Australia's Department of Industry, Science, and Technology funds a physical mapping effort in the multinational Arabidopsis genome project. In this effort, genes involved in male fertility, P450s, cytokinin metabolism, and cellulose biosynthesis are being mapped and isolated. Random expressed sequence tags (ESTs) are also being mapped to the Arabidopsis genome, either to yeast artificial chromosomes (YACs) or by restriction fragment length polymorphism (RFLP). Additionally, Arabidopsis projects are ongoing at approximately six Australian institutions; these are focused on floral mutations, male sterility, root morphology, signal transduction on anaerobiosis, and sugar transport.
Contact: Elizabeth Dennis, CSIRO
Belgium has several national programs that support Arabidopsis research, including some financed by the Flemish Community. One program aims specifically at the further development of Arabidopsis as a model plant: It includes mapping and sequencing projects and molecular biological studies of, for example, ethylene response and oxidative stress. In another project, the interaction of plant nematodes with Arabidopsis is under investigation. This effort includes (1) mapping and analysis of mutant Arabidopsis plants that are resistant to nematode infection and (2) establishment of a protein database of Arabidopsis thaliana, using two-dimensional electrophoresis and partial amino acid sequence analysis.
The Belgian Government promotes collaboration among universities. For example, the Laboratory of Genetics at Ghent works with other Belgian universities, such as those in Antwerp and Brussels, that also have a department of plant biotechnology. This
particular project involves the study of regulatory genes involved in morphological development, and study -- on the microscopic level -- of interphase nucleus, using fluorescence in-situ hybridization.
Another Belgian project entails the evaluation of a novel genetic strategy -- selective restriction fragment amplification -- to identify markers closely associated with a mutation. This new technique was developed by Keygene, a Dutch company.
Contact: Marc Van Montagu, University of Ghent
The groups of Bertrand Lemieux at York University and Peter McCourt at the University of Toronto are collaborating with the group of Ronald W. Davis at Stanford University to map EST sites by polymerase chain reaction (PCR) amplification. These groups are using DNA isolated from YAC clones of a French collaboration - comprised of the Center for the Study of Human Polymorphisms (CEPH), the Institut National de Recherche Agronomique (INRA), and the Centre National de la Recherche Scientifique (a collaboration also known as CIC) - the YAC bank at the University of Pennsylvania, and the Grill-Somerville banks as template DNA. To date, primers for the amplification of 800 ESTs have been produced by Deval Lashkari (Stanford, CA); and DNA has been isolated from 3,456 Arabidopsis clones by Gus Lagos (York, Canada). The YAC clone pooling strategy of Balding and Torney (Los Alamos, NM) is being used; this allows complete mapping of EST sites without colony hybridizations. This 2-year project has a target of 2,000 EST markers.
Contact: Bertrand Lemieux, York University
The Biotechnology Research for Innovation, Development, and Growth in Europe (BRIDGE) project, "Molecular Identification of New Plant Genes," supported by the European Community (EC), ended in June 1994. Highlights of the program included the isolation of several genes that mediate environmental effects on flowering and the isolation of two genes involved in responses to abscisic acid. The transposon tagging development program, based on the transposons Ac and En, has provided two very useful systems. Using these, several thousand families containing potential mutants have been generated, as well as a series of lines with mapped transposon "launching pads" for targeted mutations. The physical mapping work has resulted in the assembly of five contigs spanning more than 16 megabases (Mb) of chromosome IV and a large area of chromosome V. The mapping has provided the basis for a large-scale sequencing project (described in the previous section).
The EC program, together with the United Kingdom's Biotechnology and Biological Sciences Research Council, also supported the Nottingham Arabidopsis Stock Centre (NASC) and the European DNA resource center at Cologne. (In fact, future plans
for NASC may include its serving as another DNA resource center.) Some of BRIDGE's initial goals remain elusive: For example, the lack of methods for gene replacement represents a serious impediment to progress, especially in determining the function
of specific genes. On the other hand, the most enduring aspect of the BRIDGE project is the development of transnational teams, which continue to provide a significant strength to European biological research.
Future work in Arabidopsis is well-funded within the Fourth Framework Program, a 4-year plan that follows the EC's budgetary cycle. Work will continue on developing a gene replacement system; this work will be part of several efforts aimed at a large-scale search for gene function -- efforts strongly linked to the sequencing project. Research in such areas as the transition to flowering, plant growth and development, genetic and metabolic controls on carbon and nitrogen assimilation, signal perception, plant-pathogen interactions, and embryo and seedling development can all be advanced via studies of Arabidopsis. The EC is interested in sponsoring transnational research in these fields.
Contact: Michael Bevan, John Innes Centre, Norwich, UK
Fourteen French laboratories use Arabidopsis as a tool for advancing plant biology research in their studies of embryogenesis, abscisic acid signaling, dormancy, ion transport, root development, lipid metabolism, cell cycle, protein kinases,
gametogenesis, plasmalemma proteins, nitrate and carbon metabolism, transposable elements, plant-pathogen interactions, and stomata. Three collective objectives in genome analysis are being pursued; these involve sequencing, mapping, and generation
of transferred DNA (T-DNA) tagged lines.
More than 30 laboratories in Germany are using Arabidopsis as their primary experimental organism. The research projects fall into three broad categories: developmental genetics, molecular physiology, and plant-microbe interactions. Groups working on developmental problems are funded under a 6-year special research program, "Arabidopsis as a Model for the Genetic Analysis of Plant Development"; this program was set up by the Deutsche Forschunggemeinschaft, Germany's national research association, in August 1993. More than 30 participants from both Germany and abroad attended the program's first meeting, which was held in Breisach, October 28-30, 1993. (Highlights of this meeting are described in the previous section.) In 1994, the program convened a specialized workshop on pattern formation; this was held in Tubingen, October 20-21, and attracted 35 participants. Another meeting on hormones will be held in the near future.
Contact: Gerd Jurgens, University of Tubingen
Arabidopsis research is carried out at more than 20 laboratories at national, prefectural, and private universities; national institutes; and some companies. The research topics explored include mutational analyses of flower and leaf development, responses to physical and chemical stimuli in roots or stems, root morphology, virus infection, hormonal regulation, transcriptional regulation, and shoot and root regeneration from calli. Also under study are the isolation and characterization of genes involved in the heat-shock response, desiccation, lipid biosynthesis, signaling pathways, protein phosphorylation, and transcription. The molecular function of isolated genes is examined using transgenic plants. In addition, efforts focus on developing experimental techniques, including Agrobacterium-mediated transformation and a new PCR-based method for isolating the flanking region of inserted T-DNA from transformants
Several leading Arabidopsis studies were summarized in a special issue of the Japanese journal Shokubutsu Saibo Kogaku (Vol. 6, No. 3). The Japanese Arabidopsis seed stock center was established in 1993 at the Miyagi College of Education, Sendai. The center has seed collections donated by Albert Kranz and Nobuharu Goto; seeds can be ordered by mail from the center.
Arabidopsis study descriptions and findings were presented at
various symposia, including the annual meetings of the Genetic Society
of Japan, the Japanese Society of Molecular Biology, and the Japanese
Society of Plant Physiologists. Also, the Fourth Workshop on
Arabidopsis Studies, organized by Kiyotaka Okada and Yoshiro
Shimura (National Institute of Basic Biology -- NIBB) was held at NIBB,
December 3-4, 1993, with more than 90 attending. Arabidopsis
databases and communication using e-mail were demonstrated. The Fifth
Workshop was held November 25-26, 1994, at NIBB.
Contact: Kiyotaka Okada, NIBB, Okazaki The Netherlands
The Netherlands
In January 1994, research groups working at the Universities of Amsterdam,
Leiden, Utrecht, and Wageningen; and at the Dutch Center for Plant Breeding
and Reproduction Research (CPRO-DLO) in Wageningen established an
Arabidopsis network for The Netherlands. The group, called
ARANED, is chaired by Maarten Koornneef (Genetics Department, Agricultural
University, Wageningen); it will organize common research activities such as joint mutant screens and exchange of DNA from recombinant inbred lines. Also, the group will promote the exchange of information, materials, and experience through annual
meetings. An ARANED research proposal to study embryogenesis and seed development was submitted to the Dutch National Science Foundation. Currently, Arabidopsis research in The Netherlands focuses on development, signal transduction --
especially related to hormones and light -- and plant-pathogen interactions.
Contact: Maarten Koornneef, Wageningen
Approximately 10 laboratories are currently involved in Arabidopsis research. Subjects under study include the mutational and molecular analysis of leaf senescence and photomorphogenesis at Pohang University of Science and Technology (POSTECH); regulation of wound-inducible genes at Chonnam National University; function and regulation of drought-inducible and other protein kinase genes at Kyoungsang National University and POSTECH; biochemical study of photosignal transduction at Kyungbuk National University; protoplast culture at Chosun University; and T-DNA insertional mutagenesis at Kyungbuk National University and POSTECH.
A few other laboratories are also beginning to get involved in Arabidopsis studies. Additionally, there is a cDNA sequencing program on Brassica plants (Brassica napus and Brassica campestris), which may complement the Arabidopsis cDNA
project. The current number of sequences obtained by the Plant Molecular Biology and Biotechnology Center in Jinju and the Agricultural Genetic Engineering Center in Suwon is nearing 3,500. All of these studies are primarily funded by the Ministry of
Education, the Korean Science and Engineering Foundation, the Plant Molecular Biology and Biotechnology Center, the Department of Agriculture, and POSTECH.
Contact: Hong Gil Nam, POSTECH
The number of laboratories using Arabidopsis in Spain is growing. Up to 25 laboratories have expressed an interest in participating in the Arabidopsis Spanish network, and more than two-thirds of these already have projects which use this species. These projects are funded by national and European agencies. Most of the laboratories involved are at institutes of CSIC, the Spanish National Research Council; at universities in Madrid, Barcelona, and Valencia; and at INIA, the Spanish Institute for Agricultural Research in Madrid.
After a meeting held in 1993 in Valencia, efforts began which involved joint use of recombinant inbred lines and the production of additional T-DNA tagged lines. And, after the November 1994 meeting of the Spanish network in Madrid, efforts involving joint generation and screening of 10,000 T-DNA tagged lines commenced; this project constitutes a main objective for the next year.
Contact: Jose Martinez Zapater, Centro de Investigacion y Tecnologia, Madrid
Overall, Arabidopsis is a well-established and widely used tool in the United Kingdom for innovative research in fundamental and applied science.
The United Kingdom's 4-year Plant Molecular Biology II (PMBII) Program -- which includes about 20 grants from the Biotechnology and Biological Sciences Research Council (BBSRC) to universities and research institutes for Arabidopsis research -- is now half complete. (The BBSRC was formed in April 1994 by a merger of the old funding councils, the Agriculture and Food Research Council and part of the Science and Engineering Research Council.) The mid-term meeting of all PMBII participants was held at the University of East Anglia, July 6-8, 1994. At this meeting, participants reported significant advances in the understanding of floral induction, plant hormones, plant development, and plant-pathogen interactions.
In addition to research sponsored under this coordinated program, BBSRC has made numerous other grants for Arabidopsis research. Beginning in 1994, further support for Arabidopsis genome research was made available as part of a new £2 million, 3-year BBSRC program on Plant and Animal Genome Analysis. Seven of the 14 funded proposals concerned Arabidopsis; these include an effort to strengthen the physical mapping effort, one to start site-selected transposon mutagenesis, and one to assess the colinearity of the Arabidopsis and Brassica genomes. Proposals for phase two of this program are now being considered.
BBSRC support for Arabidopsis researchers in the United Kingdom also includes funding for the Nottingham Arabidopsis Center. (Further details of the stock center's activities and collaborations are provided in the previous section.)
Also, EC-funded Arabidopsis research in the United Kingdom includes genome mapping and sequencing under the ESSA program.
Contact: Bernard Mulligan, University of Nottingham
The North American Arabidopsis Science Steering Committee (NAASC) convened a workshop in June 1994 on Arabidopsis genome sequencing. The workshop report proposes establishing a program to sequence the entire Arabidopsis genome by the year 2004. The proposal was endorsed by the Multinational Science Steering Committee at its June meeting in Amsterdam. It was also widely circulated via electronic bulletin board, and has subsequently been used to present the views of the Arabidopsis community to various U.S. funding agencies.
In terms of funding Arabidopsis research in the United States, the Department of Agriculture, Department of Energy, National Institutes of Health, and National Science Foundation together provided a total of approximately US$22 million for fiscal year (FY) 1993 (i.e., the period Oct. 1, 1992, to Sept. 30, 1993) for research and related activities involving Arabidopsis as an experimental material. (These four agencies had signed in June 1990 an interagency agreement to collaborate on Arabidopsis research in the United States.) These funds supported approximately 180 individual research projects and about 35 individual post doctoral research fellowships, in addition to resource centers; tools and technology development projects; databases; conferences, workshops, courses, and graduate training programs; and equipment and facilities. The areas of research supported cover all aspects of plant biology from population biology and genetics to cell/developmental biology and physiology to biochemistry. Federal funding totaled approximately US$19 million in FY 1992, US$15 million in FY 1991, and US$7.5 million in FY 1990.
The Arabidopsis Biological Resource Center at Ohio State University has a mailing list of 1,225 U.S. individuals (out of a total of 2,102 on the list) in 47 States; this indicates the widespread use of Arabidopsis as an experimental system for plant biology research and teaching in the United States.
Contact: David Meinke, Oklahoma State University
