Source: NORTH CAROLINA STATE UNIV submitted to
TURFGRASS BREEDING AND GENETICS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
NEW
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0220358
Grant No.
(N/A)
Project No.
NC02304
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Jul 1, 2009
Project End Date
Jul 1, 2014
Grant Year
(N/A)
Project Director
Milla-Lewis, SU, R..
Recipient Organization
NORTH CAROLINA STATE UNIV
(N/A)
RALEIGH,NC 27695
Performing Department
Crop Science
Non Technical Summary
As an agricultural commodity, turfgrass is not food, fiber or animal feed. It impacts the lives of millions of people in many different ways, however. The turfgrass industry has grown into a multibillion dollar business worldwide, that includes $57.9 billion in business in the U.S alone in 2000. According to the last North Carolina Turfgrass survey (1999), the total acreage of tufgrass in the state neared 2.14 million acres. Over $1.22 billion was spent to maintain these areas. Single family dwellings were by far the largest single sector of the industry, with 69% of the turf area and 60% of expenditures. Regarding the types of grasses used, tall fescue accounted for over 40% of the total area of maintained turf. Centipede and Bermuda were the next two most common types with 16% and 11%, respectively. North Carolina is located in the transition zone for cool- and warm-season grasses. While this means that we can grow both types of grasses, no one type of grass does well in all weather conditions. Cool season grasses go dormant during hot, dry months and can turn brown and even die during extremely hot and dry conditions. Warm season grasses are very drought tolerant because of their low water requirement, which makes them ideal during the summer months. However, they go dormant and turn brown during the cool days of fall and winter, and winter kill can often be a problem in the NC region. Applied plant breeding can be used to develop turfgrass cultivars that are better adapted to environmental conditions present in North Carolina. Development of cool-season grasses with increased heat and drought tolerance, and warm-season grasses with improved cold tolerance would contribute significantly to different sectors of the turfgrass industry. Genetics and applied plant breeding can also be used to improve aesthetics, performance, and disease resistance of turfgrass cultivars which, in turn, affect the efficiency of maintenance and production. The long-term goal of the program is to improve the sustainability and economic gain of the overall turfgrass industry in the state through the development of cultivars that require reduced inputs and that are capable of tolerating biotic and environmental stresses while still maintaining good quality and overall performance. Despite the fact that large amounts of genomic information have been generated for other grasses such as rice, maize, and wheat, turfgrass species have lagged far behind. Work is needed to generate such information and utilize it to relate phenotypic traits to DNA nucleotide polymorphism.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2011620108010%
2011621108010%
2011629108010%
2021629108010%
2031620108010%
2031621108010%
2031629108010%
2111629108110%
2121621108110%
2121629108110%
Knowledge Area
212 - Pathogens and Nematodes Affecting Plants; 211 - Insects, Mites, and Other Arthropods Affecting Plants; 203 - Plant Biological Efficiency and Abiotic Stresses Affecting Plants; 202 - Plant Genetic Resources; 201 - Plant Genome, Genetics, and Genetic Mechanisms;

Subject Of Investigation
1620 - Warm season perennial grasses; 1629 - Perennial grasses, other; 1621 - Cool season perennial grasses;

Field Of Science
1081 - Breeding; 1080 - Genetics;
Goals / Objectives
a) Recurrent phenotypic selection methods will be used to develop breeding populations of tall fescue. Replicated field studies with no or limited supplemental irrigation will be conducted at the Sandhills Research Station to evaluate this material for heat and drought tolerance. b) Accessions of Cynodon transvaalensis collected from South Africa and that possess good shade tolerance will be hybridized with accessions of C. dactylon obtained from NPGS that have been previously screened for cold tolerance. Sterile triploid hybrids derived from this crosses will be evaluated in replicated field studies under a shade tent and at high elevation in the mountains of NC (for cold tolerance). c) To study the inheritance of cold tolerance, a diallel mating design will be set up with the most promising lines from cold tolerance studies in the mountains of NC. Crosses among lines will be performed without reciprocals. Progenies will be evaluated for establishment rate, winter survival rate, quality, and morphological characteristics in a replicated trial at two different locations in the mountains of NC. d) A set of PIs and cultivars of St. Augustinegrass will be evaluated in the NCSU phytotron for resistance to gray leaf spot, Pyricularia grisea. Additionally, a set of plant introductions, including accessions of Zoysia japonica, Z. matrella, and Z. pacifica, and cultivars of Zoysiagrass will be evaluated under the same conditions for resistance to brown patch, Rhizoctonia solani. The goals of this work are to: 1) identify resistance genes that can be subsequently used in development of disease resistant cultivars, 2) evaluate the effect of different isolates on disease response, and 3) elucidate genetic control of disease resistance. e) Ten thousand of each St. Augustine and Zoysiagrass EST sequences will be developed and screened for SSRs. Subsequently, SSR primer pairs will be designed from non-redundant EST sequences. Additionally, maize SSRs will be tested for transferability across different warm-season grass species. Our goal is to develop a set of 200-300 SSRs to be used in combination with AFLP and SRAP markers for creating the first linkage maps of both St. Augustine and Zoysiagrass. f) A set of 20 St. Augustine cultivars and 13 plant introductions from the National Plant Germplasm System will be analyzed for presence of molecular polymorphism with AFLP and SRAP markers. Data will be analyzed in order to assess levels of intra- and inter-specific variation present in these germplasm. Additionally, genetic distances will be used to elucidate taxonomic relationships through cluster analysis. The same procedures will be utilized to assess levels of molecular diversity among a collection of 30 cultivars and 53 plant introductions representing different species of Zoysiagrass. g) Ethyl methane-sulfonate (EMS) will be used to induce mutations in centipedegrass cultivar `Common'. Following mutagenic treatment of seeds, regenerated plantlets will be grown in the field and screened for morphological differences.
Project Methods
a) Recurrent phenotypic selection methods will be used to develop breeding populations of tall fescue. Replicated field studies with no or limited supplemental irrigation will be conducted at the Sandhills Research Station to evaluate this material for heat and drought tolerance. b) Accessions of Cynodon transvaalensis collected from South Africa and that possess good shade tolerance will be hybridized with accessions of C. dactylon obtained from NPGS that have been previously screened for cold tolerance. Sterile triploid hybrids derived from this crosses will be evaluated in replicated field studies under a shade tent and at high elevation in the mountains of NC (for cold tolerance). c) To study the inheritance of cold tolerance, a diallel mating design will be set up with the most promising lines from cold tolerance studies in the mountains of NC. Crosses among lines will be performed without reciprocals. Progenies will be evaluated for establishment rate, winter survival rate, quality, and morphological characteristics in a replicated trial at two different locations in the mountains of NC. d) A set of PIs and cultivars of St. Augustinegrass will be evaluated in the NCSU phytotron for resistance to gray leaf spot, Pyricularia grisea. Additionally, a set of plant introductions, including accessions of Zoysia japonica, Z. matrella, and Z. pacifica, and cultivars of Zoysiagrass will be evaluated under the same conditions for resistance to brown patch, Rhizoctonia solani. The goals of this work are to: 1) identify resistance genes that can be subsequently used in development of disease resistant cultivars, 2) evaluate the effect of different isolates on disease response, and 3) elucidate genetic control of disease resistance. e) Ten thousand of each St. Augustine and Zoysiagrass EST sequences will be developed and screened for SSRs. Subsequently, SSR primer pairs will be designed from non-redundant EST sequences. Additionally, maize SSRs will be tested for transferability across different warm-season grass species. Our goal is to develop a set of approx. 200 SSRs to be used in combination with AFLP and SRAP markers for creating the first linkage maps of both St. Augustine and Zoysiagrass. f) A set of 20 St. Augustine cultivars and 13 plant introductions from the National Plant Germplasm System will be analyzed for presence of molecular polymorphism with AFLP and SRAP markers. Data will be analyzed in order to assess levels of intra- and inter-specific variation present in these germplasm. Additionally, genetic distances will be used to elucidate taxonomic relationships through cluster analysis. The same procedures will be utilized to assess levels of molecular diversity among a collection of 30 cultivars and 53 plant introductions representing different species of Zoysiagrass. g) Ethyl methane-sulfonate (EMS) will be used to induce mutations in centipedegrass cultivar Common. Following mutagenic treatment of seeds, regenerated plantlets will be grown in the field and screened for morphological differences.

Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Dr. Milla-Lewis does not hold a formal extension appointment. The turfgrass breeding and genetics program, however, has a broad group of clientele. As such, Dr. Milla-Lewis’ extension activities typically include outreach interactions with sod growers, turfgrass seed companies, and other components of the turfgrass industry. Consequently, Dr. Milla-Lewis is involved in interactions with these groups through supply of genetic information and genomic resources, and also through training and research. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Participated in Creating Awareness of Agriculture and Life Science Degrees, Disciplines, and Discoveries (CAALS 3D) Program: - July 17th – 19th, 2012: Nia Crews, Angel Chukwu, Gabrielle Masseaux (North Carolina School of Science and Math, Durham, NC). Graduated two MS students: Katherine Youngs. St. Augustinegrass germplasm resistance screening against North Carolina southern chinch bug, Blissus insularis Barber, populations. (Co-Advisor, Dr. Susana R. Milla-Lewis, Department of Entomology, completed Spring 2013) Steven Mulkey. SSR Development and Mapping of Gray Leaf Spot Resistance Genes in St. Augustinegrass. (Advisor, Dr. Susana R. Milla-Lewis, Department of Crop Science, completed Spring 2013) How have the results been disseminated to communities of interest? Publications in peer reviewed journals (listed on previous page) Presentations at professional meetings: 1. Mulkey, S.E., Zuleta, M.C., Ma, B., Arellano, C., and Milla-Lewis, S.R. 2013. Resistance to gray leaf spot in St. Augustinegrass: Characterization and Mapping. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Tampa, FL. Nov 2-6. 2. Dunne, J.C., Reynolds, W.C., Miller, G.L., Arellano, C., Brandenburg, R., Schoeman, A., Yelverton, F., and Milla-Lewis, S.R. 2013. Evaluation of South African bermudagrass germplasm for shade tolerance. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Tampa, FL. Nov 2-6. 3. Harris-Shultz, K.R., Milla-Lewis, S.R., Patton, A.J., Kenworthy, K.E., Chandra, A., and Waltz, F.C. Jr. 2013. Genetic and ploidy variability within six vegetatively propagated zoysiagrass cultivars. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Tampa, FL. Nov 2-6. 4. Carbajal, E.M., Mulkey, S.E., Ma, B., Zuleta, M.C., Arellano, C., and Milla-Lewis, S.R. 2012. Identificación de resistencia al tizón foliar causado por M. grisea en genotipos diploides de cesped San Agustin [Stenotaphrum secundatum (Walt.) Kuntze]. XIII Congreso Internacional de Manejo Integrado de Plagas. Tegucigalpa, Honduras. Oct. 17-19. 5. Milla-Lewis, S.R., Zuleta, M.C., Ma, B., Kenworthy, K.E., and Tredway, L.P. 2012. Phytotron evaluation of zoysiagrass germplasm for large patch resistance. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Cincinnati, OH. Oct 21-24. 6. Mulkey, S.E., Zuleta, M.C., Ma, B., Tredway, L.P., and Milla-Lewis, S.R. 2012. Mapping gray leafspot resistance genes in St. Augustinegrass. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Cincinnati, OH. Oct 21-24. 7. Kimball, J.A., Zuleta, M.C., Harris-Shultz, K.R., Kenworthy, K.E., Lehman, V.G., and Milla-Lewis, S.R. 2012. Patterns of genetic variation suggest introgression between Zoysia species based on simple sequence repeat (SSR) markers and inflorescence traits. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Cincinnati, OH. Oct 21-24. 8. Kenworthy, K.E., Unruh, J.B., Chandra, A., Nelson, L., Schwartz, B.M., Raymer, P., Miller, G.L., Milla-Lewis, S.R., Wu, Y.Q., Martin, D.L, Munoz, P., Giurcanu, M.C. 2012. Genotype by environment interaction in zoysiagrass. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Cincinnati, OH. Oct 21-24. Presentations at field days and grower organization meetings and field days: Milla-Lewis, S.R. Turfgrass breeding at NCSU: Year 4. 2013. N.C. Sod Producers Assoc. Ann. Conf., Raleigh, NC, January 16, 2013. Speaker at 2013 Turfgrass Field Day at the Lake Wheeler Turfgrass Field Lab, Raleigh, NC, August, 2013 What do you plan to do during the next reporting period to accomplish the goals? Continue breeding efforts with the goal of developing warm- and cool season grass cultivars that possess the desired aesthetic characteristics and disease resistance, and that are especially adapted to conditions in North Carolina. * Developing crossing and selection programs to improve resistance to biotic and abiotic stresses in Zoysiagrass, Bermudagrass, St. Augustinegrass, and tall fescue. * Conduct greenhouse and phytotron evaluations for cold tolerance and disease resistance. * Investigate the application of molecular markers in grass improvement as it refers to 1) assessment of genetic diversity and 2) mapping of disease resistance genes. * Investigate the use of mutation breeding for creation of genetic variation in species for which the germplasm base is narrow. * Seek support for the program objectives from the Turfgrass Centere, The Turfgrass Council of North Carolina, and the North Carolina Crop Improvement Association. * Participate in the Turfgrass Council of North Carolina and North Carolina Sod Producers annual conferences, the North Carolina Crop Improvement Association annual meeting, and turf field days. Interact with components of the turfgrass industry to identify targets for future plant breeding efforts.

Impacts
What was accomplished under these goals? a) We're currently on our third cycle of recurrent selection. A new nursery was planted last fall and will be evaluated this summer under no irrigation. b) Due to the unseasonal weather conditions present last Spring in NC, almost no flowring was obtained from crossing blocks. Therefore, no new hybrids were generated last year. c) Last year was the second year of data collection on the diallel. Because of plots loss due to winterkill, data collection was extended one year. Data on winterkill adn spring green-up will be collected this spring. Data will be analyzed and results will be put together for publication in the fall. Preliminary analysis has identified a few lines with imporved cold tolerance and turf quality. d) Work on St. Augustinegrass has been completed. Diploid and polyploid plant introductions with high levels of resistance to the fungus has been identified. A publication summarizing this work is currently under preparation. A resistant PI was used to develop a mapping population in order to identidy QTL associated with resistance. Four putative QTL for area under the disease progress curve and 2 QT: for area under the lession expansion curve were identified. A publication summarizing this work is currently under preparation. The zoysiagrass work is still ongoing, but preliminary results indicate a few accessions appear to have good levels of resistance to Rhizoctonia. e) 200 SSR markers were synthesized and evaluated across a diverse panel os ST. AUgustinegrass germplasm. A publication outlining this work has been accepted for publication in Crop Science. f) Results of this work were published last year in Crop Science (Milla-Lewis et al, 2013) g) 150 M1 plants were grown in the field and allowed to self-pollinate. A total of 3,000 M2 plants were obtained and have been planted at the Sanhills Research Station for evaluation.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Chandi, A., Jordan, D.L., York, A.C., Burton, J.D., Milla-Lewis, S.R., Spears, J.F., Whitaker, J.R., and Wells, R. 2013. Response of herbicide-resistant Palmer amaranth (Amaranthus palmeri) accessions to drought stress. Intl. J. of Agronomy. (DOI 10.1155/2013/823913). Quesenberry, K.H., Kenworthy, K.E., Crow, W.T., Harmon, P.F., Lu, H.J., and Milla-Lewis, S.R. 2013. Lance nematode effects on rooting of two St. Augustinegrass cultivars. International Turfgrass Society Research Journal 12:357-361. Schwartz, B.M., Harris-Shultz, K.R., Contreras, R.N., Hans, C.S., Hanna, W.W., and Milla-Lewis, S.R. 2013. Creation of artificial triploid and tetraploid centipedegrass using colchicine and breeding. International Turfgrass Society Research Journal 12:327-334. Milla-Lewis, S.R., Kimball, J.A., Tuong, T.D., Claure, T.E., and Livingston, D.P. 2013. Freezing tolerance and the histology of recovering nodes in St Augustinegrass. International Turfgrass Society Research Journal 12:523-530. Mulkey, S.E., Zuleta, M.C., Van Esbroeck, G.A., Lu, H.J., Kenworthy, K.E., Milla-Lewis, S.R. 2013. Genetic analysis of a St. Augustinegrass germplasm collection using AFLP markers and flow cytometry. International Turfgrass Society Research Journal 12:281-291. Chandi, A., Milla-Lewis, S.R., Zuleta, M.C., Jordan, D.L., Burton, J.D., York, A.S. Culpepper, S. 2013. Use of AFLP markers to assess genetic diversity in Palmer Amaranth [Amaranthus palmeri (S. Wats.)] populations from North Carolina and Georgia. Weed Sci. 61: (DOI:10.1614/ws-d-12-00001.1) Chandi, A., Jordan, D.L., York, A.C., Milla-Lewis, S.R., Burton, J.D., A.S. Culpepper, Whitaker, J.R. 2013. Interference and control of glyphosate-resistant and susceptible Palmer amaranth (Amaranthus palmeri) populations under greenhouse conditions. Weed Sci. 61: (DOI 10.1614/WS-D-12-00063.1) Kimball, J.A., Zuleta, M.C., Kenworthy, K.E., Lu, H.J., Milla-Lewis, S.R. 2013. Molecular markers enable the identification of contaminants in production fields of Captiva St. Augustinegrass. International Turfgrass Society Research Journal 12:267-273.


Progress 10/01/11 to 09/30/12

Outputs
OUTPUTS: Data was generated on: Zoysiagrass: cold tolerance, brown patch resistance Bermudagrass: shade tolerance, turf quality and establishment St. Augustinegrass: gray leaf spot resistance, cold tolerance, chinch bug resistance, turf quality and establishment. Characterized 98 newly developed SSRs. Germplasm development: 400 St. Augustinegrass and 100 zoysiagrass hybrids were generated. Additionally 1000 centipedegrass mutants were generated. PARTICIPANTS: Individuals: Susana Milla-Lewis, project leader, responsible for inception, design and oversight of all research projects; Carolina Zuleta, research specialist, responsible for management of laboratory activities, execution of crossing programs, maintenance of phytotron experiments; Casey Reynolds, research specialist, responsible for management and data collection of field trials, maintenance of greenhouse experiments. Collaborators: Lane Tredway and Bangya Ma, Dept. of Plant Pathology, NCSU; Ron Qu, Dan Bowman, Matt Martin, and Grady Miller, Dept. of Crop Science, NCSU; Yasmin Cardoza and Rick Brandenburg, Dept. of Entomology, NCSU; Consuelo Arellano, Dept. of Statistics, NCSU; David Livingston, USDA-ARS, Raleigh, NC; Kevin Kenworthy, Univ of Florida; Brian Schwartz, Univ of Georgia Athens; Karen Harris-Shultz, USDA-ARS, Tifton, GA; Aaron Patton, Purdue University; Mike Richardson, Univ of Arkansas; Ambika Chandra, Texas AgriLife. Partner Organizations: Blue Moon Farms, Lebanon, OR; North Carolina Crop Improvement Association, Raleigh, NC. Training: Jennifer Kimball, PhD Student; Steve Mulkey, MS student; Esdras Carbajal, International Scholar from Honduras. TARGET AUDIENCES: Target audiences: North Carolina Sod Producers Association, Turfgrass Council of North Carolina Efforts: oral presentations were given at Turfgrass Annual Field Day in order to provide an update on the program's progress and general education on what breeding processes involve. International scholar Esdras Carbajal (Universidad Nacional de Agronomia, Honduras), continued work in the program for him to acquire hands on experience in the design and implementation of experiments related to different areas of the plant sciences. By learning the proper methods for conducting agricultural research, Esdras can contribute to the advancement of this area of science in his home country. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Molecular markers continue to be used in order to evaluate levels of genetic diversity and speciation in Zoysia, and St. Augustinegrass. In Zoysiagrass, SSR markers were used to verify the presence of interspecific hybrids supporting the hypothesis that Zoysia spp. are subpopulations within one species rather than separate species. In St. Augustinegrass, 98 newly developed SSR markers were used to identify subpopulations and the presence of admixture within the genus. A set of common bermudagrass accessions collected from South Africa was screened for shade tolerance. Two lines were identified which can perform better than the check under 63 and 80% shade were identified. Additionally, African bermudagrass with good levels of cold tolerance have been identified. These accessions will be used in breeding efforts to combine shade and cold tolerance in order to develop improved bermudagrass cultivars. Over 1500 St. Augustinegrass hybrids have been generated through artificial hybridizations. These hybrids are currently being evaluated at three locations for cold tolerance (Upper Mountain Research Station), drought tolerance (Sandhills Research Station), and overall turfgrass quality (Lake Wheeler Turfgrass Field Lab).

Publications

  • Kimball, J.A., Zuleta, M.C., Harris-Shultz, K.R., Kenworthy, K.E., Lehman, V.G., Milla-Lewis, S.R. 2012. Genetic relationships in Zoysia and the identification of putative interspecific hybrids using simple sequence repeat markers and inflorescence traits. Crop Sci. (doi: 10.2135/cropsci2012.04.0218; published online 21 Aug 2012)
  • Milla-Lewis, S.R., Zuleta, M.C., Van Esbroeck, G.A., Quesenberry, K.H., Kenworthy, K.E. 2012. Cytological and molecular characterization of genetic diversity in Stenotaphrum. Crop Sci. (doi: 10.2135/cropsci2012.04.0234; published online 21 Aug 2012)
  • Chandi, A., Jordan, D.L., York, A.C., Milla-Lewis, S.R., Burton, J.D., Culpepper, A.S., Whitaker, J.R. 2012. Interference of selected Palmer Amaranth (Amaranthus palmeri) populations in soybean (Glycine max). Intl. J. of Agronomy. (DOI 10.1155/2012/168267).
  • Milla-Lewis, S.R., Zuleta, M.C., Ma, B., Kenworthy, K.E., and Tredway, L.P. 2012. Phytotron evaluation of zoysiagrass germplasm for large patch resistance. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Cincinnati, OH. Oct 21-24.
  • Mulkey, S.E., Zuleta, M.C., Ma, B., Tredway, L.P., and Milla-Lewis, S.R. 2012. Mapping gray leafspot resistance genes in St. Augustinegrass. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Cincinnati, OH. Oct 21-24.
  • Kimball, J.A., Zuleta, M.C., Harris-Shultz, K.R., Kenworthy, K.E., Lehman, V.G., and Milla-Lewis, S.R. 2012. Patterns of genetic variation suggest introgression between Zoysia species based on simple sequence repeat (SSR) markers and inflorescence traits. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Cincinnati, OH. Oct 21-24.
  • Kenworthy, K.E., Unruh, J.B., Chandra, A., Nelson, L., Schwartz, B.M., Raymer, P., Miller, G.L., Milla-Lewis, S.R., Wu, Y.Q., Martin, D.L, Munoz, P., Giurcanu, M.C. 2012. Genotype by environment interaction in zoysiagrass. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., Cincinnati, OH. Oct 21-24.
  • Chandi, A., Milla-Lewis, S.R., Jordan, D.L., Burton, J.D., Spears, J.F., York, A.C., Whitaker, J.R. 2012. Inheritance of evolved glyphosate resistance in a North Carolina Palmer amaranth (Amaranthus palmieri) biotype. Intl J of Agronomy. (DOI 10.1155/2012/176108).


Progress 10/01/10 to 09/30/11

Outputs
OUTPUTS: Data was generated on zoysiagrass cold tolerance, bermudagrass cold tolerance, bermudagrass shade tolerance, St. Augustinegrass gray leaf spot resistance, zoysiagrass brown patch resistance, St. Augustinegrass turf quality and establishment. Developed 200 SSRs. Graduated two MS students. another 400 St. Augustinegrass hybrids were generated PARTICIPANTS: Individuals: Susana Milla-Lewis, project leader, responsible for inception, design and oversight of all research projects; Carolina Zuleta, research specialist, responsible for management of laboratory activities, execution of crossing programs, maintenance of phytotron experiments; Casey Reynolds, research specialist, responsible for management and data collection of field trials, maintenance of greenhouse experiments; George Van Esbroeck, research technician, responsible for mutation experiments in St. Augustine and Bermudagrass. Collaborators: Lane Tredway and Bangya Ma, Dept. of Plant Pathology, NCSU; Ron Qu, Dan Bowman, Matt Martin, and Grady Miller, Dept. of Crop Science, NCSU; Yasmin Cardoza and Rick Brandenburg, Dept. of Entomology, NCSU; Consuelo Arellano, Dept. of Statistics, NCSU; David Livingston, USDA-ARS, Raleigh, NC; Kevin Kenworthy, Univ of Florida; Brian Schwartz, Univ of Georgia Athens; Karen Harris-Shultz, USDA-ARS, Tifton, GA; Aaron Patton, Purdue University; Mike Richardson, Univ of Arkansas; Ambika Chandra, Texas AgriLife. Partner Organizations: Blue Moon Farms, Lebanon, OR; North Carolina Crop Improvement Association, Raleigh, NC. Training: Jennifer Kimball, MS Student; Steve Mulkey, MS student; Tito Claure, MS student, Fullbright scholar from Cochambamba, Bolivia; Bessy Raudales, Global Training Initiative Intern from Honduras; Esdras Carbajal, International Scholar from Honduras. TARGET AUDIENCES: Target audiences: North Carolina Sod Producers Association, Turfgrass Council of North Carolina Efforts: oral presentations were given at the North Carolina Crop Improvement Association annual meeting, the North Carolina Sod Producers Association summer meeting, and the Turfgrass Annual Field Day in order to provide an update on the program's progress and general education on what breeding processes involve. An international scholar from the Universidad Nacional de Agronomia in Honduras, Mr. Esdras Carbajal, was brought to work in the program for him to acquire hands on experience in the design and implementation of experiments related to different areas of the plant sciences. By learning the proper methods for conducting agricultural research, Esdras can contribute to the advancement of this area of science in his home country. Moreover, he is interested in learning how to manage a breeding program: the evaluation of materials for a trait of interest, the selection of the best individuals, the actual hybridization of the parents of choice, the decision of what breeding methods to use for the development of advanced populations. Learning the basic principles of plant improvement will be extremely useful to him because he will be able to apply those concepts to other crops in the future. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Molecular markers have been used in order to evaluate levels of genetic diversity and speciation in Zoysia, Centipede, and St. Augustinegrass. In Zoysiagrass, results have thrown light into the issue of speciation in Zoysia spp. While previous classifications separated species very firmly, marker results indicate that variation is overlapping and a classification based on leaf texture might be more appropriate. In St. Augustinegrass, molecular research is extremely limited and our results are the first ones of their kind in this species. Data indicate clear separation of accessions into ploidy levels and provides important information to breeders across the nation on the usefulness of some accessions over others in terms of sexual compatibility. In centipedegrass, our marker data indicates very low levels of genetic variation in the species. A significant number SSR markers have been developed for St. Augustinegrass. Breeders across the nation will be able to use these new tools to increase the efficiency of their breeding programs. Pathology work in St. Augustinegrass has generated useful information for breeders working with gray leaf spot. A set of accessions has been identified that are resistant to the fungus. Once this information is dissemination, breeders in areas where gray leaf spot is prevalent can use these accessions in their breeding programs. Our cold physiology work in has generated important information on proper protocols for germplasm screening.

Publications

  • Chandi, A., Milla-Lewis, S.R., Giacomini, D., Westra, P., Preston, C., Jordan, D.L., York, A.C., Burton, J.D., Whitaker, J.R. 2011. Inheritance of evolved resistance to glyphosate in a North Carolina biotype of Palmer amaranth (Amaranthus palmieri). Weed Science (pending).
  • Milla-Lewis, S.R., Ma, B., Arellano, C., Zuleta, M.C., Reynolds, W.C., Tredway, L.P. 2011 Evaluation of St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze) germplasm for gray leaf spot resistance. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., San Antonio, TX. Oct 16-19.
  • Mulkey, S.E., Zuleta, M.C., Kenworthy, K.E., Lu, H., Milla-Lewis, S.R. 2011. Genetic analysis of a St. Augustinegrass germplasm collection based on AFLP markers. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., San Antonio, TX. Oct 16-19.
  • Mulkey, S.E., Zuleta, M.C., Kenworthy, K.E., Lu, H., Milla-Lewis, S.R. 2011. Development and characterization of simple sequence repeat (SSR) markers for St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze). Proc. Amer. Soc. Agron. Intl. Ann. Mtg., San Antonio, TX. Oct 16-19.
  • Kimball, J.A., Zuleta, M.C., Martin, M., Kenworthy, K.E., Chandra, A., Milla-Lewis, S.R. 2011. Use of molecular markers for identity preservation in St. Augustinegrass: elucidating the authenticity of Raleigh samples from sod farms across the Southeastern United States. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., San Antonio, TX. Oct 16-19.
  • Kimball, J.A., Zuleta, M.C., Milla-Lewis, S.R. 2011. Characterization of warm-Season turfgrass germplasm using transferred cereal SSR markers. Proc. Amer. Soc. Agron. Intl. Ann. Mtg., San Antonio, TX. Oct 16-19.
  • Claure, T.E., Livingston, D.P., Tuong, T.D. Milla-Lewis, S.R. 2011. Histological analysis of pre- and post-freezing nodal structure of St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze). Proc. Amer. Soc. Agron. Intl. Ann. Mtg., San Antonio, TX. Oct 16-19.
  • Hassell, L.E., Villegas-Chirinos, F., Milla-Lewis, S.R., Copeland, S.C., Isleib, T.G. 2011. Segregation for branching pattern in two crosses between var. hypogaea and var. vulgaris parents. Proc. Amer. Peanut Res. Educ. Soc. 43: (in press).
  • Chandi, A., Milla-Lewis, S.R., Zuleta, M.C., Jordan, D.L., Burton, J.D., York, A.C. Culpepper, S. 2011. Use of AFLP markers to assess genetic diversity in Palmer Amaranth (Amaranthus palmeri (S. Wats.)) populations from North Carolina and Georgia. Weed Sci. Soc. Am. Online Abstr. 211.
  • Chandi, A., Milla-Lewis, S.R., Zuleta, M.C., Jordan, D.L., Burton, J.D., York, A.C. Culpepper, S. 2011. Elucidating the inheritance of evolved resistance to glyphosate in populations of Palmer Amaranth (Amaranthus palmeri (S. Wats.)) from North Carolina. Weed Sci. Soc. Am. Online Abstr. 95.
  • Chandi, A., Jordan, D.L., York, A.C., Burton, J.D., Milla-Lewis, S.R., Spears, J.F., Whitaker, J. 2011. Recovery of selected Palmer amaranth (Amaranthus palmeri) biotypes from drought stress. Proc. South. Weed Sci. Soc. Am. 64: (in press).
  • Kimball, J.A., Zuleta, M.C., Martin, M., Kenworthy, K.E., Chandra, A., and Milla-Lewis, S.R. 2011. Assessment of Molecular Variation within Raleigh St. Augustinegrass using AFLP markers. HortScience (In press).
  • Milla-Lewis, S.R., Harris-Schultz, K.R., Zuleta, M.C., Kimball, J.A., Schwartz, B.M., and Hanna, W.W. 2011. Use of sequence-related amplified polymorphism (SRAP) markers for comparing levels of genetic diversity in centipedegrass germplasm. Genetic Resources and Crop Evolution (DOI) 10.1007/s10722-011-9780-8.
  • Harris-Shultz, K.R., Milla-Lewis, S.R., Brady, J.A. 2011. Transferability of SSR and RGA markers developed in Cynodon spp. to Zoysia spp. Plant Molecular Biology Reporter (pending).


Progress 10/01/09 to 09/30/10

Outputs
OUTPUTS: A large collection of germplasm has been acquired through agreements with public and private institutions. While the turfgrass breeding program had only a very small amount (ten lines of St. Augustinegrass and nine accessions of Bermudagrass) of available germplasm at the time of its creation, it currently has amassed a significant amount of germplasm consisting of 334 families of tall fescue, 110 accessions of Bermudagrass, 250 accessions of Zoysiagrass, 226 accessions of St. Augustinegrass, and 150 lines of Centipedegrass. Fourteen trials to evaluate germplasm of all these species were planted during the summer of 2010. A total of 1250 drought tolerant selections of tall fescue were made during the fall. These plants were planted in crossing blocks to generate populations for further cycles of selection. Another 120 shade tolerant accessions of tall fescue were planted in a separate block in order to generate populations for selection. A Bermudagrass shade tolerance trial was established at two locations. Shade structures were erected at both locations, plots were established, and data collection will begin in summer 2011. Two separate Bermudagrass trials were established, one to evaluate common Bermuda for cold tolerance and yield, and another one to evaluate African Bermuda accessions for cold tolerance. Two Zoysiagrass trials were established: one at two locations in order to evaluate advanced Zoysiagrass lines developed by AgriLife for their potential for release as cultivars, and another one also at two locations for evaluation of two large Zoysiagrass collections for cold tolerance. Through artificial hybridizations and tissue culture, a total of 263 hybrids have been produced. These materials will be used to 1) study the inheritance of cold tolerance in the species, 2) map genomic regions controlling cold tolerance, and 3) understand physiological aspects of cold tolerance. Additionally, 62 genotypes of St Augustinegrass have been screened for resistance to gray leaf spot. As a result of this work, ten accessions have been identified as resistant and crosses among them have been started in order to generate hybrids for evaluation and selection. All cultivars and PIs of St. Augustinegrass have been classified with the aid of molecular markers and flow cytometry. Also, a large collection of Zoysiagrass germplasm has been classified with the use of molecular markers. A total of 150 mutant Centipedegrass plants have been generated with the use of chemical mutagen, EMS. These materials have been planted in the field in order to generate populations for selection. PARTICIPANTS: Individuals: Susana Milla-Lewis, project leader, responsible for inception, design and oversight of all research projects; Carolina Zuleta, research specialist, responsible for management of laboratory activities, execution of crossing programs, maintenance of phytotron experiments; Casey Reynolds, research specialist, responsible for management and data collection of field trials, maintenance of greenhouse experiments; George Van Esbroeck, research technician, responsible for mutation experiments in St. Augustine and Bermudagrass. Collaborators: Lane Tredway and Bangya Ma, Dept. of Plant Pathology, NCSU; Ron Qu, Dan Bowman, Matt Martin, and Grady Miller, Dept. of Crop Science, NCSU; Yasmin Cardoza and Rick Brandenburg, Dept. of Entomology, NCSU; Consuelo Arellano, Dept. of Statistics, NCSU; David Livingston, USDA-ARS, Raleigh, NC; Kevin Kenworthy, Univ of Florida; Brian Schwartz, Univ of Georgia Athens; Karen Harris, USDA-ARS, Tifton, GA; Aaron Patton, Purdue University; Mike Richardson, Univ of Arkansas; Ambika Chandra, Texas AgriLife. Partner Organizations: Blue Moon Farms, Lebanon, OR; North Carolina Crop Improvement Association, Raleigh, NC. Training: Jennifer Kimball, MS Student; Steve Mulkey, MS student; Tito Claure, MS student, Fullbright scholar from Cochambamba, Bolivia; Bessy Raudales, Global Training Initiative Intern from Honduras. TARGET AUDIENCES: Target audiences: North Carolina Sod Producers Association, Turfgrass Council of North Carolina Efforts: oral presentations were given at the North Carolina Sod Producers Association and the Turfgrass Council of North Carolina Annual Conferences in order to provide an update on the program's progress and general education on what breeding processes involve. An intern from Zamorano University in Honduras, Ms. Bessy Raudales, was brought to work in the program for her to acquire hands on experience in the design and implementation of experiments related to different areas of the plant sciences. By learning the proper methods for conducting agricultural research, Bessy can contribute to the advancement of this area of science in her home country. Moreover, she is interested in learning how to manage a breeding program: the evaluation of materials for a trait of interest, the selection of the best individuals, the actual hybridization of the parents of choice, the decision of what breeding methods to use for the development of advanced populations. Learning the basic principles of plant improvement will be extremely useful to her because she will be able to apply those concepts to other crops in the future. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Molecular markers have been used in order to evaluate levels of genetic diversity and speciation in Zoysia, Centipede, and St. Augustinegrass. In Zoysiagrass, results have thrown light into the issue of speciation in Zoysia spp. While previous classifications separated species very firmly, marker results indicate that variation is overlapping and a classification based on leaf texture might be more appropriate. In St. Augustinegrass, molecular research is extremely limited and our results are the first ones of their kind in this species. Data indicate clear separation of accessions into ploidy levels and provides important information to breeders across the nation on the usefulness of some accessions over others in terms of sexual compatibility. In centipedegrass, our marker data indicates very low levels of genetic variation in the species. These results are prompting breeders to utilize mutation breeding as a means of generating variation in Centipedegrass. Additionally, large amounts of sequence information for St. Augustinegrass have been generated. This information will be used to develop new molecular markers for St. Augustinegrass. Breeders across the nation will be able to use these new tools to increase the efficiency of their breeding programs. Pathology work in St. Augustinegrass has generated useful information for breeders working with gray leaf spot. A set of accessions has been identified that are resistant to the fungus. Once this information is dissemination, breeders in areas where gray leaf spot is prevalent can use these accessions in their breeding programs. Our cold physiology work in has generated important information on proper protocols for germplasm screening.

Publications

  • Harris-Shultz, K. R., Milla-Lewis, S.R., Zuleta, M.C., Schwartz, B.M., Hanna, W.W., and Brady, J.A. 2011. Development of SSR markers and the analysis of genetic diversity and ploidy level in a centipedegrass collection. (pending, for submission to Crop Science)
  • Milla-Lewis, S.R., Zuleta, M.C., Van Esbroeck, G.A., Quesenberry, K.H., and Kenworthy, K.E. 2011. Assessment of genetic diversity in Stenotaphrum germplasm using AFLP markers and flow cytometry. (pending, for submission to Crop Science)
  • Milla-Lewis, S.R., Ma, B., Reynolds, W.C., Zuleta, M.C., Arellano, C., and Tredway, L.P. 2011. Identification of sources of resistance to gray leaf spot in Stenoraphrum germplasm. (pending, for submission to Plant Disease)
  • Milla-Lewis, S.R., Harris, K.R., Zuleta, M.C., Schwartz, B.M., and Hanna, WW. 2010. Assessing Genetic Diversity in Centipedegrass Germplasm with Sequence-related Amplified Polymorphism (SRAP) Markers. Proceedings of the XVIII Plant and Animal Genome Conference, San Diego, CA, Jan 09-13. (available at http://www.intl-pag.org/18/abstracts/P03a_PAGXVIII_123.html
  • Milla-Lewis, S.R., Zuleta, M.C., Van Esbroeck, G.A., Quesenberry, K.H., and Kenworthy, K.E. 2010. Molecular and Cytological Assessment of Genetic Diversity in Stenotaphrum Germplasm. Proceedings of the Agronomy Society of America International Annual Meetings, Long Beach, CA. Oct 31 - Nov 4.
  • Zuleta, M.C., and Milla-Lewis, S.R. 2010. Use of AFLP Markers for Assessment of Genetic Diversity in St. Augustinegrass Germplasm. Proceedings of the International Conference on green plant breeding technologies, Vienna, Austria, Feb 02-05 (in press).
  • Kimball, J.A., Zuleta, M.C., and Milla-Lewis, S.R. 2010. Transferring informative cereal SSRs to warm-season turfgrasses for germplasm characterization and evaluation. Proceedings of the Agronomy Society of America International Annual Meetings, Long Beach, CA. Oct 31 - Nov 4.
  • Kimball, J.A., Zuleta, M.C., Kenworthy, K.E., Lehman, V.G., and Milla-Lewis, S.R. 2010. Assessment of Genetic Diversity in Zoysiagrass Germplasm Using AFLP Markers. Proceedings of the Agronomy Society of America International Annual Meetings, Long Beach, CA. Oct 31 - Nov 4.