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ESSENTIAL AND DETRIMENTAL TRACE ELEMENTS ENTERING THE FOOD CHAIN VIA PLANTS

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WELCOME

Dear Friends and Colleagues,

This is our fourth annual scientific workshop focussing on essential and detrimental elements entering the food chain via plants. We are glad to observe that more than 80 scientists from nearly 30 countries within and outside Europe will present their recent results on various topic of the workshop.

To strengthen the outcome of our workshop we have invited the leading experts from different corners of the world as keynote speakers in the fields of plant nutrition, plant biology and genomics, biotechnology, food processing, nutritional physiology and human nutrition.

We are grateful for support from our sponsors for the conduction of this workshop.

On behalf of the Organizing Committee,

Best wishes to everyone for an interesting and inspiring workshop

LOCAL ORGANIZING COMMITTEE

Bal Ram Singh, UMB

Peder Lombnæs, BIOFORSK

Anja Nieuwenhuis, UMB

Liv Korslund, UMB

Anne Elisabeth Munkeby, UMB

Signe Dahl, UMB

ORGANIZING SECRETARY

LIV KORSLUND, UMB - E-MAIL: [email protected]

4TH ANNUAL CONFERENCE - (ALL WGS) - 09TH -13TH JUNE 2013

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COST ACTION FA 0905

Mineral Improved CROP PRODUCTION FOR HEALTHY FOOD AND FEED

PROF. DR. BAL RAM SINGH, MC CHAIR NORWEGIAN UNIVERSITY OF LIFE SCIENCES

([email protected])

PROF. DR. PETER SCHRÖDER, MC VICE CHAIR

HELMHOLTZ-ZENTRUM MÜNCHEN, GERMAN

RESEARCH CENTER FOR ENVIRONMENTAL HEALTH

([email protected])

COST FA 0905 AIMS AT:

• identify bottlenecks limiting the content of bioavailable minerals (Fe, Zn, Mg, Se) in the consumable crop part

• provide solutions for 3-fold increase in bioavailable food/feed mineral content

• assess ways to limit the entry of Cd and As into the food chain

• disseminate this knowledge to stakeholders and the public. European dimension and added values COST

COST FA 0905 WILL STRIVE TO:

• maximise European synergy in research cooperation

• provide innovative ideas to improve the mineral status of food and feed

• find methods to exploit genetic variability of food and fodder crops

• enhance scientific know how to improve minerals in food under modern processing

• create training and exchange programs, for students, postdoctoral fellows and young researchers, especially women

• identify key institutes and personnel for efficient assessment of food and feed mineral quality across Europe


PROGRAMME


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SUNDAY JUNE 09TH

16:00 - 18:00 Arrival and Check-in the hotel or Pentagon

MONDAY JUNE 10TH

08:15 - 09:00 REGISTRATION – BIOTECHNOLOGY BUILDING -

09:00 - 10:00 BIOTECHNOLOGY BUILDING – AUDITORIUM ARABIDOPSIS

OPENING CEREMONY

Hans Fredrik Hoen, Rector, Norwegian University of Life Sciences (UMB)

Nils Vagstad, Director of Research, Norwegian Institute for Agricultural and Environmental Research (BIOFORSK)

Øystein Johnsen, Head of the Department, Plant and Environmental Sciences, UMB

Chair: Bal Ram Singh Cost Action FA0905

10:00 - 17:30 WORKING GROUP 1 ( WG1) Chair: Ismail Cakmak

Rapporteur: Michel Mench

10:00 - 10:30 KEYNOTE: Availability of micronutrients and undesirable trace elements in the soil-plant-microbe continuum

Zed Rengel

University of Western Australia, Perth, Australia

10:30 - 10:50 Transformation of As-species between cultivation medium and plants

Tommy Landberg and M. Greger

Department of Ecology, Environment and Plant Sciences, Stockholm University, Sweden


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10:50 - 11:10 Decreasing of cadmium in wheat grain depends on the form of the applied zinc

Zdenko Lončarić, Maja Manojlović, Brigita Popović, Krunoslav Karalić, Ranko Čabilovski, Vladimir Ivazić, Espen Govasmark, Bal Ram Singh

Faculty of Agriculture in Osijek, Kralja Petra Svačića 1d, HR–31000 Osijek, Croatia

11:10 - 11:40 Coffee Break

11:40 - 12:00 Agriculture activities and their input on keeping the entry of cadmium into the food chain

Marta Pogrzeba, Aleksandra Sas-Nowosielska, Ewa Gucwa-Przepióra, Eugeniusz Małkowski and Jacek Krzyżak

Institute for Ecology of Industrial Areas Katowice, Poland

12:00 - 12:20 Selenium cycle in field ecosystem Mervi M. Seppänen, Nashmin Ebrahimia, Anthony Owusu-Sekyere and Helinä Hartikainen

Department of Agricultural Sciences, University of Helsinki, Finland

12:20 - 14:00 Lunch

14.00 - 16:20 WORKING GROUP 1 (WG1) Chair: Rainer Schulin

Rapporteur: Elena Comina

14:00 - 14:30 KEYNOTE: Understanding zinc bioavailability to plants

Ellis Hoffland and Andreas Duffner

Wageningen University, Dept. Soil Quality, The Netherlands

14:30 - 14:50 Effects of diffusion limitations of zinc fluxes into wheat root , DGT, and PLM devices in presence of organic ligands

Anja Gramlich, Susan Tandy, Emmanual Fossard, Jost Eikenberg and Rainer Schulin

Institute of terrestrial Ecosystems, ETH, Zurich, Swtizerland


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14:50 - 15:10 Effect of different cropping systems on soil zinc content and availability

Thilo Dürr-Auster, R. Schulin, P. Mäder, J. Mayer, M. Mazzoncini and E. Frossard

Institute of Agricultural Sciences, Group of Plant Nutrition, ETH Zurich, Switzerland

15:10 - 15:30 Effect of intercropping systems and fertilizers on maize and soybean grain composition

Vesna Dragicevic, Oljaca Snezana, Dolijanovic Zeljko, Stojiljkovic Milovan, Spasojevic Igor and Nisavic Marija

Maize Research Institute,Belgrade, Serbia

15:30 - 15:50 Improved uptake of mineral nutrients by intercropped forage maize and faba beans

Eva Stoltz

The Rural Economy and Agricultural Society, Örebro, Sweden


16:20 - 18:20 POSTER SESSION 1 Chair: Maria Greger Rapporteur: Peder Lombnæs

TUESDAY JUNE 11TH

09:00 - 15:00 WORKING GROUP 2 (WG2) Chair: Danuta Maria Antosiewicz Rapporteur: Rolf Herzig

09:00 - 09:30 KEYNOTE: Cellular iron pools: localization and mobilization by specialized transporters

Catherine Curie Hannetz Roschzttardtz, Fanchon Divol, Daniel Couch, Mathilde Séguéla, Louis Grillet, Geneviève Conéjéro and Stéphane Mari

Laboratoire de Biochimie et Physiologie Moléculaire Végétale, CNRS-INRA, Montpellier, France


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09:30-09:50 Analysis of a zinc-sensor function in the Arabidopsis bZIP19/23 transcription factors

Ana G.L. Assunção, Mark G.M. Aarts, Pai Pedas, Daniel P Persson, Søren Husted and Jan Kofod Schjoerring

University of Porto, Vila do Conde, Portugal

09:50 - 10:10 Natural variation of Arabidopsis thaliana hapmap population reveals candidate genes controlling zinc deficiency tolerance

Ana Carolina Atala Lombelo, Charles Neris Moreira, David E Salt and Mark G.M. Aarts

Wageningen University, Wageningen, The Netherlands

10:10 - 10:30 Capturing unprecedented natural diversity in plant metal accumulation: a comprehensive survey of european Arabidopsis halleri populations

Stephan Clemens Stephan Höreth, Ricardo Stein and Ute Krämer

Department of Plant Physiology, University of Bayreuth, Germany

10:30 - 10:50 Translocation of copper from roots to shoots is mediated by OsHMA5 in rice

Fenglin Deng, Ikuko Yonamine, Naoki Yamaji and Jian Feng Ma

Institute of Plant Sciences and Resources, Okayama University, Japan


11:20 - 15:10 WORKING GROUP 2 (WG2) Chair: Nathalie Verbruggen Rapporteur: Sylvain Legay

11:20 - 11:50 KEYNOTE: Transporters involved in Cd accumulation in rice

Jian Feng Ma

Institute of Plant Science and Resources, Okayama University, Japan

11:50 - 12:10 Reaction of root cells to cadmium Alexander Lux

Conenius University, Bratislava, Slovakia

12:10 - 13:40 Lunch


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13:40 - 14:00 Controlling heavy metal accumulation in plants

Antonella Furini Andrea Nesler and Giovanni DalCorso,

Università di Verona, Italy

14:00 - 14:20 Modification of Fe-Zn-Cd cross-homeostasis through transformation contributes to changes in the tolerance to metals

Katarzyna Tracz, Holger Schmidt, Stephan Clemens and Danuta Maria Antosiewicz

University of Warsaw, Faculty of Biology, Warsaw, Poland

14:20 - 14:40 Silicon alleviates cadmium-induced oxidative stress in maize

Marek Vaculík, Miroslava Luxová and Alexander Lux

University of Bratislava, Slovakia


14:40 - 16.00 Core Group meeting

15:10 - 17:10 POSTER SESSION 2 Chair: Peter Schröder Rapporteur: Maria J Poblaciones

17:30 Bus from Biotech Building, UMB to the ferry to Oscarsborg

19:00 - Reception and Conference dinner – Banquet at Oscarsborg Ferry returns Drøbak at 23:00 - Bus to Ski via Ås


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WEDNESDAY JUNE 12TH

09:00 - 15:00 WORKING GROUP 3 (WG3) Chair: Richard Hurrell Rapporteur: Gerd Elisabeth Vegarud

09:00 - 09:30 KEYNOTE: Biotechnological approaches to the iron biofortification of rice

Navreet K. Bhullar Swiss Federal Institute of Technology (ETH), Zurich, Switzerland

09:30 - 10:00 KEYNOTE:Iron bioavailability in humans from plant ferritin

Bo Lönnerdal

Department of Nutrition, University of California, Davis, USA

10:00 - 10:20 Iron bioavailability from low phytic acid common beans (Phaseolus vulgaris L.) is increased in young women

Nicolai Petry, Ines Egli, Bruno Campion, Erik Nielsen and Richard Hurrell

Institute of Food, Nutrition and Health, ETH Zurich, Switzerland

10:20 - 10:40 Comparison of the speciation, digestibility, and bioavailability of iron in fortified and unfortified unleavened breads of different cultivars and extraction rates

Tristan Eagling, Anna Wawer, Fangjie Zhao, Andy Neal, Steve McGrath, Peter R Shewry and Susan Fairweather-Tait

Rothamsted Research Hertfordshire UK

10:40 - 11:00 Selenium accumulation and speciation in biofortified hard wheat (Triticum durum L.) under Mediterranean conditions: from grain to cooked pasta

María J. Poblaciones, Sara M. Rodrigo, Oscar Santamaría, Santiago Lledó, Yi Chen and Steve P. McGrath

University of Extremadura, Badajoz, Spain



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11:30 - 11:50 Genotype specific accumulation of selenium in various Allium sativum L. cloneas and association with sulphur content

Jaroslava Ovesná, Horníčková Jana, Kučera Ladislav, Stavělíková Helena, Poucghová Vladuimíra and Velíšek Jan

Crop Research Institute, Prague, Czech Republic

11:50 - 12:10 Anti-regurgitation (ar) infant formulas. expression of mineral transporters and storage proteins in caco-2 cells after in-vitro gastrointestinal digestion

Carlos Alberto González-Bermúdez, Miranda-Miranda L, Legay S, Corvisy A, Klein S, Frontela-Saseta C, Martínez-Graciá C and Evers D

Department of Food Science and Nutrition, University of Murcia. Murcia, Spain

12:10 - 12:30 Lead enters plant cell by endocytosis Aneta Basińska, Krzesłowska M., Mellerowicz E.J., Napieralska A., Rabęda I., Woźny A. and Adam Mickiewicz

University, Poznań, Poland

12:30 - 14:00 Lunch

14:00 - 16:00 WORKING GROUP 3 (WG3) Chair: Susan Fairweather-Tait Rapporteur: Sara Rodrigo

14:00 - 14:30 KEYNOTE:Algorithms for predicting iron absorption from plant foods

Manju B. Reddy and Seth M. Armah

Department of Food Science and Human Nutrition, Iowa State University, Ames, USA

14:30 - 15:00 KEYNOTE: Addressing the challenges of intrinsic labeling in iron and zinc stable isotope absorption studies with biofortified plant foods

Janet R. Hunt

International Atomic Energy Agency, Vienna, Austria

15:00 - 15:20 Iron-biofortified pearl millet provides additional bioavailable iron compared to regular-iron millet but to a lesser extent than post-harvest iron-fortified millet

Colin Cercamondi, Ines Egli, Evariste Mitchikpe, Erick Boy, Felicien Tossou, Christophe Zeder, Joseph Hounhouigan and Richard Hurrell

Laboratory of Human Nutrition, ETH Zurich, Switzerland


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15:20 - 15:40 Impact of polyphenols on iron bioavailability from potato after simulated gastro-intestinal digestion

Danièle Evers, Lisa Miranda, Aude Corvisy, Hannah Deußer and Sylvain Legay

Centre de Recherche Public-Gabriel Lippmann, Luxembourg

15:40 - 16.00 General discussion and concluding remarks Poster awards

Bal Ram Singh and Peter Schröder

16:00 - 18:00 EXCURSION TO NOFIMA, Ås

18:00 - SOCIAL EVENTS - Dinner – Tapas in the Biotech building

THURSDAY JUNE 13TH DEPARTURE


PRESENTATION – KEYNOTE SPEAKERS


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WG1 SOIL-PLANT INTERACTIONS AND PHYSIOLOGY

Professor ZED RENGEL, University of Western Australia, Perth, Australia BIOAVAILABILITY AND ROOT UPTAKE OF BENEFICIAL AND DETRIMENTAL TRACE ELEMENTS IN

FOOD CROPS

Zed Rengel is Winthrop Professor in the School of Earth and Environment at the University of Western Australia. His research interests focus on nutrient uptake and ion toxicity in the soil-plant-water-microbe continuum, covering macro (phosphorus, potassium and magnesium) and micronutrients (zinc, copper and manganese) as well as toxic ions (aluminium, arsenic, cadmium, zinc and sodium). The approaches and techniques used range from fluorescence imaging microscopy and ion-specific microelectrodes to enzyme assays and radioisotope tracking. He also works on chemistry and biology of the rhizosphere soil and computer modelling of root growth and function.

Associate. Professor ELLIS HOFFLAND, Wageningen University, The Netherlands UNDERSTANDING ZINC BIOAVAILABILITY TO PLANTS

She is personal professor at the Department of Soil Quality of Wageningen, the Netherlands. Her field of expertise is soil fertility and soil-plant interactions and she is particularly interested in how plants improve soil fertility through rhizosphere effects. She tries to integrate biology, chemistry and physics in her research.

WG2 BIOLOGICAL FEATURES IN THE RELATIONSHIP BETWEEN PLANTS AND MINERALS

Dr. CATHERINE CURIE, CNRS Montpellier, France CELLULAR IRON POOLS IN PLANTS: LOCALIZATION AND MOBILIZATION BY SPECIALIZED

TRANSPORTERS

Catherine Curie obtained her PhD in 1992 in the University of Toulouse, France, during which she studied transcriptional regulation in Arabidopsis. From 1993 to 1996, she joined the lab of Sheila McCormick in Berkeley, California, to study pollen-specific regulatory pathways in tomato. In 1997, she went to the Biochemistry and Molecular Physiology lab in Montpellier, France, to work on iron homeostasis in plants in the group of Jean-François Briat. Since 2008, she has been leading an independent research group focusing on the mechanisms of transport of iron and manganese and their regulation. Right now, she is on sabbatical at the James Cook University of Townsville, Australia, to study iron/Mn in microalgae.


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Professor JIAN FENG MA, Okayama University, Kurashiki, Japan TRANSPORTERS INVOLVED IN CD ACCUMULATION IN RICE

Professor at Institute of Plant Science and Resources, Okayama University, Japan.

Major interests: Plant transporter identification of minerals including essential, beneficial and toxic minerals.

Published more than 160 papers in international journals including Nature, PNAS, Plant Cell, Plant Journal. etc.

Received JSPS Prizes and Japan Academy Medals in 2006, Awards of Japanese Society of Soil Science and Plant Nutrition in 2007, and Kihara Award in 2012.

Associate Editor of Functional Plant Biology and Section Editor of Plant and Soil.

WG3 FOOD PROCESSING AND HUMAN NUTRITION

Dr. NAVREET BHULLAR, ETH Zurich, Switzerland BIOTECHNOLOGICAL APPROACHES TO THE IRON BIOFORTIFICATION OF RICE

Dr. Bhullar is a research group leader at Swiss Federal Institute of Technology (ETH) in Zurich, Switzerland since 2010. She obtained her PhD in 2008 from the Institute of Plant Biology at University of Zurich and then undertook postdoctoral research at the same institute. Her research interests include crop biofortification, genetic diversity and mining for novel genes in the crop genepools. She has published several research articles, review articles, book chapters and recently, co-edited a book entitled ‘Agricultural sustainability’. Dr. Bhullar received distinctive appreciations to her research work, including the Gatersleben Research Award in 2010.

Professor BO LÖNNERDAHL, Distinguished Professor of Nutrition & Internal Medicine, Department of Nutrition & Program in International and Community Nutrition University of California, Davis, USA IRON BIOAVAILABILITY IN HUMANS FROM PLANT FERRITIN

He received his master’s and doctorate degrees in biochemistry from University of Uppsala in Sweden, and has been professor at UCD since 1980. Dr. Lönnerdal’s research has focused on bioactive components in breast milk, the effect of breast milk on the recipient infant and mechanisms underlying the protection against infection. His research includes the micronutrients iron and zinc; how they are secreted into milk and how they are utilized by the infant. He is also studying the bioavailability of iron and zinc from plant sources.

He has recieved many honours and awards for example: Borden Award, American Institute of Nutrition, International Award for Modern Nutrition 2000 - Honorary Doctorate (Honoris causa) in Medicine, University of Uppsala, Sweden, and 2004 - Gabriel Bertrand Prize and Medal for research on trace


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element metabolism

Dr. Lönnerdal has published more than 500 scientific articles, book chapters and books and is currently a member of the American Society of Nutrition (ASN), and the European Society of Pediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN).

Professor MANJU B. READDY, Iowa State University, Ames, USA ALGORITHMS FOR IRON PREDICTING IRON ABSORPTION FROM PLANT FOODS

Manju Reddy is a Professor at Iowa State University, Ames, Iowa, in the department of Food Science and Human Nutrition. She graduated from Texas A&M University, College Station, Texas with a PhD in Nutrition and worked at Kansa University Medical Center, Kansas City, Kansas, prior to joining Iowa State University. Her research focuses on strategies to improve iron bioavailability and iron status using in vitro models and humans. She also studies the iron excess related diseases, especially Parkinson’s and cardiovascular diseases and how nutritional factors prevent those diseases.

Dr. JANET HUNT, IAEA, Vienna, Austria ADDRESSING THE CHALLENGES OF INTRINSIC LABELING IN IRON AND ZINC STABLE ISOTOPE

ABSORPTION STUDIES WITH BIOFORTIFIED PLANT FOODS

Janet R. Hunt, Ph.D., R.D. is a nutrition specialist working for the International Atomic Energy Agency, an agency of the United Nations, in Vienna, Austria. She formerly worked as a research leader of the Mineral Utilization and Bioavailability Project at the U.S. Department of Agriculture/Agricultural Research Service (USDA/ARS) Human Nutrition Research Center in Grand Forks, ND. Dr. Hunt received her Ph.D. in nutrition from the University of Minnesota. She has been active in the American Society for Nutrition and the Academy of Nutrition and Dietetics. She also served on USA Institute of Medicine, National Academy of Sciences committees on Interpretation and Use of Dietary Reference Intakes and on Mineral Requirements for Military Personnel. Her research interest is human iron and zinc requirements as influenced by dietary bioavailability, and she has published over 100 papers on these topics


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ABSTRACTS – KEYNOTE SPEAKERS


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K1

AVAILABILITY OF MICRONUTRIENTS AND UNDESIRABLE TRACE ELEMENTS IN THE SOIL-PLANT-MICROBE CONTINUUM

Zed Rengel

The University of Western Australia, Faculty of Science, Perth, Australia

Micronutrient availability in the rhizosphere is controlled by soil and plant properties, and interactions of roots with microorganisms and the surrounding soil. Plants exude a variety of organic compounds (carboxylate anions, phenolics, carbohydrates, amino acids, enzymes, etc.) and inorganic ions (protons, phosphate, etc.) to change chemistry and biology of the rhizosphere. Acidification of the rhizosphere soil increases mobilisation of micronutrients (eg. for Zn, 100-fold increase in solubility for each unit of pH decrease) as well as potentially toxic trace elements, such as Cd and Al.

Decreased availability of micronutrients and undesirable trace elements is due to complexation with humic acids, lignin and other high-molecular-weight compounds. In contrast, increased availability may result from solubilisation and mobilisation by short-chain organic acids, amino acids and other low-molecular-weight organic compounds.

For diffusion-supplied micronutrients, the uptake rate is governed by the soil nutrient supply. Fertilisation with micronutrients (more so in case of Zn than Fe) can be effective in increasing the concentration of micronutrients at the soil-root interface. In addition, micronutrient-efficient crops and genotypes can increase an available nutrient fraction and hence increase micronutrient uptake.

Our understanding of the physiological processes governing exudation and the soil-plant-microbe interactions in the rhizosphere is currently inadequate, especialy in terms of spatial and temporal variability in root exudation as well as the fate and effectiveness of organic and inorganic compounds in increasing availability of soil micronutrients and undesirable trace elements. The interactions between microorganism and plants at the soil-root interface are particularly important as well obscure.

Keywords: cadmium, iron, microorganisms, rhizosphere, zinc


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K2

UNDERSTANDING ZINC BIOAVAILABILITY TO PLANTS

Ellis Hoffland and Andreas Duffner

Wageningen University, Dept. Soil Quality, The Netherlands

Objectives Most soils contain sufficient amounts of zinc (Zn) to support crop growth for many years, but most of this Zn is not bioavailable. So understanding soil Zn bioavailability to plants is relevant to mineral-improved crop production for human health. I will give an overview of our work on processes in the soil-plant continuum that are relevant to Zn bioavailability.

Methods We performed field and pot experiments with wheat and other crop species on low Zn soils collected around the world. Root box experiments allowing for rhizosphere sampling were done to understand the effect of soil characteristics and root exudates on Zn bioavailability. Chemical speciation modeling was combined with plant bioassays.

Results Biogeochemical modelling showed that dissolved organic anions and pH are important determinants of Zn bioavailability. We showed that chemical conditions in the rhizosphere, which may deviate from the bulk soil, can impact Zn bioavailability, depending on the soil. Adsorption of Zn on the root surface was pH dependent and nonlinear and a useful proxy for bioavailable Zn.

Conclusions Rhizosphere conditions are relevant to understand mobilization of Zn from low Zn soils. Zinc speciation models develloped for Zn-contaminated soils appeard to be valid for low Zn soils, too, and a usefull tool to improve prediciton of Zn bioavailability to crops.

Keywords: bioavailability, root exudates, rhizosphere, soil, zinc


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K3

CELLULAR IRON POOLS: LOCALIZATION AND MOBILIZATION BY SPECIALIZED TRANSPORTERS

Hannetz Roschzttardtz, Fanchon Divol, Daniel Couch, Mathilde Séguéla, Louis Grillet, Geneviève Conéjéro, Stéphane Mari, Catherine Curie

Laboratoire de Biochimie et Physiologie Moléculaire Végétale, CNRS-INRA, Campus SupAgro, Montpellier, FRANCE

To acquire the essential heavy metal iron (Fe), plants have to overcome its low bioavailability in most soils by activating a now well-described high affinity machinery whose main actor in dicots is the Fe transporter IRT1. The next challenge to our community is to understand, once in the root, how Fe is distributed to plant organs and within cellular organelles. Parcels of knowledge are starting to emerge thanks to the recent development of elemental imaging techniques. In our lab, we have implemented a powerful histochemical staining method and used it both as a tool to study Fe movement within the plant, and to describe the Fe pools at the subcellular level within each plant organ. Doing so, and together with X-ray based elemental analyses, we identified a new unexpected abundant iron pool in the nucleolus of plant cells. By combining reverse genetics, Fe imaging and biochemistry, we have established the role of the efflux of citrate, a Fe ligand, in the distribution of iron to pollen grains, as well as characterized the role of two metal transporters, YSL4 and YSL6, in preventing Fe toxicity by mediating its efflux from the chloroplasts.


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K4

TRANSPOTERS INVOLVED IN CD ACCUMULATION IN RICE

Jian Feng Ma

Institute of Plant Science and Resources, Okayama University, Japan

Cadmium (Cd) in foods is a major health concern worldwide. Rice, an important staple food for nearly a half of the world’s population, is a major source of Cd intake. Therefore, it is necessary to limit Cd into the food chain from soil to reduce potential health risks to humans. We have identified three transporters involved in Cd accumulation in rice. OsNramp5 is a major transporter for Cd uptake in the roots. It is localized in the distal side of both exodermal and endodermal cells. OsHMA3 is mainly expressed in the tonoplast of root cells. It is responsible for sequestration of Cd into the vacuoles in the roots. On the other hand, OsHMA2 is a plasma membrane-localized transporter and localized in the pericycle of the root, which is responsible for translocation of Cd from the roots to the shoots. Knockout of OsNramp5 and OsHMA2 resulted in significant reduction of Cd accumulation in the grain, but also caused decreased yield. By contrast, overexpression of OsHMA3 selectively decreased accumulation of Cd in the grains without affecting the growth, while mutation of this gene resulted in increased Cd accumulation. Our results provide insight into the strategies for reducing Cd accumulation in rice grain.

Keyword: Accumulation, cadmium, rice, root, transporter


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K5

BIOTECHNOLOGICAL APPROACHES TO THE IRON BIOFORTIFICATION OF RICE

Navreet K. Bhullar

Institute of Agricultural Sciences, Department of Biology, Swiss Federal Institute of Technology (ETH), Zurich, Switzerland

Food security and healthy nutrition is of critical global importance. Around two billion people suffer from iron deficiency, particularly affecting women and children in developing countries. Monotonous diets based on staple cereals are frequently associated with such human micronutrient malnutrition. Cereal grains constituting most important basis of human food, are in fact a poor source of iron. Biofortification of cereal grains, such as rice, has therefore emerged as a promising strategy. However, the variability for most micronutrients is very low in the rice germplasm and this does not leave traditional breeding alone to be a valid option for rice biofortification in many circumstances. Complementing the breeding efforts, gene technology offers perspectives for efficiently improving iron content in rice grain. The biotechnological strategies used to date in order to improve rice for iron content, and the opportunities for future research will be presented.

Keywords: biofortification, iron, metal homeostasis, cereals


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K6

IRON BIOAVAILABILITY IN HUMANS FROM PLANT FERRITIN

Bo Lönnerdal

Department of Nutrition, University of California, Davis, USA

Meeting iron requirements is difficult for populations dependent upon plant foods, and iron deficiency is common. Beans contain ferritin in low concentrations, but it is possible to enhance this content by plant breeding or by inserting the gene for ferritin into plants, e.g., soybeans. Because each ferritin molecule can bind to thousands of iron atoms, this may be a sustainable means to increase the iron contents of plants. Before such efforts are launched, it is important to determine whether iron in ferritin is bioavailable. This was assessed in vitro by using human intestinal (Caco-2) cells and by using radiolabeled ferritin and whole-body counting in human subjects. Dietary factors affecting iron absorption had limited effect on iron uptake from intact ferritin by Caco-2 cells, suggesting that ferritin-bound iron is absorbed via a mechanism different from that of nonheme iron. In an in vitro digestion system, ferritin was shown to be relatively resistant to proteolytic enzymes. Binding of ferritin to Caco-2 cells was saturable, and kinetics for binding were characteristic of a receptor-mediated process. In human subjects, iron from soybean ferritin given in a meal was as well absorbed as iron from ferrous sulfate. In conclusion, iron is well absorbed from ferritin and may represent a means of biofortification of staple foods.

Keywords: ferritin, iron, iron absorption, iron bioavailability, plant ferritin


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K7

ALGORITHMS FOR PREDICTING IRON ABSORPTION FROM PLANT FOODS

Manju B. Reddy and Seth M. Armah

Department of Food Science and Human Nutrition, Iowa State University, Ames, IA 50014, USA

Various algorithms have been developed taking into consideration the combined effect of dietary factors on nonheme iron absorption using single meal data. However, accuracy of predicting iron absorption from complete diets using those algorithms raised some concerns. We developed a new algorithm based on complete diets using previous data where nonheme iron absorption was measured for 3 different dietary periods when each subject (n=53) consumed diets containing either high, low or typical amounts of the dietary factors (meat, vitamin C, calcium and tea) for 5-days. Our algorithm was validated with measured absorption reported in the literature. Compared to 16% in the single meal algorithm, only 3% of the variation in absorption was explained by dietary factors in our algorithm. Ferritin and interpersonal variation explained a large proportion of the differences. Significant correlation was found when our algorithm was validated with single meal (R2=0.57; P<0.001) and complete diet (R2=0.84; P<0.0001) data. Using our algorithm with the NHANES data, total iron absorption from the US diet was found to be 15% which is lower than the current 18%. In conclusion, the algorithm based on complete diets will be useful for predicting nonheme iron absorption from the diets of different populations.

Keywords: absorption, algorithms, bioavailability, nonheme iron


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K8

ADDRESSING THE CHALLENGES OF INTRINSIC LABELING IN IRON AND ZINC STABLE ISOTOPE ABSORPTION STUDIES WITH BIOFORTIFIED PLANT FOODS

Janet R. Hunt, Ph.D., R.D.

International Atomic Energy Agency, Vienna, Austria

Biofortification of foods with Fe and Zn requires enhanced mineral content plus demonstrated bioavailability (absorption and utilization) for humans. Bioavailability can be sensitively determined from retention of isotopes that label the respective mineral in the foods. Intrinsic labeling involves incorporation of the isotope into the food during growth under controlled (e.g., hydroponic) conditions emulating field conditions. Intrinsic labeling of Fe and Zn has often validated extrinsic labeling, the addition of isotope in the final stages of food preparation. One challenge of isotopic labeling is matching breeding or growing conditions that may result in Fe or Zn compounds that don’t fully interchange with other ingested nonheme Fe or Zn (heme Fe from animal foods is a primary example, but other examples may include ferritin iron). Another labeling challenge is to use tracer quantities that do not disturb absorptive efficiency or the ratios of mineral to components that affect bioavailability, such as phytate (which may also be affected by biofortification). Extrinsic labeling is economical and can be useful when added stable isotopes do not alter the amount or relative ratios of mineral to known enhancers and inhibitors. Substantial changes in the organic form of biofortified minerals may require tests using intrinsic labeling.

Keywords: extrinsic, intrinsic, iron, isotope, zinc


ABSTRACTS - ORAL PRESENTATION


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A1

TRANSFORMATION OF AS-SPECIES BETWEEN CULTIVATION MEDIUM AND PLANTS

Tommy Landberg1 and M. Greger1,2

1)Department of Ecology, Environment and Plant Sciences, Lilla Frescati, Stockholm University, SE-10691 Stockholm, Sweden email: [email protected]

2)Faculty of Applied Ecology and Agricultural Sciences, Hedmark University College, Blæstad, NO-2418 Elverum, Norway

Arsenic exists in several organic and inorganic forms. Environmental conditions such as redox potential and bacterial activity influence the As-speciation. The various As-species are differently available and toxic and their properties in cultivation medium and plants are necessary to understand to reduce the As risk in food. Therefore, the dynamics in As speciation during uptake in lettuce from cultivation medium was investigated.

Various cultivars of lettuce (Lactuca sativa L.) were cultivated either in soil or hydroponically. Treatment concentrations of As in soil were 2 - 140 mg kg-1 and 0 – 100 µM in hydroponics. Arsenic-species used were arsenite, arsenate, monomethylarsonic acid (MMA) and dimethylarsenic acid (DMA). Plants were treated 3 hours - 7 weeks. Total As and As-species were analysed in plants and cultivation medium.

The results show that arsenic is transformed from arsenate, MMA and DMA to arsenite (which is the most toxic species). It is not due to selective uptake because treatment with either arsenate, MMA or DMA results in high arsenite in the plant. Arsenic translocated to the shoot have further higher propoportion of arsenite. The reduction from arsenate to arsenite in plants is known, while not the transformation from MMA and DMA to arsenite.

Keywords: Arsenic, food risk, plant, soil, speciation


26

A2

DECREASING OF CADMIUM IN WHEAT GRAIN DEPENDS ON THE FORM OF THE APPLIED ZINC

Zdenko Lončarić1, Maja Manojlović2, Brigita Popović1, Krunoslav Karalić1, Ranko Čabilovski2, Vladimir Ivazić1, Espen Govasmark3 and Bal Ram Singh3

1)Faculty of Agriculture in Osijek, Kralja Petra Svačića 1d, HR–31000 Osijek, Croatia 2)Faculty of Agriculture Novi Sad, Trg Dositeja Obradovića 8, 21000 Novi Sad, Serbia 3)Norwegian University of Life Sciences (UMB), Ås, Norway

The field experiment with three wheat cultivars was conducted on fertile calcareous soil with low zinc availability. The aim was to determine the effect of the applied zinc form on zinc, iron and cadmium in the grain. Therefore three following treatments were conducted: 1. control, 2. Zn sulfate foliar, 3. Zn EDTA foliar. Fertilization treatments didn’t affected iron concentration in wheat organs, but significant differences were determined among cultivars (Divana>Srpanjka>Simonida) and organs (leaves>=glumes>spike axis>=grain>straw). The highest zinc concentration as average of all treatments was determined for cultivar Divana, and significantly lowers for Simonida and Srpanjka. As it was expected, the lowest zinc concentration in grain was in control treatment (24,4 mg kg-1), higher after Zn-EDTA (33,4) and highest after Zn-sulfate application (48,9). All cadmium concentrations were very low, and additionally affected by zinc application. Highest average cadmium was in straw, than leaves and spike axis and lowest in glumes and grains. Zn-sulfate significantly decreased cadmium concentration only in spike axis (85 vs. 128 µg kg-1), and not significantly in grain (53 vs. 62) and straw (91 vs. 110). The Zn-EDTA decreased cadmium in grain 66% compared to Zn-sulfate and 72% compared to control. Cadmium was also decreased in spike axis (22-48%), but significantly increased in leaves (20-37%) and straw (20-57%).

Keywords: cadmium, foliar application, iron, zinc-EDTA, zinc-sulfate


27

A3

AGRICULTURE ACTIVITIES AND THEIR IMPUT ON KEEPING THE ENTRY OF CADMIUM INTO THE FOOD CHAIN

Marta Pogrzeba1, Aleksandra Sas-Nowosielski1, Ewa Gucwa-Przepióra2, Eugeniusz Małkowski3 and Jacek Krzyżak1

1)Department of Environmental Biotechnology, Phytoremediation Team, Institute for Ecology of Industrial Areas (IETU), 6 Kossutha Street, 40-833 Katowice, Poland, mail:[email protected]

2)Department of Plant Systematic, Faculty of Biology and Environmental Protection, University of Silesia, Jagiellońska 28 St, 40-032 Katowice, Poland

3)Department of Plant Physiology, Faculty of Biology and Environmental Protection, University of Silesia, Jagiellońska 28 St, 40-032 Katowice, Poland

Barley in Poland is cultivated mostly for beer production, however about 4% is used for direct human consumption. Like other plants, when grown on contaminated soils it tends to accumulate heavy metals like cadmium. Cadmium accumulation and distribution within plants is strongly affected by the cultivar and the soil type. Recent data show that it can be also impacted by plant care activities.

Approximately 95% of plant species form mycorrhizal associations. It was found that AM hyphe are able to accumulate 10–20 times higher rates of Cd relative to the plant roots. Therefore AM fungi may be responsible for limiting Cd translocation to shoots via soil bioaugmentation. Cadmium uses zinc transport pathways to enter the plant, thus zinc application may be helpful in diminishing of Cd accumulation. Foliar application of zinc is increasingly used to elevate micro nutrient deficiencies of this element.

Barley roots colonization with AM fungi for different cultivars was investigated on clean soil and soil contaminated with cadmium and other heavy metals. AM attendance and Cd accumulation were revealed under the application of plant care and physiological treatments. Moreover reduction in cadmium uptake by barley after Zn foliar application was investigated. Physical and chemical soil properties were analyzed using ISO methods. The pot experiment with two barley cultivars was performed in natural conditions. Total cadmium concentration in contaminated soil was about 12.39 mg kg-1, while total Zn concentration, about 2174.5 mg kg-1. Cadmium bioavailable fractions were about 4.6% of total concentration, while for Zn about 2.2%.

Correlation between AM root colonization and Cd concentration in grains was assessed. The parameters of mycorrhizal development were evaluated microscopically based on frequency of mycorrhization of barley root fragments (F%), intensity of root cortex colonization (M%) and arbuscule abundance in the root system (A%). Cd concentration in grains was studied after application of two different Mono-Zn concentrations during the stages of ear formation and flowering.

The significant negative correlation was found between frequency of mycorrhization of root fragments (F%) and Cd concentration in grains. F% for particular barley cultivars decreased with increasing concentration of bioavailable Cd in soil, thus suggesting that accumulation of Cd in barley grains depends both, on plant physiological properties of cultivars and AM fungi. Application of Mono-Zn on leaves surface of some barley cultivars resulted in decrease of cadmium concentration in grain and straw. The results were related to the type of barley cultivar and kind of soil (clean, Cd-contaminated).

In conclusion our results highlight the necessity of reconsidering the usage of fungicydes, as this practice may actually open the gate for heavy metals to the human food chain and point to the possible utility of Zn foliar application.

Keywords: arbuscular mycorrhiza (AM), Zn foliar application, barley grains, Cd concentration, health risk


28

A4

SELENIUM CYCLE IN FIELD ECOSYSTEM

Mervi M. Seppänen1, Nashmin Ebrahimi1, Anthony Owusu-Sekyere1 and Helinä Hartikainen2

1)Department of Agricultural Sciences, University of Helsinki 2)Department of Food and Environmental Sciences, University of Helsinki

Selenium (Se), an essential micronutrient for humans and animals, is circulated to food chain via plants. In Finland, an annual Se fertilization is required in order to acquire adequate Se content in locally produced food and feed. Only 5 to 10 % of the applied Se is translocated to seeds and removed from fields whereas the fate of most Se in field ecosystem is unknown. We have conducted 2 to 3-year field and supplementary greenhouse experiments to study the Se cycle in field ecosystems where wheat, oil seed rape or forage grass-clover mixtures are cultivated. Se uptake and translocation to various plant parts were monitored during growing period and the interaction between Se and N fertilization was assessed. Up to 60 % of applied Se was taken up by plant but majority, 30 %, was left in plant depris, mainly in stems. Over 10 % is lost maybe due to volatilization or, as indicated by greenhouse experiments, metabolized by soil microbes. The annual variation between years was substantial; in a wet growing season 2012 Se uptake was significantly lower than in other studied years. There also seemed to be an interaction between Se uptake and plant N-status since foliar N fertilization at booting stage increased Se uptake.

Keywords: clover, forage grass, oil seed rape, translocation, uptake, wheat


29

A5

EFFECTS OF DIFFUSION LIMITATIONS ON ZINC FLUXES INTO WHEAT ROOTS, DGT AND PLM DEVICES IN PRESENCE OF ORGANIC LIGANDS

Anja Gramlich1, Susan Tandy1, Emmanuel Frossard2, Jost Eikenberg3 and Rainer Schulin1

1)Institute of Terrestrial Ecosystems, ETH Zurich, Switzerland 2)Institute for Plant, Animal and Agroecosystems Sciences, ETH Zurich, Switzerland 3)Paul Scherrer Institute (PSI), Villigen, Switzerland

Organic ligands increase metal mobility in soils. The extent to which they enhance plant metal uptake depends besides other factors on diffusion limitations in the soil solution. In this study, we investigated the influence of diffusion layer thickness (δ) on zinc (Zn) uptake by wheat in the presence of EDTA, citrate and histidine from hydroponic solutions with equal free Zn concentrations by measuring 65Zn uptake from stirred, non-stirred and agar-containing nutrient solutions. Analogous experiments were performed using permeation liquid membranes (PLM) and ‘diffusive gradients in thin films’ (DGT) probes instead of plants in order to identify the potential role of diffusion limitation in root Zn uptake. In treatments with EDTA or ligand-free Zn solution an increase in δ reduced the fluxes of Zn into roots, PLM and DGT probes to a similar extent, suggesting that they were limited by diffusion. In presence of citrate the flux of Zn into the roots was similar as in presence of EDTA for the same total and free Zn concentrations under stirred conditions, but did not decrease when δ was increased by not stirring the solution and by adding agar. This indicates that diffusion and complex dissociation were both rate limiting. This explanation is supported by the fact that similar δ effects were found in PLM influxes where uptake of negatively charged complexes can be excluded. Zn influx into roots was much higher in the histidine treatments than in all other ligand treatments and also not affected by increasing δ. PLM measurements suggested that dissociation kinetics for Zn-histidine were fast and could explain the higher uptake rate by the plants partially. However, as Zn root uptake in presence of histidine was very high, it is probable that the neutral or positive Zn-histidine complexes contributed directly to metal uptake.

Keywords: diffusion limitations, Zn bioavailability, Zn-citrate, Zn-histidine, wheat


30

A6

EFFECT OF DIFFERENT CROPPING SYSTEMS ON SOIL ZINC CONTENT AND AVAILABILITY

Thilo Dürr-Auster1, R. Schulin2, P. Mäder3, J. Mayer4, M. Mazzoncini5 and E. Frossard1

1)Institute of Agricultural Sciences, Group of Plant Nutrition, ETH Zurich, Eschikon 33, Lindau (ZH), 8315, Switzerland ([email protected])

2)Institute of Terrestrial Ecosystems, Group of Soil Protection, ETH Zurich, Universitätstrasse 16, Zuerich, 8092, Switzerland ([email protected])

3)Research Institute of Organic Agriculture FIBL, Group of Soil Science, Ackerstrasse, Frick, 5070, Switzerland ([email protected])

4)Agroscope Reckenholz-Tänikon Research Station ART, Reckenholzstrasse 191, Zürich, 8046, Switzerland ([email protected])

5)Department of Agronomy, University of Pisa, Via S. Michele degli Scalzi 2, Pisa, 56124, Italy ([email protected])

Zn deficiency in crops can be linked to two major causes: i) low Zn content of the soil and ii) low plant-available Zn in the soil. The present project is investigating the impact of fertilization strategies, with emphasis on organic fertilizers, on the Zn transfer from the soil to the plant. On the one hand organic fertilizer can be considered as an important Zn source, and on the other they can affect the exchangeable Zn pools of the soil.

In the present part of the project, the effect of different fertilization strategies on the soil Zn status are being compared by analysing soils from long-term field trials for their total-Zn (XRF) and available-Zn (DTPA-extractable zinc). Three sites have been chosen: i) the “DOK” which was set up in 1978 (Therwil near Basel, Switzerland), the “ZOFE” which was established in 1949 (Zurich-Reckenholz, Switzerland) and the “MASCOT” field trial set up in 2001 (Pisa, Italy). The applied organic fertilizers are specific to each field trial and are covering the most common organic fertilizers used in agriculture: manure, compost, sewage sludge, dry manure pellets, dry bovine blood but also mineral fertilizer, all added in agronomically relevant quantities. Archive-samples will allow us to analyse the evolution of the total Zn-status in the treatments of the two oldest long-term field trials.

Preliminary results suggest that there is a significant increase of plant available and total Zn in the organic treatments compared to the control with mineral or no fertilization. Furthermore, these results indicate that changes of the zinc status only happen in the upper layer of the soil. Further analysis is currently in progress. Data evaluation will consider the long term Zn input-output budgets and soil properties of the different treatments. These results will be available at the conference.

Keywords: cropping system, long-term field trials organic fertilizer, zinc


31

A7

EFFECT OF INTERCROPPING SYSTEMS AND FERTILIZERS ON MAIZE AND SOYBEAN GRAIN COMPOSITION

Dragicevic Vesna1, Oljaca Snezana2, Dolijanovic Zeljko2, Stojiljkovic Milovan3, Spasojevic Igor1 and Nisavic Marija2

1)Maize Research Institute “Zemun Polje”, Slobodana Bajica 1, 11185 Zemun Polje-Belgrade, Serbia 2)University of Belgrade, Faculty of Agriculture, Nemanjina 6, 11080 Zemun-Belgrade, Serbia 3)Vinca Institute of Nuclear Sciences, PO Box 522, 11001 Belgrade, Serbia

Ecological cropping systems include combination of different crops at the same field and application of organic fertilizers, what could enable better utilization of space and nutrients, with minimal inputs. Trial was conducted during 2011 and 2012, with varieties of red grain maize and black grain soybean, grown as: single crop (SC), alternating rows of both crops (AR) and alternating strips (3 rows of each species - AS). Fertilization regimes included incorporation of: urea, Ofert (organic fertilizer) and Uniker (microbiological fertilizer). After harvest, grain yield, mass of 1000 grains, content of Mg, Fe, Zn, P, including ratio between phytic and inorganic P (Pphy/Pi), as well as antioxidants: phenolics and glutathione were determined in grains.

There were minor variations in grain yield between treatments, while Uniker and Ofert increased mass of 1000 grains. In SC cropping in maize, Uniker increased Zn, while Ofert increased Fe content. Irrespective to several times higher contents of Mg, Fe, Zn and P in soybean grain, compared to maize, Ofert increased Mg and Zn content in soybean in AS cropping. Moreover, Ofert increased accumulation of antioxidants in both crops, mainly in AR treatment, while the both organic fertilizers decreased Pphy/Pi ratio, underlining their positive effect on nutritive quality of produced crops.

Keywords: antioxidants, cropping systems, fertilization, grain, mineral composition


32

A8

IMPROVED UPTAKE OF MINERAL NUTRIENTS BY INTERCROPPED FORAGE MAIZE AND FABA BEANS

Eva Stoltz

The Rural Economy and Agricultural Society, Hushållningssällskapet/HS Konsult AB, Box 271, S-701 45 Örebro, Sweden email: [email protected]

Intercropping may have advantages such as higher yield, reduced presence of plant diseases and increased uptake of mineral nutrients. The aim was to investigate the effect of intercropping on uptake of micronutrients and sulphur in organically produced faba bean (Vicia faba L.) and maize (Zea mays L.). Three field experiments were performed with faba bean and maize cultivated in pure stands or intercropped. Disease severity index (DSI) of leaf spots in faba beans was determined. At harvest, shoot samples were taken and analysed for mineral nutrients. Intercropping significantly increased the concentrations of Cu (from 4.1 to 4.6 mg/kg) and Zn (from 17.2 to 20.8 mg/kg) in maize and B (from 15.3 to 17.8 mg/kg) and S (from 0.14 to 0.15 % of DM) in faba bean compared with sole crops. Leaf spot DSI was significantly lower in intercropped faba beans (mean 25) than in sole faba bean (mean 45). A significant negative relationship between Cu concentration in faba bean and DSI of leaf spots was found, and the shoot Cu concentration tended to increase by intercropping. Intercropped maize and faba bean improved forage quality by increased concentration of mineral nutrients and reduced DSI of leaf spots in faba beans.

Keywords: faba bean, intercrop, leaf spots, maize, mineral nutrients


33

A9

ANALYSIS OF A ZINC-SENSOR FUNCTION IN THE ARABIDOPSIS BZIP19/23 TRANSCRIPTION FACTORS

Ana G.L. Assunção1, Mark G.M. Aarts2, Pai Pedas3, Daniel P. Persson3, Søren Husted3 and Jan K. Schjoerring3

1)Plant Evolution Group, Research Center in Biodiversity and Genetic Resources, University of Porto, R. Padre Armando Quintas, 4485-661 Vairão, Vila do Conde, Portugal

2)Laboratory of Genetics, Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands

3)Section for Plant and Soil Sciences, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark

As a plant micronutrient, zinc is of major interest in agronomy and for human and cattle nutrition. Zinc deficient soils are widespread all over the world and the risk of inadequate diet and zinc malnutrition is estimated to affect one third of the global human population, i.e. around 2 billion people. Developing countries, where people depend on cereal-rich diets for sustenance, are the most affected. Improvement of crop efficiency in zinc deficient environments, and bio-fortification to alleviate human nutrition are plant-based solutions as source of micronutrients.

An incomplete understanding of zinc homeostasis mechanisms in plants is however a limitation. Recently the first transcription factors regulating zinc homeostasis in Arabidopsis were identified. The AtbZIP19 and AtbZIP23 were shown to be essential for the adaptation to zinc deficiency. Transcript profiling revealed only a small set of genes, to be induced in wild-type plants in response to zinc deficiency, but not in the bzip19bzip23 double mutant line, which shows a zinc deficiency hypersensitive phenotype1. Anchored to this knowledge, it is now possible to elucidate, upstream to these transcription factors, the molecular mechanisms that allow plants to sense zinc deficiency and adapt to this stress. We hypothesised that the characteristic His/Cys-rich domains in the bZIP19 and bZIP23 transcription factors2 are involved in the direct sensing of zinc.

In order to test this hypothesis, we are analysing metal-binding properties of bZIP19/bZIP23 by SEC-ICP-MS (SizeExclusion Chromatography/Inductively Coupled Plasma Mass Spectrometry). Additionally, amino acid substitutions on bZIP19/23, targeting the His/Cys-motif2, are being prepared for similar metal-binding analysis. The results obtained so far are discussed.

Keywords: Micronutrients, Plant nutrition, Transcription factors, Zinc deficiency, SEC-ICP-MS References:

[1] Assunção, A. G. L. et al. (2010) Arabidopsis thaliana transcription factors bZIP19 and bZIP23 regulate the adaptation to zinc deficiency. Proceedings of the National Academy of Sciences of the United States of America 107, 10296–301.

[2] Jakoby, M. et al. (2002) bZIP transcription factors in Arabidopsis. Trends in plant science 7, 106–11.


34

A10

NATURAL VARIATION OF ARABIDOPSIS THALIANA HAPMAP POPULATION REVEALS CANDIDATE GENES CONTROLLING ZINC DEFICIENCY TOLERANCE

Ana Carolina Atala Lombelo1, Charles Neris Moreir a1, David E. Salt2 and Mark G.M. Aarts1

1)Laboratory of Genetics, Wageningen University, Wageningen, The Netherlands 2)Institute of Biological and Environmental Sciences, University of Aberdeen, Aberdeen, UK

The low availability of Zn in calcareous and high pH soils result on severe growth and yield reduction for plants. Zn deficiency affects 30% of the world soils, including many agricultural lands in Australia, South-east Asia, Central and South America, Africa, India, Spain, USA, among others (Alloway 2004). In nature, some plants are able to overcome this nutrition limited gift by uptaking and utilizing Zn more efficiently. In this study we used natural variation to investigate which genes are involved in Zn deficiency tolerance or sensitiveness of Arabidopsis thaliana. The effect of Zn deficiency in the phenotype of different A. thaliana accessions was clear with plants showing chlorosis, stunted growth and necrotic spots. Several candidate genes were identified by Genome Wide Association (GWAS) mapping. Traits analysed were, shoot dry biomass, root total length, shoot chlorosis, Zn deficiency tolerance index (%), and difference between treatments (Zn Control – Zn Deficiency). Shoot and root complete elemental profile is being analysed in collaboration with the group of Professor David E. Salt at the University of Aberdeen with an STSM funding from the Cost Action office. Manhattan plots showed many peaks of small and moderate significance suggesting that the traits studied are polygenic (Figure 1). An average of 5 QTLs (peaks) per trait distributed over the 5 chromosomes was observed. Candidate genes were selected based on the LOD score, gene function and co-expression with other genes related to Zn homeostasis.

Keywords: biofortification, genome wide association mapping, nutrient homeostasis


35

A11

CAPTURING UNPRECEDENTED NATURAL DIVERSITY IN PLANT METAL ACCUMULATION: A COMPREHENSIVE SURVEY OF EUROPEAN ARABIDOPSIS

HALLERI POPULATIONS

Stephan Höreth1, Ricardo Stein2, Ute Krämer2 and Stephan Clemens1

1Department of Plant Physiology, University of Bayreuth, Germany 2Department of Plant Physiology, University of Bochum, Germany

As part of a project aiming at the genome-wide molecular analysis of variation in A. halleri metal hyperaccumulation we conducted a large survey covering the natural range of A. halleri in Europe. We have analyzed in total over 2000 field-collected leaf and rhizosphere soil samples of A. halleri individuals from 195 populations. Herbivory was scored to test the elemental defense hypothesis. About 800 individuals were transferred to the lab for phenotyping under controlled, uniform conditions. The field data for 20 elements reveal associations between soil factors and elemental profiles as well as between metal acquisition pathways. We found, for instance, an extreme extent of variation and regional patterns in Cd hyperaccumulation. The initial phenotyping under controlled conditions confirmed these results, suggesting recent evolution of the Cd hyperaccumulation trait. Moreover, not only variation in Cd and Zn accumulation was observed, but also substantial variation for other micronutrients such as Fe and Mn to a degree that has rarely been documented to date. Thus, through the use of next-generation sequencing technologies, this collection of 800 genotypes will enable the molecular dissection of many aspects of plant microelement nutrition in addition to the evolution of Zn/Cd hyperaccumulation.

Keywords: Cd accumulation, field studies, genome-wide analysis, natural variation, Zn accumulation


36

A12

TRANSLOCATION OF COPPER FROM ROOTS TO SHOOTS IS MEDIATED BY OSHMA5 IN RICE

Fenglin Deng, Ikuko Yonamine, Naoki Yamaji and Jian Feng Ma

Institute of Plant Sciences and Resources, Okayama University, Japan

Objectives: Copper (Cu) is an essential element for both plants and animals. Daily copper intake for a normal healthy adult is between 0.97-3.0 mg/day. Both deficiency and excess of Cu will cause healthy disorders. Rice is a staple food and provides most part of Cu intake. The objective of this study is to identify a transporter involved in root-shoot translocation.

Methods: The expression pattern of OsHMA5 was determined by quantitative RT-PCR. Immunostaining using antibody against OsHMA5 was performed. Phenotypic analysis was carried out by using the knockout lines.

Results: OsHMA5 was mainly expressed in the rice roots and its expression was up-regulated by excess Cu, but not by the deficiency of Cu, Zn, Fe and Mn. OsHMA5 was localized to the plasma membrane and expressed in the pericycle cells of the roots. Knockout of OsHMA5 resulted in increased Cu accumulation in the roots, and decreased Cu concentration in the shoots and xylem sap especially at higher external Cu concentrations. The Cu concentration in the grain was lower in the knockout lines compared with the wild-type rice.

Conclusions: All results show that HMA5 localized at the root pericycle cell is responsible for Cu translocation from the roots to shoots in rice, especially at high Cu concentrations.

Keywords: copper translocation, heavy metal P-type ATPase 5 (HMA5), rice (Oryza sativa), xylem loading


37

A13

REACTION OF ROOT CELLS TO CADMIUM

Alexander Lux

Department of Plant Physiology, Faculty of Natural Sciences, Comenius University in Bratislava, Mlynská dolina B2, SK 84215 Bratislava, Slovakia

The effect of cadmium on root cells and tissues is reviewed in this contribution. Root is usually the first organ in contact with Cd and it regulates the translocation of Cd ions into aboveground plant parts. “The firewall” system of root defense in reaction to various abiotic stresses is a complex system, with participation of both symplasmic and apoplasmic components. Reactions in the cell walls and intercellular spaces (apoplasm) especially those in the root peripheral parts (epidermis, cortex) represent in many cases the key stage in defense. Protection of vascular cylinder, including pericycle, is crucial for root principal functions – transport of water and solutes and to defend pericycle for formation of lateral roots and thus maintaining root absorption..

Conclusions: Apoplasmic suberin, lignin and other cell wall components including antioxidant defense system occurring in peripheral root parts form barrier preventing uncontrolled uptake of toxic ions.

The support by the Slovak Research and Development Agency under contracts APVV-0140-10 and APVV SK-FR-0020-11 and by grant VEGA 1/0817/12 is acknowledged.

Keywords: apoplasm, cadmium, root


38

A14

CONTROLLING HEAVY METAL ACCUMULATION IN PLANTS

Andrea Nesler, Giovanni DalCorso, Antonella Furini

Università di Verona, Dipartimento di Biotecnologie, Strada Le Grazie 15, 37134 Verona, Italy

The objective of this study is to modulate the accumulation of heavy metals, particularly of cadmium (Cd), in tobacco and food plants. To this goal, A. thaliana, N. tabacum and L. esculentum plants transformed with the membrane transporter CzcCBA, a member of the Czc family (cobalt/zinc/cadmium) efflux transporter, isolated from a Pseudomonas putida strain collected in a heavy metal polluted site, are under investigation. The organization of the czc-like systems is generally conserved in all organisms studied, and confers resistance to Cd, Zn and Co. The three open reading frames, czcA, B and C, were independently cloned under the control of the CaMV 35S promoter and the three constructs were used for plant transformation. At the same time the chimeric genes czcA::GFP, czcB:: DsRED and czcC::YFP, cloned downstream of the CaMV 35S were used for tobacco transformation to detect subcellular protein localization. Transgenic plants over-expressing 35S::czcA, 35S::czcB and 35S::czcC were then crossed in order to test the effect of the tripartite protein complex in plant heavy metal accumulation. Results so far achieved indicate that the accumulation of Cd is lower in transgenic plants compared to wild-type, suggesting that the bacterial membrane transporter CzcCBA can be functionally expressed in plant cells, and that it may be useful for the development of crop plants that are safe from heavy metal contamination.

Keyworks: cadmium, CzcCBA, heavy metals, transgenic plant


39

A15

MODIFICATION OF FE-ZN-CD CROSS-HOMEOSTASIS THROUGH TRANSFORMATION CONTRIBUTES TO CHANGES IN THE TOLERANCE TO

METALS

Katarzyna Tracz1, Holger Schmidt2, Stephan Clemens2 and Danuta Maria Antosiewicz1

1)University of Warsaw, Faculty of Biology, Institute of Experimental Plant Biology and Biotechnology, Miecznikowa 1, 02-096 Warsaw, Poland

2)University of Bayreuth, Department of Plant Physiology, Universitätsstrasse 30,

95449 Bayreuth, Germany e-mails: [email protected]; [email protected]

The aim of the research was to evaluate the usefulness of transforming tomato with AhNAS2 under its own promoter to enhance a plant performance under low Fe and modify Cd metabolism.

We have used tomato plants expressing AhNAS2p::AhNAS2 from A.halleri encoding nicotianamine synthase (NAS), which is involved in NA biosynthesis. Transgenic and wild-type plants were exposed to control and Fe-deficient conditions without and with 0,1µM Cd. Plant tolerance to growth conditions, Fe, Zn, Cd accumulation, expression of AhNAS2, LeNAS, LeIRT1, LeFRO1, level of NA and metal speciaction in plant parts were evaluated.

Until 10-12 days under low Fe transformants and WT plants were chlorotic. However, on 13th day in AhNAS2-expressing plants Fe-deficiency symptoms have receded and their leaves turned to green. Higer tolerance of transformed plants to low Fe was accompanied by higher Fe level in leaves and expression of LeIRT1 and LeFRO1 genes. Expresion of AhNAS2 and LeNAS in transformants under Fe-deficiency was higher in roots than in leaves, which is consistent with NA concentrations. Transgenic plants also exhibited a higher tolerance to cadmium accompanied by higher level of Fe and lower level of Cd in leaves.

Results indicate that AhNAS2p::AhNAS2 expression in tomato is developmentally and organ-specifically regulated, and suggest the role of NA not only in Fe-deficiency tolerance but also in reducing Cd root-to-shoot translocation.

Keywords: cadmium, iron deficiency, nicotianamine, tomato Supported by: FP6 EU PHIME, BW no. 501/86 - 102354 intramural grant; STSM no. 0905-11474 from COST Action 0905.


40

A16

SILICON ALLEVIATES CADMIUM-INDUCED OXIDATIVE STRESS IN MAIZE

Marek Vaculík1, Miroslava Luxová2 and Alexander Lux1

1)Department of Plant Physiology, Faculty of Natural Sciences, Comenius University in Bratislava, Mlynska dolina B2, SK 842 15 Bratislava, Slovakia

2)Institute of Botany, Slovak Academy of Sciences, Dúbravská cesta 19, SK 845 23 Bratislava, Slovakia

Accumulation of cadmium (Cd) – a dangerous environmental pollutant in agricultural plants represents a serious threat for food production. The aim of the research community is to find out the mechanisms how to decrease the uptake of this metal into crops and make food safer. Silicon (Si) was shown to decrease the harmful effect of various abiotic and biotic stresses, including heavy metals. In our study the effect of Si application on hydroponically grown maize (Zea mays L.) plants exposed to Cd was investigated. The results indicate that silicon actively improves the growth of young maize plants exposed to cadmium and influences the uptake and subcellular distribution of Cd as well as other elements, which might be related with the root and shoot anatomical changes. Moreover, the alleviation of Cd toxicity can be partially attributed to Si mediated decrease in production and accumulation of reactive oxygen species, as well as to the activation of defense mechanisms against oxidative stress in young maize plants.

Keywords: Cadmium, maize, oxidative stress, root anatomy, silicon

Supported by the Slovak Research and Development Agency under contract No. APVV-0140-10, APVV SK-FR-0020-11, and the Slovak Grant Agency VEGA No. 1/0817/12, and is a part of COST FA 0905 Action.


41

A17

IRON BIOAVAILABILITY FROM LOW PHYTIC ACID COMMON BEANS (PHASEOLUS VULGARIS L.) IS INCREASED IN YOUNG WOMEN

Nicolai Petry1, Ines Egli1, Bruno Campion2, Erik Nielsen3 and Richard Hurrell1

1)Laboratory of Human Nutrition, Institute of Food, Nutrition and Health, ETH Zurich, Switzerland 2)Consiglio per la Ricerca e la Sperimentazione in Agricoltura- Unità di Ricerca per l'Orticoltura,

Montanaso Lombardo, LO, Italia 3)Dipartimento di Biologia e Biotecnologie, Università di Pavia, Pavia, Italy

Background: Iron bioavailability from common beans is negatively influenced by phytic acid (PA) and polyphenols (PP). Newly developed low phytic acid (lpa) beans with 90% less PA and variable PP might improve iron bioavailability.

Objective: Evaluate the influence of lpa beans with high or low PP on iron bioavailability.

Design: In an iron absorption study using a paired crossover design, 20 women consumed four different beans (lpa beans high and low in PP; wild type (wt) parents with normal PA, high and low in PP).

Results: Iron absorption and total amount of iron absorbed was significantly higher from the lpa bean high in PP compared to the wt parents high in PP (P< 0.001) and low in PP (P< 0.001) and from the lpa line low in PP compared to the wt parent low in PP (P< 0.001). We did not observe significant differences between the lpa beans high or low in PP and between the wt parents high or low in PP.

Conclusions: A 90% reduction in PA leads to a significant improvement of iron bioavailability from beans, independent of the PP concentration. The lpa mutation could be a key tool for increasing bioavailable iron from beans.


42

A18

COMPARISON OF THE SPECIATION, DIGESTIBILITY, AND BIOAVAILABILITY OF IRON IN FORTIFIED AND UNFORTIFIED UNLEAVENED BREADS OF DIFFERENT

CULTIVARS AND EXTRACTION RATES

Tristan Eagling1, Anna Wawer2, Fangjie Zhao1, Andy Neal1, Steve McGrath1, Peter R Shewry1 and Susan Fairweather-Tait2

1)Rothamsted Research Hertfordshire AL5 2JQ, UK 2)Norwich Medical School, University of East Anglia, Norwich, Norfolk, NR4 7TJ, UK

We have shown previously that Fe is present in different chemical forms in the endosperm and outer layers of wheat grain. Therefore it is possible that after gastrointestinal digestion the Fe in white and wholemeal flour products will be chemically different.

To explore this we have used size exclusion chromatography coupled with inductively coupled plasma-mass spectrometry (SEC-ICP) to determine speciation changes in two wheat cultivars during in vitro digestion, in both unfortified and fortified (ferrous sulphate 30 mg Fe/kg flour) unleavened white and wholemeal bread samples. We also determined the bioavailability of Fe in the digested samples using an in vitro Caco-2 cell model system.

The speciation of Fe differed between white and wholemeal bread at all stages of digestion. Overall, the bioavailability of Fe assessed using a Caco-2 cell model system appeared to be much higher in white bread than in wholemeal bread. Addition of ferrous sulphate was therefore a more effective fortificant in white bread compared to wholemeal bread. The quantity of total or soluble Fe did not appear to be related to predicted bioavailability from the Caco-2 cell model, indicating that the effects of Fe speciation and the presence of absorption inhibitors are of greater importance than total or soluble iron when determining Fe.

Keywords: Bioavailability, Iron, Fortification, Wheat bioavailability


43

A19

SELENIUM ACCUMULATION AND SPECIATION IN BIOFORTIFIED HARD WHEAT (TRITICUM DURUM L.) UNDER MEDITERRANEAN CONDITIONS: FROM GRAIN TO

COOKED PASTA

María J. Poblaciones1, Sara M. Rodrigo1, Oscar Santamaría1, Santiago Lledó1, Yi Chen2 and Steve P. McGrath2

1)Department of Agronomy and Forest Environment Engineering, University of Extremadura, Avda. Adolfo Suárez s/n, C.P. 06007, Badajoz, Spain

2)Sustainable Soils and Grassland Systems, Rothamsted Research, Hertfordshire AL5 2JQ UK email: [email protected]

Selenium (Se) is an essential trace element for humans being feeding the main route of Se intake. Durum wheat is widely used for human consumption in the Mediterranean area, principally as pasta. To improve its nutritional value, two foliar Se fertilizers (sodium selenate and sodium selenite) at four rates (0-10-20-40 g ha-1) were tested in the 2010-2011 and 2011-2012 growing seasons in a field experiment conducted under semiarid Mediterranean conditions. Sodium selenate was much more effectively taken by plants than sodium selenite, and there was a strong and linear relationship between total Se and Se-Met content and Se rate for both. For each gram of Se fertilizer, applied as sodium selenate or sodium selenite, the increases of total Se in grain were 138 and 38 and 35 and 11 μg Se kg-1 dry weight in 2010-2011 and 2011-2012, respectively. The effect of milling and further processing of semolina into pasta cooking produced a lost of Se of about 40% and the main Se form was selenomethionine (≈ 90 %). Thus, cooked pasta, and therefore hard wheat, would be a very good candidate to be included in Se biofortification programs.

Keywords: Agronomic biofortification, Cereal, HPLC-ICP-MS, Selenate, Selenite


44

A20

GENOTYPE SPECIFIC ACCUMULATION OF SELENIUM IN VARIOUS ALLIUM SATIVUM L. CLONEAS AND ASSOCIATION WITH SULPHUR CONTENT

Ovesná Jaroslava1, Horníčková Jana2, Kučera Ladislav1, Stavělíková Helena1, Poucghová Vladuimíra1 and Velíšek Jan2

1)Crop Research Institute, Drnovská 507, 161 06 Prague 6 – Ruzyně, Czech Republic 2)Institute of Chemical Technology, Technická 3, 160 00 Prague 6, Czech Republic

Garlic, Allium sativum (L.), has been cultivated for thousands years and has been used for both culinary and medicinal purposes. Already in ancient Egypt garlic health benefits were described based on previous experiences. Garlic is known to act against viruses, bacteria. Garlic is also claimed to help prevent heart disease, high cholesterol, high blood pressure and cancer. Garlic is rich in organically bound sulphur namely in form of cysteinsulphoxides.

We have analysed set of 136 garlic genetic resources by AFLP (amplified fragment lenght polymorphism – PCR based DNA analysis) and HPLC (high performance liquid chromatography). Association between cysteinsulphoxides content and genetic basis of genotypes was found (Ovesná et al., Hort.Sci, 2011).

As selenium can play important role in human metabolism, especially in its organic form, we investigated whether selenium can be accumulated by garlic genotypes. We have studied five genotypes of genetically different type (bolting, non-bolting, semibolters). All genotypes were able to accumulate selenium in both anorganic and organic form, however repeatedly in different extend. Association with garlic type was found. We concluded that garlic can be used as a source of organically bound selenium and that investigation of ability of different genotypes to accumulate selenium should continue.

Keywords: garlic, selenium, suplhur, cysteisulpoxides


45

A21

ANTI-REGURGITATION (AR) INFANT FORMULAS. EXPRESSION OF MINERAL TRANSPORTERS AND STORAGE PROTEINS IN CACO-2 CELLS AFTER IN-VITRO

GASTROINTESTINAL DIGESTION

Carlos Alberto González-Bermúdez, Miranda-Miranda L2, Legay S2, Corvisy A2, Klein S2, Frontela-Saseta C1, Martínez-Graciá C1, Evers D2

Department of Food Science and Nutrition, University of Murcia. Murcia, Spain 1Department of Food Science and Nutrition. Faculty of Veterinary Sciences. University of Murcia.

30100 Murcia,Spain 2Department Environmentand Agro-biotechnologies (EVA). Centre de Recherche Public Gabriel

Lippmann. L 4422, Belvaux,Luxembourg

Objective: To study the effect of three thickeners (Locust Bean Gum, LBG; Modified Corn and Rice Starch, MCS/MRS), used as ingredients in anti-regurgitation infant formula, on different transporters and storage proteins at intestinal level: CalD9K (calcium), DMT-1, FTH1 and FTL (iron) and Zip4, ZnT1 and MT-1(zinc).

Methods: Eleven different combinations of LBG, MCS and MRS were added up to a total concentration of 3.5 g per 100 g of a commercial infant formula (reference). After in-vitro digestion, soluble fractions were added to a Caco-2 cell monolayer. After 6h-exposure, relative expression of genes of interest and ferritin content were measured using qPCR and ELISA respectively. As controls, Serum Free Media and 5 M CaCl2, 200 µM FeSO4 and 50 µM ZnSO4 were used.

Results: Only significant differences for ZnT1 and MT-1 were founded in samples with (1.75 g LBG + 1.75 g MRS) and (1.15 g LBG + 1.15 g MCS + 1.15 g MRS), showing a significantly lower expression than for the Zn positive controls and the other combinations. These findings could indicate reduced zinc availability. Regarding iron, whereas the relative expression of ferritin genes (FTH1 and FTL) was stable and independent from iron extracellular content, ferritin protein content was significantly higher for the cells treated with the FeSO4 control. This indicates that iron status might influence ferritin content at a post transcriptional level. No significant differences were found between other samples although a slightly higher trend was observed for the (1.75 g LBG + 1.75 g MCS) treatment.

Conclusion: Study of these protein expressions could clarify the possible interactions between thickeners and minerals. Due to the complexity of infant formula composition and the low mineral specificity of some proteins, other analysis as in-vitro mineral availability or quantification of other ingredients would be necessary.

Keywords: Caco-2 cells, digested infant formula, mineral transporters, storage proteins, thickeners.


46

A22

LEAD ENTERS PLANT CELL BY ENDOCYTOSIS

Krzesłowska M.1, Basińska Aneta1, Mellerowicz E.J.2, Napieralska A.1, Rabęda I. and Woźny A.1

1)Laboratory of General Botany, Faculty of Biology, Adam Mickiewicz University Umultowska 89, 61-614 Poznań, Poland,

2)Ůmea Plant Science Center, Department of Forest Genetics and Plant Physiology Swedish University of Agricultural Sciences, SE-90183 Ůmea, Sweden

Pb deposits were often observed in transport vesicles (Vs). However, it was not clear if observed vesicles belonged to secretion or endocytic pathway. Therefore the aim of our study was to estimate if endocytosis was the really entrance pathway for Pb.

The material was Funaria hygrometrica protonemata and root tip of poplar. The exposure conditions were similar for both plant species: Pb 1000µM 4h; control H2O, 4h. The Vs transport was studied with FM1-43 (in LSM). To estimate if Pb enter the protoplast by endocytosis brefeldine A (BFA) which allows endocytosis and inhibits exocytosis was used (observations in TEM).

After Pb exposure cells of both plant objects indicated the intensification of vesicle trafficking. The vesicles were more numerous and additionally larger than in control. Many of them contained lead deposits. Inhibition of exocytosis by BFA, caused that Pb deposits (often extremely large) occurred in BFA compartments, large vacuoles and vesicles.

The results show that Pb enter plant cell via endocytic pathway. It occurs in different cells and different plant species. Moreover, formation of extremely large Pb deposits suggests that BFA inhibited Pb removing from the protoplast via secretion pathway and exocytosis, e.g. to the cell wall.

Keywords: endocytosis, Pb, plant cell, trace metal, uptake


47

A23

IRON-BIOFORTIFIED PEARL MILLET PROVIDES ADDITIONAL BIOAVAILABLE IRON COMPARED TO REGULAR-IRON MILLET BUT TO A LESSER EXTENT THAN

POST-HARVEST IRON-FORTIFIED MILLET

Colin Cercamondi1, Ines Egli1, Evariste Mitchikpe2, Erick Boy3, Felicien Tossou4, Christophe Zeder1, Joseph Hounhouigan2 and Richard Hurrell1

1)Laboratory of Human Nutrition, ETH Zurich, Switzerland 2)Laboratoire de Physiologie de la Nutrition, Université d’Abomey Calavi, Benin 3)HarvestPlus, Washington, District of Columbia 20006 4)Zone Sanitaire Natitingou, Ministère de la Santé, Benin

Iron biofortification of pearl millet (Pennisetum glaucum) is a promising approach to combat iron deficiency in millet consuming communities in developing countries. To evaluate the potential of iron-biofortified millet to provide additional bioavailable iron in comparison to regular-iron and post-harvest iron-fortified millet, an iron absorption study was conducted in 20 Beninese women with low iron status. Iron absorption from multiple composite meals was measured as erythrocyte incorporation of stable iron isotopes. Fractional iron absorption from meals based on regular-iron millet (7.5%) did not differ compared to iron-biofortified millet meals (7.5%; P = 1.0) resulting in a higher quantity of total iron absorbed from the iron-biofortified millet meals (527 µg vs. 1125 µg; P < 0.0001). Fractional iron absorption from post-harvest iron-fortified millet meals (10.4%) was higher than from regular-iron (P < 0.05) and iron-biofortified millet meals (P < 0.01). Total iron absorbed from the post-harvest iron-fortified millet meals (1500 µg) was higher than from the regular-iron (P < 0.0001) and iron-biofortified millet meals (P < 0.05). Results indicate that iron-biofortified millet has potential to provide additional bioavailable iron compared to regular millet and could be a promising approach for populations with limited access to post-harvest fortified foods.

Keywords: Iron biofortification, Iron bioavailability, Iron fortification, Pearl millet


48

A24

IMPACT OF POLYPHENOLS ON IRON BIOAVAILABILITY FROM POTATO AFTER SIMULATED GASTRO-INTESTINAL DIGESTION

Lisa Miranda, Aude Corvisy, Hannah Deußer, Sylvain Legay and Danièle Evers

EVA Department, Centre de Recherche Public-Gabriel Lippmann, Luxembourg, [email protected]

The aim of this work was to evaluate iron bioavailability for human consumption from different potato cultivars after simulated gastro-intestinal digestion and to study the impact of enhancers of iron bioavailability (e.g. vitamin C) or inhibitors commonly present in potato, such as chlorogenic acid and rutin, polyphenols present in high amounts in potatoes.

Caco-2 cells were used as model for intestinal iron uptake and transport. Abundance of ferritin protein was measured by a human ferritin ELISA test and used as a marker of iron uptake. Polyphenol composition was determined by UPLC-DAD.

Chlorogenic acid inhibited iron uptake induced by vitamin C. Rutin significantly increased iron uptake in a dose dependent manner. Chlorogenic acid was the most abundant polyphenol in both potato cultivars before and after in vitro digestion and absorption by Caco-2 cells. Treatment with digested extracts from potato cvs. Nicola and Vitelotte led to a decreased ferritin synthesis compared to controls. Vitelotte containing more polyphenols than Nicola induced a more pronounced inhibition of ferritin synthesis.

These results highlight the influence of antioxidant compounds from potato on iron uptake. They also point out the complexity of potato as a matrix containing both enhancers and inhibitors of iron bioavailability.

Keywords: chlorogenic acid, iron bioavailability, potato, rutin


ABSTRACTS - POSTERS


49

P1

INCLUSION OF UREA IN THE FOLIAR 59FEEDTA SOLUTION STIMULATED LEAF PENETRATION AND TRANSLOCATION OF 59FE WITHIN THE WHEAT PLANTS

Seher Bahar Aciksöz Özden, Levent Ozturk, Atilla Yazici and Ismail Cakmak

Sabanci University, Faculty of Engineering and Natural Sciences, 34956, Istanbul, Turkey

In this study, role of urea in translocation of 59Fe from the 59FeEDTA-treated leaves was studied in durum wheat (Triticum durum L.) which was grown for 2 weeks in nutrient solution or until grain maturation in soil culture. Five-cm long tips of the first leaf of the young wheat seedlings or the flag leaves at the early milk stage were immersed daily for 10 second in the 59FeEDTA solutions containing increasing amounts of urea (0, 0.2, 0.4, 0.8% w/v) over 5 days. In the experiment with young wheat seedlings, urea inclusion in the 59Fe-EDTA solution significantly increased translocation of 59Fe from the treated leaf into roots and the untreated part of shoots. In the experiment conducted in soil culture until grain maturation, translocation of 59Fe from the flag leaves into grains was also strongly promoted by urea, whereas 59Fe translocation from flag leaves into untreated shoot was very low, and not affected by urea. The urea treatments resulted in an increase in soluble free amino acids in the treated leaf sections. In conclusion, urea contributes to transportation of the leaf-absorbed Fe into sink organs. Probably, the nitrogenous compounds formed after assimilation of foliarly-applied urea (such as amino acids) contributed to Fe chelation and translocation in wheat.

Keywords: biofortification, foliar spray, iron, urea, wheat


50

P2

ACCUMULATION OF TRACE ELEMENTS IN EDIBLE PLANTS PRODUCED IN THE VICINITY OF ABANDONED MINING AREAS: THE CASE STUDY OF MINES

LOCATED IN THE IBERIAN PYRITE BELT

Paula Alvarenga1,2, Isabel Simões1, Patrícia Palma,3, Olga Amaral1 and João Xavier Matos4

1)Departamento de Tecnologias e Ciências Aplicadas, Escola Superior Agrária, Instituto Politécnico de Beja, Beja, Portugal, [email protected]

2)UIQA – Unidade de Investigação Química Ambiental, Instituto Superior de Agronomia, Universidade Técnica de Lisboa

3)CIMA - Centro de Investigação Marinha e Ambiental, CIMA, FCT, Edifício 7, Piso 1, Universidade do Algarve, Campus Universitário de Gambelas, 8005-139 Faro

4)Laboratório Nacional de Energia e Geologia, UI Recursos Minerais e Geofísica, Beja, Portugal

The development of agriculture activities in soils potentially affected by mine workings is a risk scenario for the transference of detrimental trace elements (TE) to the food chain. Therefore, it is important to evaluate the contamination of soils with potentially toxic TE and ascertain their accumulation in edible plants produced in these soils. Eighteen different sampling sites were selected in small farms from three mines located in the Portuguese sector of the Iberian Pyrite Belt (IPB) (São Domingos, Aljustrel and Lousal) and their soils were analyzed considering: soil nutritional status, total TE concentrations and bioavailable fractions (As, Cu, Pb, and Zn). The same TE were analyzed in three different species of edible plants: lettuce (Lactuca sativa), coriander (Coriandrum sativum), and cabbage (Brassica oleracea), collected at the same locations.

The soils could be considered contaminated with trace elements, since the total As, Cu, Pb, and Zn concentrations surpassed the Canadian Soil Quality Guideline Values for agricultural use in 100, 72, 94 and 75%of the sampling sites, respectively. The maximum total concentrations for those TE were extremely high in some of the sampling sites (e.g. 1851 mg As kg-1 in São Domingos, 1126 mg Cu kg-1 in Aljustrel, 4946 mg Pb kg-1 in São Domingos, and 1224 mg Zn kg-1 in Aljustrel). However, the soil pH(H2O) were mainly neutral, a factor that contributes to their low bioavailable fractions. As a result, the plants showed levels of these elements characteristic of uncontaminated plants, and transfer coefficients for all elements <1, typical of excluders plants. The estimated daily intake (EDI) of Cu and Zn, through the consumption of these vegetables, falls below the recommended limit for daily intake of these elements by the Institute of Medicine, Food and Nutrition in the USA. The sampling site that stood out from the others was located at Aljustrel, where bioavailable Zn levels were higher, a consequence of the slight acidity of the soil. Therefore, the Zn content in vegetables were also higher, characteristic of contaminated plants, yielding Zn EDI values that exceed the maximum permissible daily intake for children with 1 to 3 years.

Keywords: edible plants, Iberian Pyrite Belt, mining activities, soil contamination, trace elements


51

P3

ALTERATIONS OF THE TRANSCRIPTION PROFILES OF TOMATO ROOTS EXPOSED TO ZN DUE TO EXPRESSION OF ATHMA4

Kendziorek Marie1, Borg Søren2 and Antosiewicz Danuta Maria1

1University of Warsaw, Faculty of Biology, Institute of Experimental Plant Biology and Biotechnology, Miecznikowa 1, 02-096 Warsaw, Poland e-mails: [email protected];

2University of Aarhus, Faculty of Agricultural Sciences, Department of Genetics and Biotechnology, 4200 Slagelse, Denmark. e-mails: [email protected]

The heterologous expression of HMA4 resulted in enhancement of Zn root-to-shoot translocation, but in a Zn-supply dependent manner [1; 2]. It suggests that HMA4-expression under a range of Zn-supply differentially modifies the expression of endogens, which contributes to different phenotypes. Understanding the interplay between the transgene and the endogens is crucial for planning genetic modification for Zn-biofortification purpose. The aim was to identify genes differentially regulated in 35S::AtHMA4-expression and wild-type tomato under a range of Zn supply. As a first step, the transcription profiles were compared between transgenic and WT-plants with enhanced Zn root-to-shoot translocation.

RNA was isolated from the LAM-excised epidermis+cortex and the stele of the roots of transgenic and WT-plants grown at 2µM Zn. Selected indicative genes will be used further to analyze the difference between transgenics with different phenotypes under low-to-high Zn conditions.

The AtHMA4 expression modified four important metabolic categories of genes specifically for the examined tissues:

1) Up-regulation of transcription factors involved in Fe/Zn homeostasis:

2) FER essential for regulation of multiply Fe/Zn homeostasis genes

3) bZIP known to be involved in the Zn metabolism

4) WRKY (response to abiotic stress)

5) Up-regulation of metal transporters (Nrapms and ABC)

6) Up- and down-regulation of the expression of genes involved in the cell-wall modifications (HMA4 expression overloads the apoplast with Zn [2; 3])

7) Down-regulation of genes from the ethylene pathway

Keywords: AtHMA4, tomato, zinc, transformation, transcriptomics Supported by: MNiSW 814/N-COST/2010; EU COST Action 0905; FP6 EU PHIME. References:

[1] Siemianowski et al., (2011), Pl Biotechnol J 9:64-74

[2] Barabasz et al., (2012), Physiol Plantarum 145: 315-331.

[3] Siemianowski et al. (2013), doi: 10.1111/pce.12041


52

P4

ARE LEGUMES ACCUMULATORS OF BARIUM IN SANDY MINERAL SOIL?

Mona Bakke, Marina A. Bleken, Michael Heim, Tore Krogstad and Elin Gjengedal

Dept of Plant and Environmental Sciences, Norwegian University of Life Sciences, Aas, Norway

Plant uptake of barium (Ba) in natural vegetation has shown large variation among plant species and plant parts. In spite of the toxicity of soluble Ba compounds to animals and humans, the mechanisms controlling Ba-availability and -uptake in plants are scarcely investigated. The aim of this study was to investigate interspecies differences, and treatment effect of lime and nitrogen (N-) fixation on Ba-uptake in agricultural plants. Representatives from grass, corn, vegetables, and herbs were grown in a fixed, pH regulated, sandy growth medium in a greenhouse experiment. Hydrofluoric acid soluble content of Ba in the sand was 350 ± 12 mg kg-1. Each species was fertilized for optimal growth conditions and harvested at a mature or flowering state. Plant samples were divided into respective plant parts, dried, ground and acid digested using microwave technique prior to analysis by means of ICP-MS. Content of Ba in the different plant species ranged from 60 (grass) to 700 (legumes) mg kg-1 dry matter. Regarding plant parts Daucus carota L. spp (carrot) had lowest Ba content in roots, while Spinacia oleracea (spinach) and Pisum sativum (peas) showed tendency of highest content in roots. Statistical interpretations by liming effect, N-fixation and Ba uptake versus nutrients will be presented.

Keywords: Barium, plant uptake


53

P5

SECOND-LEVEL FACTORS AS QUANTITATIVE PROCESS EVALUATION METHOD FOR ESSENTIAL AND DETRIMENTAL ELEMENT UPTAKE AND EXCLUSION

Edita Baltrėnaitė1*, Pranas Baltrėnas2, Donatas Butkus3 and Arvydas Lietuvninkas4

1),2),3)Department of Environmental Protection, Vilnius Gediminas Technical University Saulėtekio al. 11, LT–10223 Vilnius, Lithuania

4)Tomsk State University, Lenin Av. 36, 634050 Tomsk, Russia *Corresponding author: [email protected], phone: +370687 52299, fax: +370 52744726

One of the objectives of the COST FA0905 is related to acquiring the knowledge helping to control the content of the essential and detrimental elements in the plant-derived food. When agronomical and plant physiological technologies are being developed to reach the objective, the quantitative process evaluation of element uptake or exclusion by plant is beneficial.

The interface of the soil and a plant is the main criterion when evaluating the peculiarities of the concentrative function of the plant for a particular element. In order to evaluate the process (the uptake or exclusion of elements by plant) it is more essential to compare the process changes rather than element concentrations.

The second-level factors of bioaccumulation, biophilicity, translocation and phytoremediation allow comparing changes in the processes of element uptake or exclusion by plants. The factors help to evaluate the influence of both the agronomical technologies as well as physiological technologies effect on the participation of elements in the metabolism of plants (Baltrėnaitė et al. 2012).

Keywords: biogeochemistry, element bioavailability, second-level factors References:

[1] Baltrėnaitė, E.; Lietuvninkas, A.; Baltrėnas, P. (2012) Use of dynamic factors to assess metal uptake and transfer in plants - Example of trees.Water, Air, & Soil Pollution 223: 4297–4306


54

P6

PHOSPHORUS AND ALUMINUM INFLUENCE ANTIOXIDANT COMPOUNDS IN TEA

Roser Tolrà, Roghieh Hajiboland, Charlotte Poschenrieder and Juan Barceló

Lab. Fisiologia Vegetal, Facultat de Biociencias, Universitat Autònoma de Barcelona

Tea is after water the most consumed beverage. Obtained from dried leaves of Camellia sinensis it is rich in healthy bioactive compounds. Tea is commonly grown in acid soils and large Al concentrations be found in tea. Aluminium has been affirmed as a food contaminant by the Joint FAO/WHO Expert Committee on Food Additives. Although no danger from tea consumption has been shown, lowering tea leaf Al but maintaining the beneficial antioxidant properties of the beverage is an objective of interest. Our aim here was to investigate whether supply with phosphorus, a typical limiting factor on acid soils, has an impact on both Al concentrations and phenolic compounds of young tea leaves. Tea plants were grown in hydroponics with and without Al and two levels of P. Leaves were analysed for phenolic compounds using HPLC-MS (LC-ESI-QTOF). Surplus P did not affect leaf Al. The major catechins in tea young leaves were epicatechin gallate (ECG) and epigallocatechin gallate (EGCG). These catechins deserve most attention as they are highly abundant in hot water extracts and appear to be the major anti-oxidant substances. Both Al and P supply increased the levels of these two compounds in young tea leaves.

Keywords: Aluminium, bioactive compounds, phosphorus, polyphenols, tea Supported by: Project BFU2010-14873


55

P7

STUDYING THE ANTIOXIDANT SYSTEM IN ARABIDOPSIS THALIANA UNDER ZINC AND CADMIUM STRESS

András Bittsánszky1 and Peter Schröder2

1)Plant Protection Institute Centre for Agricultural Research Hungarian Academy of Sciences 2)Helmholtz Zentrum München, Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH)

The present study aimed at quantifying the effects of environmentally realistic concentrations of zinc and cadmium in the model plant Arabidopsis thaliana. It is well known that excess of heavy metals will cause the generation of reactive oxygen species (ROS) inside plant tissues. The most common response of plants to ROS is the increase of the antioxidant enzyme activities.

Various concentrations of zinc-sulphate and cadmium-sulphate were applied to mature plants through sub-irrigation. One week after treatments protein were extracted from the aerial parts and subjected to biochemical analysis. Enzyme activities related to reactive oxygen species were measured spectrophotometrically.

Our results revealed that the synergistic effect of zinc and cadmium on plants is much more different compared to the treatment with single elements alone. Whereas single metal ions alone seem to be taken up at similar rates, the combination has synergistic side effects. Therefore to improve the success rates on valuable food and feed production, breeding programs for better ability to exclude Cd in parallel with higher capability for Zn accumulations should be started. It should be investigated if crops are available that show the same traits.

Keywords: antioxidant enzymes, Arabidospis thaliana, cadmium, zinc


56

P8

TOXIC AND ESSENTIAL ELEMENTS CONTENT IN THE SEED OF SELECTED VARIETIES OF LINSEED AS AFFECTED BY SELENIUM FERTILIZATION

Marie Bjelková1, Miroslav Griga2 and Martina Větrovcová2

AGRITEC Plant Research, Ltd., 1)Department of Legumes and Industrial Crops 2)Plant Biotechnology Department, Zemědělská 2520/16, CZ-787 01 Šumperk, Czech Republic

email: [email protected]

The study monitors the impact of selenium (Se) fertilization on the metal elements content in linseed. Selenium supplementation may be the way to increase the content of selenium in food and feed. Selenium influences the formation of the enzyme glutathione peroxidase, which protects cell membranes from damage caused by lipid peroxidation.

The field experiment was represented by variants without/with Se fertilization: 0 (control), 10 and 30 kg Se ha-1 in the form of sodium selenite. Linseed varieties Amon, Raciol, Natural and Kaolin were studied. Seeds from mature plants were analyzed for toxic/essential elements content by AAS.

Seed yield was influenced by Se fertilization: 10 kg Se. ha-1 (123.25 g.m2), 30 kg Se.ha-1 (110.87 g.m2), control (102.37 g.m2), but linseed varieties responded to fertilization differently. A higher dose of selenium fertilization influenced the increase of its content in the seeds, but decreased the content of other microelements (Zn, Mo, Cu). 10 kg Se. ha-1

increased seed Cd and Co content in all cvs, in contrast, 30 kg Se.ha-1 decreased seed content of both elements. Se fertilization increased Pb and decreased Fe accumulation. The relationships between both toxic and essential elements content in the seed as affected by Se fertilization is discussed.

Keywords: essential elements, linseed, selenium fertilization, seed, toxic elements

Acknowledgement: This research was financially supported by Ministry of Education CR, grant No. LD 11053


57

P9

A COMPREHENSIVE ANALYSIS OF CU INDUCED CHANGES IN THE MINERAL PROFILES OF CU SENSITIVE AND TOLERANT POPULATIONS OF

SILENE PARADOXA L.

Ilaria Colzi1, Sara Pignattelli1, Antonella Buccianti2, Ilenia Cattani3, GianMaria Beone3, Henk Schat4 and Cristina Gonnelli1

1)Department of Biology, Università di Firenze, via Micheli 1, 50121 Firenze, Italy. 2)Department of Earth Science, University of Florence, via La Pira 4, 50121 Firenze, Italy. 3)Institute of Agricultural and Environmental Chemistry, Università Cattolica del Sacro Cuore, Via

Emilia Parmense 84, Piacenza, Italy. 4)Department of Genetics, Faculty of Earth and Life Sciences, Vrije Universiteit, De Boelelaan 1085,

1081 HV Amsterdam, The Netherlands

Objectives

This work investigates the Cu induced changes in element profiles in contrasting ecotypes of Silene paradoxa L.

Methods

A copper tolerant population and a sensitive population were grown in hydroponics and exposed to different CuSO4 treatments. Shoot and root concentrations of Ca, Cu, Fe, K, Mg, Mn, Mo, Na, P, S and Zn were evaluated through ICP-OES.

Results

In the tolerant population Cu treatment induced a higher element accumulation in roots and had minimal effects on the shoot element profile, thus resulting in a progressively decreasing shoot-to-root ratio for each element. In the sensitive population element concentrations in root and shoot were much more affected and without a consistent trend. Copper treatment also affected the correlations between the elements more in the sensitive population than in the tolerant one.

Conclusions

Cu exposure strongly disturbed element homeostasis in the sensitive population, but barely or not in the tolerant one, probably mainly due to a higher capacity to maintain proper root functioning under Cu exposure. Differences in element profiles were also observed in the absence of toxic Cu exposure, reflecting divergent population-specific adaptations to differential nutrient availability levels prevailing in the populations’ natural environments.

Keywords: compositional data analysis, copper tolerance, mineral profile, Silene paradoxa


58

P10

IRON UPTAKE IN LACTUCA SATIVA: WATER IRRIGATION TREATMENT BY PHYTOREMEDIATION

Elena Comino, Adriano Fiorucci, Stefania Menegatti and Vincenzo Riggio

Department of Environment, Land and Infrastructure Engineering, C.so Duca degli Abruzzi, 24 - 10128 - Politecnico di Torino; email: [email protected]

Trace elements presence in living organisms are positive and necessary. If they are accumulated by autotrophy, in tolerated concentration, they can contribute to the plant bio fortification. If they are translocate to the heterotrophy, by the food chain, they can contribute to the organisms improvement.

For these two reasons the main transport vehicle for the trace elements to the humans are food and water. Several time water contains elements that are in high concentration.

Water depuration has heavy costs and request good technologies.

There are some situation (poor areas, or agricultural lands, or damaged tube-well water) in which is not easy to provide the best available technologies. Directly consequence is often the use of wastewater and industrial water as irrigation water without any treatment.

The goal of our research is to find out a balanced solution to treat “contaminated” irrigation water with the best and cheapest available technologies.

The experimental study was focused to show the results rising out from years of experiments. We use a phytoremediation plant to treat wastewater highly polluted by iron. (Phytoremediation is a suitable technology with high performance rates, low cost and easy management). After the pre-treatment the water iron rate removal has been calculated, and the treated water was used to irrigate Lactuca sativa. Several analyses has been performed in order to calculate the bio concentration factor (BCF) and the efficiency of the phytoremediation plants.

Laboratory test analysis, give confidence that phytoremediation water treatment could be really a good “tuning system” in establish the right iron supply to the plants from irrigation water.

Keywords: iron, Lactuca sativa, irrigation water, phytoremediation, plant uptake


59

P11

RECYCLED BY-PRODUCTS CAN MAKE POSITIVE CONTRIBUTIONS TO THE YIELD AND NUTRITIONAL COMPOSITION OF GRASS-CLOVER LEYS

A Sigrun Dahlin1, Atefeh Ramezanian2, Colin D Campbell3,1, Stephen Hillier3,1 and Ingrid Öborn2,4

1)Swedish University of Agricultural Sciences (SLU), Dept. Soil & Environment 2)SLU, Dept. Crop Production Ecology 3)The James Hutton Institute 4)World Agroforestry Centre (ICRAF)

Four selected by-products (biogas digestate, pot ale, wood ash and rockdust) were tested for their fertiliser value to a mixed perennial ryegrass–red clover crop in terms of crop performance (yield and botanical composition) and mineral quality. To render the results relevant to production systems on marginal land, two inherently low-fertility soils were used for the 14 months pot experiment. A number of nutrients of interest in high-yielding dairy production were determined in plants and soils: N, P, K, Ca, Mg, Na, Co, Cu, Mn, Mo, Ni and Zn. All tested by-products increased overall yield and affected nutrient concentrations of the individual plant species. The effects differed between the grass and clover, though, which lead to changes in botanical composition indirectly affecting the nutrient concentrations of the mixed crop. Individual by-products increased concentrations of Cu, Mg, Na and/or Zn to close to or over the recommended minimum concentrations. There is thus potential to enhance agricultural productivity on marginal land through improved forage production and quality by matching of by-products, soils and forage species/mixtures. However, to secure sustainable use of by-products as fertilisers or soil amendments, soil quality must be considered and monitored in addition to crop quality, e.g. by using element balances.

Keywords: botanical composition, crop growth, crop quality, macro- and micronutrient concentrations, by-product recycling


60

P12

CAN SOIL GRAIN SIZE DETERMINE THE AVAILABILITY OF CD AND AS IN SOIL?

M. Greger1, 2, M. Dvorsak1, T. Motrøen1 and T. Landberg2

1Faculty of Applied Ecology and Agricultural Sciences, Hedmark University College, Blæstad, NO-2418 Elverum, Norway, email: [email protected];

2Department of of Ecology, Environment and Plant Sciences, Lilla Frescati, SE-106 91 Stockholm, Sweden.

The availability of cadmium (Cd) and arsenic (As) to plants depends on the content and how much is firmly bound to soil particles. The aim was to find out if some grain sizes had higher available Cd and As concentrations than others.

Soils of various kinds and origin were collected from a depth of 40 cm at seven agricultural fields in Norway. Three were collected on sites with slate/limestone bedrock, one on sandstone and three on granite bedrock. Soils were dried and sieved into various sizes; <63, 63-80, 80-125, 125-180, 180-250, 250-355, 355-500, 500-630, 630-1000, 1000-2000, 2000-4000µm. The size fractions were analyzed on mass and total and exchangeable (available) Cd and As.

The total as well as extractable Cd concentration in all investigated soils was highest in grain size 180-500µm. Except for one soil, the total and extractable As concentration was highest in fractions 63-630µm. The lowest concentrations of both As and Cd were found in size fraction 1000µm and higher. The conclusion so far is that soils with high percentage of the grain size 63-630µm (corresponding to sand of fine to medium size), may create a higher risk for high uptake of Cd and As by plants than soils with lower percentage of these grain sizes.

Keywords: Arsenic, availability, cadmium, soil grain size


61

P13

IN VITRO TESTING OF POTENTIAL FOR SELENIUM ACCUMULATION BY LINSEED (LINUM USITATISSIMUM L.)

Magdalena Cvečková, Martina Větrovcová and Miroslav Griga

Plant Biotechnology Department, AGRITEC Plant Research, Ltd., Zemědělská 16, CZ-78701 ŠUMPERK, Czech Republic, email: [email protected]

Selenium is an importatnt essential element to human health. As a constituent of selenoproteins selenium plays structural and enzymatic roles. The potential capacity of linseed cultivars to accumulate specific microelements brought an interest in their use for mineral improvement of healthy food and feed in selenium deficient areas.

In this work, linseed cultivars and GM lines transformed with αMT1a subunit of mammalian metallothionein for overproduction of heavy metal-binding peptide were used. In vitro testing of selective concentrations for callus cultures in liquid media and multiple shoot cultures on solid media was performed. Selenium in the form of Na2SeO3 was added into the medium as follows: A=0 µM; B=10 µM; C=50 µM; D=100 µM; E=150 µM. Furthemore, an influence of selenium on growth parameters and the estimation of selenium content in the biomass by atomic absorption spectroscopy were determined.

Preliminary results show the growth stimulation by very low concentration of selenium. The response was not uniform in different cultivars, nevertheless, GM lines showed higher tolerance and accumulation capacity for selenium than non-GM cultivars tested.

Based on the results, quick screening methodology for selenium accumulation potential by linseed cultivars may be proposed. Further benefits of transformation for agricultural plants in terms of human nutrition improvement are demonstrated.

Keywords: αMT1a, GM lines, linseed, mammalian metallothionein, selenium Acknowledgement: This research was financially supported by Ministry of Education CR, grant No. LD 11053.


62

P14

MOBILIZATION OF BARIUM IN CARBONATITE ROCK POWDER BY ADDITION OF ORGANIC MATTER

Mari Haugene, Mona Bakke, Tore Krogstad and Elin Gjengedal

Department of Plant and Environmental Sciences, Norwegian University of Life Sciences, Aas, Norway

Apatite-biotite-carbonatite (ABC) rock powder from Stjernøy, Norway, has shown great potential as a fertilizer or soil amendment for agricultural purposes. A potential but barely investigated problem area is its elevated levels of barium (Ba), which can be up to 50 times higher than the average Ba content in Norwegian soil. In spite of the toxicity of soluble Ba compounds to organisms, the mechanisms controlling Ba availability in soil are not well known. However, it is acknowledged that addition of rock powder to organic matter (OM) generally causes increased mineralization of OM and weathering of the rock powder. Continuous flow-through experiments will be conducted on various mixes of an inert growth medium (silica), ABC rock powder, a water soluble Ba salt, and matured compost. A final size fractionation of Ba in pore water and leachate will be conducted in order to understand its behavior and fate. Each soil treatment combination will be given three flow-through treatments with deionized water, artificial precipitation representative of coast sites, and organic acid, respectively. The results will give decisive input to the design of further greenhouse experiments on Ba uptake in plants cultivated on growth media supplied with ABC rock powder.

Keywords: Barium availability, carbonatite rock powder, organic matter, size fractionation of Ba


63

P15

RARE EARTH ELEMENT (REE) DISTRIBUTION IN BEDROCK, SOIL AND PLANTS – RESULTS OF TWO RECENT NORWEGIAN STUDIES

Michael Heim, Mona Bakke, Maria H. Hillersøy, Marina A. Bleken, Karl A. Jensen and Elin L.F. Gjengedal

Dept. of Plant and Environmental Sciences, Norwegian University of Life Sciences

For the last decades there has been an increasing agricultural interest in the rare earth elements, i.e. scandium, yttrium and the 15 lanthanides (La57 to Lu71). Positive crop responses (up to several 10%) to REE-enriched fertilizers have been reported, especially from China. Various physiological and biochemical effects of REEs to plants, like their interaction with calcium are well documented, but still without understanding their overall synergism, resulting in claimed growth and yield increases (1, 2).

With today’s analytical methods REE-contents are easily quantified as bonus of standard investigations of all types of materials. From a first study we present REE-contents in bedrock, mineral soil and vegetation at three sites on the island of Stjernøy (N-Norway), an area with exceptional high contents (∑REE up to 2000 mg/kg) in silico-carbonatite bedrock, and enriched in especially light REE. Concentration variations in bedrock and soil are only partly reflected in plants, the variations among REE, plant species and plant parts being more determining. Likewise, REE-contents in soil and plants from a second study are presented and discussed. In a pot experiment agricultural plants were grown in a sandy soil with low REE-content (∑REE <100 mg/kg), and showed large species and plant part dependant variations of REE-concentrations.

Keywords: Agricultural plants, rare earth elements, sandy soil, silico-carbonatite bedrock, vegetation References:

[1] Hu,Z.Y., Richter,H., Sparovek,G. and Schnug,E. (2004) Physiological and biochemical effects of rare earth elements on plants and their agricultural significance: A review. Journal of Plant Nutrition 27, 183-220.

[2] Tyler,G. (2004) Rare earth elements in soil and plant systems - A review. Plant and Soil 267, 191-206


64

P16

BIOFORTIFICATION OF ZINC AND IRON GRAIN CONTENT OF EFFICIENT CADMIUM EXCLU-DING WINTER WHEAT GENOTYPE BATIS BY NON-GM-

BREEDING AND MUTANT SELECTION FROM LABORATORY TO FIELD CONDITION

Rolf Herzig1, Arturo Ricci1, Erika Nehnevajova2 and Satish K. Gupta3

1Phytotech-Foundation (PT-F), 3013 Berne, Switzerland 2Institute of Biology/Applied Genetics, Free University of Berlin, Germany 3ETH Zürich Institut f. Terrestrische Ökosysteme, Zürich, Switzerland e-mail: [email protected]

In the past decades, both crop breeding and production has largely focused on maxi-mising yield in terms of biomass production per hectare and on securing food and feed production. As a negative side effect of this strategy concentration of essential macro/micro-nutrients for man and animals have been dramatically diluted. Estimates says, that over three billion people are currently micronutrient malnurished.

The team of Phytotech Foundation introduced in 2011 a mutagenesis approach with the best Cd-excluding winter wheat cultivar BATIS from Germany. The aim is to biofortify the grain content for desired Zn and Fe (and possibly Se, Mg) and exclude toxic elements such as Cd. Based on experiances with sunflower a modified EMS mutagenesis was used to produce a significant amount of putative mutants. About 20'000 mutagenized grains (0.08M of 3, 4, 7, 11h of EMS treatment) were sown in autumn 2011 for direct field screening on our phyto-remediated and formerly zinc contaminated site of Bettwiesen (CH). A few thousand of fertile mutant spikes and it’s controls were harvested in august 2012, and mutant screening of the grain content was done on a sensitive XRF instrument. For the better11-h EMS-treatment the relative relative iron and zinc gain, compared to controls reached up to a factor of 8 for iron, and 5 for zinc, whereas cumulative iron and zinc grain biofortification was enhanced up to a factor of 5.7. Best individual mutant lines with a cumulative gain factor >1.9 for iron and zinc were sown again in field for next M3-generation in 2013. The project is ongoing and pro-mising results will be shown.

Keywords: iron, zinc biofortification, bread-wheat BATIS, mutagenesis, freeland mutant screening References:

[1] Herzig, R, et al., 2012. Biofortification of Zinc, Iron and Selenium content of an efficient Cd-excluding wheat variety by non-GM fast track breeding and mutant selection from laboratory until field condition. COST Action Meeting WG3 + WG4: Modifying the composition of plant food for better mineral nutrition, ETHZ, 4.-5.6.2012.

[2] Nehnevajova E, Herzig R, Federer G, Erismann KH, Schwitzguébel JP. 2007. Chemical mutagenesis – an effi-cient technique to enhance metal accumulation and extraction in sunflowers. Int J. Phytorem 9, 149-165.

[3] Herzig, R., Nehnevajova, E., 2007. Establishing of improved cultivars of high yielding food crops by means of conventional in vitro-breeding, mutagenesis and selection techniques (non-GMO) with special emphasis of accumulation, extraction and exclusion characteristics for toxic metals. Final Project Report of Cost Action 859 State Secretariat for Education and Research SER, Berne.


65

P17

NUTRIENTS AND TRACE ELEMENTS IN THE SEED OF SELECTED VARIETIES OF QUINOA (CHENOPODIUM QUINOA WILLD.) UNDER MEDITERRANEAN

CONDITIONS

Theodore Karyotis and Ch. Noulas

Hellenic Agricultural Organization-DG Research-Institute for Soil Mapping and Classification, 1, Theophrastou Str., 41335, Larissa, Greece, email: [email protected]

Quinoa (Chenopodium quinoa Willd.) belongs to the pseudo-cereals and is an example of healthy diet. Quinoa is rich in proteins and contains minerals, vitamins, fatty acids and antioxidants, which are valuable for human nutrition. Eight varieties obtained from the International Centre of Potato of Peru were cultivated in two different soils. A clay and a loamy soil located in Central Greece were selected for the field experiments. After harvesting, whole seed analyses were performed for macro nutrients and trace elements. The varieties originated from South America accumulated higher protein in the seeds at both locations. Protein ranged between 12.1 and 15.7 % and significant variation was recorded for macronutrients and micro elements. Results indicated that mineral composition of quinoa was higher in comparison to that of the cereals. Quinoa stands out for its high content of calcium, magnesium, phosphorus and micronutrients, in comparison to wheat, corn, rice, barley and oats. Furthermore, its deep rooting system enables quinoa to extract nutrients left by previous crops. Quinoa can be an important new crop for greek agriculture as can provide a number of food products. It can be characterised as a promising crop due to its nutritional potential and its adaptability to a wide range of soil types. It may be suggested for cultivation in areas where the degraded land and ground water scarcity severely hamper production of other crops. Taking into consideration the xeric moisture regime and thermic temperature regime in Central Greece, this crop can be adapted well under the prevailing unfavourable climatic conditions in Greece.


66

P18

SELENIUM IN THE SOIL-PLANT SYSTEM FROM A HEAVY METALS POLLUTED AREA

Radu Lacatusu1;2, Mihaela Monica Stanciu-Burileanu1, Mihaela Lungu1, Anca-Rovena Lacatusu1 and D. Popa2

1)National Research and Development Institute for Soil Science, Agrochemistry and Environment Protection, Bucharest, Romania

2)„Al. Ioan Cuza” University, Iassy

The researches were conducted in Copsa Mica, one of the most heavy metals polluted areas in Romania and even Europe. Emissions from a non-ferrous ore processing plant have polluted, over time, an area of about 21,800 ha. Soil samples were taken from 41 points within an area of 1,200 hectares, very strong polluted. Samples from maize were collected in their first part of the growing season, at 5-6 leaves stage, and, grain samples at their maturity. Analyses revealed heavy metal content in soil, which exceed up to 44 times (Cd), 36 times (Pb) and 9 times (Zn) maximum allowable limit values. Total selenium concentrations in the soil exeed up to 2.5 times the normal concentration of selenium in soils without reaching the maximum allowable limit value. Mobile selenium content in soil was double than usual. Maize plants accumulated up to 4 times more Cd, up to 10 times more Pb, and up to 22 times more Zn than normal content values. As a result, the absorption of selenium in plants was also higher, registering double values compared to normal. Between selenium contents in plant and soil have been calculated directly proportional relationship, statistically assured. Tendency distribution maps of Se in plants are almost identically with those for Se in soils. However, at maturity, in grain and other plant parts useful in human and animal nutrition, normal levels of selenium were recorded, but cadmium contents were higher than normal.

Keywords: maize, selenium, soil, tandency maps


67

P19

A BROAD RANGE OF MINERAL CONTENTS AND POTENTIAL GENES LINKED TO HIGH IRON ACCUMULATION IN POTATO TUBERS FROM LUXEMBOURG

Sylvain Legay, Cédric Guignard, Johanna Ziebel and Danièle Evers.

Centre de Recherche Public - Gabriel Lippmann, Department EVA, 41, rue du Brill, L-4422 Belvaux, Luxembourg.

The lack of iron constitutes the main form of micronutrient deficiency in the world, namely iron defiency anemia, which strongly affects pregnant women and children from developing countries. Iron biofortification of major staple crops such as potato could thus be a way to improve the iron status of populations from these countries.

To better understand mechanisms leading to iron accumulation in potato, we harvested, in a multiplication field located in the north of Luxembourg, 21 commercial and old varieties.

First, mineral contents in the flesh (Fe, Zn, Mn, Mg, K, Na, Ca and Cu) were investigated using ICP-MS. As expected, principal component analysis performed on the complete data set displayed a strong correlation between iron and zinc content. Additionnally, we observed a broad range in tuber iron contents (13.66 µg.g DW-1 to 32.02 µg.g DW-1). Three varieties presented a tuber iron content above 25 µg.g DW-1, thereby constituting a promising germplasm.

Secondly, genes being involved in iron uptake, homeostasis and storage such as the IRT1 and NRAMP1 transporters, the FER-like transporters or the ferritins, were investigated at the gene expression level.

Both datasets were finally analysed in parallel in order to find some potential correlation between iron content in tubers and specific gene expression.

Keywords: Gene expression, iron, mineral content, potato


68

P20

WHEAT ORIGIN INFLUENCES GRAIN ZINC AND CADMIUM CONCENTRATION

Andrijana Rebekić1, Zdenko Lončarić1, Sonja Petrović1, Sonja Marić1

1)Faculty of Agriculture in Osijek, Kralja Petra Svačića 1d, HR–31000 Osijek, Croatia

Almost half of world population uses wheat as a major foodstuff, therefore wheat is important source of essential (zinc) and detrimental (cadmium) elements in their nutrition. Objective of this study was to examine difference in zinc and cadmium grain concentration between wheat varieties of different origin and year of recognition. Experiment was set up as completely randomized design with 52 winter wheat varieties (34 Croatian, 6 Austrian, 5 Hungarian, 3 French, 2 German, 1 Russian and 1 Italian), two levels of cadmium soil contamination (0 and 20 mg Cd kg-1 soil) and four replicates. According to year of recognition wheat varieties were classified into six groups. Results of Kruskal-Wallis test showed a significant difference in the distribution of the zinc and cadmium between the country of origin and year of recognition as well. In general, older varieties had higher zinc and cadmium concentration in compare to newer varieties. Croatian varieties had significantly different distribution of cadmium at contaminated and uncontaminated soil in compare to other examined varieties. Wheat origin and year of recognition influences zinc and cadmium in grain. Even they are not in commercial use anymore, older varieties could be used in biofortification since they have preferable zinc-cadmium ratio.

Keywords: Cadmium, origin, zinc, winter wheat


69

P21

THE USE OF THICKENERS AS INGREDIENTS. EVALUATION OF THEIR EFFECT ON MINERAL AVAILABILITY USING AN IN VITRO

DIGESTION MODEL

Carlos Alberto González-Bermúdez1, Frontela-Saseta C1, López-Nicolás Rubén1, Gomez-Gomez VP1, Miranda-Miranda L2, Legay S2, Corvisy A2, Klein S2, Evers D2,

Rincón-León F3 and Martínez-Graciá C1

1Department of Food Science and Nutrition. Faculty of Veterinary Sciences. University of Murcia. 30100 Murcia (Spain).

2DepartmentEnvironmentandAgro-biotechnologies (EVA). Centre de Recherche Public Gabriel Lippmann. L 4422, Belvaux (Luxembourg).

3Department of Bromatology and Food Science, University of Cordoba (Spain)

Objective: With the aim of increasing food viscosity, thickeners as Locust Bean Gum (LBG), Modified Corn Starch (MCS) and Modified Rice Starch (MRS) are used as ingredients. Nevertheless, some studies have indicated that these ingredients could affect mineral availability. The objective of this work was to clarify their effect, alone or combined, on Ca, Fe and Zn availability after in vitro digestion.

Methods: 10 different combinations of thickeners were evaluated. For each 100 g of thickeners mixture, a homogenised combination of CaCl2 (3 g), FeSO4 (0.03 g) and ZnSO4 (0.015 g) were. For solubility, 152.19 mg of each sample was in vitro digested and centrifuged in order to obtain the soluble fraction. For dialysability, 76.1 mg of each sample was in vitro digested with a dialysis bag during intestinal stage. The mineral content in the soluble fraction and the dialysate was determined by AAS, expressing the result as percentage respect the mineral content in the original sample.

Results: Whereas dialysability was similar among the different samples, the addition of thickeners seemed to have a negative effect on mineral solubility. When Ca was studied, all the combinations showed a decrease on its solubility respect the sample without thickeners, which was higher for the combination in which LBG was the only ingredient added. Regarding Fe and Zn, the effect on solubility showed a higher variability than the Ca solubility values, depending on the combination used.

Conclusions: The use of thickeners as LBG, MCS and MRS seems to affect mineral in vitro availability, affecting solubility and dialysability in a different way depending on the combination studied. In order to minimize the effect on mineral availability complementary studies should be done looking for the optimal combination.


70

P22

ARSENIC PHYTOEXTRACTION BY PTERIS VITTATA L. IN LINE WITH FROND CONVERSION BY SOLVOLYSIS FOR RESTORING HEALTHY CROP PRODUCTION

Michel Mench1, Fadila Kechit1, Marion Carrier2, Anne Lopinet-Serani2, Nathalie N. Caille3, Fang-Jie Zhao3 and Jaco Vangronsveld4

1)INRA, UMR BIOGECO 1202, University of Bordeaux 1, Talence, France 2)CNRS, University´of Bordeaux, ICMCB, IPB-ENSCBP, FR-33608 Pessac Cedex, France 3)Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, UK 4)Environmental biology, Hasselt university, B-3590, Diepenbeek, Belgium

The Reppel small-scale field experiment was set up in 1997, i.e. 10 lysimeters (surface area: 0.25 m2, depth: 0.5 m, surrounded by a 0.25-m compensation zone) filled with a Belgian soil polluted by atmospheric fallout from a former As smelter and 2 lysimeters with an uncontaminated soil, all placed in a greenhouse. Beringite (B, 5% w/w), iron grit (1% w/w) and their combination (BZ) were incorporated into the soil for immobilizing metals and As. Since 2004, Pteris vittata L. was cultivated for bioavailable As stripping. This aims at demonstrating As phytoextraction in line with frond conversion by sub- and supercritical water treatments to green fine chemicals and the restoration of ecosystem services, notably safety of following crop productions. Fronds were harvested in spring and autumn. Leachates were analysed for trace elements, pH and EC. Their phytotoxicity was assessed by radish germination. Restoration of the safety of crop productions was investigated with lettuce, radish and French bean. Root DW yield of lettuce was higher in BZ-treated soils and lower in the B-treated ones, whereas shoot DW yield did not differ across soil treatments. Shoot As concentration (in µg/g DW) varied from 1.28±0.25 to 2.5±0.5 and was lower in BZ-, B-, and Z-lettuces. Frond As and metal removals and As and metal leaching were periodically monitored. Frond biomass was converted by sub- and supercritical water treatments and organic products were identified.

Keywords: arsenic, food safety, lettuce, metal, phytoremediation Acknowledgements: This work was supported by ADEME, Dept Urban Landfills and Polluted Sites, Angers, France, the Aquitaine Region Council, and the European Commission under the Seventh Framework Programme for Research (FP7-KBBE-266124, GREENLAND).


71

P23

HOW TO PROCEED TO ACHIEVE THE HIGHEST POSSIBLE LEVELS OF RESVERATROL IN WINE

Juan P. Navarro-Aviñó, Juanjo Fernández Navarro, F. L., V. Vargas Castro, I. Ilzarbe Ripoll and M.J. Márquez Sahuquillo.

ABBA Gaia, Albert Einstein 100, 46980,Valencia,Spain Dagón Bodegas S.A.T, C/ Cooperativa 4, 46310 Los Marcos (Utiel-Requena) España

Objectives: The aim of this writing is to propose ways forward, in Agriculture, pollution and human health. Starting first by defining a way, of getting harmonize these three aspects. To this end it is proposed a philosophy of behavior, and a conceptual way of addressing the possible improvement of biotechnology as a tool. Finally, it is shown a way, to increase technical production of resveratrol, in a non-biotechnological form.

Methods: HPLC coupled to two different detectors and independently UV spectrophotometry. The resveratrol and other stilbenes content were identified and quantified in skins and pulp of “Bobal” grapes, and resveratrol also in wine.

Results: Versus 14,3 mg/l, maximum in the world reported, wines from “Bodegas Dagón” contain 1611,73 ± 72,66 mg/l of resveratrol.

Conclusions:

1) The Agrodynamic Agriculture implemented by "Bodegas Dagón" is capable of producing a remarkable biofortification process (understood in concentrations of stilbenes), in both, grapes and in the wine investigated.

2) The wine, bottled by "Bodegas Dagon", has a concentration of resveratrol, 100 times greater than the maximum reported up to date, and almost 1000 times higher than the median of all the world's wines studied in this work.

Key words: Agrodynamic, resveratrol, biofortification, bioavailability, neuroprotective


72

P24

INFLUENCE OF SELENIUM ON CARBOHYDRATE METABOLISM AND NODULATION IN ALFALFA (MEDICAGO SATIVA L.)

Anthony Owusu-Sekyere1, Juha Kontturi1, Helinä Hartikainen2, Roghieh Hajiboland3, Naser Aliasgharzad3 and Mervi M. Seppänen1

1)Department of Agricultural Sciences, University of Helsinki 2)Department of Food and Environmental Sciences, University of Helsinki 3)Department of Plant and Soil Sciences, University of Tabriz

At trace concentrations, selenium (Se) is an essential micronutrient for humans and animals. But it’s role in plants remains debatable. Studies have reported of beneficial Se effects in plants including growth, tolerance to abiotic stresses and energy metabolism. However, studies on Se effects on plant-microbe interactions are very limited. This study investigated Se effects on carbohydrate metabolism and nodulation in alfalfa. Pot and hydroponic experiments (for carbohydrates studies) with varying Se (0, 1, 5, 10 and 15 µmol L-1 Na2SeO4) and N (2 and 10 mmol L-1 Ca(NO3)2) concentrations were conducted. Seedlings in perlite were inoculated with Sinorhizobium meliloti for nodulation studies. Data were analyzed at different time points. Selenium increased F1,6-BPase activity and also, soluble sugars and starch in the shoots and roots of plants. Low Se (1 and 5 µmol L-1) increased nodule number about 40 % and 62 % respectively in young plants. Nodulation, however, decreased with growth. Nodule fresh weights and nitrogenase activity were not affected. Selenium also did not alter dry matter (DM) accumulation in plants. The results suggest that the up-regulation of carbohydrate metabolism may have synergistic effects on nodulation. More detailed studies are needed to fully understand the role of Se in N2-fixation.

Keywords: Selenium, Medicago sativa, Nodulation, Carbohydrates


73

P25

ANTIMONY ACCUMULATION IN TRIFOLIUM SPECIES

Isabel Corarles, Juan Barceló and Charlotte Poschenrieder

Plant Physiology Laboratory, Bioscience Faculty, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain

Natural concentrations of antimony (Sb) in the environment are low. Higher concentrations of Sb in soil are usually a result of human activities. Soil contamination by antimony has become an environmental problem in recent years due to increasing mining and industrial use. Antimony is a nonessential element for both plants and animals. Antimony is toxic to humans at chronic uptake rates exceeding 100 mg/d. Severe Sb pollution poses a great threat to food safety and human and animal health. In the present study, we investigated the effect of 0, 50, 100 and 200µM SbV in the form of KSb(OH)6 on growth and Sb uptake of clover (Trifolium pratense cv. Milvus and Trifolium repens). In spite of Sb accumulation both in roots and shoots of this fodder plant, any negative effect on root growth, cellular viability (Evans blue and vital staining) and lipid peroxidation (MDA content) was observed. Antimony tolerance

seems not related to phytochelatins because supply of the ɣ-glutamylcysteine synthetase

inhibitor, L-buthionine-[S,R]-sulphoximine (BSO) did not enhance toxicity. Due to the absence of toxic visual effects in plants, Sb can be inadvertently incorporated into the trophic chain causing important toxic effects both in animals and humans. (Supported by project BFU2010-14873).

Keywords: Antimony, clover seedlings, tolerance, trophic chain, uptake


74

P26

ASSESSMENT ON THE NEGATIVE EFFECTS OF EXCESSIVE ACCUMULATION OF MN IONS BY SELECTED LETTUCE CULTIVARS

Arkadiusz Przybysz, Wrochna Mariola, Yahya Noor Ain Binti, Pietrzyk Sylwester and Gawroński Stanisław Waldemar

Laboratory of Basic Research in Horticulture; Faculty of Horticulture, Biotechnology and Landscape Architecture; Warsaw University of Life Sciences – SGGW; 159 Nowoursynowska Street, 02-776 Warsaw, Poland; email: [email protected]

Objective: In this study, an attempt was made to evaluate effects of Mn on lettuce (Lactuca sativa L.) cv. ‘Satine’ (red-leafed) and ‘Locarno’ (green-leafed).

Methods: Plants were grown in hydroponic culture and treated with Mn in concentrations of 0.5 (control), 5, 25 and 50 mg/dm3. During plants growth (i) photosynthetic apparatus efficiency and (ii) transpiration rates were examined. At harvest, data on (iii) biomass accumulation, (iv) levels of ROS, (v) activity of anti-oxidative system enzymes and (vi) ions content was recorded.

Results: Obtained results showed increased biomass accumulation at lowest Mn concentration and decreased at higher. Chlorophyll content was higher for plants grown in presence of elevated Mn. Effect of Mn on photosynthesis intensity was negative when plants were exposed to higher metal concentrations. Mn had no effect on Fv/Fm. Increased values of P.I. and Yield were noted for ‘Satine’ plants treated with Mn. Red-leafed plants in presence of Mn had stomatal conductance leading to greater transpiration rates but the opposite was true for green-leafed. Impact of Mn on ROS (anion-radical, hydroxyl radical) and activity of enzymes of anti-oxidative system (catalase, ascorbate peroxidase) was diverse and dependent on Mn concentration, examined cv. and plant organ. Evaluation of ions content is in progress.

Keywords: biomass accumulation, lettuce, manganese, oxidative stress, photosynthetic apparatus


75

P27

APPLES BIOFORTIFIED WITH CALCIUM

Vânia Ribeiro1, Cláudia Sánchez2, Maria Paula Duarte1, Paula Vasilenko2, Mário Santos2 and Fernando Lidon1

1)Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal 2)INIAV, I.P., Quinta do Marquês, Oeiras, Portugal

Calcium is one of the most important mineral elements in any healthy human diet. Adequate calcium intake from childhood to the end of the life span is essential for the formation and retention of a healthy skeleton. The objective of this study was to analyze the ability of apples Malus domestica, varieties Jonagold and Golden Delicious, for calcium biofortification by three different pre-harvest calcium treatments. Laboratory and field experiments were carried out to assess the efficacy of different treatments. Golden variety showed higher calcium assimilation than Jonagold variety. The results provide evidences that Ca2+ represent an important contribution to reduce the incidence of Ca-related disorders over the postharvest cold-storage period and increased the calcium content in apples. The results showed that apple is a good matrix for calcium biofortification.

Keywords: Apples, biofortification, Calcium, Malus domestica


76

P28

CONCENTRATION AND SPECIATION OF SELENIUM (SE) IN BIOFORTIFIED CHICKPEA (CICER ARIETINUM L.) UNDER MEDITERRANEAN CONDITIONS

Sara M. Rodrigo1, María J. Poblaciones1, Oscar Santamaria1, Teodoro García-White1, Yi Chen2 and Steve P. McGrath2

1)Department of Agronomy and Forest Environment Engineering, University of Extremadura, Avda. Adolfo Suárez s/n, C.P. 06007, Badajoz, Spain

2)Sustainable Soils and Grassland Systems, Rothamsted Research, Hertfordshire AL5 2JQ UK email: [email protected]

Selenium (Se) is an essential dietary component for animals including humans. Low intake of Se can cause health disorders including oxidative stress-related conditions, epilepsy, fertility reduction, immune deficiency, etc. To improve the nutritional value of chickpea, two foliar Se fertilizers (sodium selenate and sodium selenite) at four rates (0-10-20-40 g ha-1) were tested in the 2010-2011 and 2011-2012 growing seasons in a field experiment conducted under semiarid Mediterranean conditions. Sodium selenate was much more effectively taken by plants than sodium selenite, and there was a strong and linear relationship between total Se content and Se rate for both. For each gram of Se fertilizer, applied as sodium selenate or sodium selenite, the increases of total Se concentration in grain were 126 and 87 and 25 and 19 μg Se kg-1 dry weight in 2010-2011 and 2011-2012, respectively. Se was found to be incorporated in chickpea grains mainly (>70%) as selenomethionine. Thus, Se-enriched chickpeas would be a potentially good way of introducing Se into the human diet, and hence may be a very good candidate to be included in Se biofortification programs.

Keywords: Agronomic biofortification, HPLC-ICP-MS, Legumes, Selenate, Selenite


77

P29

ASSESSMENT OF ARSENIC IN HYBRID AND LOCAL RICE CULTIVARS GROWN IN AN ARSENIC HOTSPOT OF BANGLADESH

Arifin Sandhi1, Maria Greger2,3, Gunnar Jacks1 and Prosun Bhattacharya1

1)Department of Land and Water Resources Engineering, KTH Royal Institute of Technology, Teknikringen 76, SE-100 44 Stockholm, Sweden

2)Faculty of Applied Ecology and Agricultural Sciences, Hedmark University College, Blæstad, N-2418 Elverum, Norway

3)Department of Ecology, Environment and Plant Sciences, Stockholm University, Lilla Frescati, 106 91 Stockholm, Sweden

Cultivation of high yielding varieties (HYV) of rice (Oryza Sativa) in Bangladesh is considered as one of the major agricultural activities compared with local rice cultivars due to its high production rate. The high level of arsenic (As) in the groundwater and soil of Bangladesh is already well-documented. Most of the rice cultivars require irrigation applications for their growth. The aim of this study was to investigate the arsenic accumulation in the HYV and local rice grain those cultivated in an As hotspot (Matlab) in Bangladesh. The ICP-OES based analysis of irrigation water of those particular areas was found the concentration (>200 µg As l-1) and the soil was (5.75-36.65 mg As kg-1). The total As concentration in the grain of the HYV rice varieties was lower than in the local rice cultivars. However, the As concentration in husk from HYV grains were higher than the rest of the grain while the opposite was found in the local rice cultivars. Our preliminary findings have suggested that the husk of the rice grain could play an important role for arsenic enrichment in both the HYV and the local rice cultivars when they have grown in an As contaminated area.

Keywords: Accumulation, arsenic, Bangladesh, husk, hybrid rice


78

P30

INFLUENCE OF CADMIUM EXPOSURE ON SORGHUM PLANTS

Petrová Šárka, Soudek Petr, Vaněk Tomáš

Laboratory of Plant Biotechnologies, Joint Laboratory of Institute of Experimental Botany AS CR and Crop Research Institute, Rozvojová 263, 165 02 Prague 6, Czech Republic

Increased concentrations of essential elements in agricultural soils mean higher plant uptake. However, soil also contains toxic elements, such as heavy metals, and the higher concentration, the higher uptake of nonessential elements is. What’s more, the concentrations of these metals are so toxic they can cause death of plants or reduce their production. Its growth on contaminated soil can reduce the pollution through a transfer of the contaminant from soil to harvestable plant parts. Sorghum bicolor L. is an important crop due to its wide use. Due to a high biomass production and known agronomic techniques, sorghum can be used as an energy crop and at the same time it can clean up the environment. The aim of this study was to expand knowledge about protection mechanisms of different species of sorghum (S. bicolor var. eusorghum, S. bicolor x S. sudanese) under cadmium stress. The effect of combination of Cd with essential metals such as Cu, Fe, and Zn was also studied.

Keywords: abiotic stress, Sorghum bicolor, heavy metals, toxicity Acknowledgement: This work was supported by project FR/Tl3/778.


79

P31

DIFFERENTIAL SUFUR METABOLISM EFFICIENCY IN CADMIUM ACCUMULATION CAPACITY

Souhir Sghayar1,2, Chedly Abdelly2 and Gian Attilio Sacchi1

1)Dipartimento di Produzione Vegetale, Università degli studi di Milano (Italy) 2)Laboratory of Extremophiles Plants, Biotechnology Centre of Bordj Cedria (Tunisia)

Corresponding author: [email protected]

Since mobile in the soil-plant-system, cadmium can be accumulated in the shoots and in the edible parts of crops grown on contaminated soils. A short term approach for reducing Cd risk for human health could consists in the selection of cultivars efficiently excluding the metal from the shoot. The aim of this work is to compare two barley (Hordeum vulgare L.) cultivars found with relatively high (Lemsi) and low (Manel) Cd accumulations in shoot and grain based on the implication of sulfur metabolism on cadmium detoxification. When grown in

hydroponic solution with Cd (0-1 μM), the influx of the metal into the roots was higher for Lemsi, but its total concentration resulted practically similar to that of Manel. It was due to a higher level of phytochelatins in Manel and to a higher xylem loading of Cd in Lemsi. These finding were also correlated to differential accumulation in total sulphur, sulphate and glutathione levels.

Keywords: cadmium, phytochelatins, glutathione, Hordeum vulgare.


80

P32

COMPOST DECREASES HEAVY METAL ACCUMULATION IN VEGETABLES

Stefan Shilev1, Todor Babrikov2 and Rossitsa Petrova3

1)Dept. of Microbiology and environmental biotechnologies, E-mail: [email protected] 2)Dept. of Horticulture, Agricultural University – Plovdiv, Plovdiv 3)Dept. of Soil science, University of Forestry, Sofia, E-mail: [email protected], Bulgaria

Objectives

The main objective of present study was to immobilize the bioavailable fractions of heavy metals in soil, decreasing their content in spinach and radish plants grown on industrially contaminated soil.

Methods

Heavy metal contaminated soil was used mixed with compost in two experiments: growth chamber immobilization experiment mixing soil and compost (1 or 10 %); greenhouse immobilization experiment, growing spinach or radish plants in containers with contaminated soil with or without compost. Total and DTPA-extractable Cd in soil was studied before, during and in the end of experiments. Heavy metals content was analized also in plant tissue. During the experiments different plant and soil parameters were observed.

Results

Generally, the incorporation of compost increased in folds the soil microbial enzyme activity and respiration and promoted the growth of spinach plants. A good immobilization effect in case of spinach plants was observed for cadmium, while in case of radish plants, significant immobilization was reached for cadmium and lead.

Conclusions

The incorporation of organic matter end-product of pharmaceutical industry could be an interesting and cost-effective way to protect plants in low contaminated soils, but also could provide important nutrients.

Keywords: compost, food safety, cadmium, lead, immobilization, vegetables


81

P33

TESTING SINGLE AND COMBINATIONS OF AMENDMENTS FOR ASSISTED PHYTOEXCLUSION OF POTENTIALLY TOXIC TRACE ELEMENTS

Grzegorz Siebielec1, Petra Kidd2, Monika Pecio1, Rafal Galazka1, Michel Mench3, Vanessa Álvarez-López2, Beatriz Rodríguez-Garrido2, Jaco Vangronsveld4,

Wolfgang Friesl-Hanl5, Andy Cundy6 and Markus Puschenreiter7

1)Institute of Soil Science and Plant Cultivation, Pulawy, Poland, email: [email protected] 2)CSIC, Santiago de Compostela, Spain 3)UMR BIOGECO INRA, University of Bordeaux, France, France 4)Universiteit Hasselt, Diepenbeek, Belgium 5)AIT Austrian Institute of Technology GmbH, Tulln, Austria 6)University of Brighton, United Kingdom 7)BOKU, University of Natural Resources and Applied Life Sciences, Vienna, Austria

Soil amendments may limit the risk related to the soil contamination through reduction of metal transfer to the food chain. The objective of our studies was to compare effectiveness of various materials in stabilizing metals in Zn/Pb/Cd/As contaminated soils and reducing their transfer to crops. Within the EU FP7 Greenland project we compared the impact of novel soil amendments and their combinations with traditional materials on metal solubility and the response of plants, contaminant uptake, behaviour of soil organisms and microbial activity.

A greenhouse pot experiment was established on agricultural soil contaminated through long-term Zn/Pb smelter emissions in Poland (pH 7.0, total Zn, Pb and Cd: 2670, 690 and 30 mg kg-1, respectively). The following treatments were tested as single amendments and in different combinations: compost, drinking water residue, iron grit, Ca-phosphate, LD slag, Thomas basic slag, gravel sludge, siderite, and cyclonic ash. Reagent grade CaCO3 was added to soils if needed in order to establish soil pH at comparable level in all treatments. The soil was seeded with lettuce. The presentation will summarize effectiveness of the broad range of soil amendments in assisted phytoexclusion of trace elements.

Keywords: bioavailability, contaminated soil, phytoexclusion, soil amendments, trace elements


82

P34

ASSESSEMENT OF ZINC BIOAVAILABILITY IN HUMANS FROM ZINC BIOFORTIFIED WHEAT INTRINSICALLY LABELLED WITH 67ZN

Coralie Signorell, Diego Moretti, Christophe Zeder, Rita Wegmüller, Ines Egli, Aciksöz Özden Seher Bahar, Emmanuel Frossard and Michael Zimmermann

ETH, Zürich, Switzerland

Background: Zinc bioavailability in humans can be determined with a double isotopic method. It is assumed that the extrinsic label given orally will homogeneously mix and be similarly absorbed as the naturally embedded zinc in the food matrix. Confirmation of this is of particular interest for bio fortified varieties where the totality of the Zn is embedded in the food matrix.

Objectives:

1) Characterize wheat grown on stable isotope enriched mediums (intrinsically labeled wheat)

2) to assess the bioavailability from intrinsically vs. extrinsically labeled wheat with different zinc concentrations.

Methods: Eighteen subjects will consume 3 different test meals in randomized order of which the fractional absorption of Zn will be measured. Extrinsic zinc will be added to bread rolls as 67ZnSO4, while hydroponically grown wheat will contain two concentrations of 67Zn. An intravenous dose 70Zn label will be administered in concomitance with the test meals.

Results: The Zn concentration in the wheat grains at the milk maturing stage (2 weeks before full maturation) wheat was quantified by AAS.

The grains contained 26.29, 18.47, 40.90 μg Zn/g for control, intrinsically labeled and intrinsically bio-fortified wheat, respectively.

Conclusion: The intrinsically labeled wheat is suitable for the conduction of a human study which is currently being planned.


83

P35

EFFECT OF BISPHOSPHONATES ON METAL ACCUMULATION IN THE METAL HYPERACCUMULATOR PLANT NOCCAEA CAERULESCENS

Alanne Aino1, Tuomainen Marjo2, Turhanen Petri1, Peräniemi Sirpa1, Vepsäläinen Jouko1, Kärenlampi Sirpa2 and Tervahauta Arja2

1)School of Pharmacy and 2)Department of Biology, Kuopio Campus, University of Eastern Finland

Bisphosphonates (BPs) have many pharmaceutical and chemical applications. They have two phosphonate groups and two substituents bound to the central carbon. The longer hydrocarbon side-chain may contain nitrogen (N-BP), affecting the chemical activity. BPs chelate metal cations effectively and, as diphosphate analogues, can interfere with several enzymatic reactions. In mammals and microbes N-BPs inhibit farnesyl-diphosphate synthase, a key enzyme in mevalonate pathway that produces e.g. phytohormones in plants.

In the present study the effects of BPs on leaf biomass and metal uptake was studied in greenhouse-grown Noccaea caerulescens. Three-week-old seedlings were exposed to metals (0.05 mmol/kg Cd or Ni; 1 mmol/kg Zn or Pb) with or without BPs (three non-N-BPs, four N-BPs; 0.25, 5 mmol/kg) in molar ratio 1:5 (metal: BP). After 8-week-exposure the shoots were weighed and roots were washed and treated with 0.5 mM EDTA/0.5 mM citrate. Samples were analysed with AAS.

Low BP concentration increased and high concentration decreased leaf biomass. Metal exposures decreased leaf biomass, except when combined with one of the N-BPs. Some bisphosphonates increased metal uptake. Leaf Ni content increased with one N-BP, while Pb content was the same as in Pb-exposed plants. As a conclusion, BPs can increase plant biomass and metal uptake.

Keywords: Accumulation, biomass, bisphosphonate, metal, Noccaea caerulescens


84

P36

CHARACTERIZATION OF HUMAN GASTROINTESTINAL JUICES INTENDED FOR USE IN IN VITRO MODEL DIGESTION.

Ellen K. Ulleberg1, Irene Comi1, Halvor Holm2, Espen B. Herud3 and Gerd E. Vegarud1

1)Norwegian Univ Life Sci, Dept Chem Biotechnol & Food Sci, N-1432 As, Norway 2)University of Oslo, Department of Nutrition, Institute for Basic Medical Sciences, 0316 Oslo,

Norway 3)Oestfold Hospital Trust, Fredrikstad, Norway E-mail: [email protected]

Human gastric (HGJ) and duodenal juices (HDJ) have been characterized individually and as a pooled batch to be used in in vitro model digestion for the comparison with commercial enzymes of non-human origin. In addition storage stability of the human gastrointestinal juices was examined. Gastric and duodenal juices were aspirated from healthy adults using a triple lumen tube at the Hospital of Oestfold. Individual variations in enzymatic activities as well as total volumes, pH, bile acids, protein and bilirubin concentrations were recorded. Individual pepsin activity in the gastric juice varied by a factor of 10, while individual total proteolytic activity in the duodenal juice varied by a factor of five. The duodenal amylase activity varied from 0 to 52.6 U/ml and the bile acid concentration varied from 0.9 to 4.5 mM. Pooled gastric and duodenal juices from 18 volunteers were characterised according to pepsin activity, total proteolytic activity, lipase activity, amylase activity and bile acids. The stability of the main enzymes in frozen or freeze dried batches with or without additions of glycerol or protease inhibitors was studied for 6 month and up to 1.5 year.

We designed an in vitro digestion study of one milk protein, lactoferrin (LF), simulating intralumenal conditions and by using human gastrointestinal (GI) enzymes. Differences in the concentration of LF, gastric pH and the amount of GI enzymes affected the degradation of LF and the released peptides.

All the human aspirations were approved by the Norwegian Ethical Board.

Keywords: In vitro model digestion, human gastrointestinal enzymes


85

P37

IDENTIFICATION OF CANDIDATE GENES ASSOCIATED WITH QTLS FOR CADDMIUM TOLERANCE IN ARABIDOPSIS HALLERI

Claire-Lise Meyer1, Elena Chaves-Rodriguez1, Piétro Salis1, Cécile Godé2, Pierre Saumitou-Laprade2 and Nathalie Verbruggen1

1)Laboratoire de Physiologie et Génétique Moléculaire des Plantes, Université Libre de Bruxelles, Campus Plaine Bat BC Boulevard du Triomphe, 1050 Brussels, Belgium

2)Laboratoire Génétique et Evolution des Populations Végétales, UMR CNRS 8198, Université de Lille 1, Cité Scientifique, 59650 Villeneuve d’Ascq, France

On sols highly polluted by trace metallic elements the majority of plant species are excluders, limiting the entry and the root to shoot translocation of trace metals. However a rare class of plants called hyperaccumulators possess remarkable adaptation because those plants combine extremely high tolerance degrees and foliar accumulation of trace elements. In the Zn hyperaccumulator A. halleri, Cd tolerance seems constitutive in the species and some populations also hyperaccumulate cadmium (Cd). In one of the Cd hyperaccumulating population of A. halleri, three QTLs for Cd tolerance were found using an interspecific cross (BC1) between this species and a non-tolerant relative [1]. The major QTL was shown to co-localize with the Heavy Metal ATPase 4 gene which was functionally validated. Nevertheless, it was also demonstrated that Cd detoxification in A. halleri requires expression of additional genes. Therefore we have fine-mapped the two other QTL (CdTol2 and CdTol3, which are different from the QTls for Zn tolerance) by using a larger mapping population (BC1 progeny) and increasing the number of markers in this region. This allowed identifying a potential candidate gene in the QTLCdTol2. This gene is over-expressed in A. halleri compared to non-tolerant relatives and its expression is correlated to Cd tolerance in the BC1 progeny. Investigation of candidate genes for the QTL CdTol3 is still ongoing.

In addition, we have observed an important variability of Cd tolerance and accumulation in A. halleri populations, which suggests that only a part of the genetic architecture of these traits has been revealed

Keywords: cadmium, fine-mapping, hyperaccumulator, tolerance


86

P38

NUTRITIONAL QUALITY OF SELECTED LETTUCE CULTIVARS ENRICHED WITH ZN

Mariola Wrochna, Szymon Jabłoński, Arkadiusz Przybysz and Stanisław Waldemar Gawroński

Laboratory of Basic Research in Horticulture; Faculty of Horticulture, Biotechnology and Landscape Architecture; Warsaw University of Life Sciences – SGGW; 159 Nowoursynowska Street, 02-776 Warsaw, Poland; email: [email protected]

Objective:

In this work an attempt was made to assess (i) possibility to enrich selected Lactuca sativa L. cv. ‘Amadeusz’ and ‘Omega’ with Zn ions and (ii) effect of increased Zn concentrations on selected parameters/processes in treated plants.

Methods:

Plants were grown in hydroponic culture in concentrations of 0.1 (control), 0.2, 0.3 and 0.4 mg Zn/dm3. During plants growth (i) photosynthetic apparatus efficiency and (ii) transpiration rates were examined. At harvest data on (iii) biomass accumulation, (iv) levels of ROS, (v) activity of enzymes of anti-oxidative system and (vi) selected ions content was recorded.

Results:

Addition of Zn had positive effect on biomass accumulation, efficiency of photosynthetic apparatus (chlorophyll content, photosynthesis intensity, stomatal conductance and chlorophyll a fluorescence) and transpiration rate in plants of cv. Amadeusz. In Omega cv. these parameters were usually lowered, especially in higher Zn concentrations. Effect on ROS level depended on Zn concentration, cultivar and plant organ. Activity of antioxidative system enzymes decreased in cv. Amadeusz, while in Omega was usually higher. Level of ROS and antioxidative compounds, beside Zn content, is critical parameter characterizing quality of functional food, as external source of these compounds may influence human health. Evaluation of ions content are in progress.

Keywords: biofortification, lettuce, oxidative stress, photosynthetic apparatus, zinc


87

P39

ROOT STRUCTURAL VARIABILITY IN NOCCAEA CAERULESCENS

Ivan Zelko1, Lišková Desana1, Sterckeman Thibault2 and Alexander Lux1,3

1)Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-84538 Bratislava, Slovakia

2)Laboratoire Sols et Environnement ENSAIA-INRA, 2, av. de la Forêt de Haye TSA 40602, 54518 Vandoeuvre-lès-Nancy, France

3)Department of Plant Physiology, Faculty of Natural Sciences, Comenius University in Bratislava, Mlynská dolina B-2, SK-84215 Bratislava, Slovakia

Noccea caerulescens, a hyperaccumulator of cadmium is known for the variability between its populations. Another feature of this species is the presence of peri-endodermal layer. The goal of this study was to compare root structure of several populations from France and Slovakia. Influence of certain environmental factors (Cd, osmoticity) was also studied. Hydroponically and aeroponically grown roots were fixed with methanol and embedded in agarose. Free-hand sections were stained by Flourol yellow 088 and observed under UV light. The root structure of N. caerulescens (including peri-endodermal layer development) varies across the populations. The reaction of peri-endodermal layer to the environmental factors seems to be independent from suberin lamellae development.

This study was supported by the Slovak Research and Development Agency under the contract No. APVV-0140-10 and COST Action FA-0905.

Keywords: Noccaea caerulescens, peri-endodermal layer, root structure, suberin lamellae


88


LIST OF PARTICIPANTS


89

Aciksöz Özden, Seher Bahar Switzerland [email protected] P1, P34

Alvarenga, Paula Portugal [email protected] P2

Antosiewicz, Danuta Maria Poland [email protected] A15, P3

Assunção, Ana Portugal [email protected] A9 Atala Lombelo Campos, Ana Carolina Netherlands anacarolina.atalalombelocampos@

wur.nl A10

Bakke, Mona Norway [email protected] P4, P14, P15

Baltrėnaitė, Edita Lithuania [email protected] P5

Barceló, Juan Spain [email protected] P6, P25

Basińska, Aneta Poland [email protected] A22

Bhullar, Navreet Switzerland [email protected] K5

Bittsánszky, András Hungary [email protected] P7

Bjelková, Marie Czech Republic [email protected] P8

Borg, Søren Denmark [email protected] P3

Burgos Zapata, Gabriela del Pilar Peru [email protected]

Cakmak, Ismail Turkey [email protected] P1

Cercamondi, Colin Switzerland [email protected] A23

Clemens, Stephan Germany [email protected] A11, A15

Colzi, Ilaria Italy [email protected] P9

Comino, Elena Italy [email protected] P10

Curie, Cathrine France [email protected] K3

Dahl, Signe Norway [email protected]

Dahlin, Sigrun Sweden [email protected] P11

Deng, Fenglin Japan [email protected] A12

Dragicevic, Vesna Serbia [email protected] A7

Dürr-Auster, Thilo Switzerland [email protected] A6

Eagling, Tristan United Kingdom [email protected] A18

Evers, Danièle Luxembourg [email protected] A21, A24, P19, P21

Fairweather-Tait, Susan United Kingdom [email protected] A18

Frontela-Saseta, Carmen Spain [email protected] A21, P21

Furini, Antonella Italy [email protected] A14

Gonnelli, Cristina Italy [email protected] P9 González-Bermúdez, Carlos Alberto Spain [email protected] A21, P21

Gramlich, Anja Switzerland [email protected] A5


90

Greger, Maria Sweden [email protected] A1, P12, P29

Griga, Miroslav Czech Republic [email protected] P8, P13

Haugene, Mari Norway [email protected] P14

Heim, Michael Norway [email protected] P4, P15

Herzig, Rolf Switzerland [email protected] P16

Hoffland, Ellis Netherlands [email protected] K2

Hunt, Janet Austria [email protected] K8

Hurrell, Richard Switzerland [email protected] A17, A23

Karyotis, Theodore Greece [email protected] P17

Kendziorek, Maria Poland [email protected] P3

Korslund, Liv Marit Norway [email protected]

Krzeslowska, Magdalena Poland [email protected]

Kärenlampi, Sirpa Finland [email protected] P35

Lăcătușu, Radu Romania [email protected] P18

Landberg, Tommy Sweden [email protected] A1, P12

Legay, Sylvain Luxembourg [email protected] A21, A24, P19, P21

Lombnæs, Peder Norway [email protected]

Lončarić, Zdenko Croatia [email protected] [email protected]

A2, P20

López-Nicolás, Rubén Spain [email protected] P21

Lux, Alexander Slovakia [email protected] A13, A16, P39

Lönnerdal, Bo United States [email protected] K6

Ma, Jian Feng Japan [email protected] K4, A12

Manojlović, Maja Serbia [email protected] A2

Mench, Michel France [email protected] P22, P33

Moretti, Diego Switzerland [email protected] P34

Munkeby, Anne Elisabeth Norway [email protected]

Navarro-Aviñó, Juan Spain [email protected] P23

Nieuwenhuis, Anja Norway [email protected]

Ovesna, Jaroslava Czech Republic [email protected] A20

Owusu-Sekyere, Anthony Finland [email protected] A4, P24

Petrová, Šárka Czech Republic [email protected] P30

Petry, Nicolai Switzerland [email protected] A17

Poblaciones, María Spain [email protected] A19, P28


91

Pogrzeba, Marta Poland [email protected] A3

Poschenrieder, Charlotte Spain [email protected] P6, P25

Przybysz, Arkadiusz Poland [email protected] P26, P38

Reddy, Manju USA [email protected] K7

Rengel, Zed Australia [email protected] K1

Ribeiro, Vânia Portugal [email protected] P27

Rodrigo, Sara Morales Spain [email protected] A19, P28

Sandhi, Arifin Sweden [email protected] P30

Santamaría, Oscar Spain [email protected] A19, P28

Schröder, Peter Germany [email protected]

P7

Schulin, Rainer Switzerland [email protected]

Seppänen, Mervi M. Finland [email protected] A4, P24

Sghayar, Souhir Italy [email protected] P31

Shilev, Stefan Bulgaria [email protected] P32

Siebielec, Grzegorz Poland [email protected] P33

Signorell, Coralie Switzerland [email protected] P34

Singh, Bal Ram Norway [email protected] A2

Stolz, Eva Sweden [email protected]

Tervahauta, Arja Finland [email protected] P35

Tracz, Katarzyna Poland [email protected] A15

Vaculík, Marek Slovakia [email protected] A16

Vegarud, Gerd Elisabeth Norway [email protected] P36

Verbruggen, Nathalie Belgium [email protected] P37

Wrochna, Mariola Poland [email protected] P25, P38

Zelko, Ivan Slovakia [email protected] [email protected]

P39

zum Felde, Thomas Peru [email protected]


92


NOTES


93


94


95


96


MAP – TRAIN – BUS SCHEDULES


97

MAP –FROM ÅS STATION, CENTRUM TO UMB AND KORSEGÅRDEN


98

MAP – CAMPUS ÅS


99


100

TIME SCHEDULE FOR TRAIN

OSLO - SKI - ÅS - MOSS PERIOD 09.06 – 23.06.2013 (‘Mandag – fredag’ = Monday – Friday; ‘Lørdag’ = Saturday; ‘Søndag’ = Sunday) (‘Første avg.’ = First departure; ‘Deretter min. over hel time’ = From then minutes past each hour: ‘Til’ = To; ‘Siste avg.’ = Last departure)


101

MOSS - ÅS - SKI – OSLO PERIOD 09.06 – 23.06.2013


102

TIME SCHEDULE FOR BUS

SKI - ÅS STATION - UNIVERSITETET I ÅS = UMB – BRØNNERUD SKOLE - DRØBAK (BRØNNERUD SKOLE = CLOSE TO KORSEGÅRDEN - FOR BUS F11 TO OSL


103

DRØBAK – BRØNNERUD SKOLE (= CLOSE TO KORSEGÅRDEN, BUS F11 FROM OSL) – UNIVERSITETET

I ÅS – AUD MAX (= SAMFUNNET = LUNCH) – ÅS STATION – SKI


104

F11 FLYBUSSEKSPRESSEN

FROM OSLO LUFTHAVN = OSL GARDERMOEN TO KORSEGÅRDEN, ÅS AND FREDRIKSTAD D = Dayly; X67= exception, not on Saturday(6) & Sunday(7)

F11 FROM FREDRIKSTAD TO KORSEGÅRDEN, ÅS - TO OSLO LUFTHAVN = OSL GARDERMOEN


105


FROM OSLO LUFTHAVN = OSL GARDERMOEN TO SKI STATION, THON HOTEL


106


FROM SKI TERMINAL, STATION, THON HOTEL, TO OSLO LUFTHAVN = OSL GARDERMOEN

€¦ · essential and detrimental trace elements entering the food chain via plants 3 welcome dear...

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