Project review

 Description of the project MOBITAG

General objective

Ever-increasing human population accompanied by a decrease of arable land, and the replacement of depleted fossil resources by biomaterials, call for a higher and sustainable agricultural production. The importance of agriculture, forestry, and fisheries, which are essential components of economics in the region of South Bohemia, is likely to increase. Since there are no reserves in available land, any increase of agricultural production must be based on technological improvements. The Biology Centre of the Academy of Sciences (BCAS) of the Czech Republic, which is located in the South-Bohemian political district (about half of the region CZ-NUTS 2 Southwest), is determined to promote development and implementation of modern technologies in agricultural production. This is well understood by the regional administration that supports the plans of BCAS. It must also be mentioned that BCAS shares the campus and collaborates very closely with the University of South Bohemia and has fruitful contacts with representatives of the business community. BCAS regularly communicates with the Ministry of Agriculture and the Ministry of Environment. All this administration back-up and active participation in student education have a very positive impact on BCAS activities supporting regional. Regional plans include construction of a new Biotechnological Institute of Applied Ecology as a component of BCAS. Support for the construction will be sought from the Research and Development for Innovations Operational Programme. The results of MOBITAG project will form a core of the Biotechnological Institute.

MOBITAG project proposes advancement of the current research potential in the convergence region to a level allowing development and deployment of novel biotechnologies for a highly productive, environment-friendly, and sustainable agriculture. The project is focused on selected topics that are of importance for the region. Emphasis is given to biotechnologies linked to the production and deployment of GMOs, including the safety aspects.
Project goals

MOBITAG proposes training of 22 researchers in the methods and conceptual approaches needed for the investigations and problem solutions in four areas that are essential for most modern agricultural biotechnologies.

They include:

• Genome analysis
• Exploitation of natural products
• Transgenosis
• Safety aspects of GMO

Selected members of the current staff will be responsible for building the teams with inclusion of newly recruited researchers (WP1). Collaboration with the experienced colleagues from other EU countries and enhanced mobility of the researchers are important parts of MOBITAG (WP2). Improvement of the technical setup of BCAS through the purchase of scientific instruments is another aspect of MOBITAG (WP4) that will contribute to bridging the gap in the research potential between the founding and the new EU countries.

Enhancement of the managerial skills, especially in respect to FP7, is another goal of MOBITAG. The administrative staff of BCAS manages nearly two hundred national and international projects and can handle most aspects of MOBITAG implementation but there is room for improvement. Our plans include training of selected BCAS personnel in the management of EU projects from their conception to the final report (WP3). MOBITAG will also be used for the build-up of pan-European research network. BCAS is represented in the Technological Platform EPSO (European Plant Science Organization) and in IOBC (International Organisation for the Biological Control of Noxious Animals and Plants). We shall send representatives to the EPSO meeting in the first project year and organize a conference in collaboration with IOBC in the third year. Our conference in 2010 will have a global dimension and will be organized jointly with the International Society for Invertebrate Development and Reproduction (WP5).

Project goals will be achieved through:

• Recruitment of experts who are working in BCAS on temporary basis,
• Return of experienced researchers working in USA),
• Training of BCAS staff in EU laboratories,
• Participation of EU scientists in BCAS research,
• Mobility of researchers,
• Training of research managers,
• Acquisition of research equipment,
• Organization of meetings,
• Several types of dissemination activities.

Social aspects

The newly built research potential, the collaborative projects with partners across Europe, and the results of performed investigations will have social impact within the region and beyond. New research capacities will provide for the development of modern biotechnologies, strengthen the ties between investigators, farmers, and other entrepreneurs in agricultural businesses, and eventually enhance economic potential of the region. The collaborations with EU partners will promote mobility with all its benefits: rapid information exchange, combination of expertise to the solution of European problems, and, last but not least, appreciation of diverse cultures. Organisation of the workshops and conferences is an important integrative component of European science and represents an indispensable part of MOBITAG (WP5). General public will be informed about the project through integrative web pages and the results of accomplished investigations on novel biotechnologies will be disseminated in several ways (WP6). Our aim is to reach a broad audience, ranging from students and farmers to the top politicians of the region and the country. This will contribute to critical but rational evaluations of new agricultural methods in the Czech Republic and also internationally. A third goal is to increase awareness of the farmers, students, and general public about the problems of contemporary agriculture, the role of GM crops, and the necessity of thorough risk assessments whenever a new technology is introduced.

Research plan

Practical training of researchers can be effectively accomplished only through their participation in targetted investigations. The chosen topics, which are specified below, are consistent with the aims of European research declared in Decision No. 1982/2006/EC of the European Parliament and the Council (7th Framework Program). They are in the focus of contemporary biology and therefore ideal for international collaboration and training, and are of pivotal importance for the advance of agricultural biotechnologies. Following paragraphs describe briefly the contents of each topic.

Genome analysis

Genome composition is a precise characteristic of each individual. Various methods of DNA analysis allow a rapid identification of species and their genotypes, as well as of introduced transgenes. Research conducted in the frame of MOBITAG will focus on the detection and identification of the viral and viroid infections in plants and on the identification of insect pests and their genotypes. Genome analysis will be used in the tracing and combating pest insects.Viroid diagnosis based on real-time PCR and RT PCR in combinations with molecular hybridization methods will focus on the monitoring of viroids that cause great economic losses in Central Europe: potato viroids in the potato, tomato, and ornamentals, hop viroids, grapevine viroids, and fruit trees viroids. New methodology based on the oriented suspension array technology will be developed. This technique can employ either nucleic acid hybridization or antigen-antiserum reaction and allows simultaneous detection of up to 100 different molecules. Many plant pathogens are transmitted by aphids that can rapidly adapt to environmental changes. Species which spread to new territories deserve special attention. MOBITAG research will focus on the recognition of genotypes that differ in economic importance in the old, already adapted immigrants (Adelgidae and Lachnidae) as well as in the newly arrived Russian wheat aphid (Diuraphis noxia), a serious wheat pest. Last application of the genome analysis in MOBITAG will be in the sterile insect technique (SIT). Adults of both sexes are sterlized and released in the current SIT programmes for the management of lepidopteran pests but the male-only release would be more efficient and better acceptable by farmers. MOBITAG program includes identification of the sex-determining genes (by means of molecular genetics and cytogenetics) as a background for the development of a method of female elimination. Most of the research will be done on the codling moth, Cydia pomonella, which is the key pest of most pome fruit and some walnut orchards in temperate regions around the world.

Exploitation of natural compounds

Natural compounds have been widely exploited in plant protection for centuries but modern techniques open new possibilities. Proteinase inhibitors, which curb digestive proteinases and thereby reduce insect growth and reproduction, have been cloned into plants and rendered them partly resistant to the pest attack. New types of inhibitors, identified in the digestive system of cockroaches, will be analyzed in MOBITAG. Attention will also be paid to adipokinetic hormones (AKH) that affect energy metabolism and the behaviour of insects. Application of AKH agonists on the locusts, aphids, and other pests is envisaged. The Imaginal Disc Growth Factors (IDGFs) represent a third class of potentially insecticidal compounds to be examined. These compounds are related to the Adenosine Deaminase Growth Factors (ADGFs). Preliminary data obtained with IDGF gene disruption using RNA interference suggest that metamorphosis is a critical period for their function. Since IDGFs and ADGFs differ from the homologous vertebrate growth factors, they have a potential to be developed for insect pest control as a new type of insect growth regulators.

Transgenosis and transgene expression

Crop production depends to great extent on the plant genotype. The methods of bioengineering permit targeted changes of the genotype by which the plant acquires desired properties, such as resistance to the pests or pathogens. Such genetic modifications (GM) have opponents but rapid spreading of the GM crops (grown on more than 100 million ha in 2007) testifies to their advantages and justifies further research. GM rendering spruce resistant to the Norwegian spruce bark beetle, Ips typographus, would be of enormous economic importance for European forestry. Proposed research will therefore include reconstruction of the natural Cry3A gene for use against I. typographus and subsequent gene cloning into the spruce. Standard methods will be used for spruce transgenosis along with simultaneous development of new biotechnology based on the transcription factors (TF) Myb, bHLH and bZip. The main goal of TF biotechnology research is to prepare systems for plant transgenosis that would modify desired parameters, for example the production of drugs such as prenylflavonoids, or to increase the plant yield, due to modified expression of the endogenous plant genes. Regulatory factors Myb, bZIP and bHLH will be isolated and analyzed in the heterologous transgenotes. Another modern and promising research topic is the use of plant nucleolytic enzymes for the suppression of human tumours. Large scale production of apoptotic nucleases in plants will be mediated via the system of Agrobacterium tumefaciens infiltrations.

Safety aspects of GM crops

The deployment of genetically modified crops is a revolutionary step that must be carrefully monitored. Scientists should observe the precautionary principle and evaluate the risks and benefits of GM crops and compare them with the risks and benefits of other agrotechniques. MOBITAG will concentrate on the environmental safety of the Bt crops that dominate the GM crops market. The insect-resistant Bt crops carry a Cry gene from Bacillus thuringiensis and express the corresponding Cry protein. The mode of action of these proteins and the cause of their specificity are not known. We chose to examine the cause of specificity using Cry3Aa and the tenebrinoid beetles. Some of them, such as Tenebrio molitor, are sensitive, but their close relatives, for example Tribolium confussum, are resistant to this insect toxin. We shall search for the cause of this difference at the level of digestive proteinases that may destroy the toxin and at the level of toxin receptors in the gut wall. Precise quantification of Cry toxins is another problem associated with the use of Bt crops. Up to fifty fold differences in the results of Cry measurements by ELISA have been reported. It is assumed that differences in the antiserum, extraction buffer, and sample preparation are responsible. The method will be standardized as part of MOBITAG research.