Progress 10/27/01 to 11/10/04
Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Our society is becoming increasingly urbanized. As more and more cropland is converted to houses, office parks, shopping centers, etc., the acreage of turfgrass is increasing exponentially. Currently, the value of the turfgrass industry is estimated at $35 billion annually in the United States. As a result, turfgrasses impact more than 90 percent of all people in the U.S. through exposure to home lawns, business landscapes, roadsides, parks, or recreational turf on a daily basis. However, with the increasing urbanization comes a greater demand on resources, such as potable water. Also, with the general public experiencing heightened awareness of the environment and its protection, use of inputs such as fertilizer, pesticides, and water on turfgrass areas is coming under greater scrutiny. In some
jurisdictions, use of these inputs will either be banned or severely restricted for turfgrass use. In addition, the urbanization of America is leading to an overuse of current recreational facilities such as parks, athletic fields, and golf courses. Therefore, there is a great demand to identify novel, unique turfgrasses with the ability to withstand drought, cold, heat diseases, insects, and excessive wear. This project will identify genes and stress tolerance mechanisms in turfgrasses and other species and will transfer those desirable genes and/or traits to improve and enhance turfgrass germplasm. The objective of this research is to use genetic and biotechnology approaches to identify and develop turfgrass germplasm with improved biotic and abiotic stress resistance. The two broad goals for the next 4 years are: 1) to develop molecular marker systems suitable for genetic map development and rapid diversity assessment in important turfgrass species. 2) develop tissue culture
procedures for the production of haploids, doubled haploids, and embryo rescue of wide species crosses. This program falls within Component 2 (Plant Resources) of NP 205. The project focuses primarily on 2.1 (Lack of available germplasm) 2.2 (Plant Biology and Gene Discovery) and 2.3 (Overcoming limitations to plant growth and development). This project also contributes to Component 2 (Genomic Characterization and Genetic Improvement) of NP 301, Genetic Improvement. 2. List the milestones (indicators of progress) from your Project Plan. Year 1 (FY 2004) Establish DNA extraction from seed methodology and screen selected Tall Fescue populations using SSR markers. Continue self-fertilization of annual ryegrass material to develop inbred plants for RI population development. Score AFLP markers on Wisconsin creeping bentgrass population and establish chromosome-pairing behavior. Year 2 (FY 2005) Establish SSR marker allele size range and develop multiplex sets to improve throughput. Submit
manuscript on DNA extraction from grass seed. Initiate anther culture procedures and establish the potential for anther culture in Lolium and Agrostis. Initiate wide crossing efforts with Festuca and Lolium germplasm and develop procedures for embryo rescue in collaboration with Dr. Bughrara at Michigan State University. Year 3 (FY 2006) Submit manuscript on chromosome pairing behavior in Agrostis. Use isozymes to screen for inbred Agrostis germplasm. Begin screening Agrostis germplasm for juvenile to adult phase change mutants. Submit manuscript of genetic diversity of turf type Tall fescue. Continue RI line development in Lolium using both inbreeding and anther culture. Begin mapping F2 populations developed from inbred parents. Begin screening fescue-lolium wide cross hybrids for improved abiotic stress resistance in collaboration with Dr. Bughrara at Michigan State University. Year 4 (FY 2007) Begin RI line development in creeping bentgrass using selected inbred germplasm. Cross
identified phase change mutants to develop segregating populations. Begin screening selected germplasm for brown patch disease resistance and cross selected material. Year 5 (2008) Complete RI line production and distribute population to selected locations for replicated phenotyping. Distribute fescue-lolium material that shows good abiotic stress resistance and turf characteristics to industry breeders for evaluation. Begin mapping F2 generation of RI population and phase change population using AFLP, SSR, and RFLP markers and identify genome regions influencing phase change. 3. Milestones: A. The milestones listed below were scheduled to be completed under Year 1. All milestones were completed. Establish DNA extraction from seed methodology and screen selected Tall Fescue populations using SSR markers. Continue self-fertilization of annual ryegrass material to develop inbred plants for RI population development. Score AFLP markers on Wisconsin creeping bentgrass population and
establish chromosome-pairing behavior. B. Year 2 (2005) Establish SSR marker allele size range and develop multiplex sets to improve throughput. The SSR marker allele sizes are being screened now and muliplex sets will be established for evaluation on an ABI 3730 sequencer. Submit manuscript on DNA extraction from grass seed. The seed DNA extraction is being modified currently to improve the reliability of the procedure. Initiate anther culture procedures and establish the potential for anther culture in Lolium and Agrostis. If suitable experienced personnel can be identified another culture procedures will be started. Initiate wide crossing efforts with Festuca and Lolium germplasm and develop procedures for embryo rescue in collaboration with Dr. Bughrara at Michigan State University. The germplasm is currently in the process of being collected. Submit manuscript on chromosome pairing behavior in Agrostis. The data collection for this manuscript is near completion. Use isozymes
to screen for inbred Agrostis germplasm. Begin screening Agrostis germplasm for juvenile to adult phase change mutants. Year 3 (FY 2006) Assuming the completion of the above described milestones the following objectives should be completed. Submit manuscript of genetic diversity of turf type Tall fescue. Continue RI line development in Lolium using both inbreeding and anther culture. Begin mapping F2 populations developed from inbred parents. Begin screening fescue-lolium wide cross hybrids for improved abiotic stress resistance in collaboration with Dr. Bughrara at Michigan State University. Year 4 (FY 2007) Begin RI line development in creeping bentgrass using selected inbred germplasm. Cross identified phase change mutants to develop segregating populations. Begin screening selected germplasm for brown patch disease resistance and cross selected material. Year 5 (FY2008) Complete RI line production and distribute population to selected locations for replicated phenotyping.
Distribute fescue-lolium material that shows good abiotic stress resistance and turf characteristics to industry breeders for evaluation. Begin mapping F2 generation of RI population and phase change population using AFLP, SSR, and RFLP markers and identify genome regions influencing phase change. 4. What were the most significant accomplishments this past year? The most significant accomplishments this year was the generation of molecular marker data from creeping bentgrass and the utilization of this marker information to establish the genome wide chromosome pairing behavior of creeping bentgrass. The molecular marker data has also been used in the construction of the first genetic linkage map of creeping bentgrass that will lead to the localization of chromosomes influencing disease resistance in this important cool-season turfgrass. In addition molecular marker screening of Tall Fescue has been initiated and these markers will be a useful resource for genetic mapping and
diversity analysis of cool-season turfgrasses especially those in the Lolium/Fescue species complex. A. The most significant accomplishment during FY 2004 is the generation of approximately 400 molecular markers for the analysis of chromosome pairing and genetic map development in creeping bentgrass. B. Two miniature inverted repeat transposable elements (MITEs) have been isolated from the genus Agrostis. One of the MITEs was found in creeping bentgrass and the other in colonial bentgrass. The MITEs belong to the Stowaway family that are often associated with genes of both Monocotyledonous and Dicotyledonous plants. The MITE sequence is being used to develop primers for monitoring presence/absence polymorphisms of these sequences. MITE sequences are often found associated with important biotic and abiotic stress resistance genes therefore these sequences may speed up our ability to clone importance stress responsive genes. D. Progress Report opportunity to submit additional
programmatic information to your Area Office and NPS (optional for all in-house ('D') projects and the projects listed in Appendix A; mandatory for all other subordinate projects). This portion of the report serves to document a Specific Cooperative agreement #58-1230-3-182, CRIS #1230-21000-035-01S, entitled 'Identification of Rhizoctonia brown patch resistant Tall Fescue germplasm' in collaboration with Dr. Andrew Hamblin at the University of Illinois to study brown patch resistance in tall fescue. Unfortunately Dr. Hamblin left the University of Illinois and the project has been terminated without reportable accomplishment. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. This is only the second year of the project, however, we have been able to provide genetic marker data for the construction of a genetic linkage map of creeping bentgrass that will be used to locate genes influencing disease resistance in this
important cool-season turfgrass species. The identification of disease resistance loci in creeping bentgrass could result in a significant reduction is fungicide use by golf courses throughout the U.S. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? AFLP marker data has been developed and shared with other scientists for use in the construction of a genetic linkage map in Agrostis. SSR marker data is being developed that can be used in cultivar identification by the grass seed industry in the U.S. Five scientific meetings were attended. Plant and Animal Genome Meeting, San Diego, CA 1-09-04 to 1-14-04 National Lawns and The Environment Stakeholder Meeting, San Antonio, TX 3-14-04 to 3-17-04 United States Golf Association Research Committee
Meeting, Basking Ridge, NJ 3-23-04 to 3-25-04 Grass Breeders Conference, Auburn, AL 5-16-04 to 5-20-04 United States Golf Association Research Committee Meeting, Madison, WI 8- 03-04 to 8-05-04 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. Jung, G., Chakraborty, J., Bae, J., Curley, J., Sim, S., Warnke, S.E., Barker, R.E. Construction of an Agrostis linkage map and comparative genome analysis with Lolium using RFLP anchor and creeping bentgrass cDNA probes. Plant and Animal Genome Conference 2004. Abstract p. 24. Sim, S., Chang, T., Curley, J., Warnke, S.E., Barker, R.E., Jung, G. Comparative genome analysis between ryegrass and other Poaceae species via the large scale chromosomal rearrangements. Plant and Animal Genome Conference 2004 p. 35. Curley, J., Chakraborty, N., Sung, S., Warnke, S.E., Jung, G. Construction of cDNA libraries of two creeping bentgrass clones, sequence analysis and
preliminary characterization of expressed sequence tags. Plant and Animal Genome Conference 2004 p45. Curley, J., Sim, S.C., Jung, G., Leong, S., Warnke, S.E., Barker, R.E. QTL mapping of gray leaf spot resistance in ryegrass, and synteny-based comparison with rice blast resistance genes in rice. Molecular Breeding of Forage and Turf Conference Proceedings. P. 37. Warnke, S.E., Barker, R.E., Jung, G. Comparative relationships of flowering control QTLs between Poaceae species. BARC poster Day. Warnke, S.E., Chakraborty, N., Jung, G. Characterization of the mode of inheritance in tetraploid creeping bentgrass using AFLP mapping. ASA- CSSA-SSSA annual meeting abstract.
Impacts
(N/A)
Publications
- Warnke, S.E., Barker, R.E., Jung, G., Mian, M.A., Saha, M.C., Brilman, L.A. , Dupal, M.P., Forster, J.W. 2004. Genetic Linkage Mapping of an Annual x Perennial Ryegrass Population. Theor. Appl. Genet. v:109, pp. 294-304.
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Progress 10/01/02 to 09/30/03
Outputs
1. What major problem or issue is being resolved and how are you resolving it? Our society is becoming increasingly more urbanized. As more and more cropland is converted to houses, office parks, shopping centers, etc., the acreage of turfgrass is increasing exponentially. Currently, the value of the turfgrass industry is estimated at $35 billion annually in the United States. As a result, turfgrasses impact more than 90 percent of all people in the U.S. through exposure to home lawns, business landscapes, roadsides, parks, or recreational turf on a daily basis. However, with the increasing urbanization comes a greater demand on resources, such as potable water. Also, with the general public experiencing heightened awareness of the environment and its protection, use of inputs such as fertilizer, pesticides, and water on turfgrass areas is coming under greater scrutiny. In some jurisdictions, use of these inputs will either be banned or severely restricted for
turfgrass use. In addition, the urbanization of America is leading to an overuse of current recreational facilities such as parks, athletic fields, and golf courses. Therefore, there is a great demand to identify novel, unique turfgrasses with the ability to withstand drought, cold, heat diseases, insects, and excessive wear. This project will identify genes and stress tolerance mechanisms in turfgrasses and other species and will transfer those desirable genes and/or traits to improve and enhance turfgrass germplasm. 2. How serious is the problem? Why does it matter? The use of inputs such as fertilizers, pesticides, and water on turfgrass areas is coming under increasing scrutiny by the general public. Therefore, the need for grasses that produce acceptable quality turf stands with reduced pesticides, fertilizer, and water is increasing. Many states have or are considering legislation that limits the use of these various inputs on turfgrass or require only certain types of
turfgrass be utilized. Severe drought situations in several states have resulted in mandatory water restrictions on turf areas. In addition, soil erosion and stabilization on construction sites, slopes, etc. is a major concern of public administrators. 3. How does it relate to the National Program(s) and National Program Component(s) to which it has been assigned? This research relates directly to the production and management of grass areas to improve quality while reducing management inputs, thereby reducing environmental impact. National Program Area 205 (100 percent). 4. What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2003: Determination of the mode of inheritance of disease resistance has a significant impact on germplasm collecting and breeding strategies. An Agrostis stolonifera mapping population developed by Dr. Mike Casler, USDA-ARS and Dr. Geunhwa Jung, University of Wisconsin was established at the U.S.
Arboretum. This population is segregating for disease resistance (gray snow mold) and several morphological characteristics. The population will be screened with AFLP markers and the mode of inheritance (polysomic vs disomic) will be evaluated. B. Other significant accomplishment(s), if any: None C. Significant activities that support special target populations: None 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. This new project is linked with the on-going National Turfgrass Evaluation Program (NTEP) (CRIS 1230-21000-036-00D) which designs, develops, and conducts uniform evaluation trials of turfgrass cultivars in the United States and Canada. These trials have resulted in identification of superior grasses for various uses and regions. 6. What do you expect to accomplish, year by year, over the next 3 years? In 2003-2004 AFLP and SSR marker data will be collected on an Agrostis stolonifera population developed by
Dr. Mike Casler, USDA-ARS and Dr, Geunhwa Jung, University of Wisconsin. Marker data will be used to establish the genome wide chromosome pairing behavior in this important turf species. Procedures for DNA extraction from turfgrass seed will be developed and used to collect SSR marker data in tall fescue. The SSR maker data from tall fescue will be used to study the genetic diversity of turf-type cultivars of this species. In 2004-2005 tissue culture protocols will be initiated to develop haploid plants through another culture and to regenerate wide cross hybrids through embryo rescue. Fescue-ryegrass hybrids will be developed and confirmed using molecular markers. Studies on phase change in creeping bentgrass will be initiated to determine the time of transition from juvenile to adult vegetative phase in this species. In 2005-2006 recombinant inbred lines of ryegrass will be developed using inbreeding procedures and another culture to produce doubled haploids. In addition wide
cross hybrid and tissue culture work will continue. 7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Molecular markers and desirable germplasm will be made available to public and commercial grass breeding programs for use in developing and identifying superior varieties. 8. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: This does not replace your peer-reviewed publications listed below). Warnke, S.E. Creeping Bentgrass (Agrostis stolonifera L.). Casler, M.D., Duncan, R.R., editors. John Wiley Sons, Inc, Hoboken, New Jersey. Turfgrass Biology, Genetics, and Breeding. 2003. p. 175-185. Warnke, S.E. Gave a talk at the Maryland Turfgrass Conference describing
the new turfgrass program at the National Arboretum. 1/21/2003 Warnke, S.E. Presented a poster for the BARC technology showcase. April 15, 2003.
Impacts
(N/A)
Publications
- Warnke, S.E., Barker, R.E., Jung, G., Mian, R., Brilman, L.A. Chromosomal regions influencing flowering time in ryegrass. Crop Science Society of America. 2003. Paper No. 419192.
- Warnke, S.E., Barker, R.E., Sim, S-C., Jung, G. Genetic map development and syntenic relationships of an annual X perennial ryegrass mapping population. Plant and Animal Genome. 2003. Abstract p.46.
- Warnke, S.E. In search of new plants. Maryland Turfgrass Conference 2003. Paper No. 29.
- Warnke, S.E., Barker, R.E., Sim, S-C., Jung, G., Mian, R. Identification of flowering time QTL's in an annual x perennial ryegrass mapping population. Molecular Breeding of Forage and Turf Conference. 2003. p. 122.
- Sim, S-C., Chang, T., Curley, J., Warnke, S.E., Barker, R.E., Jung, G. Comparative genomics of the Lolium and Poaceae species. Molecular Breeding of Forage and Turf Conference. 2003. p. 96.
- Brown, R., Warnke, S.E., Barker, R.E. Towards a DNA-based screening test for annual ryegrass (Lolium multiforum) contamination of perennial ryegrass (L. perenne) seed. Molecular breeding of forage and turf conference. 2003. p. 158.
- Curley, J., Sim, S-C., Leong, S., Warnke, S.E., Barker, R.E., Jung, G. Comparison of putative orthologous Magnaporthe grisea resistance QTL in rice, barley and ryegrass. Molecular Breeding of Forage and Turf Conference. 2003. p. 4.
- Warnke, S.E. Identification of flowering time QTL's in an annual x perennial ryegrass mapping population. BARC poster day. 2003. p. 25.
- Barker, R.E., Warnke, S.E., Davidson, S.K., Brown, R.N. Combining morphological and isozyme markers to predict growth type contamination in high quality ryegrass turf. Crop Science Society of America. 2003. Paper No. 315223.
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