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Climate change: challenge for training applied plant scientists

Final Report Summary - AGRISAFE (Climate change: challenge for training applied plant scientists)

Project context and objectives:

Climate variability has a fundamental influence on agro-ecosystems. For a substantial part of Europe, climate change scenarios forecast significant decreases of up to 20 % in plant productivity, coupled with a general decline in the stability of agricultural ecosystems. This is particularly true of the Carpathian Basin. Agriculture and food safety are extremely sensitive to climate changes, so adaptability to stress is likely to gain priority over the quantitative aspects of yield. This will demand new approaches both to plant breeding and crop production, and in research strategy. The Agricultural Research Institute of the Hungarian Academy of Sciences (ARI HAS) is one of the leading centres for crop research and breeding in Central and Eastern Europe, in the Central Transdanubian convergence region, with a profile involving complex, interdependent, basic, methodological and applied research projects culminating in practical applications. Based on its international recognition, accumulated knowledge and close contacts with farmers and processors, the institute aims to develop into a regional Research Training and Service Centre to train and develop researchers, breeders and producers capable of offering practical help to farmers in Central and Eastern Europe in countering the unfavourable effects of predicted climate change. An increase in research potential is envisaged through strategic partnerships based on existing international contacts, whereby young scientists could gain valuable know-how and experience abroad, while experienced colleagues would be invited to work at ARI HAS. The results will be published both in scientific papers, and in talks and pamphlets aimed at farmers and food consumers in general, to raise awareness of the likely effects of climate change and of how these can be mitigated in the interests of achieving secure food supplies.

Project results:

WP1: Exchange of know-how and experience by developing strategic partnerships (including twinning) with well-established research teams in the European Union (EU)
Coordinator: Dr Beáta Barnabás

The clarification of the biological background to the interactions between the plant and its environment is important for the following reasons:

- The negative effects of climate change on the life processes of crop plants has a great influence on yield potential, thus exerting an effect on the ability of the world to achieve sustainable food and fodder production.
- The study of the genes responsible for the morphological traits and metabolic processes that determine plant responses and of how they are expressed helps scientists to breed plant varieties resistant to environmental stress factors.
- The achievement of optimum production conditions greatly promotes the exploitation of the genetic potential latent in various crops.

Special attention is paid in the Martonvásár Institute to the complex investigation of the environmental stress tolerance of cereals, from the molecular level to the whole plant level. The system of chambers available in the institute for artificial plant growth (phytotron) provides excellent facilities for this research, as it allows drought and temperature stress to be simulated in an exact manner. A considerable number of young scientists from the institute are involved in research on this topic. The AGRISAFE project provided an excellent opportunity for them to develop their knowledge in foreign laboratories working on similar subjects.

All of those who made study trips for training abroad carried out tasks related to their research in the institute on the effect of climate change. The partner institutions also had the opportunity to send experienced or young scientists to Martonvásár, not only to attend the training courses and symposiums, but also to exploit the special facilities available in the phytotron and to learn various experimental methods developed in the institute.

Scientists received training in the following fields:

1. Study of stress tolerance using molecular genetic tools and cell biology

Tibor Kiss PhD student spent seven months (16 November 2009 - 17 December 2009 and 11 January 2010 - 30 June 2010) at the Dipartimento di Scienze Biomolecolari e Biotecnologie (DSBB), Milano, Italy, where his project leader was Prof. Mirella Sari Gorla.

He is a young scientist working at the Cereal Breeding Department of ARI HAS, who needed to learn new experimental techniques in the field of molecular breeding. He joined a running project in DSBB: the isolation of pollen-specific genes and the examination of phylogenetic relationships between the Ga1 gene in maize inbred lines and other Zea species.

Katalin Jäger PhD spent almost six months (1 October 2009 - 11 March 2010.) in Spain at the Department of Plant Biochemistry, Cell and Molecular Biology, Estación Experimental del Zaidín of the Consejo Superior de Investigaciones Científicas (CSIC), Profesor Albareda 1, Granada, where her project leader was Dr Adela Olmedilla.

The Department of Plant Cell Biology at the Agricultural Research Institute of HAS and the Department of Plant Biochemistry, Cell and Molecular Biology at EEZ CSIC Granada have cooperated in research on the effect of environmental stress on in vitro microspore development and plant reproduction for the past ten years. Both partners are involved in work on the in situ hybridisation technique, and are interested in learning and adapting immunogold labelling methods for cereal species. The continuing exchange of experienced scientists is necessary to ensure methodological progress.

The aim of her stay at EEZ CSIC was to study:

- in situ hybridisation on acrylic resin-embedded semithin sections; and
- immunogold labelling for electron microscopy.

Ambrus Bakó PhD student visited the Institute of Plant Genetics and Biotechnology (IPGB SAS), Nitra for two months (1 February 2010 - 11 April 2010). His project leaders were Dr Anna Pretova and Dr Bohus Obert.

His work as a research associate in the Applied Genomics Department of ARI HAS includes the quantitative analysis of gene expression in the transgenic maize lines used in the institute's breeding programmes. In the temperate zone, low temperatures (near or below 0 °C) in spring can be a significant threat to crop production. At IPGB SAS there is an ongoing maize biotechnology project which can provide embryogenic calli of maize for molecular studies.

His task at IPB SAS focused on the study of gene expression changes caused by cold treatment in embryogenic calli of maize, including:

1. semi-quantitative RT-PCR for determining fold changes in the mRNA levels of selected genes;
2. 2D-PAGE analysis of changes in the proteome of the samples.

Annamaria Mészáros PhD spent a month in the Department of Experimental Biology, Babes-Bolyai University, Cluj-Napoca, Romania (18 March 2010 - 3 April 2010 and 16 April 2010 - 30 April 2010).

The Plant Biotechnology group directed by Prof. E. Rakosy deals with the development of tissue culture methods to establish propagation protocols, the investigation of abiotic stress effects on cultures of potato and tobacco, the creation of resistant plants via genetic engineering, the investigation of stress-induced genes by molecular methods and the somatic hybridization of potato to transfer quantitative resistance traits to biotic and abiotic stresses from wild relatives. The work was focused on studying the drought stress response of the model plant Arabidopsis thaliana cv. Columbia.

On two other occasions (2 December 2009 - 22 December 2009 and 12 January 2010 - 21 January 2010) Annamaria Mészáros visited the Department of Technical and Natural Sciences, Sapientia University, Mircurea-Ciuc, Romania. The laboratories, covering a total area of 220 m2, specialise in microbiology, biochemistry and molecular biology, enzymology and bioinformatics, and also include two preparative laboratories. Although plant sciences did not previously form part of the curriculum, due to the growing interest in this subject it has been decided to establish a new field of specialisation, general biotechnology.

Andrea Szenzenstein PhD student paid a one-month visit (1 March 2010 - 31 March 2010) to the Laboratory of Growth Regulators, Palacký University and Institute of Experimental Botany AS CR, lechtitel 11, 783 71 Olomouc, the Czech Republic.

One of the main topics in the Plant Cell Biology group, where she works in Martonvásár, is the improvement of the in vitro androgenic ability of elite lines of maize and the inheritance of this ability. The physiology of the donor plant greatly influences the androgenic response, but the mechanism is not precisely known. It is thought that hormonal levels may have an important role. In this connection, the aim of her stay was to learn techniques for the detection of plant hormones. In the partner laboratory a UPLC-MS method for plant hormone detection was developed and is used as a routine procedure. Very few data can be found in the literature about the hormonal levels of maize tassels. Her purpose was to determine the cytokinin and auxin levels in different parts of the central spike of the tassel and to analyse the differences between them.

Attila Fábián PhD student spent 12 days in the Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Czech Republic (8 November 2010 - 20 November 2010).

During his visit, he worked on the adaptation of fluorescent cytoskeleton visualising techniques to the plant materials used in experiments in Martonvásár, such as maize microspores and maize suspension cultures. The aim of the research training was to study the indirect immunofluorescencent labelling of microtubules and the fluorescent labelling of F-actin by fluorescent phalloidin.

Visiting scientist working in this field:

Adela Olmedilla PhD from the Spanish Council for Scientific Research (CSIC), spent a month at the Agricultural Research Institute HAS, in the Department of Plant Cell Biology (15 October 2010 - 15 November 2010).

The main purpose of her stay was to share her experience in light and electron microscopy techniques with the members of Dr Barnabas' group.

This stay also provided an opportunity to prepare an I-LINK-project proposal with CSIC that, if successful, will facilitate new collaboration including not only the Spanish (CSIC) and Hungarian (HAS) labs, but also a Slovak lab belonging to the Institute of Plant Genetics and Biotechnology of the Slovak Academy of Sciences (SAS), in order to introduce proteomics in the study of plant embryogenesis (identification of major proteins in early embryo development, in zygotic, somatic and gametic embryogenesis).

2. Identification of candidate genes for stress tolerance by means of functional genomics

András Bálint, PhD, spent the period from 1 November 2008 - 31 January 2009 as a post-doctorate fellow in Gatersleben, with the aim of learning the targeting induced local lesions in genomes (Tilling) method. This is a new reverse genetic method with which lines with mutations for a given gene can be identified rapidly and cheaply. It can also be used to identify allelic variants of a given gene, e.g. in variety collections (this method is known as Eco-Tilling). The original method uses artificially induced mutants, while Eco-Tilling examines naturally occurring mutations. It is planned to apply this method for a variety of purposes in the Department of Genetics and Plant Physiology of ARI HAS.

Fruzsina Szira, PhD student, spent the period from 1 November 2008 - 31 January 2009 in Gatersleben, participating in the work of two research teams: the Genome Diversity team headed by Dr Nils Stein, in which she examined gene expression changes during the cold hardening of cereals, and the Resources Genetics and Reproduction team headed by Dr Andreas Börner, where she tested methods for measuring the osmotic stress tolerance of barley and the mapping of loci influencing this trait.

Ildikó Karsai DSc spent four months in the School of Agriculture, Policy and Development, University of Reading, Whiteknights, P.O. Box 237, Reading, Berkshire, RG6 6AR, Great Britain (15 September 2010 - 15 January 2011).

She worked on two research topics during this time:

(1) elaboration of a non-destructive methodology for establishing plant developmental patterns in cereals;
(2) studying the effect of heat stress on the early plant development in wheat.

The type and intensity of negative changes caused by heat stress during flowering and seed development showed strong dependence on the developmental stages of the developing seed. To study this phenomenon, the progenies of plants stressed with heat at different times after anthesis were included in an experiment in which the initial plant development was studied under control and heat stress conditions. The aim was to evaluate how applying heat stress to developing seeds affects the early vigour and heat stress tolerance of the next generation of seedlings developing from the stressed seeds.

Imre Majláth PhD student spent three months in the School of Biological Sciences, University of Bristol, United Kingdom (10 January 2011 - 20 April 2011).

He worked with the Arabidopsis Research team, who had already demonstrated the different growth habits and distinct morphology of Landsberg erecta (Ler) and Cape Verde Islands (Cvi) natural accessions and of Ler-Cvi Near Isogenic Lines (NIL) both at low and high temperatures under different levels of red:far-red (R:FR) illumination. A QTL was found on chromosome 2 and Erecta, which encodes a putative receptor protein kinase with a pivotal role in cell-cell communication and plant morphogenesis, was identified as the candidate gene.

The aim of his work was to explore the QTL region on chromosome 2 and confirm the gene(s) responsible for the light forage strategy.

Ildikó Vashegyi PhD student spent 3.5 months in the Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park, United Kingdom (3 January 2011 - 30 April 2011).

The knowledge gained from experiments on genetic model organisms, combined with the use of the well-constructed experimental systems designed for cereals at the institute could lead to a better understanding of the functioning of complex signal transduction mechanisms and genetic regulation processes in monocots in relation to their abiotic stress responses. Therefore, the aim of her visit to the Department of Cell and Developmental Biology, John Innes Centre, Norwich was to become involved in the research there, to become acquainted with new approaches and scientific views, to learn new experimental methods and techniques, and to work with model plant species (Arabidopsis thaliana and Brachypodium distachyon).

Visiting scientists working in this field:

Kerstin Neumann, a PhD student from the Leibniz Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK), Gatersleben, Germany spent six months (1 September 2009 - 28 February 2010) at the Department of Genetics and Plant Physiology ARI HAS, Martonvásár.

The subject of her work as a PhD student at IPK is the mapping of drought tolerance loci in barley. In addition to a DH population for classical QTL mapping, a diverse barley collection for association mapping was also phenotyped in IPK and in ARI HAS. All statistical procedures for dealing with the population structure were examined in IPK with a set of Diversity Array Technology markers (DArT).The main aim of the visit was:

1. to teach the Hungarian research partners to use the statistical procedures and software necessary for association mapping;
2. to evaluate the data from a rain shelter experiment using the association mapping population for barley;
3. to conduct a small germination experiment with this population to evaluate differences in root length between the genotypes.

Jean-Marc Ducruet PhD from INRA/Universite Paris-Sud, Orsay, spent a month at ARI HAS in the Department of Plant Physiology (13 February 2011 - 19 March 2011).

The aim of his visit to ARI-HAS was:

A. to improve the thermoluminescence set-up used in ARI-HAS;
B. to train scientists in the use of the thermoluminescence technique.

Due to the extreme continental climate, there are few years when Hungary does not face severe economic losses related to environmental factors. These include long periods of drought, excessive quantities of rainfall in the wrong place at the wrong time, extremely cold winters or hot summers, or the sudden appearance of pests or pathogens in epidemic proportions, all of which are capable of destroying the efforts of a whole year, or even a whole decade. Both plant breeders and crop producers have an interest in finding crops capable of tolerating environmental changes with as little damage as possible. In order to develop such crops, knowledge of plant defense mechanisms and regulatory processes is essential.

As the scientist from the Faculty of Agriculture of Babes Bolyai University was unable to participate, Frantiek Bezunk PhD student, working on the molecular background of drought stress, was delegated by the University of South Bohemia in Czech Republic to ARI HAS. He spent two months (19 January 2009 - 19 March 2009) in ARI HAS in order to learn new molecular methods. He participated in work on the selection and characterisation of smoke-induced genes and in cloning and vector construction. He became acquainted with the microarray and quantitative PCR methods. Possibilities have been outlined for long-term cooperation between the two institutes, in which the experience gained by both partners could be used to mutual advantage. The first project is likely to be the development of transgenic flax lines with drought tolerance and insect resistance, involving joint construct preparation and transformation experiments. In some regions of the Czech Republic flax production is of great significance. The use of drought-tolerant transgenic lines could be one way of improving yield stability and obtaining a higher quantity of good quality oil.

Éva Tamás, PhD student from Sapientia University, Romania spent two months (1 November 2008 - 20 December 2008 and 5 - 16 January 2009) in ARI HAS. Her PhD work involves enhancing the synthesis of terpenoids in tissue cultures of Ligularia, partly by selection at cell level and partly by genetic transformation. She already had satisfactory knowledge on molecular methods, so the aim of her visit was to learn plant regeneration and transformation methods. Although the duration of her visit was not sufficient to carry out a complete transformation cycle, she was able to follow the different parts of the process on various plants (tobacco, marrow, maize). She was able to learn the advantages of both the Agrobacterium-mediated and biolistic methods of transformation, including the various factors that influence their efficiency. Staff from the Cell Biology Department were also involved in teaching these methods. The knowledge acquired during her visit will allow her to carry out transformation experiments independently and carry out checks on the transformed plants. The Sapientia University has indicated an interest in further cooperation. The department in question deals principally with nature protection and ecology, and aims to elaborate new technologies and products for horticulture, which has a long tradition in the region. The joint development of plants resistant to drought or other stress factors could thus be of outstanding importance.

Lenka Fraterova, PhD student from the Institute of Genetics and Biotechnology, Slovak Academy of Sciences, Nitra, Slovakia spent two months (23 February 2009 - 30 April 2009) at ARI HAS, during which period she also took part in the course on 'Climate change: Challenge for the training of applied plant scientists. Biotic and abiotic stresses' organised in the framework of WP4.The subject of her PhD work is the induction of somatic embryogenesis in pine species and the measurement of extracellular protein activity (chitinases, glucanases) in the embryos. As the necessary analytical background is available in her home laboratory, the aim of her visit was to learn various molecular techniques. She became acquainted with the bacterial cloning technique, and with methods for the isolation and identification of plant nucleic acids. She was also able to follow the process of Agrobacterium-mediated transformation. Many of the Martonvásár staff have good personal and professional contacts with staff from the Nitra institute, and there is frequent cooperation between the two institutes. ARI HAS would be interested in the method used for the induction of somatic embryogenesis. As this is a pathway in which new plants are guaranteed to originate from a single cell, it would allow transgenic plants to be selected with better efficiency after genetic transformation, eliminating somatic mosaicity in the plants. The Applied Genomics Department possesses transformation constructs with which alien catalase (CAT) or superoxide dismutase (SOD) genes can be incorporated into transformed plants. These two oxidative enzymes are involved in the general defense mechanism of plants, and their over-expression in pine species is expected to result in increased heat or drought tolerance. The Nitra research team is currently working on the development of plant material that could be used for re-afforestation in the wake of the destruction of forests in Slovakia. This system provides the possibility of selecting and multiplying lines with better resistance to climate change.

3. Agro-ecological research

Balázs Varga PhD spent three months in the Institute of Biodiversity, VTI-Braunschweig, Johann Heinrich von Thünen-Institut, Germany (6 September 2010 - 3 December 2010).

Free air carbon dioxide enrichment (FACE) is one of the best ways to investigate the effects of elevated carbon dioxide concentration on the phenological and physiological properties of plants. The aim of the visit to VTI-Braunschweig was to study the combined effects of drought and elevated CO2 in a FACE experiment and to learn methods for root system analysis. Roots are often more stimulated by the CO2 concentration than leaves, stems and reproductive structures, making accurate investigations on the root system essential. Previous studies showed that the roots become more numerous, longer and thicker in a CO2-enriched environment. The root system of CO2-enriched crops is often highly branched, especially at shallower soil depths, compared to the roots of crops grown at the ambient CO2 concentration.

4. Long-term crop production experiments

Györgyi Micskei PhD student spent three months in the Sustainable Soils and Grassland Systems Department, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, United Kingdom (17 January 2011 - 15 April 2011).

She did her training in soil and plant analysis as a visiting scientist under the supervision of Dr Andy Macdonald in the Department for Sustainable Soils and Grassland Systems at Rothamsted Research from 17 January 2011 to 15 April 2011. The aim of her stay was to become acquainted with the Rothamsted Classical Experiments, which are the oldest continuous agronomic experiments in the world. She received training in soil and crop sampling and sample preparation, grain quality measurements, general plant and soil analytical techniques and data compilation and analysis, the collection and preparation of drainage water samples and the general management of long-term and other field experiments.

5. Phytotron research

Szilvia Bencze, PhD, spent six months in the United Kingdom (Plant Science Department, Rothamsted Research, Harpenden) as a post-doctorate fellow from 1 October 2008 to 31 March 2009. Dr Bencze has worked on problems related to global climate change since joining the staff of ARI HAS in 1998. Her special field has been the effect of enhanced CO2 concentrations and climatic extremes on cereals, and the results of this work formed the basis of her PhD thesis in 2007. During her study trip Dr Bencze took part in the research in progress in the team led by Martin Parry, and also participated in experiments in the framework of a new EU project (Bioenergy and Climate Change) under the leadership of the biochemist John Andralojc.

6. Breeding of field crops

István Molnár PhD spent three months in the John Innes Centre, Norwich Research Park, Colney, Norwich, United Kingdom (3 January 2011 - 30 March 2011).

The wild relatives of wheat represent a large reservoir of useful genes and alleles for wheat improvement, which can be transferred by interspecific and intergeneric hybridisation. Within the Aegilops genus, the allopolyploid species Ae. biuncialis Vis. (2n = 4x = 28, UbUbMbMb) and Ae. geniculata Roth. (2n = 4x = 28, UgUgMgMg) originated from hybridisation between the diploid species Ae. comosa Sm. in Sibth. and Sm. (2n = 2x = 14, MM) and Ae. umbellulata Zhuk. (2n = 2x = 14, UU). Several genes for resistance to rusts and powdery mildew (Lr9, Lr57, Sr34, Yr8, Yr40, Pm 29) have been transferred into wheat from these species and tolerance of salt, drought, frost and heat stress was observed in these genotypes. The use of PCR-based molecular markers significantly accelerates the selection of wheat-Aegilops introgression lines, the mapping of stress tolerance QTLs in the Aegilops species and the positional cloning of genes responsible for these traits. Unfortunately, few markers specific for these Aegilops species are available, if at all.

The aim of the short visit was to determine the chromosomal locations of the conserved orthologue set (COS) markers developed by the research team headed by Dr Simon Griffiths on the allotetraploid species Ae. biuncialis and Ae. geniculata, together with their diploid progenitors Ae. umbellulata and Ae. comosa.

Klaudia Kruppa PhD student spent three months in the Gene and Genome Mapping group of the Department of Cytogenetics and Genome Analysis, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Germany (10 January 2011 - 10 April 2011).

The aim of this visit was to study the microsatellite marker technique and to analyse wheat x barley and wheat x Agropyron introgression lines with SSRs. Various types of biological material, all produced in Martonvásár, were used for SSR marker analysis: the wheat x barley translocation lines 6B.4H 4D.5H and 7D.5H the homozygous wheat / barley centric fusion line 4HL.5D BE-1 × wheat progenies and wheat x Agropyron glael introgression lines. Two Thinopyrum species, Thinopyrum ponticum and Thinopyrum intermedium were also tested with wheat SSR markers.

Rakszegi Mariann PhD spent three months in the Plant Science Department of Rothamsted Research, Harpenden, Herts, AL5 2JQ, United Kingdom (17 January 2011. - 15 April 2011).

The aim of the visit was to compare the amount, solubility and structure of wheat and barley glucans, including variation between wheat varieties. This included the determination of the total glucan in wheat and barley flour and in wheat fractions extracted with water at increasing temperatures (Megazyme kit), the determination of the ratio of (1-3) to (1-4) bonds by enzyme digestion (HPAEC), the determination of the molecular weight distribution by SE-HPLC and the determination of the structure by NMR. The determination of the effects of heat and drought stress on the properties of wheat glucan and the properties of a number of oat and barley lines was also planned.

7. Crop protection

Emese László PhD student spent two months in the Department for Agrobiotechnology, IFA-Tulln, Institute of Biotechnology in Plant Production, Austria (8 October 2010 - 3 December 2010).

The aim of her stay at IFA was to study:

- classical methods in Fusarium taxonomy;
- species-specific markers for fungal identification;
- identification of Fusarium isolates derived from naturally infected kernels.

As a result of her visit to the Department of Agrobiotechnology, Institute of Biotechnology in Plant Production, IFA Tulln she became familiar with classical methods in Fusarium taxonomy and the use of species-specific markers for fungal identification, which will allow ARI-HAS to monitor changes in the pathogen composition or changes in the toxin profiles of the most predominant species.

The exchange visits made possible by the project contributed greatly to improving the already high level of methodology used in the Martonvásár Institute for research on the abiotic stress tolerance of cereals. The mutual exchange of the most up-to-date experimental approaches and analytical methods, and the forging or reinforcing of relationships between individual scientists will greatly increase the range of joint research projects and grant applications in the future. The acquisition of new methods and their application in ongoing research can open up new vistas for solving problems related to climate change.

The experience gained in foreign laboratories will also be of great service in advancing the careers of the young scientists. The forging of new professional contacts and friendships will ensure a continuous exchange of opinions in the fields of both science and culture.

WP2: Recruitment of experienced researchers working in the field of environmental stresses
Coordinator: Dr Gábor Galiba

From the four persons whose recruitment was planned only Balázs Tóth accepted the offered position while the others rejected for different reasons. Actually through advertisement and networking we could fill all of the remaining 3 positions in 2009.

It was not possible to carry out Task 2.1. as it was originally planned, because Dávid Kszegi received meanwhile a research position in the Institut für Pflanzengenetik und Kulturpflanzenforschung in Gatersleben, Germany. Instead of Dávid Kszegi another scientist, Gergely Gulyás (Hungarian) was recruited. Gergely Gulyás works in the laboratory of Prof. László Láng at the Department of Cereal Breeding in the Agricultural Research Institute of the Hungarian Academy of Sciences. He returned from Japan to Martonvasar in 2009 to begin the analysis of wheat genes related to adaptability and tolerance of biotic or abiotic stresses.

His personal background

His master and doctoral degrees were obtained at the Tokyo University of Agriculture and Technology (TUAT), Tokyo, Japan. During the two-year master and the three-year doctoral courses, he worked on cytoplasmic male sterility (CMS) in chilli pepper. Even though the profile of this lab was basic plant research, it was an excellent place to acquire molecular techniques which can be used also in cereal research. During his stay at TUAT, he investigated the nuclear-mitochondrial interaction using cytoplasmic male-sterile chilli pepper lines. The available restorer, maintainer and CMS lines allowed him to identify the segregation ratio of restorer gene. In addition, the comparison of the molecular marker information linked to restorer gene and the phenotypic data revealed important information on physical distance between the marker and the gene. Moreover, during the search for the candidate gene for CMS phenotype in the mitochondria he was able to identify an extended transcript of a sterility-related gene named orf507 in the mitochondria of chilli pepper.

Experimental work

Adaptation of the new varieties to the changing environmental conditions caused by the global climate change is essential to ensure food supply. Rainfall and temperature extremes in Central and Eastern Europe are getting more frequent. According to the climate change forecasts, plant productivity will suffer a significant loss in great part of Europe. Therefore, it is extremely important to improve varieties for better adaptability to the changing environment. Studies on the molecular background of adaptation, genes that control flowering including vernalisation and the response to photoperiod genes, are essential to utilise such abilities of plants.

Analysis of adaptive characteristics of plants

Flowering time is one of the most important adaptive characteristics of plants. Genetic regulation of physiological processes acts to ensure that flowering occurs at seasonal optima for pollination, fertilisation, and seed development. Photoperiod and vernalisation regulate the vegetative to reproductive phase transition, and photoperiod regulates the expression of key vernalisation genes.

Differences in the rate of development (ear emergence and ripening time) of cultivars of common wheat (Triticum aestivum L.) are the main components contributing to their adaptation to a wide range of environmental conditions. The Vrn genes (growth habit) and the Ppd genes (response to photoperiod) contribute to these differences.

Analysis of genetic background of extreme earliness

The control of flowering is fundamental to reproductive success in plants, and has a major impact on grain yield in crop species. Temperate environments with a long growing season allow cereal crops to flower late in the year and thus exploit an extended vegetative period for resource storage. Conversely, early flowering has evolved as an adaptation to short growing seasons.

The aims are to explore the genetic components of the extreme earliness of the winter wheat variety 'Mv Toborzó' and to identify the environmental factors causing this unique reaction. For this reason Mv Toborzó was crossed with eight different wheat varieties selected from diverse ecological conditions. In their F1 and F2 populations, the earliness showed continuous frequency distribution and was determined by the PPD-D1 allele constitution of the given parents. In the insensitive x sensitive crosses the earliness of F1 and F2 populations was the same as the average heading of the parents. In this experiment, the extreme early heading of Mv Toborzó might be caused by one or more recessive gene(s) that is / are down regulated by the PPD-D1 sensitive allele. When Mv Toborzó, the photoperiodic insensitive variety, was crossed with the sensitive one called Tommi, the PPD-D1 allele structure was determined by gene-allele specific markers in the segregating F2 population. Since the segregation ratio was not significantly different from the suggested 1:2:1 ratio, the effect of PPD-D1 gene on plant height, on heading date and some other agronomical characteristics could be analysed. The results showed that the PPD-D1 gene significantly influenced plant height and heading while the insensitive allele caused reduced height and earlier heading. In this population, the heterozygotes flowered later than the homozygotes. The homozygous insensitive individuals in this populations showed late heading and the same insensitive allele (Mv Toborzó allele) in different genetic background showed different heading dates. These results suggest that some other minor genes of earliness can be segregating as well.

In the last year of the AGRISAFE project, the effect of the insensitive allele of the PPD-D1 gene on plant height was further analysed in order to examine the interaction with some plant height determinant genes. In wheat, 21 genes with major effects on reducing plant height have been identified and assigned to Rht designations. These are grouped into two categories, insensitive and sensitive to exogenous gibberellic acid (GA). The GA insensitive alleles Rht-B1b (Rht1) and Rht-D1b (Rht2), derived from Norin 10, reduced plant height by 15 % and increase yield by 24 %. The GA-responsive Rht genes, Rht8 and Rht9, derived from the Japanese cultivar (cv.) Akakomugi, were introduced into Italian germplasm and spread into South European countries where high temperatures and drought are normal in summer. It was reported that Rht8, in close association with the photoperiodic insensitivity gene Ppd-D1a, reduced plant height by around 10 % without significant negative effects on yield. The Rht8 gene also has the potential to increase the early vigour of semi-dwarf wheat although its effect on plant height was not as significant as those of Rht-B1b and Rht-D1b. A 192 bp allele of wheat microsatellite marker Xgwm261-4A, in combination with the pedigree information, is diagnostic for Rht8.

The homoeologous genes Rht-B1 and Rht-D1 were molecularly characterized and both mutations involve single base-pair changes leading to a TAG stop codon shortly after the start of translation. Furthermore, PCR-based specific markers were developed to discriminate between the dwarf genes Rht-B1b and Rht-D1b and their wild type tall alleles Rht-B1a and Rht-D1a. Therefore, both seedling tests for the lack of responsiveness to GA and molecular markers can be used to detect the presence of these two genes in the breeding program. The presence of RhtB1b and RhtD1b was determined in both F2 and F3 generations of Mv Toborzó x Tommi. Determining the distribution of the Rht8 gene in the segregating population (F2 and F3) will be the next step in order to get closer to the understanding of the molecular background of the adaptation ability.

Better understanding of the genetic components of the flowering time regulation will help to improve the agricultural productivity in new production zones or in conventional regions which are subjected to greater climatological fluctuations.

Summarising his research activities:

He has analysed the adaptive characteristics of flowering time in some varieties and in their offspring generations at the Department of Cereal Breeding in the Agricultural Research Institute of HAS. Finding new regulating genes, identification of new regulatory processes, a better utilization of genetic resources in the development of new varieties and the elaboration of a marker assisted selection system can be the possible outcome of this research.

Balázs Tóth works in the laboratory of Prof. Gábor Galiba at the Department of Plant Molecular Biology, in the Agricultural Research Institute of the Hungarian Academy of Sciences. He returned from the United States (US) to Martonvasar in 2008 to begin the analysis of different lipid classes which play crucial roles in the environmental stress tolerance (especially cold and drought stress) in cereals.

His personal background

Balázs Tóth obtained his PhD degree at the Szent István University, Gödöll in 2004. His PhD thesis was based on the genetic studies of frost tolerance in cereals. As a postdoc he spent four years (2004 - 2008) in the laboratory of Dr Tamas Balla at the National Institute of Child Health and Human Development (National Institutes of Health (NIH), Bethesda, Maryland, US). Although the profile of this lab was basic medical research, it was an excellent place to acquire a lot of scientific techniques for the mammalian system which can also be used in plant research. During his stay at the NIH, he took part in several projects while working on lipid signalling and lipid research. He investigated the regulatory role of phosphatidylinositol 4-kinases in the ceramide transport from the endoplasmic reticulum to the Golgi. Moreover, he performed subcloning, purification and kinase assay of a phosphatidylinositol 3-kinase, He was involved in studies with pleckstrin homology domains, in experiments investigating the STIM1-Orai1 complex and in designing the mutant PtdIns4Kinase. He carried out also other experiments to characterize a newly discovered protein family which is involved in the glycosphingolipid metabolism of the cell.

Experimental work

Since lipids are structurally (as components of the lipid membranes) and functionally (as components of the lipid signalling pathways) in the front line of the cells being exposed to different environmental changes, their role in the response to the challenging growing conditions during the climate change appears to be obvious. This is the reason why he designed a long-term experimental plan to introduce lipid research in Martonvásár.

Identification of lipid signal transduction components

In the last year, he identified signal transduction components involved in cold acclimation and frost tolerance in barley and einkorn wheat using the pharmacological approach. He investigated the possible utilization of callus cultures as an experimental system to study frost tolerance. He observed that barley callus has altered the cold response because of its different hormone content, so he concluded that it is an inappropriate object for this kind of experiments.

In barley seedlings, he proved the role of calcium in the recovery following freezing injury. In the gene expression level in barley he found that the cold induction of CBF14 and COR14b appears to be dependent on the intracellular calcium release, while CBF9 seems calcium independent and the whole system is regulated by phospholipase C.

In einkorn wheat, he observed that the cold induced expression of CBF12 and COR14b was calcium dependent, while the expression of CBF14 was not affected by the decrease in the calcium response.

Using microarray techniques, he identified a series of calcium dependent cold responsive genes including the components of the antioxidant defence system. He also investigated the effect of increased MAPK kinase activity on the cold response and noticed that the CBF-COR system was independent of MAPK induction.

Two PhD students (Ildikó Vashegyi, Zsuzsa Tóth) are involved in this project. The methodology and the scientific background of this work provide them with important knowledge, experience and new scientific results for the completion of their PhD studies.

Summarising his research activities:

He made significant progress in the identification of lipid signalling components during cold stress. In the third research year he obtained new results on the involvement of the phospholipase C pathway in cold acclimation and frost tolerance.

Additional benefits generated by the AGRISAFE project

The financial support of his employment by the AGRISAFE opened the door to applications for additional financial resources to support his research project. As a participant, he has received a research grant ('Identification of lipid components suitable for the improvement of freezing tolerance in cereals'; No. K68894) from the Hungarian Scientific Research Found. As a project leader he obtained financial support for the identification of lipid signalling components in cold stress ('Identification of signal transduction pathways involved in cold stress by gene expression studies in cereals'; No. NNF78866) from the Hungarian Scientific Research Found and the Norwegian Research Found. This project involves also a position for a new PhD student, Zsuzsa Tóth. Moreover, a highly talented PhD student, Ildikó Vashegyi, is also working under his supervision in the lipid research area and these projects will be the base for her PhD thesis.

It was not possible to accomplish Task 2.3. as it was planned because Péter Szcs received a position in the SYNGENTA Hungary, meanwhile. He did not intend to return back to ARI HAS. So, instead of Péter Szcs, Robert Dóczi accepted the open position from 1 September 2009. He initiated a new research programme to study the MAP kinase-substrate networks at the Plant Cell Biology Department (Head: Prof. Beáta Barnabás) when he returned back from London. This goal is also supported by a Marie Curie Reintegration grant of the European Commission. His Task is 2.3 within WP 2 to facilitate the return of experienced researchers working in the field of environmental stresses to Hungary.

Personal background

Róbert Dóczi has a long-term research interest in signal transduction. He carried out his PhD studies on characterising the regulation of a drought-specific gene which can be found only in the potato crop plant and its close relative species. Subsequently he became a postdoctoral researcher in Prof. Heribert Hirt's lab at Max F. Perutz Laboratories, Vienna Biocenter, Austria (see http://www.heribert-hirt.info online). Here he got involved in a project that unravelled the cold / salt MEKK1 MKK2 MPK4 signal transduction pathway. His most significant result in Vienna was the identification and characterisation of the H2O2 MKK3 MPK7 PR1 pathway. These results were achieved within the framework of his own project and revealed connections between previously unknown plant MAP kinase components. Meanwhile he took also part in the describing of the molecular interactions of a MAP kinase regulatory phosphatase. These results contribute to our understanding of the importance and complexity of MAPK signalling in plant stress responses. He then successfully applied for a Marie Curie Intra-European Mobility Fellowship of the European Commission (EC) to join Prof. Laszlo Bogre's lab at Royal Holloway, University of London (see http://www.rhul.ac.uk/Biological-Sciences/AcademicStaff/Bogre/index.html online). Here he characterised the role of the MKK7/9 MPK(3)/6 pathway in meristem function and proved its negative regulatory role. An important goal of the Marie Curie fellowship application was to receive training in bioinformatics and systems biology. Accordingly he successfully joined the first systems analysis project of the Arabidopsis MAP kinase network, and got familiar with a range of the relevant methods.

Experimental work

The mitogen-activated protein kinase (MAPK) phosphorylation cascades are conserved signalling modules in all eukaryotes and known to have pivotal roles to regulate cell division and cell growth as well as stress responses in animals and plants. How can the same MAPKs regulate both defence and developmental responses is poorly understood. One of the major pitfalls in our understanding is the paucity of known target proteins (substrates) in plants through which MAPK signalling pathways connect to physiological responses. Most well-characterised MAP kinase proteins of yeast and animals have multiple substrates, for example human ERK1/2 have over 100 different substrates. Our knowledge of plant MAP kinase substrates lags far behind and the identified substrates so far have been isolated on a more or less random basis, mostly by yeast two-hybrid screens. This research programme aims changing this situation by systematically finding and characterizing novel plant MAP kinase substrates.

IIdentification of novel putative MAP kinase substrates by a bioinformatics-based approach

Kinase-substrate recognition specificities within complex networks are often provided by dedicated interaction surfaces outside the kinase active sites and target phosphorylation sequences. Protein interaction specificities thus remain highly evolvable without compromising the strict stereo chemical requirements for efficient catalysis performed at the active site.

MPKs possess an evolutionarily conserved common docking (CD) domain as a docking site for MKKs, phosphatases and protein substrates that contain the corresponding MAP kinase docking site. Our knowledge of actual MAP kinase substrates in Arabidopsis is rather limited, nevertheless all five characterised substrates contain putative MAP kinase docking domains with minor variations to the consensus motif13-17. Remarkably, the predicted MAPK docking motif (amino acids 244-252) of EIN3 was mutated in the ein3-3 mutant (K244D) that diminished EIN3 function18. Robert Dóczi took advantage of the conservation of MAP kinase docking sites in a bioinformatics screen to identify novel potential MAP kinase substrates. He searched for the presence of potential MAP kinase docking sites in Arabidopsis protein sequences, and filtered the resulting candidates for the presence of the MPK phosphoacceptor motif [S/T]P.

He has successfully set-up all the required experimental techniques, such as cDNA cloning, protoplast transfection, protein immunoblotting, etc. One PhD student (Magdolna Dry) has been involved in the new project since May 2010. She has made an impressive progress by learning all the techniques for the execution of the experimental programme within this time-frame.

Summarising his research activities:

He has made a significant progress in establishing a new research project aimed to characterise molecular connections between stress-activated MAP kinase pathways and developmental regulation. During his 19-month period under AGRISAFE employment, he initiated a project successfully via adopting new experimental techniques. Preliminary results indicate that at least one predicted MAP kinase substrate is genuinely modified by MAP kinase in planta.

Additional benefits generated by the AGRISAFE project

Since his employment at the Agricultural Research Institute, Dr Dóczi's goal has been to initiate the new research programme. To achieve this he prepared applications to granting agencies and commenced establishing molecular biology and protein biochemistry methods required for the project.

He has received an EU FP7 Marie Curie Reintegration Grant (ERG 256554; support: EUR 45 000, duration: three years). He has also successfully applied for an infrastructure development grant at the National Office for Research and Technology (MVNOVPRO). Support: HUF 20 000 000, one-off payment for the establishment of new proteomics laboratory facilities with the participation of the members of the Department of Applied Genomics. Currently, he has two grant applications pending.

Later this year, a methodological book chapter, written together with former co-workers, on assaying MAP kinase activities in plant samples will be published:

Dóczi, R., Hatzimasoura, E. and Bögre, L. Mitogen-activated protein kinase activity and reporter gene assays in plants. Methods in Molecular Biology (in press), preliminary view: http://www.springer.com/life+sciences/plant+sciences/book/978-1-61779-263-2?changeHeader.

It was not possible to execute task 2.4. as it was planned because Mariyana Georgieva arrived to Martonvásár on 16 April 2009 for six months, instead of Dr Svetlana Landjeva, who could not come because of health problems. The original plan was that Dr Landjeva would bring wheat genetic materials developed from wheat-alien hybridisation in Bulgaria and the plants would be analyzed by up-to-date molecular cytogenetic techniques in Martonvásár. As Dr Landjeva had serious health problems since the end of 2008 it was suggested by Prof. Gecheff, director of the Institute in Sofia, and Dr Landjeva that Mariyana Georgieva, a young research fellow from the Institute of Genetics in Sofia should come instead and carry out the planned experiments for six months.

Personal background

Mariyana Stamova Georgieva was born in 1980 and she has been working at the Department of Molecular Genetics in the Institute of Genetics in Sofia, as a junior research fellow since 2003. She has good experience in the field of molecular genetics of cereals. She carried out research in the field of DNA damage, DNA repair, Comet assay, oxidative stress. She has several publications at conferences and in scientific journals. As she is trained in the field of molecular genetics she is supposed to be able to learn fluorescent in situ hybridisation (FISH) techniques soon to carry out the planned experiments.

Experimental work

The wheat-Agroypron intermedium amphidiploids produced in Sofia have several useful agronomical characteristics but the genome structure is not known. The amphiploids have high protein content (19 - 22 %). The parental Agropyron intermedium has good adaptability to extreme environmental conditions, so it is hoped that the abiotic stress tolerance of the parental Agropyron accession is inherited in the amphidiploids. This will be studied later. The main purpose of Georgieva's visit was to detect the Agropyron chromosomes in wheat background in the amphiploid and identify them using FISH / GISH techniques with the help of repetitive DNA probes. Besides, another aim of her stay is to learn up to date molecular cytogenetic techniques.

Summary of the achieved results during the six months training

One direct benefit of Georgieva's stay in the Agricultural Research Institute was the acquisition of the following useful methods and techniques which she will be able to apply in her work in Bulgaria:

1. Mitotic chromosome preparations using Pseudoroegneria spicata, Thinopyrum intermedium, Triticum aesivum and two partial amphiploids (H95 and 55(1-57))
2. Probe labelling- pTa71, Afa family, pSc119.2. In order to study the genome composition of H95 and 55(1-57) genotypes a large collection of species were used from the Genebank, USDA, US and Martonvásár Cereal Gene Bank, Hungary.
3. Multicolor fluorescence in situ hybridisation (McFISH).
4. Genomic in situ hybridization (GISH).

Additional benefits generated by the AGRISAFE project

The visit of Mariyana Georgieva was the first step to establish long-term research collaboration between the Department of Molecular Genetics, Department of Cytogenetics of the Institute of Genetics, Bulgaria and the Department of Plant Genetic Resources and Organic Breeding in the Agricultural Research Institute, Hungary.

During Mariyana Georgieva's stay in Martonvásár, she also took part in the symposium and the training course III: Impact of climate change on crop production within an AGRISAFE programme held on 7 - 11 September 2009. Her participation in this course gave her an opportunity to expand her international contacts with other scientists mainly from Central and Eastern European research teams.

WP3: Development of scientific and other equipment for training courses on environmental stress research connected with global climate change
Coordinator: Dr. Béla Kszegi

Prior to the start of the project, the equipment available to the research institute was in part outdated, due to lack of funds, thus seriously hindering competitive research.

As WP3 was concerned with the purchase of the scientific and IT equipment required to achieve the aims of the project, it was intended that the vast majority of these items should be purchased as quickly as possible after the granting of the funds, so that they could be put into operation as soon as possible.

The laboratory instruments and the IT equipment purchased in the framework of WP3, and the improvements made in the IT structure, network and software were exploited by both basic and applied research to achieve the aims of the project.

The most important tasks were:

- the establishment of an automatic meteorological station;
- the creation of an audiovisual lecture hall;
- the purchase of new laboratory instruments, or in some cases the improvement of existing equipment;
- the purchase of IT equipment and software licences, and the expansion and improvement of the institute's IT network.

Information on the implementation of these improvements, including details of the public procurement procedure, was provided in the first annual report.

In the course of the project, regular calculations were made on the use of the funds available to WP3, and after the first round of purchases had been completed, it became clear how much of these funds had been utilised. Thanks to the public procurement procedure the sum actually spent was less than the funds made available on the basis of the original budget. This allowed equipment costing a total of around 8.4 million forints to be purchased during the second period, based on the experience gained during the operation of the new equipment purchased during the first year.

Since only the amortisation of the purchased equipment can be funded from the grant, a calculation of the amortisation rate of the equipment already purchased revealed that further equipment could be purchased during the third year of the project, thus expanding or renovating the IT background of the institute in order to promote the implementation of the aims of the project.

The benefit gained from the improvements can be summarised as follows:

Equipment purchased for basic research

The equipment detailed in the first annual report, including microscopes, microtomes and the software required for data processing continues to be used by the staff of the Plant Cell Biology Department to investigate changes in plant tissues as the result of environmental stress, especially drought and heat. The evaluation, processing and publication of the results are carried out with the help of the notebooks and computers purchased from the grant.

The instruments were used to study the effects of drought and heat stress on the sexual processes of wheat, particularly on tissue development in the anther, pistil and developing grain. Investigations were also made on the effect of water withholding and increased CO2 concentration on the epidermal structure of wheat. Simultaneous water withholding and elevated temperature were found to cause alterations in embryo and endosperm development.

Supplementation of the existing instruments to allow plant stress responses to be analysed at the molecular level

The equipment purchased from project funds was used to continue existing research and also to initiate investigations on new topics, all involving the analysis of crop responses to environmental stress at the molecular level.

In the Cereal Resistance Breeding Department the effects of various abiotic (heat and drought, geographical adaptation) and biotic (various leaf diseases) stress factors on the physiology, phenology and yield production of wheat and barley genotypes were studied under phytotronic and field conditions.

Little work has been done in cereals on how non-vernalising temperatures influence the genetic regulation of flowering. Work is underway to discover the effect of temperatures lower or higher than the optimum, and of daily fluctuations in these factors. Research topics include the sensitivity of individual genotypes to such temperatures, the vernalisation- and daylength-independent regulation of major plant development genes and interactions between these genes, other chromosome regions involved in temperature sensing, the genetic background of differences in the regulation of flowering in wheat and barley, and the relationship between the haplotypes of major plant development genes and the geographical spread of different genotypes. For this purpose, phytotron experiments and molecular genetic analyses are carried out on two-parent populations and on large numbers of barley and wheat varieties. This work is expected to lead to the identification of new regulatory genes and processes, the more efficient use of genetic resources in variety development and the elaboration of methods for marker-assisted selection.

Research is also underway on the effect of higher temperature or drought combined with an increase in the atmospheric CO2 level, the effect of enhanced atmospheric CO2 concentration on host plant-pathogen relationships, and the joint effect of nutrient supplies, temperature and enhanced CO2 level.

Equipment purchased for applied research

Purchase of additional instruments and software for analysing the effect of drought on the leaf area and the intensity of photosynthesis and for processing the data of field experiments

In the Crop Production Department, the equipment purchased from the AGRISAFE project was used mainly in analysing the effect of treatments in the long-term fertilisation experiments set up fifty years ago. The portable leaf area meter was used for growth analysis on the rate and intensity of maize shoot development, while the up-to-date root washing equipment was essential for characterising root development in order to determine the effect of nutrient treatments and the damage caused by the larvae of the western corn rootworm. Differences in the N supplies of wheat varieties and maize hybrids were determined on the basis of numerous measurements using the chlorophyll meter.

The Rotina 420R refrigerated centrifuge was used to prepare extracts from maize stalk tissue samples taken from plants artificially inoculated with Fusarium strains in the field. The cellulase enzyme activity of the extracts was then determined using the Colim 4.0 image analysing program. The results were published in a number of scientific and non-scientific journals, indicating funding from AGRISAFE.

The DU 730 Life Science spectrophotometer purchased from the project is used in marker-assisted analysis. It enables the DNA concentration of different genotypes to be determined after DNA isolation. In order to optimise PCR reactions, the DNA extracted from different samples must be adjusted to approximately the same concentration, and the data obtained using the spectrophotometer allow the necessary dilutions to be made. The spectrophotometer can also be used to check the concentrations of primers and ALF sizers.

Establishment of a meteorological station for the collection and analysis of local data on meteorological changes

In the framework of a cooperation agreement, the new meteorological station was linked up with the data-collecting network of the Hungarian National Meteorological Service, which means that not only local data, but also up-to-date national meteorological data are constantly available. The data provided by the new station are continuously uploaded to the institute website (see http://www.mgki.hu online), where they can be accessed by institute staff and PhD students. These data can be used for planning field experiments, for analysing the effect of climate change on the experimental results and in writing papers. If required, the meteorological data from the station can also be utilised by those attending the training courses organised in the framework of the project. The agreement reached with the National Meteorological Service also ensures the regular, expert maintenance of the station. Construction and installation of an up-to-date audiovisual lecture room as a venue for the courses planned in the project This air-conditioned lecture room, seating 40 people and containing up-to-date audiovisual equipment, is located in the phytotron building and is used not only for the training sessions held as part of the project, but also for the regular meetings required in preparation for these. The room is equipped with microphones, projection equipment and a digital video camera. Wireless internet and access to the electricity network are available to the course participants both in the lecture room and throughout the institute. Notebooks were also purchased from project funds for the use of course participants, allowing both internet access and the possibility of recording and replaying the lectures. The courses held as part of the project provided not only theoretical, but also practical knowledge for the participants. The equipment purchased in the framework of tasks 3.3 and 3.4 was used during the practical sessions to give the participants detailed knowledge on the stress tolerance of cereals and on their physiological responses to abiotic stress. In the framework of WP3, special efforts were made to improve the IT structure of the institute and to purchase the IT equipment required to achieve the project aims, thus significantly improving the bio-informatics background of the AGRISAFE project. The IT equipment purchased from AGRISAFE funds (servers, work stations, notebooks and improvements to the institute's IT network) makes a day-to-day contribution to achieving the aims outlined in the project by ensuring a satisfactory IT background.