EPSCoR funding will be used to fund the first two years of salary for a new Assistant Professor of Molecular Cytogenetics. Start-up funds for this position will be provided by the state as part of the match. This person will forge essential linkages with the plant genome evolution group (Blake, Lavin McCoy and Talbert) and will also interact with the other two groups. It is expected that this individual will conduct research using in situ hybridization and other appropriate technologies to determine critical elements of genome organization in the species targeted in this proposal. After the first two years this position will be supported by state funds as part of the match.
As part of the building of a quality unified group we are also reqesting NSF funding for a new Assistant Professor of Plant Virology. This person will be hired in year 3 and supported by NSF for the first two years. In the fifth year of this project the molecular virologist would be picked up on state funds as part of the match. Also startup funds for this faculty member will be provided as part of the match. This person will provide essential expertise, and assist our building of a comprehensive genetic manipulation of plants research center.
We are also asking for NSF funds for the new support scientist in the Plant Gene Transfer Facility (See below).
Justification for the Development of a Core Facility for Gene Transfer in Plants.
Five of the eleven PIs in this proposal are already actively involved in Agrobacterium tumefaciens-mediated plant transformation research, and two more are ready to begin transformation activities. Research to date has been carried out in individual laboratories, or in small groups. As such, there has been an unnecessary duplication of equipment, expertise, and efforts. Likewise, none of the individual laboratories engaged in this research is sufficiently equipped to offer hands-on graduate and undergraduate instruction in plant tissue culture and transformation. Therefore, the overall goal of this section is to propose creation of the MSU Core Facility for Plant Transformation. This facility will contribute substantially to plant biology infrastructure development in Montana by fostering collaboration among PIs and providing a common research/teaching resource for student instruction.
Current plant transformation research can be categorized under two common themes:
1) gene expression in transgenic plants, and 2) plant protection through gene transfer. The McCoy laboratory studies the expression and inheritance of Manduca sexta insect protease inhibitor genes in transgenic alfalfa plants. The effects of promoter strength, targeting sequences, and host plant developmental stage on gene expression levels are being evaluated. The goals of the Sharrock laboratory are to identify the molecular receptors for light regulation of growth and development in plants, to define their functions, and to develop methods to manipulate these genes in transgenic plants, thus altering the responses they control. Studies are in progress to identify the physiological effects of disruption or alteration of specific phytochrome genes in transgenic A. thaliana. In related research in the Stout laboratory, the role of free cytosolic Ca++ levels in the intracellular signal transduction of red- and blue-light mediated responses will be quantified in transgenic A. thaliana plants carrying genes for the Ca++-sensitive luminescent protein aequorin. The Dyer laboratory is isolating genes whose expression regulates seed dormancy, which, when transferred to other plants, may allow specific manipulation of this fundamental plant developmental process.
A second common theme of research carried out by six PIs involves the identification and transfer of genes conferring protection against plant pests. The molecular basis of plant defense against insect and pathogen attack is being studied in A. thaliana and Brassica campestris in the Mitchell-Olds laboratory, using a combination of physiological (myrosinase activity) and genetic (RFLP and RAPD mapping) approaches. Genes thus identified will be available for transfer into agronomically important plants. The Anderson laboratory has identified genes that, when introduced into barley, may confer resistance to barley yellow dwarf virus. Likewise, the Sherwood laboratory is cloning genes involved in pheromone synthesis in fungal pathogens of barley: transfer of these genes into plants may interrupt fungal mating and thus severely reduce disease incidence. Genes conferring resistance to a nonselective herbicide are being transformed into safflower in the Dyer laboratory, thus permitting the use of environmentally safe weed management practices.
Formation of a Plant Transformation Core Facility will create a valuable teaching resource for modern training in plant biology. Use of this facility for exercises in plant tissue culture, transformation, and detection of transgenes will nicely complement current course offerings in plant molecular biology. We propose that the facility will not only augment ongoing research programs by stimulating collaboration, but will also provide first-hand experience in the procedures of plant biotechnology for graduate and undergraduate students. The facility will likewise be available for research/instruction of Research Opportunity Awardees. Thus, the plant science infrastructure will be greatly improved and expanded by this facility.
EPSCoR funding for this facility will be used to support a PhD level laboratory manager, who will carry an adjunct faculty appointment in the Plant and Soil Science Department. Responsibilities of this position will include: technical research assistance and advice, maintenance and operation of the particle gun, development and teaching a plant biotechnology course, and collaboration with ongoing research projects, including participation in extramural funding efforts. This individual will be expected to carry out transformation experiments for the Mitchell-Olds laboratory, since they are located in Missoula. It is anticipated that after five years salary for the position and funding for all supplies will be provided by the state. During the five-year funding period state funds will be used for purchasing routine tissue culture and plant transformation supplies. In addition, facility space and the particle gun lease arrangement will be provided as the institutional match. EPSCoR PROJECT: Plant Biosciences (P.I. Tom McCoy)