERPLANT 181 MATEI, M.

MINERAL NITROGEN ASSESSMENT USING POROUS CERAMIC CUPS AND 15N TRACER 182 MOUTONNET, P.

GEOGRAPHICAL CONCENTRATION PATTERNS OF TRACE ELEMENTS IN EASTERN ROMANIAN CARPATHIANS STUDIED BY MOSS BIOMONITORING TECHNIQUE 183 OPREA, C.

THE ANTISEPTICAL AND STABILIZER EFFECT OF PINE'S RESINE AS AN SUSBTITUTE OF SULPHUR DIOXIDE IN RETSINA TYPE WINES 184 RADOI, F., POMOHACI, N., KOUROUKU, E.

FACTORS AFFECTING THE FORMATION OF OLIGOSACCHARIDES FROM WHEY PERMEATE BY ENZYMATIC HYDROLYSIS 185 SHEHATA, A. E., ALI, A. A., KHORSHID, M. A., FODA, M. I.

THE EFFECT OF PHYSIOLOGICALLY ACTIVE COMPOUNDS UNDER Pb POLLUTION ON WINTER WHEAT {TRITICUM AESTIVUM L.)186 SKOPETSKA, O., SVETLOVA, N.

STUDY OF Fe PRESENCE IN THE ECOSYSTEM USING THE MOESSBAUER EFFECT (HIGH - FIDELITY DETERMINATION) 187 VACARU-OPRIS, M., VACARU-OPRIS, I., GHEORGHE, P.

TOMATO FERTILIZATION WITH SLUDGE COMPOST AND HEAVY METALS TRANSLOCATION TO THE SOIL - PLANT SYSTEM 188 VAJIALA, M., DUMITRU, M., CIOFU, R., STAN, S., GAMENT, E., GEORGESCU, M.

168 Working group No. 5 NOTES RESEARCH AND RESULTS REGARDING THE CORNICHON - TYPE CUCUMBER CULTURE ON RE-USED ORGANIC SUBSTRATE IN GREENHOUSES ATANASIU, N. University of Agricultural Sciences and Medicine Veterinary, 59 Marasti Blvd. Bucharest - 71331, Romania

In Romania, the comichon - type cucumber culture in greenhouses is practiced on large areas. In the traditional cultures on soil, the cornichon - type cucumber culture is reduced, but it requires significant expenses for the soil disinfection works. Non-conventional cultures on rooting substrata represents future alternative also for cucumbers, due to the high production potential, and the partial decrease of soil and production - pollution with pesticide residues. Under the present economic conditions, the organic substrata made up of peat mixtures with forest residues (tree bark, sawdust) are relatively expensive for the traditional areas of cucumber culture. Due to these considerations, the re-use of the organic rooting substrata represents a useful research direction both from the technological and economic. This paper presents the method of using various type of rooting mixtures in the second year after preparation, in soilles culture as compared to newly - prepared mixtures. The production data reflect its high level, the particular response of all the hybrids, as well as several elements of production quality that prove both the conclusions and the diversification of future research.

169 Working group No. 5 NOTES TOMATO SOILLESS CULTURES WITH NUTRIENT SOLUTIONS ON RECYCLING ROOTING SUBSTRATE IN AN OPEN CIRCUIT ATANASIU, N., LUCHIAN, V., CHIRA, A., POPESCU, V., DOBRESCU, A. University of Agricultural Sciences and Medicine Veterinary, 59 Mara§ti Blvd., Bucharest - 71331, Romania

Up to the present, Romania has accumulated a great volume of experimental data regarding the non-conventional tomato culture. This has opened the way to new research aiming to re-use the organic rooting substrata for one more cycle. This new research direction was established as a result of the high prices of the materials necessary for mixture preparation (peat and forest residues). Under the technological conditions presented in this paper, the re-use of these substrate determines a significant decrease in the production costs and, at the same time, it assures a satisfactory production level that makes this tomato culture system extremely profitable. The production data underline an increase in its volume and earliness, and a technological quality level superior to the soil-cultivated control. At the same time, certain differences occur with respect to the behaviour of the hybrids experimented on these culture substrata, as shown by the production results. The conclusions emphasize on the results applicable in future research, as well as for those farmers who intend to use modern culture system.

17fl Working group No.5 NOTES RESEARCHES ABOUT THE RECYCLE OF SOME ORGANIC WASTE IN GREENHOUSE VEGETABLE CULTURE DAVIDESCU, V., NEATA, G., DIMA, I. University of Agricultural Sciences and Veterinary Medicine, Bd. Marasti 59, section 1, 71331 Bucharest, Romania

The accumulation of some organic wastes in high quantities resulting from agriculture, industry and household activities involve some problems from the point of view of depositing; utilisation that could be made after their decomposition and transformation in an organic substratum and then cultivated with vegetables. These materials must be tested to identify their content in nutritive elements and eventually toxic elements. The composts must be tested in culture technology to establish the possibility of their use on some cultures and the effects on the quality of the crop. The researches wanted to establish the use of mixed composts: waste compost resulted from decomposition of litter and forest compost resulted from forest exploitation. The experimental plant was bean culture, variety Lavinia, and the results obtained showed that the accumulation of nitrates, nitrites and lead quantities in plants were different with the components of substrates. Also, the crops were different in direct dependence with the components of culture substrates.

171 Working group No.5 NOTES NEW TYPES OF NITRIC FERTILIZERS AND BIOLOGICALLY ACTIVE SUBSTANCES AS ELEMENTS OF ECOLOGICALLY PURE TECHNOLOGY OF GROWING WINTER WHEAT DRAGA, M., SHUMIK, S., MUSIENKO, M. Taras Shevchenko University, Dept. of Plant Physiology and Ecology, Volodymyrska 64, 252033 Kiev, Ukraine

Developing new, non-traditional ways of agricultural production has become very important recently due to significant pollution of the environment. To meet those ends we used a new type of non-nitrate nitric fertilizer, i.e.CARBON-AMMONIUM SALTS (CAS), a mixture of carbonate of ammonium with hydrocarbonate of ammonium of the general formula NH4HCO3 (N-17%, CO2 - 50%). We also used the following anti-stress physiologically active substances: TRiMAN, AGROSTIMULIN and EMISTIM. Triman is a synthesised substance derived from N-oxides of pyridin. Agrostimulin and Emistin, growth regulators, are produced in vitro in the Institute of Bio-organic Chemistry and Petrochemistry of the Academy of Sciences of Ukraine. They are products of metabolism of the epiphytes of roots of Panax ginseng and Hippopho rhamnoides. We studied the impact of those substances on growth of wheat in the open-air field. Due to application of these substances the percentage of photosynthetic pigments in flag leaf increased by up to 25-30%, especially during the crucial period of plant development which is blooming. Also the assimilating surface of the leaf expanded, especially when Triman was applied in combination with CAS during all stages of plant development. Under the effect of substances of natural origin, Agrostimulin and Emistin, the percentage of water in the assimilating system of wheat plants increased, especially at the stage of blooming. Application of any of the substances resulted in improvement of such adaptation ability of the plants as their ability to retain water, especially at the stage of milk-wax ripeness. Agrostimulin, Emistin and Triman activated the functions of the nitrate-reduction system in the flag leaf and ability of the plants to use soil N and fertilizers. Application of CAS or Triman, or their combination, and Agrostimulin and Emistin, increased the soluble protein fractions in the flag leaf. This is especially true for glutelines and albumins. This effect, in turn, gave better quality of grains and increased quantity. Productivity of wheat improved due to augmentation of the quality of spikelets in ear and quantity of grains in ear. The weight of 1000 grains also increased, as well as the weight of grains in each ear. The effect of substances of natural origin was less significant relative to the effect of Triman and CAS. Nevertheless, application of substances of natural origin is very promising, especially in unfavourable ecological conditions. Therefore, the technology of application of those substances of natural origin needs to be further improved to increase the efficiency of their application.

172 Working Group No.5 NOTES DYNAMIC OF 1015N-URE, A IN SILT LOAM AND CLAY SOILS DURING TWO YEARS ROTATION GREGO, S.\ BADALUCCO, L.1, MOSCATELLI, M. C.1, MARINARI, S.1, FALCHINI. L.2 1Dipartimento di Agrobioiogia ed Agrochimica, Universitf delta Tuscia, 01100 Viterbo, Italy 2Dipartimento di Scienza del Suolo e della Nutrizione delle Piante, Firenze, Italy This work is part of a wider study devoted to investigate transformations and sharing out of 15N-urea into various soil nitrogen pools (plant, microbial biomass, organic matter, etc.) in order to draw a fertiliser-N balance. The following treatments were set up in two different soils (silt loam and clay) in the presence of a 2 year-rotation (1993 sorghum; 1994 wheat): 1) no crop + 15N-urea; 2) sorghum + 15N-urea; 3) no urea + sorghum-„-...., in 1993; 1) no crop. , *Vresidual effect; 2) wheat, 15N-residual effect; 3) no crop + 15N-urea; 4) wheat + " N-urea, in 1994. The amount of fertiliser added was always 100 kg N/ha and N-urea was enriched at 10% in N. Soils were sampled and analysed at the crop harvest. Microbia! biomass held only a small percentage of N deriving from urea, confirming its fast turnover. Soil microbial biomass immobilized more N in the presence of crops than without, probably due to rhizodepositions released from roots. Moreover, in both soils sorghum seemed more competitive than wheat for uptaking available N.

173 Working group No. 5 NOTES PROTECTION METHOD OF MQLLIC ALLUVIAL SOIL STRUCTURE IN VEGETABLE AGROECOSYSTEMS CULTIVATED WITH CARROT CHIVULETE, S.\ CIOFU, R.2, DITU, D.3, CHITANU, G.4, MATEI, M.1, MOTELICA, M.1 institute of Pedological and Agrochemical Research, 69 Mara§ti Blvd., Bucharest - 71331, Romania zUniversity of Agricultural Sciences and Medicine Veterinary, 59 Marasti Blvd., Bucharest - 71331, Romania 3Station of Vegetable Research and Production, 23 Mesteacanului Street, Buz2u - 5100, Romania 4"Petru Poni" Institute of Macromolecular Chemistry, 41 A Grigore Ghica Voda Alley, iasi - 660, Romania

Intensive cultivation of vegetable agroecosystems inevitably result in the worsening of some physical soil characteristics. Soil structure degradation at the seed bed surface determines an intensification of the crust formation process and plant density reduction. Among the modern methods of soil structure protection and reduction of the crust formation process, several synthetic soil conditioners are mentioned (De Boodt, M., 1971, Chivulete, S., Carpov, A. et al'., 1990). The paper presents experimental results on the protection of moilic alluvial soil structure cultivated with carrot for the obtaining of seed - stock roots, using a Romanian polyelectrolyte - PONILIT GT 1. The application of this product as a concentrated solution of 0,1 and 0,2 % resulted in the improvement of soil structure at the seed bed surface, as well as of other of its physical, chemical and biological characteristics. Thus, there was an increase from 3 % to 14 % in the content of aggregate hydrostability elements (0greater than 0,20 mm), and a decrease in the index of structure instability from 3,06 to 0,65. As compared to the control, there were also noticed increases in the initial humidity, hydraulic saturated conductivity and total porosity, as well as decreases in the bulk density and resistance to penetration. Important increases were also noticed at several chemical elements (40-52 % for total nitrogen, 7-8 % for total phosphorus, 43-59 % for mobile potassium, 31-53 % for anions, and 42-67 % for cations). From the biological point of view, the treatment determined an increase in soil respiration, bacteria number, and microbia! biomass (13-19 %), while the fungi number decreased by about 27-28 %, as compared to the control. PONILIT GT - 1 determined an increase in the number of germinated plants and total production of carrots. The standard production of seed - stock roots increased by 30-34 %, as compared to the control, resulting in a decrease of about 33 % of the cost price per root.

17/1 Working group No.5 NOTES A BIOMONITORING SYSTEM FOR URBAN TRAFFIC POLLUTION IERADI, LA1, CRISTALDI, M.2, SABELLI, S.2, PALMI, I.1, GROSSI, R.3, CAMPANELLA, L.3 1,,La Sapienza" University, Department of Genetic and Molecular Biology, Nucleic Acid Center, A. Moro 5, 00185 Rome, Italy. E-mail: [email protected] 2,,La Sapienza" University, Department of Animal and Human Biology, A. Borelli 50, 00161 Rome, Italy. E-mail: [email protected] .it 3 nLa Sapienza" University, Department of Chemistry, A. Moro 5, 00185 Rome, Italy.

The aim of the present work is to realise an environmental biomonitoring system through the use of the outdoor house mice (Mus musculus domesticus) as monitors for contamination related to motor vehicle traffic. This study was carried out in the city of Rome (Italy). The trapping sites were selected in two locations exposed to different flows of motor vehicles: Site 1 (Muro Torto Avenue) exposed to high traffic flow (on average 5000 motor vehicles/h in 1991 and 3000/h in 1995) and Site 2 (a Zoo) practically closed to traffic. Animals were collected from 1991 to 1995. The following analyses were applied on the collected animals (n=110): two mutagenetic tests (bone marrow and peripheral blood micronucleus test and sperm abnormality assay) and heavy metal (lead, cadmium, copper and zinc) determination in liver, kidney and bone in order to ascertain possible genetic damage and a correlation with heavy metal concentration. Furthermore, anomalies of the five inter-molar palatal ridges were studied on the same animal groups because it is known that the intermolar palatal ridges are subjected to changes of the epigenetic type, which are determined during embryonic development and remain fixed. Data were statistically analysed by ANOVA and t-test. Results show that the frequency of micronuclei and abnormal sperm cells and the content of lead and cadmium measured in liver, kidney and bone are significantly higher in Sitel mice in comparison with Site2 mice. A significant decrease of these parameters was observed in Sitel mice collected in 1994 in comparison with mice collected in 1991. No significant differences were observed between mice from Site2 collected in 1992 and 1995. Parallely the number of palatal ridges anomalies was significantly higher in Sitel mice, collected in 1991, in comparison with other mice groups, and a significant decrease of anomalies was observed in mice from Muro Torto (1994) in comparison with mice collected in the same site in 1991. These results may be explained by the decrease of traffic flow on Muro Torto Avenue (from 1991 to 1996), by general improvement (from 1993) of air quality, measured in the city of Rome, introduction of unleaded petrol and control of motor vehicles exhausts. In conclusion the efficacy of the use of the house mouse as monitor for urban pollution was confirmed. Also it is suggested that physical, chemical and biological monitoring be ensured within our laws.

175 Working group No. 5 NOTES TRANSGENIC CROPS AND ENVIRONMENTAL RISKS: WHAT TO MONITOR JAVORNIK, B. University of Ljubljana, Biotechnicai Faculty, Agronomy Department, Centre for Plant Biotechnology and Breeding, Jamnikarjeva 101, Ljubljana 1000, Slovenia, E-mail: [email protected]

After a decade of small-scale field trials with transgenic crops, they are now being commercially produced on a large-scale in some countries. Transgenic soybean, corn, cotton, canola and potato were grown on more then 12 m ha in 1997. Paralel! to the rapid development of genetically modified plants and releases into the environment, their biological safety has been investigated and countries have been setting up regulations to control work on genetically modified organisms. There are considerable differences in approaches to regulating transgenic plants, and the results are clearly seen in the extent to which they are commercially grown, particularly in North America and Europe. The main purpose of regulations is a requirement for an assessment of risks before a transgenic crop is released into the environment. The first step in risk assessment analysis is to identify any hazards (characteristics of an organism which have potential for harm, or potential to cause an adverse effect) and whether they can be realized and to what magnitude in the receiving environment. Risk assessment of the transgenic crop involves characterization of the ecology and toxicology of the transgene and transgene product and evaluation of the consequences of the agronomic crop being transgenic in current agricultural practice. The most important potential hazards for the release of transgenic plants are the capacity of the plant to survive, establish and spread, the potential of gene transfer to wild relatives, and possible pathogenic effects on other non-target organisms. How do we monitor releases? A significant problem in defining responsibility for harm is the difficulty in deciding whether a change has occurred and to what extent, and further, what impact a released transgenic crop has on existing changes. Anything obvious will almost certainly be discovered during the monitoring associated with the development of the transgenic plant, but low frequency events in which harm might occur are unlikely to be detected. Risk assessment can also involve broader criteria which are covered in some regulations, such as sustainability, socioeconomics and ethics. In this context, full assessments (and monitoring) of all aspects of risk potentially posed by transgenic crops are very demanding and difficult tasks, especially in terms of interpretation of what is harmful in the relative values of different (socio)environments. This regulatory approach stresses dissatisfaction with current intensive agricultural practices, and uses agrobiotechnology to tackle the problem, but at the same time, has a major impact on the acceptance of agrobiotechnology and, consequently, on the existing freedom of farmers to choose the most suitable crop on economic and environmental grounds.

176 Working group No. 5 NOTES STATION NETWORK DATA: REPRESENTATIVE OF DATA AND THEIR SPATIAL INFORMATION KAJFEZ-BOGATAJ, L, BERGANT, K. University of Ljubljana, Biotechnical Faculty, Department of Agronomy 1000 Ljubljana, Jamnikarjeva 101, Slovenia E-mail: [email protected], [email protected]

The paper relates aspects of measurement and observation representativity and spatial interpolation of data in environment sciences. Spatial distribution of data is major concern in detecting the changes in different areas. There are some steps that need to be taken before comparing the data, using the data for spatial interpolation or in forecasting models. Verifying the quality of data is necessary before using them in models, because of GIGO problem: garbage in, garbage out. To have quality data we need to have good measurements and observations and for them we need good instruments, suitable locations, time and highly skilled observers. If we want to compare the data from different locations, we need to have standardized instruments, measurements and terrain. Interpolating the data on locations, where measurements are not available, can help us to get more global view on changes in some region. Portraying these data on maps is the clearest way of presenting the data if we are interesting in spatial distribution and spatial variability of data. For reasonable spatial interpolation of data, we need to have enough data for specific area. Corresponding to processes we would like to detect, we need to choose the density of measurements. Needed density of data for spatial interpolation in aspect of changes in different scales is discussed in second part of paper. The need for choosing the scale of changes we are interested in, regarding the needed density of data, is pointed out with simple example. With increasing number of station are also increasing the costs of our experiment. That is the reason that we can not dense the station network without limitations. There are also technics for spatial interpolation of data where the auxiliary variable beside measurements of primary variable, can be used to improve the maps. Lost of spatial information with decreasing number of data is shown on example of meteorological data at the end of the paper together with example of improving the precipitation map with information about relief.

177 Working group No.5 NOTES COMPARISON OF THE PRIMARY AROMA COMPOUND MAPS OF MUSCAT OTTONEL MUST AND WINE BY GASCHROMATOGRAPHY/ MASS SPECTROMETRY KOVACS, T., KALLAY, M., SZABO, S. A., KORANY, K. University of Horticulture and Food Industry, Department of Food Chemistry and Nutrition, Somloi str. 14-16, Budapest, H 1118 Hungary, E-mail: [email protected]

Elaboration a new sample preparation method the separation of the aroma constituents have been solved on a Supelcowax 10 stationary phase coated (WCOTT) capillary. Having identified as much substances as possible by mass selective detector the map of the aroma compounds has been constructed by a recently developed evaluation method applying principals used in relative mass spectrum creation.

178 Working group No.5 NOTES PROCESSING OF ORGANIC SECONDARY AND RENEWED RAW MATERIAL FOR ALTERNATIVE POWER-CARRIERS MANUFACTURE

LAGNO, YU., ISHKOV, V.t SOBOLEV, W. State Enterprise of Defense Ministry of Ukraine "UkrEcoBusiness", 59 Artyom St., 252053 Kiev 53, Ukraine.

Pyrolitical products from wood, industrial organic waste, rubber-technical (autocover) and polymeric materials - gas, liquid fuel, the pitches, pyrocarbon - can find selling not only in Ukraine, but also behind its borders. Practically from each ton of waste it is possible to receive about 500 kgs of liquid fuel, up to 100 e3 of gas and 300-400 kgs of half- coke. The carried out elaborations are perspective not only for the purposes of reception of alternative kinds of fuel, but also as a means for environment protection. Doubtless advantage of the developed technology is its reserved cyclic character distinguished by absence of constant emissions in an atmosphere, which are characteristic for technologies of a generating type. A number of innovations, ensuring increase of speed and efficiency of pyrolise processes and ecological cleanliness of system is stipulated: preliminary training of raw materials in an acoustic-vortical flow; development and use a new filtering materials for catalytical deep clearing of pyrolisis gases from toxic oxides and linkage of heavy metals cations and radionuclides with the subsequent burialing in the capsulated kind. The designing and construction of a modular factory for processing of raw materials with productivity 100-300 tons per day and also a pilot installation (10-20 tons per day) on a mobile platform for work in a Chernobyl zone of alienation is planned. The calculating profit from factory's work (with volume of processing up to 300 tons per day), planned on the conversion basis, will make about USD 3,5 mln. per one year. The introduction in action only one pilot installation with volume of processing 5000 tons of rubber-technical goods per one year will allow to give a work for 35 persons. Thus will be received of fuel: gas - 350 tons, liquid - 2250 tons, firm product - 2400 tons per one year.

179 Working group No. 5 NOTES IMPROVEMENT POSSIBILITIES OF SEVERAL PHYSICAL CHARACTERISTICS OF NUTRIENT POTS AND THEIR EFFECT ON THE QUALITY OF CABBAGE TRANSPLANTS LUCHIAN, V.1, CIOFU, R.1, CHIVULETE, S.2, MOTELICA, M.2 1University of Agricultural Sciences and Medicine Veterinary, 59 Marasti Blvd., Bucharest - 71331, Romania institute of Pedological and Agrochemical Research, 69 Marasti Blvd., Bucharest - 71331, Romania

Within vegetable agroecosystems, the obtained of transplants represents a complex and expensive technological system. It must be sustained through new methods resulting in a highly superior material. The nutrient substrata frequently used for pot formation are made up of various components (garden-mould, peat, composts, sand), whose structure and chemical composition sometimes produce disturbances in the growth of young plants. The paper presents the results of the research on the improvement possibilities of physical properties of these substrata by means of addition of several hydrophilic synthesis products obtained in Romania. The effect of products was materialized in the increase of water-retention capacity, the decrease of absorption time, and the extension of water-yielding duration up to 144 hours. These influences were reflected by the quality of cabbage transplants, where the best results were obtained by the usage of the product FITPOL C in concentrations of 0,5-1,00 % of the mixture volume used. These doses determined an increase in the root-system volume and mass, height, folliar surface and transplant mass. There were also noticed economic effects of the use of the studied products, expressed by a increase of 17-37 % in the number of nutrient pots obtained / m3 of mixture, and a corresponding increase in the quantities of substratum necessary to transplantation.

180 Working Group No. 5 NOTES ENVIRONMENTAL IMPACT ASSESSMENT FOR A BIOMASS FUELLED POWERPLANT MATEI, M. RENEL-GSCI, 8, Blvd Energeticienilor, Bucharest, 79619, Romania, E-mail: [email protected]

Former ICEMENERG, nowadays Studies Research and Engineering Group (GSCI) of RENEL was involved in three R&D projects in the frame of PECO'93 program (extension of J0U2 Programme to the Central and East European countries), partially supported by European Union, referring at the biomass use from electricity (and heat) production. Environmental impact of biomass fueled power plant and advantage analyses of this versus conventional fuelled powerplant shows that biomass conversion in electricity and heat is one of the alternatives for clean electricity generation and is one of the components of a ..sustainable development" policy. Environmental impact can be assessed as: local impact, regional impact and global impact. Local environmental impact has to be assessed for: land use; atmospheric emissions; noise and vibrations; visual and landscape use; traffic and infrastructure; soil hydrogeology and water resource; terrestrial ecology; socio economics. The global environmental impact due to powerplant emissions has to be discussed in relationship with global warming, carbon cycle and acid rain. Biomass fueled power plant has a different impact versus a conventional fueled power plant because of biomass low sulphur content and C02 balance. Biomass utilization for power generation has the advantage of C02 balance. C02 emission is higher than the CO2 emission for a conventional fuel, due to low efficiency of a small power plant, but, in the meantime, the C02 obtained from biomass burning is fixed by the energy plantation. Also, by the biomass use instead of conventional fuels, the higher hydrogen to carbon ratio contributes to C02 emission reduction. The biomass fueled systems are an option in a ..sustainable development" policy.

181 Working group No.5 NOTES MINERAL NITROGEN ASSESSMENT USING POROUS CERAMIC CUPS AND 15N TRACER MOUTONNET, P. IAEA P.O. Box 100, A 1400 Vienna, Austria

Traditionally, porous ceramic cups are used for monitoring ion concentration in soil solutions with time and depth. A new device, the Tensionic, has been derived from tensiometer and allows measurement of soil water potential and mineral nitrogen in soil water solution. Field experiments have been done under maize for assessing NO3-N, NO2-N, and NH4-N through a set of Tensionics working either under passive diffusion or active vacuum.. Results are shown and interpreted. A field study allowing the comparison of different techniques of soil water sampling and determination of nitrate concentration was conducted in an apple orchard, drip irrigated and fertigated, near Vienna. Suction cups and Tensionics were used to obtain soil water samples in the root-zone at different depths (15, 45, and 75 cm). Percolation and nitrogen leaching were measured by small iysimeters and suction cups below the root zone (105 cm). Ammonium Sulphate (5% N atom excess) was applied as fertiliser during a period of ten weeks in spring 1995.The comparison of the different techniques will be presented.

182 Working group No.5 NOTES GEOGRAPHICAL CONCENTRATION PATTERNS OF TRACE ELEMENTS IN EASTERN ROMANIAN CARPATHIANS STUDIED BY MOSS BIOMONITORING TECHNIQUE OPREA, C. Joint Institute for Nuclear Research, Department of Activation Analysis, Frank Lab, for Neutron Physics, 141980 Dubna, Russia, E-mail: [email protected]

The first systematic monitoring survey from Eastern Carpathians was carried out in 1995, using Hylocomium splendens, Pleurozium schreheri and Hypnum cupressiforme mosses as biomonitors for trace -element air pollution. The moss samples were analysed by neutron activation analysis. Copper, lead and cadmium were determined by atomic absorption spectroscopy. Comparison of data for the three moss species showed the relationship of the geographical concentration patterns with the possible pollution sources. The results achieved for the nine most pollutant elements indicated the important industrial emission sources of heavy metals, the Baia Mare, Bistrita Nasaud, Bacau and the Valley of Prahova, and the areas influenced. The long-range atmospheric transport showed weak north -south gradients, especially pronounced in the case of lead, copper, vanadium and nickel. Also, some peculiarities of rare earths distribution were determined. Statistical and graphical interpretation of results obtained from the moss data and coloured contour maps GIS (Geographical information System) allowed a realistic estimation of geographical distribution patterns of trace elements.

183 Working group No. 5 NOTES THE ANTISEPTICAL AND STABILIZER EFFECT OF PINE'S RESINE AS AN SUSBTITUTE OF SULPHUR DIOXIDE IN RETSINA TYPE WINES RADOI, F., POMOHACI, N., KOUROUKU, E. University of Agronomical Sciences and Veterinary Medicine Bucharest, Faculty of Biotechnology, Marasti, 59, 71331, Bucharest, Romania, E-mail: [email protected]

Beginning with the fact that the technology of obtaining the Retsina type wines uses small doses of sulfur dioxide, in this paper we try to explain the substitution of a chemical compound with a natural product (pine's resin). Thus, starting from crops without sulfur dioxide, we supervise in time the evolution of alcoholic fermentation in the presence or in the absence of pine's resin. We ascertained that the length of alcoholic fermentation in the presence of the resin is longer and the increase curve of the selected yeast population is different in the sense of a slower development of the microorganisms. In addition, the variants with sulfur dioxide in mixture with pine's resin, after condition, proved to be stable from microbiological point of view.

184 Working group No.5 NOTES FACTORS AFFECTING THE FORMATION OF OLIGOSACCHARIDES FROM WHEY PERMEATE BY ENZYMATIC HYDROLYSIS SHEHATA, A. E.\ ALI, A. A.1, KHORSHID, M. A.2, FODA, M. I.2 1 Food Science Dept, FacAgric, Ain Shams Univ., Cairo, Egypt Food Science Dept., National Research Center, Giza, Egypt

Factors affecting the formation of oligosaccharides from whey permeate by three commercial 6-D-galactosidases {A.oryzae, Maxilact L2000 and Lactozym 3000L) were investigated. Oligosaccharides were produced using batch and continuous process techniques. Factorial design was used to arrange and study the effect of initial lactose concentration, enzyme concentration, pH and temperature on the formation of oligosaccharides at Lab scale. Increasing the initial lactose concentration significantly increased the concentration of oligosaccharides and significantly decreased the yield of oligosaccharides with A.oryzae 6-D-galactosidase as compared to Maxilact L2000 and Lactozyme 3000L. High concentration (0.01) of A.oryzae and Lactozyme 3000L 8-D-galactosidase decreased all responses insignificantly, except reaction time, which increased insignificantly with Lactozyme 3000L. Maximum formation of oligosaccharides was 3.7, 5.0 and 4.9% for A.oryzae, Maxilact L2000 and Lactozyme 3000L 6-D-galactosidase, resp. Amicon type as membrane factor showed effective results, while Hallow fibre membrane reactor was simple and easy to use for continuous formation of oligosaccharides. With Hallow fibre, low enzyme concentration was more effective on the continuous formation of oligosaccharides that higher concentration.

185 Working group No.5 NOTES THE EFFECT OF PHYSIOLOGICALLY ACTIVE COMPOUNDS UNDER Pb POLLUTION ON WINTER WHEAT (TRITICUM AESTIVUM L.) SKOPETSKA, O., SVETLOVA, N. Taras Shevchenko University, Department of Plant Physiology and Ecology, 252033 Kiev, Ukraine

Technical pollution of the environment and irrational use of plant protectors on corn cultivars affect negatively plant development and harvest quality. We studied the protective abilities of new natural physiologically active compounds to the production process of Pb-polluted wheat. Winter wheat Poleska 90 (Triticum aestivum L.) grown in an Ukrainian forest steppe region on loamy light black soil (pH = 7.0 with 3.1% of humus in the top 20 cm of the profile) was used in experiments. Pb salt, i.e. Pb(NO3)2, was applied in tillering stage as 1% solution of the N120P90K90 background. Chlorophyll and carotenoid content, peroxidase activity were determined spectrophotometricaliy. Samples were taken in the stages of flowering and milk ripeness. The physiologically active compounds were Vitalin and Proton (intermediates of vitamin B12 synthesis). Vitalin was applied in quantity of 500 ml/ha and Proton 20 ml/ha in the stage of stooling and ear formation. It is known that the plant reaction to stress factors firstly develops in changing the physiological-biochemical indices such as pigment content and fermentation system activity. The decreasing chlorophyll content can serve as bioindicator of environmental pollution. We discovered that the chlorophyll a abd b content in plants treated with lead nitrate Pb(NO3)2 decreased during flowering (by 37.7%) and milk ripeness (by 32.9%). Under the action of physiologically active substances the negative lead influence decreased and the summary content of chlorophyll came to the control level. The most effective protective action of Vitalin and Proton was observed during flowering, the most critical period for wheat, when the flag leaf gave 80% of assimilates for the formation of reproductive organs. A well known fact is that the totality and great diversity of ferment reaction underlines the cell organism vital activity. The changing ferment activity determined the intensity and character of functionality of the cells and organism on the whole and also their vitality under conditions of environmental pollution. Peroxidase activity was studied during flowering and milk ripeness and it was determined that wheat peroxidase activity decreased (in these stages) by 30-34% in variants treated with lead, but in the case of plants treated with Vitalin the peroxidase activity increased by 32% compared to the control in the stage of milk ripeness. We suppose the data obtained could allow us to conclude that physiologically active compounds could be a successful strategy for Pb-polluted fields as protectors. Working group 5 NOTES STUDY OF Fe PRESENCE IN THE ECOSYSTEM USING THE MOESSBAUER EFFECT (HIGH - FIDELITY DETERMINATION) VACARU-OPRIS, M.\ VACARU-OPRIS, I.1, GHEORGHE, P.2 1"lon lonescu de ia Brad" University of Agronomy, lasi - Romania institute of Physics and Nuclear Engineering, Magurele - Bucuresti, Romania

For Romania there was specific not only is characterized not only by a process of intensification of agriculture, but also by a process of excessive industrialization; all these resulted in modifications of some ecosystems that influence human and animal health. Within the areal of influence of the Agronomical University - lasi, Romania, where we are working, there were registred values over the compulsory limits of noxes in some "hot" points. So, we frequently register an increase of the concentrations of heavy metals, mainly of Fe, on a ray of many kilometers around the Heavy Metal Equipments Plant - located around the city, but in a zone having a developed agriculture and husbandry. Considering those data we purposed to analyzethe degree of contamination with Fe, using the Moessbauer effect (high fidelity determinations). In order to achieve the goal, we took into account many problems such as: -technique and the instruments used to collect the samples (first stage 1997 - soil samples, second stage 1998 - samples of plants cultivated in the region); -method of preparing them for analyzing the absorbtion and the retrodispersion; -technique of work by Moessbauer spectometry or absorbtion at resonance; -technique of results interpretation.

187 Working group No. 5 NOTES TOMATO FERTILIZATION WSTH SLUDGE COMPOST AND HEAVY METALS TRANSLOCATION TO THE SOIL - PLANT SYSTEM VAJIALA, M.1, DUMITRU, M.2, CIOFU, R.1, STAN, S.\ GAMENT, E.2, GEORGESCU, M.1 1University of Agricultural Sciences and Medicine Veterinary, 59 Marasti Blvd., Bucharest - 71331, RomSnia 2lnstitute of Pedological and Agrochemical Research, 69 Mara§ti Blvd., Bucharest - 71331, Romania

Organic products of various origins are acknowledged as real sources of nutrient elements with positive effects on soil and vegetal yield. Yet, at the same time, they can present negative effects, due to their extremely complex composition, as well as to the properties of the different components. The negative effects may occur not only on soil, but also on vegetal yield, from the quantitative and qualitative point of view. The agricultural compost use at random is limited because of the potential and actual danger of soil pollution with heavy metals, toxic organic compounds, nitrates, and their transfer to the plant, particularly to its edible parts. Our research was carried out on tomato culture, on a typical brown-reddish soil in the solarium. The results were positive with respect to the compost used as a source of organic matter and nutrient elements on the tomato yield, which justifies its use as a fertilizer. The highest tomato yield was obtained in the case of fertilization with 70 t/ha compost, as compared to the other doses used (30; 50 and 90 t/ha). Particularly at the large doses of compost (70 and 90 t/ha), the soil analysis reflected a tendency of pH decrease, humus and total nitrogen increase (up to the doubling point), and a maintenance under the maximum accepted limits of heavy metals (Cu, Zn, Pb, Co, Ni, Mn), with the exception of chrome. At tomato fruit, no analyzed heavy metals exceeded the maximum accepted limits, as they were accumulated particularly in root and stem. Abstracts presented in working group No. 6

Chairmen: Prof. S. Ignatowicz and Dr. Marec INVESTIGATION OF CHLORPYRIFOS RESIDUE IN TOMATOES AND TOMATO PRODUCTS BY RADIOTRACER TECHNIQUE 192 AYSAL, P., GOZEK, K., ARTIK, N., TUNCBILEK, A. S.

RESISTANCE OF THE VENTURIA INAEQUAUS (CKE.) WINT. TO METHYLTHIOPHANATE AND BIOLOGICAL POSSIBILITIES TO CONTROL THE RESISTANT FORMS 193 DELIAN, E., CHIRA, L, CHIRA A.

MECHANISM OF RESISTANCE TO INSECTICIDES IN LARGE PINE WEEVIL (HYLOBIUS ABIETIS L.) 194 DOBROWOLSKI

THE FUMIGATION TREATMENT OF STONE FRUITS WITH ACETIC ACID TO PREVENT POSTHARVEST DECAY 195 CHIRA, A.

MODIFIED - ATMOSPHERE PACKING OF APPLES AND STRAWBERRIES TREATED BY FUMIGATION WITH ACETIC ACID 196 CHIRA, A., CHIRA, L, DELIAN, E.

THE PREVENTING AND CONTROL OF FIRE BLIGHT CAUSED BY ERWINIA AMYLOVORA USING MECHANICAL AND BIOLOGICAL METHODS 197 CHIRA, L

RESULTS OF THE FAO/IAEA PROGRAM ON ..IRRADIATION AS A QUARANTINE TREATMENT OF MITES, NEMATODES AND INSECTS OTHER THAN FRUIT FLY" 198 IGNATOWICZ, S.

EFFECTS OF GAMMA RADIATION ON THE MELANIZATION PROCESS IN LARVAE OF STORED PRODUCT MOTHS 199 LUPA, D.

ESTABLISHMENT POTENTIAL OF THE WESTERN CORN ROOTWORM {DIABROTICA VIRGIFERA VIRGIFERA LA CONTE) IN ROMANIA200 ROSCA, I.

191 Working group No. 6 NOTES INVESTIGATION OF CHLORPYRIFOS RESIDUE IN TOMATOES AND TOMATO PRODUCTS BY RADIOTRACER TECHNIQUE AYSAL, P.1, GOZEK, K.2, ARTIK, N.3, TUNQBILEK, A. S.4 1 Turkish Atomic Energy Authority, Department of Nuclear Agriculture, Kazan, 06983 Ankara, Turkey, E-mail: [email protected] 2 Turkish Atomic Energy Authority, Department of Radiation Treatments 3 Ankara University Faculty of Agriculture, Department of Food Engineering, Diskapi, Ankara, Turkey 4 Erciyes University Faculty of Science, Department of Biology, Kayseri, Turkey

!n this study, which chlorpyrifos residues in tomatoes and tomato products were investigated by radiotracer technique, tomato plants were grown in outdoor conditions and applied with C14-Chlorpyrifos at a dose rate of 86.39 a.i./da twice as foliar application. The chlorpyrifos residue levels of tomatoes were above the national maximum residue limits with an average value of whole growing season of 0.23 ppm, but below the international limits (Codex Alimentarius). During the processing of harvested tomatoes to tomato juice, paste and catchup, total residue losses ranged between 21-39 per cent and the most important amount of chlorpyrifos residue was removed with peel, seed and flesh in palper. Samples were extracted in methanol both by cold and supercritic fluid extraction, and then extracts were chromatographed on thin layers of silica by using toluen+methanol+n-hexane (18:1:1 v/v) solvent system. Most of the chlorpyrifos residues in tomatoes and tomato products were determined to be translocated in the forms of extractable conjugates bound to polar components of tomatoes.

192 Working group No. 6 NOTES RESISTANCE OF THE VENTURIA INAEQUALIS (CKE.) WINT. TO METHYLTHIOPHANATE AND BIOLOGICAL POSSIBILITIES TO CONTROL THE RESISTANT FORMS DELIAN, E., CHIRA, L, CHIRAA. University of Agricultural Sciences and Veterinary Medicine, Mara§ti Biv., 59, Sector 1, Bucharest, Romania

Investigation performed confirmed occurrence in Romania of Venturia inaequalis forms resistant to benzimidazole fungicides, especially to metoben (methylthiophanate). In this work we present the aspects regarding the resistance of some monoisolates of the fungus with different origin (obtained from different regions of our country, different varieties of apple tree, different control treatments) and possibilities to control the resistant forms. The monoisolates obtained from eight areas were tested "in vitro" using methylthiophanate than these were classified in function of DL 50, in three groups: sensitive monoisolates (DL 5010); medium resistance (DL 50: 10 - 100) and resistant (DL 50100). With find out that the monoisolates variability, regarding the fungus resistance it is not depend only variation of the region in fact of the fungus control program, but this depend in the same time of variety. For instance, Starkrimson variety developed resistant forms to methylthiophanate in proportion of -18%, while as Golden Delicious - 5%. Among the fungicides which we tested "in vitro" to control the resistant forms, the best results were obtained with triazolic product - trifmine and the tests regarding the possibility of biological control, emphasised the efficacity of the fungus Chaetomium globosum.

193 Working group No.6 NOTES MECHANISM OF RESISTANCE TO INSECTICIDES IN LARGE PINE WEEVIL (HYLOBIUS ABIETIS L.) DOBROWOLSKI Forest Research Institute, ul. Bitwy Warszawskiej 1920 r. No 3,00-973 Warsaw, Poland

Two populations from different sites in Poland were tested to find potential mechanism of resistance to pyrethroid insecticides. The alfamethrin, deltamethrin, zetacypermethrin and lambdacyhalothrin were used in comparison to chlorpiryfos, carbosulfane, DDT and HCH. To reveal the oxidative and the hydrolitic mechanism of resistance to pyrethroids the oxidases inhibitor, piperonyl butoxide, and esterase inhibitor, DEF, were used. All tested insects were treated using topical application method. Results showed that mixed function oxidases are the main reason of decreased susceptibility of large pine weevil to pyrethroids. Mechanism of hydrolitic metabolism is not selected in tested populations.

1Q/1 Working group No. 6 NOTES THE FUMIGATION TREATMENT OF STONE FRUITS WITH ACETIC ACID TO PREVENT POSTHARVEST DECAY CHIRA, A. University of Agricultural Science and Veterinary Medicine, MaYas.ti Blv., 59, Sector 1, Bucharest, Romania

Acetic acid (AA) as a vapor at low concentration was effective in preventing fruits decay by postharvest stone fruit pathogens Monilinia laxa and Rhizopus stolonifer. Fruit samples of peach and apricot were inoculated through induced lesions with Monilinia laxa and Rhizopus stolonifer and divided into two lots : one exposed to acetic acid for 30 hours and the other not (control). The AA concentration were 3000, 6000, 9000 and 12000 ppm and the treatments were run at room temperature (22 - 26°C) by using a fumigation chambers (0,5 m3) which were constructed in our laboratory. After the treatment the fruits were stored at 20°C and 75% relative humidity. A fungi - inhibition effect, which increased over time as the amount of AA increased, was observed for both pathogen. A stasis in fungi growth that was observed at 9000 - 12000 ppm varied depending on species between 54 and 78 hour after inoculation.

195 Working group No. 6 NOTES MODIFIED - ATMOSPHERE PACKING OF APPLES AND STRAWBERRIES TREATED BY FUMIGATION WITH ACETIC ACID CHIRA, A., CHIRA, L, DELIAN, E. University of Agricultural Science and Veterinary Medicine, Mara§ti Blv., 59, Sector 1, Bucharest, Romania

A brief exposure of fruits to a low concentration of acetic acid (fumigation treatment) was combined with modified - atmosphere packing to reduce storage rots and increase shelf life of apples and strawberries. The fruits were inoculated with spores of Botrytis cinerea before fumigation with acetic acid, packing and stored in modified - atmosphere conditions. The treatment with acetic acid (7500 ppm) followed by modified - atmosphere packing for 60 days at 2°C reduced the percentage of rotted apples from 90% to 6%. Strawberries fumigated with acetic acid at 5000 ppm were lower rotted (3%) compared to 85% rotted for the control fruits stored for 10 days at 4°C. Working group No. 6 NOTES THE PREVENTING AND CONTROL OF FIRE BLIGHT CAUSED BY ERWINIA AMYLOVORA USING MECHANICAL AND BIOLOGICAL METHODS CHIRA, L University of Agricultural Sciences and Veterinary Medicine, Marasti Blv., 59, Sector 1, Bucharest, Romania

Fire blight caused by Erwinia amylovora (Burr.) Winslow et al. is the most dangerous bacterial disease of pome fruit trees especially for pear, apple and quince trees. The research were carried out to the southern area of Romania, and they showed the efficacity of some methods for preventing and control of fire blight pathogen, such as: 1. the biological methods: - the anthagonistic activity of Erwinia herbicola 122 s and 127 strains; 2. the mechanical methods: - pruning the attacked branches at 30 - 35 cm under the infected site, followed by their burning; - summer pruning should be done only during dry weather and when storms are not expected to occur within 24 hours; - desinfecting the pruned wounds with mineral oil and the tools with 10% natrium hypochlorit for 2-3 seconds after each tree which was pruned; - tools desinfection and lubrication with mineral oil at the end of each working days; 3. the chemical methods (with protective chemicals which are accepted in the biological agriculture): 2 - sprays during bloom period with CuSO4 - 0,5% and also during growing season after each rain above 10 mm/m . By combination of this methods with routine prophylactic treatment it is possible to reduce the spread of this dangerous disease.

197 Working group No. 6

RESULTS OF THE FAO/IAEA PROGRAM ON ,,IRRADIATION AS A QUARANTINE TREATMENT OF CZ9928649 MITES, NEMATODES AND INSECTS OTHER THAN FRUIT FLY" IGNATOWICZ, S. Warsaw Agricultural University, Department of Applied Entomology, Nowoursynowska 166, 02-787 Warszawa, Poland

The FAO/IAEA Program on ..Irradiation as a Quarantine Treatment of Mites, Nematodes and Insects other than Fruit Fly" has been implemented in 1992, and lasted up to the end of 1997. More than dozen scientists from 10 countries participated in the Program. The Coordination Research Program (CRP) put emphasis on the following aspects of research: (1) Determine criteria, e.g. inability to reproduce, for accepting irradiation as a quarantine treatment against quarantine pests; (2) Determine the effect of irradiation on the most resistant stage of these quarantine pests at the time of treatment; (3) Evaluate the quality of agricultural commodities irradiated at 2-3 times the dose(s) required to meet quarantine requirements; (4) Develop method(s) for identifying insects/other pests which were subjected to irradiation at a dose required for quarantine purposes. The followings are the most important achievements of the CRP: Generic dose for sterilization of both males and females of spider mites (Tetranychidae) was determined to be 320 Gy. With regards to insects other than fruit flies, it appears that a minimum dose of 300 Gy would cause either no adult emergence or sterility of most species of insects studied. Radiation doses required to cause complete mortality to various infective stages of plant parasitic nematodes appeared to be higher than 6 kGy. In most cases, the minimum dose required to prevent gall development and reproduction of these nematodes is over 2 kGy which is too high for most fresh plant materials to tolerate. Thus, irradiation should be considered as an alternative to methyl bromide fumigation to control nematodes in non-perishable materials. Although many fresh fruits and vegetables could tolerate radiation doses required for quarantine purposes, the response of various types of cut-flowers to irradiation varied widely. Some cut-flowers and ornamentals such as ferns, phoenix leaf, narcissus, tulips, carnation, red ginger, etc. appeared to be tolerant to irradiation up to at least 700 Gy, others such as chrysanthemum, rose, lily, anthurium, dendrobium, gerbera cannot tolerate radiation dose above 200 Gy. It was found that the tolerance of cut-flowers to irradiation is proportional to their initial quality. Melanization test appears to be a simple and reliable marker of irradiated larvae of various species of fruit flies subject to irradiation at egg or early larval stages. This method, however, does not appear to be reliable as a marker for irradiated insects of other species. Other methods such as histological changes in the midgut and electrophoretic separation of proteins and other macromolecules should be explored as markers of other species of insects and mites.

198 Working group No. 6 EFFECTS OF GAMMA RADIATION ON THE MELANIZATION PROCESS IN LARVAE OF STORED CZ9928650 PRODUCT MOTHS LUPA, D. Warsaw Agricultural University, Department of Applied Entomology, ul. Nowoursynowska 166, 02-787 Warszawa, Poland

Non-irradiated larvae of the Indian meal moth, Plodia interpunctella Hbn., the Mediterranean flour moth, Ephestia (Anagasta) kuehniella Zell., and the almond moth, Cadra cautella Wlk., showed a strong melanization after killing by freezing. However, there were some insects which showed a clear lack of melanization or meianized only partially, sometimes like those that have been irradiated. A part of larval body was dark black, while the rest body was of natural color or only slightly gray. Also, black and gray patches were observed in the larvae. After the irradiation treatment, the number of non-melanized larvae and larvae exhibiting a slight melanization usually increased. The degree of melanization in treated larvae was significantly different from untreated insects. Generally, it decreased with increasing dose and time elapsed after the treatment. The melanization test for detecting irradiated moth larvae may produce sometimes inconsistent results because (a) irradiation does not completely prevent melanization in mature moth larvae, and (b) the untreated larvae, killed by freezing and examined at room temperature, often show incomplete melanization. An ideal method for detection of irradiated insects should be: (1) specific for irradiation and not influenced by other processes, (2) accurate and reproducible, (3) have a detection limit below the minimum dose likely to be applied to agricultural commodity as a quarantine treatment, (4) applicable to a range of pests, (5) quick and easy to perform, and (6) capable of providing an estimate of irradiation dose. The melanization test to detect irradiated larvae of the stored product moths fulfills only some of these requirements. Therefore, the further detailed studies were performed to improve this test before it is recommended for quarantine inspection. Because the visual assessment of the effects of irradiation on melanization of the moth larvae is very subjective and difficult to perform, a trial to determine the activity of phenoloxidase enzyme in the control and irradiated larvae was performed using the 2-methyl-DOPA substrate in the spot on cellulose acetate transparency film. Results of the trials indicate that the lowest dose tested (0.1 kGy) had no effect on the production of enzyme(s) responsible for melanin formation, but the higher doses inhibited the process, and almost all larvae failed to produce the color with the substrate. This test may be used for detection of irradiated larvae of stored product moths. We found that the activity of the enzyme increases with the age of larvae, and the enzyme amount is highest in the 1_5 instars. The irradiation treatment inhibits activity of the enzyme, and this effect is the most profound in the young larvae (U- L3). At the ESNA Meeting, results of the studies on the effects of gamma irradiation on activity of phenoloxidase will be presented in detail.

199 Working group No. 6 NOTES ESTABLISHMENT POTENTIAL OF THE WESTERN CORN ROOTWORM (DIABROTICA VIRGIFERA VIRGIFERA LA CONTE) IN ROMANIA ROSCA, I. University of Agricultural Sciences and Veterinary Medicine, Bucharest, Romania

The Western corn rootworm (Diabrotica virgifera virgifera La Conte) (WCR), which was first reported from Yugoslavia in 1972, has spread rapidly into other European countries. This most serious insect pest on maize in the USA and Canada has great economic importance for European maize growing countries, too. The EPPO included this pest in its quarantine list as A2 pest, and it is assumed to become a quarantine pest for the EC in 1998. Besides France and Germany, Romania has the largest maize-growing area in the Europe. The pest was recorded for the first time in Romania in 1996, and established during 1997 in the southwestern part of the country. As for other countries, the established of D. virgifera virgifera in Romania would have an impact on maize yield and pest management. Therefore, the evaluation of its establishment potential as a part of pest risk assessment is necessary. The emergence of the different developmental stages of D. virgifera virgifera was calculated for calculated for 6 areas in Romania by means of temperature sums bases on the lower development threshold of 11°C for larvae and of 9°C for the other stages. In 1997, a countrywide system was worked out for records of presence of the pest in Romania. This system is based on the Integrated Forecasting System. The application of pheromone traps proved a head spreading of WCR. However, no damage was yet registered in Romania, even at places with high number of trapped beetles. The WCR ability to adapt itself to the new conditions and absence of its natural enemies makes especially important an international study on the ecology and ethology of this potentially most dangerous pest for corn in the Centra! and Eastern Europe. The actual IPM strategies which have been adopted in Romania for attempt to control the pest will be presented. Working group Nr. 5 NOTES: AMMONIUM RECOVERY FROM CLAY FIXED POOL - A STABLE 15N ISOTOPE STUDY Stopar, D., Mahne, I., Megusar, F. Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia

A sample of arable soil (0 -28 cm, brown post carbonic sandy loam) with pronounced capacity to fix cations (31 % clay particles) was studied for additional fixation and defixation od added ammonium. To follow the external nitrogen in the course of changes during incubation (fixation, defixation, mineralisation, immobilisation, nitrification, denitrification) external supplement was enriched with stable 15N isotope. Air dried soil samples were ground to pass a 2mm sieve and stored at 4°C. The 250 ml wide neck flasks were utilized for fixation/defixation study in an incubation experiment with soil/silica sand mixture in the ratio 3:1, amended with 100 and 200 iug/g soil ammonium sulphate nitrogen enriched with 10% atom excess 15N. Soil/sand mixtures were rewetted to50% WHC. In a destructive sampling procedures 2N KCI extraction was performed and steam distillation procedure utilized to measure ammonium, nitrite and nitrate nitrogen after 1, 3, 5, 7, 10, 14, 21 and 28 days incubation from triplicate subsamples. The 30 ml aliquots of three replicates for reduced and oxidized mineral nitrogen forms were utilized for isotope ratio analyses. A diffusion procedure was applied for transfer the labelled ammonium to Watman glass filter. Excess water was removed in a dessicator. Mass spectrometer VG 602 was utilized for 14N/15N isotope ratio analysis. Absolute amounts of labelled 15N wee made by calculation from 14N/15N isotope ratios and from the data of total amounts of 2N KCI exchangeable ammonium, nitrite and nitrate. 50 - 60% of ammonium added to the soil sample at rewetting was removed from exchangeable pool of mineral nitrogen within 10 minutes. The fraction removed did not reappear in exchangeable pool during 28 days incubation under constant moisture, pH, temperature and aeration. All abstracts included in this book are based on the authors' manuscripts. No major amendments have been made.

Printed: Edicni stfedisko MZLU Brno 180 copies August 1998

XXVIIIth Annual ESNA Meeting Mendel University of Agriculture and Forestry, Brno, 1998

ISBN 80-7157-316-7

'S8071 E73166