Progress 07/01/08 to 06/30/13

Outputs
OUTPUTS: The most significant outputs of this project are developing diverse buffalograss germplasm for improved turfgrass quality and performance, chinch bug resistance, and enhanced seed production. To date nine cultivars have been released from the project and two more are in the process of being released. Management practices including improved seed and vegetative establishment, mowing and fertilization practices, and use of mixtures to enhance spring and fall performance of buffalograss stands were developed. This outputs have been share with the scientic community through peer-reviewed journals and professional society meetings, and with the turfgrass industry through field days, workshops, state and regional meetings, industry publications, and electronically through various web-based information sources. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: The target audience of this project is portions of the turfgrass industry interested in reduced input and management grasses. Specific aspects of the industry are lawns, parks, cemeteries, roadsides, sod production, seed production and golf course roughs and fairways. PROJECT MODIFICATIONS: Released nine cultivars that had an impact on turfgrass selection options for the turfgrass industry.

Impacts
Buffalograss turfgrass breeding programs need diverse germplasm to use as parental material, and to develop diverse progeny and improved cultivars. Studies conducted with germplasm collection indicated that turfgrass performance varied among genotypes for quality, color, density, spring green up and seed yield. Thirty two percent of the genotypes evaluated equaled or exceeded turfgrass quality ratings of the standard entries. Seed yield ranged from 460 to 4964 kg ha-1, which was a 10 fold increase between the lowest and highest yielding genotypes. Based on this research, it can be concluded that excellent diversity exists for turfgrass performance and seed yield potential in germplasm available to this program. Hybridization and selection have been one of the methods used to generate turfgrass cultivars in buffalograss improvement. Our studies found differences among crosses and parents for all the traits studied. Best linear unbiased prediction (BLUP) indicated a high improvement potential for turfgrass lateral spread, spring density and turfgrass genetic color. Hybridization breeding is a worthwhile approach for generating and identifying transgressive segregants for specific buffalograss traits.

Publications

• Abeyo, B., and R. C. Shearman. 2009. Buffalograss (Buchloe dactyloides) turfgrass performance and seed yield characteristics. Int. Turfgrass Soc. Res. J. 11(Part 1):p. 519-532.
• Shearman, R. (B.) C., and B. G. Abeyo. 2010. Buffalograss germplasm improvement and management. Turfgrass Environ. Res. Summ. p. 12.
• Severmutlu, S., N. Mutlu, R. C. Shearman, E. Gurbuz, O. Gulsen, M. Hocagil, et al. 2011. Establishment and turf qualities of warm-season turfgrasses in the mediterranean region. HortTechnology. 21(1):p. 67-81.
• Amundsen, K. L., and R. "Bob" Shearman. 2011. Buffalograss germplasm improvement and management. USGA Turfgrass Environ. Res. Summ. p. 18.
• Heng-Moss, T., K. Amundsen, R. Shearman, F. Baxendale, P. Twigg, B. Abeyo, et al. 2011. Molecular characterization of chinch bug-resistant buffalograsses. USGA Turfgrass Environ. Res. Summ. p. 24.
• Peterson, K. W., R. C. Shearman, R. E. Gaussoin, G. L. Horst, and W. H. Schacht. 2010. Growing degree-day influence on sprigged establishment of two buffalograss cultivars. HortScience. 45(2):p. 293-298.
• Heng-Moss, T., O. Gulsen, T. Eichkoff, R. Shearman, and F. Baxendale. 2010. The role of peroxidases in the defense response of warm-season turfgrasses to chinch bugs. [Online]USGA Turfgrass Environ. Res. Online. 9(11):p. [1-10].

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

Outputs
OUTPUTS: Developing chinch bug, Blissus occiduus Barber (Hemiptera: Lygaeidae) resistant cultivars is one of the objectives of buffalograss breeding program. Fifteen buffalograss genotypes comprising diploids, tetraploids, a pentaploid, and hexaploids and 94 diploid full-sib progeny were evaluated for chinch bug resistance in separate experiments. The experiments were laid out in a completely randomized design with five and six replications for the genotypes and progeny, respectively. Ten and eight adult chinch bugs were introduced into caged single clone genotypes and progeny in the green house, respectively. Chinch bug damage was assessed using a 1-5 visual damage rating scale with 1= ≤10%; 2= 11-30%; 3= 31-50%; 4= 51-70%; and 5= ≥70% of buffalograss leaf area with severe discoloration, or dead tissue. Marker assisted selection (MAS) is of interest to enhance breeding progress for turfgrass quality and chinch bug resistance of buffalograsses. The objectives of this study were to construct a framework genetic linkage map of diploid buffalograsses as a prelude to the application of MAS, and to study genome organization of buffalograss. Ninety-four F1 progeny generated by crossing two heterozygous diploid parents were genotyped using polymorphic SRAP and SSR markers. Linkage analysis was performed using OneMap linkage analysis software at maximum recombination fraction of 0.48 and a minimum Logarithm of Odds (LOD) score of 3.0. Results from these studies have been shared at field days, workshops and scientific meetings. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Highly significant differences were observed among the genotypes and progeny for chinch bug damage ratings. Among the genotypes, Legacy, Prestige, 184, 196, Bowie, NE 3297, NE2769 and NE 2768 were moderately resistant with damage ratings of >1, but <3, while NE 2990, NE 2838, and 1-57-19 were moderately susceptible with damage ratings of &#8805;3, but <4. One progeny (MP45) was highly resistant with a chinch bug damage rating of 1.0, 78 progeny (83 %) had moderate resistance, with damage rating of >1.0 and < 3.0, 13 progeny (14 %) were moderately susceptible with damage ratings ranging from 3.0 to 3.9, while only two were highly susceptible with damage ratings of >4.0. The significant variability among genotypes and progeny for chinch bug resistance guarantee improvement through selection or hybridization of selected genotypes. Co-segregation analysis placed 42 markers into nine discrete linkage groups covering 355.10 cM, with linkage group sizes ranging from 10 cM to 119.78 cM. A range of 2 to 18 loci per linkage group were mapped with an average map distance between two consecutive markers of 12.68 cM. This linkage map is a first for buffalograss and is a logical starting point for further linkage mapping with more markers. These results provide a foundation for a new direction for buffalograss breeding that will aid further study and improvement of turfgrass quality and pest resistance.

Publications

• No publications reported this period

Progress 10/01/08 to 09/30/09

Outputs
OUTPUTS: Research was completed on advanced germplasm in our buffalograss breeding program, which investigated turfgrass performance and seed yield potential. Data were collected from 3 trials initiated in 2002, 2003, and 2004, respectively; and terminated in 2009. In addition, 13 genotypes wre selected and twenty one single crosses were established in a field study in 2007 in an effort to identify parents that possess desirable turfgrass quality and seed yield potential. Parents with diverse genetic backgrounds were used to set the single crosses.Crosses were matched by ploidy level and isolated in space. Information obtained from these studies was shared at national and international meetings involving researchers and turfgrass seed producers. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: The target audience of this research is grass seed producers and turfgrass managers. This research is designed to identify superior seed yielding genotypes with excellent turfgrass performance. End users will profit from this research by allowing seed producers to harvest greater seed yields for increased profitability, while end users will have larger quantiities of quality seed available to meet their needs. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Turfgrass performance varied among genotypes for quality, color, density, spring green up, and seed yield. Thirty two percent of the genotypes evaluated equaled or exceeded the turfgrass performance ratings of the stadard entries. Seed yield ranged from 460 to 4964 kg per hectare, which was a 10 fold increase between the lowest and highest yielding genotypes. Based on these results, it can be concluded that excellent diversity exists among the advanced germplasm in this program for turfgrass performance and seed yield potential. Significant correlations between total seed yield, bur size, bur hundred weights, and caryopses weight, indicating that selection for one will improve the other. By selecting high seed yielding parents and using optimum seed production practices combined with efficient harvest and cleaning technologies, it is possible to double the seed yield potential of buffalograss compared to industry standard cultivars.

Publications

• Abeyo, B. and R.C. Shearman. 2009. Buffalograss (Buchloe dactyloides) Turfgrass Performance and Seed Yield Characteristics. Int. Turf. Soc. Res. J. 11:519-532.
• Abeyo, B.G., R.C. Shearman, R.E. Gaussoin, L.A. Wit, D.D. Serba, and U. Bilgili. 2009. Blue Fescue Overseeding Improves Performance of Fairway Height Buffalograsses. HortScience. 44: 1-3.

Progress 07/01/08 to 09/30/08

Outputs
OUTPUTS: This is a new project started in July, 2008 and extending to June, 2013. The objectives of this study are to: Develop seed propagated cultivars of buffalograss with improved turf quality; Develop vegetatively propagated cultivars of buffalograss with improved turf quality; Develop protocols for establishing vegetative and seeded buffalograss cultivars ; Develop management programs for use with vegetative and seeded buffalograsses to provide acceptable turf at significantly reduced levels of input; and Use molecular marker tools to identify genes which can be used in the buffalograss breeding and improvement program. Nine seeded and seven vegetative genotypes were established at nine locations in eight states. Data from each location were analyzed separately for seeded and vegetative genotypes. Significant differences were observed among genotypes tested for establishment rate, seedling vigor, lateral spread, turfgrass color, turfgrass cover, and spring green up. In 2008, germplasm was enriched through collection and hybridization. New selections and advanced lines established. Elite genotypes were evaluated under a wide range of environments. Significant differences were observed among genotypes were tested for most traits of interest. These trials will continue and seed from some genotypes will be increased. Results from management studies indicated significant differences among N rates, mowing heights and genotypes, but not for their interactions. These trials will continue for several more seasons. A two year study on growing degree effects on preharvest of sprigs and their subsequent reestablishment was completed. A frame work map of a diploid population of buffalograss is in the process of development. This work will assist in identifying markers and enhance our general understanding of the buffalograss genome. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Since this project has been on going for less than six months, there are no real outcomes or impacts to assess at this time.

Publications

• No publications reported this period

Progress 10/01/06 to 09/30/07

Outputs
These results demonstrate that primers targeting the peroxidase gene family can be used to study genotypic diversity and evolutionary relationships on an intraspecific and interspecific basis. The PCR-based peroxidase markers may also have potential for linkage mapping and differential gene expression studies in grasses.

Impacts
Plant peroxidases are a family of related proteins possessing highly conserved domains. Degenerate oligonucleotide primers based on these conserved domains can be used to amplify DNA sequences coding for peroxidases from plants with unsequenced genomes. Polymorphisms in peroxidase genes among buffalograss [Buchloe dactyloides (Nutt.) Engelm.] genotypes and eight other grasses were evaluated, and potential evolutionary relationships were deduced using this approach. Fourteen peroxidase specific primers with alternative forward and reverse primers using 34 rice peroxidase cDNAs were designed based on conserved motifs of this gene family. Targeted-PCR amplification of genomic DNA from 28 buffalograss, four C4 and four C3 grass genotypes yielded polymorphisms, differentiating diploids from polyploids within buffalograss, and C3 and C4 grass species from each other. A total of 11 peroxidase gene fragments, seven belonging to buffalograss and four to the other grass species were sequenced. Five of these sequences were clustered with rice ascorbate peroxidase known to have chloroplast origin. The objective of this study was to identify and quantify differentially expressed genes in response to rust infection (Puccinia sp.) in fescue species (Festuca rubra var. rubra) using mRNA differential display technique. Three differentially induced genes were identified as homologues of mitogen-activated protein kinase 3 of Arabidopsis thaliana, stem rust resistance protein Rpg1 of barley and HSP70 of Oryza sativa. The change in the steady state expression level of these three genes in response to rust infection was further confirmed by Northern blot analysis and quantified by real time PCR. The time-dependent change in the expression of all genes proved a steady accumulation of transcript in the course of rust infection. The whole transcript of mitogen-activated protein kinase 3 of Festuca rubra var. rubra was obtained by RACE PCR. The protein encoded was 369 amino acid in length with a predicted MW of 42,5 kDa. This is the first study on rust infection of Festuca rubra var. rubra which is aimed to provide new insights into the rust-dependent metabolic pathways in fescue.

Publications

• Ergen, N.Z., G. Dinler, R.C. Shearman and H. Budak. 2007. Identifying, cloning and structural analysis of differentially expressed genes upon Puccinia infection of Festuca rubra var. rubra. Gene. 393(1):145-152.
• Gulsen, O., R.C. Shearman, T.M. Heng-Moss, N. Mutlu, D.J. Lee, and G. Sarath. 2007. Peroxidase gene polymorphism in buffalograss and other grasses. Crop Science 47: 767-772.

Progress 10/01/05 to 09/30/06

Outputs
Peroxidases play an important role in stress related interactions. This study investigated the role of peroxidase enzymes response to the chinch bug (Blissus occiduus Barber) in buffalograsses [Buchloe dactyloides (Nutt.) Engelm.]. In the first study, twenty-eight chinch bug resistant and susceptible buffalograsses were used to investigate the correlation among chinch bug injury, total protein content, basal peroxidase levels, and ploidy level. In the second study, changes in peroxidase activity and total protein content were evaluated in four chinch bug resistant and two susceptible buffalograsses at 7, 14, 21, and 28 day after chinch bug exposure (DAE) by using spectrophotometer and native gel electrophoresis. Correlation analyses indicated that buffalograss total protein content and chinch bug injury were correlated (r = 0.47, P = 0.01), while basal peroxidase levels and chinch bug injury were not (r = 0.19, P = 0.29), suggesting upregulation of peroxidases in response to chinch bug feeding. At 14 DAE, peroxidase activity levels were found to be greater in three of the four resistant genotypes, while activity levels were similar between infested and non-infested plants of the two highly susceptible genotypes. Native gel analysis indicated Prestige and 196 had higher peroxidase expression in infested plants, and PX3-5-1 had a different peroxidase mobility, suggesting buffalograss peroxidases have variation in both their expression level and isozyme mobility. Results from this study indicate that peroxidases have the potential to be used as markers for selecting chinch bug resistant buffalograsses. Plant peroxidases are a family of related proteins possessing highly conserved domains. Degenerate oligonucleotide primers based on these conserved domains can be used to amplify DNA sequences coding for peroxidases from plants with unsequenced genomes. Polymorphisms in peroxidase genes among buffalograss genotypes and eight other grasses were evaluated, and potential evolutionary relationships were deduced using this approach. Fourteen peroxidase specific primers with alternative forward and reverse primers using 34 rice peroxidase cDNAs were designed based on conserved motifs of this gene family. Targeted-PCR amplification of genomic DNA from 28 buffalograss, four C4 and four C3 grass genotypes yielded polymorphisms, differentiating diploids from polyploids within buffalograss, and C3 and C4 grass species from each other. A total of 11 peroxidase gene fragments, seven belonging to buffalograss and four to the other grass species were sequenced. Five of these sequences were clustered with rice ascorbate peroxidase known to have chloroplast origin. These results demonstrate that primers targeting the peroxidase gene family can be used to study genotypic diversity and evolutionary relationships on an intraspecific and interspecific basis. The PCR-based peroxidase markers may also have potential for linkage mapping and differential gene expression studies in grasses.

Impacts
Buffalograss is a native turfgrass species that is adapted to the Great Plains of North America. Chinch bugs are a native insect pest that cause turf and seed production injury to buffalograss. This research identifies chinch bug resistant buffalograss genotypes and mechanisms associated with the expressed resistance. These responses can be used to expedite the development of chinch bug resistant cultivars and reduce the need for pesticides in buffalograss turf management.

Publications

• Shearman, R.C. 2006. Fifty years of splendor in the grass! Crop Sci. 46:2218-2229.

Progress 10/01/04 to 09/30/05

Outputs
Germplasm resources provide genetic variability in plants for insect resistance that may impact insect infestation. The goal of this project was to detect variation among selected buffalograss genotypes [Buchloe dactyloides (Nutt.) Engelm.] for chinch bug resistance, and relate resistance to ploidy level, chinch bug number, and pubescence. The forty-eight buffalograss genotypes from diverse geographic locations were evaluated for chinch bug resistance in replicated studies under greenhouse conditions. Of the genotypes studied, 4 were highly resistant, 22 were moderately resistant, 19 were moderately susceptible and three were highly susceptible to chinch bug damage. The mean number of chinch bugs was significantly different among the 48 genotypes. There was no significant correlation between chinch bug resistance and ploidy level or chinch bug resistance and pubescence. It is apparent that considerable variation in chinch bug resistance exists among buffalograss germplasm. As interest in using the new turf-type buffalograss cultivars has increased, the need for nitrogen rate and mowing height recommendations supported by research have become necessary. Cody, Texoka, 378, and Scout buffalograss cultivars were planted at sites located in Nebraska, Kansas, and Utah to determine nitrogen rate and mowing height effects on buffalograss quality, color, density, and biomass production. There were significant cultivar x nitrogen rate interactions at all sites for quality, color, and density. Regardless of cultivar, there were significant nitrogen rate x year interactions at all sites that revealed the 98 kg N ha-1 rate sustained quality, color, and density over the 3 yr research period, while lower nitrogen rates had decreased quality, color, and density over time. Regardless of the cultivar selected, the data from this research support the general mowing height recommendation for turf-type buffalograss of 5.0 to 7.5 cm. The data also support that if the desired use is for golf course fairways maintained at a mowing height of 2.5 cm, the vegetatively established cultivars, in this case Scout and 378, are the best choice to achieve high turfgrass quality. Dormant buffalograss turfs, grown under field conditions, were treated with a colorant and evaluated for turfgrass color, quality, and cover. In addition, turfgrass canopy and soil temperatures were measured. Colorant treatments improved turfgrass color and quality when compared to the untreated control, and resulted in a color response that appeared similar to cool season turfgrasses growing in areas adjacent to the studies. Colorant treatments increased canopy and soil temperatures, and enhanced spring green-up. These results support the use of colorants as a means of extending the green appearance, and enhancing dormant buffalograss turf performance.

Impacts
Buffalograss does not have many pest problems currently. However, chinch bugs have caused turf and seed production problems. This research was the first to identify resistant cultivars and experimental lines. This plant material offers opportunities to recommend resistant cultivars for turf use and lines that can be used in breeding programs designed to develop chinch bug resistant cultivars. Management of these lines is also important.

Publications

• Gulsen, O., T. M. Heng-Moss, R.C. Shearman, P.S. Baenziger, D.J. Lee, F.P. Baxendale. 2004. Buffalograss germplasm resistance to Blissus occiduus. J. Econ. Entomology. 97:2101-2105.
• Budak, H., R.C. Shearman, O. Gulsen, and I. Dweikat. 2005. Understanding ploidy and geographic origin of the Buchloe dactyloides genome using cytoplasmic and nuclear marker systems. Theor. Appl. Genet. 111:1545-1552.
• Gulsen, O., R.C. Shearman, K.P. Vogel, D.J. Lee, P.S. Baenziger, T. M. Heng-Moss, and H. Budak. 2005. Nuclear genome diversity and relationships among naturally occurring buffalograss genotypes determined by sequence-related amplified polymorphism markers. HortScience. 40:537-541.
• Gulsen, O. R. C. Shearman, K.P. Vogel, D. J. Lee, and T. Heng-Moss. 2005. Organelle DNA diversity among Buffalograsses from the great plains of North America determined by cpDNA and mtDNA RFLPs. Crop Sci. 45:186-192.
• Shearman, R.C., L.A. Wit, S. Severmutlu, H. Budak, and R.E. Gaussoin. 2005. Colorant effects on dormant buffalograss turf performance. HortTechnology. 15:246-249.
• Shearman, R. C., H. Budak, S. Severmutlu, and R.E. Gaussoin. 2005. Bur Seeding Rate Effects on Turf-type buffalograss establishment. HortTechnology. 15:246-249.
• Severmutlu, S., R.C. Shearman, T.P. Riordan, R.E. Gaussoin, and L.E. Moser. 2005. Overseeding buffalograss turf with fine-leaved fescues. Crop Sci. 45:704-711.
• Severmutlu, S., C. Rodgers, T.P. Riordan, and R.C. Shearman. 2005. Registration of Bowie buffalograss. Crop Sci. 45:2120.
• Severmutlu, S., T.P. Riordan, and R.C. Shearman. 2005. Registration of Cody buffalograss. Crop Sci. 45:2122.

Progress 10/01/03 to 09/30/04

Outputs
Dormant buffalograss turfs, grown under field conditions, were treated with a colorant and evaluated for turfgrass color, quality, and cover. In addition, turfgrass canopy and soil temperatures were measured. Colorant treatments improved turfgrass color and quality when compared to the untreated control, and resulted in a color response that appeared similar to cool season turfgrasses growing in areas adjacent to the studies. Colorant treatments increased canopy and soil temperatures, and enhanced spring green-up. These results support the use of colorants as a means of extending the green appearance. Little or no research information exists in the literature regarding recommended seeding rates of the improved turf-type buffalograsses cultivars, like Bowie. This study was conducted to determine bur seeding rate effects on turfgrass establishment of Bowie buffalograss. Two experiments were initiated on 21 July 2002 near Mead, NE. Bur seeding rate effects on turfgrass quality, shoot density and cover, and seedling density were evaluated. Burs were seeded at 2.5, 5.0, 10, 20 and 40 g m-2 of pure live seed (PLS). Turfgrass quality ratings increased linearly with bur seeding rate, during the first growing season. However, by early in the second growing season, the response was quadratic with little difference in quality between 10 and 40 g m-2. Turfgrass cover ratings responded in a similar manner. Buffalograss is reported to establish slowly, taking more than one growing season to establish an acceptable level. In this study, Bowie, a turf-type cultivar had acceptable turfgrass quality (> 5.0) and cover (> 75%) ratings by three months at bur seeding rates of 5 to 40 g m-2 of PLS, and acceptable quality and cover ratings were obtained at slightly over one month at rates of 20 to 40 g m-2. These results indicate that bur seeding rates of 20 to 40g m-2 are advisable where rapid establishment of turf-type buffalograss is desired, and rates as low as 5 g m-2 can be used when establishment with in two growing seasons is deemed reasonable. Use of the new turf-type buffalograss cultivars has increased the need for nitrogen and mowing recommendations that are supported by research. Cultivars Cody, Texoka, 378, and Scout were planted at sites located in Nebraska, Kansas, and Utah to determine nitrogen rate and mowing height effects on quality, color, density, and biomass. There were significant cultivar x nitrogen interactions at all sites for quality, color, and density. There were significant nitrogen x year interactions at all sites that revealed the 98 kg N ha-1 rate sustained quality, color, and density over the 3 yr research period, while lower nitrogen rates had decreased quality, color, and density over time. The data from this research support the general mowing height recommendation for turf-type buffalograss of 5.0 to 7.5 cm. The data also support that if the desired use is for golf course fairways maintained at a mowing height of 2.5 cm, the vegetatively established cultivars, in this case Scout and 378, were the best choice.

Impacts
New seeded and vegtative cultivars of buffalograss that use less water, fertilizer, pesticides and require less mowing than other turfgrasses have been developed. Management systems will be developed that will encourage society to use this environmently friendly species. Use of overseeded fine fescues or colorants can extend turfgrass quality and color by as much as two months. Bur seeding rates of 20 to 40g m-2 are advisable where rapid establishment of turf-type buffalograss is desired, and rates as low as 5 g m-2 can be used when establishment with in two growing seasons is deemed reasonable. The general mowing height recommendation for turf-type buffalograss of 5.0 to 7.5 cm. If the desired use is for golf course fairways maintained at a mowing height of 2.5 cm, the vegetatively established cultivars were the best choice.

Publications

• Shearman, R.C., T.P. Riordan, and P.G. Johnson. 2004. Buffalograss. In Warm Season Grasses. (Ed.) Lowell Moser. Agronomy Monograph 45. American Society of Agronomy. Madison WI. Pp. 1003-1026.
• Budak, H., R.C. Shearman, I. Parmasksiz, R.E. Gaussoin, T.P. Riordan, and I. Dweikat. 2004. Molecular characterization of buffalograss germplasm using sequence-related amplified polymorphism markers. Theor. Appl. Genet. 108:328-334.
• Frank, K.W., R.E. Gaussoin, T.P. Riordan, R.C. Shearman, J.D. Fry, E.D. Miltner, P.G. Johnson. 2004. Nitrogen rate and mowing height effects on turf-type buffalograss. Crop Science. 44:1615-1621.
• Budak, H., R. C. Shearman, R.E. Gaussoin, I. Dweikat. 2004. Application of sequence-related amplified polymorphism markers for characterization of turfgrass species. HortScience. 39:955-958.
• Budak, H., R.C. Shearman, and I. Dweikat. 2004. Cloning and characterization of resistance gene like sequences in warm season turfgrass species. Bioinformatics. CSREA Press. Las Vegas, NV. Pp. 225-230.

Progress 10/01/02 to 09/30/03

Outputs
This project was renewed July 1, 2003. Although the project is a continuation of a long term effort with buffalograss, the new project is now under the leadership of Robert C. Shearman. This change was due to internal reallocations of manpower at UNL. There are several new areas of research that are currently being conducted; however, one area that has been recently completed is the work to evaluate the use of cool season fine fescues to lengthen the period of time that buffalograss and the mixture of buffalograss-fine fescue is green and functional. Results indicate that blue fescue (Festuca ovina L, glauca Lam.) seeded at a 20 g m-2 seeding rate provided an attractive, uniform, weed-free, and high quality turf with an extended growing season. The color retention of the buffalograss plots was improved in both the spring and fall. All fine fescue mixture plots exhibited above 65 % green cover when buffalograss was dormant, which extended the growing season from less than five months with the monostand buffalograss to eight months with the fine fescue/buffalograss mixture. The buffalograss:fine fescue composition stabilized at around 30:70. In a second trial bullalograss was overseeded with fine fescues in the spring. Establishment, turfgrass quality, color, and green cover at all seeding rates were inferior to the results obtained in the fall trial. A study was conducted to: (1) use sequence-related amplified polymorphism (SRAP) markers in evaluation of genetic diversity and phenetic relationships in a diverse collection of 53 buffalograss germplasm, and (2) identify buffalograss ploidy levels using flow cytometry. Based on DNA contents, buffalograss genotypes were grouped into four sets, corresponding to ploidy level. Thirty-four SRAP primer combinations were used. This is the first report of differentiating diploid, tetraploid, pentaploid and hexaploid buffalograss genotypes, representing diverse origin of locations, using SRAP markers. Cluster analysis by the unweighted pair-group method with arithmetic averages (UPGMA) based on genetic similarity matrices indicated there were eight clusters. The coefficients of genetic distance among genotypes ranged from 0.33 up to 0.99 and averaged D = 0.66. The genetic diversity estimate, He, averaged 0.35. These results demonstrated genotypes with potential traits for turfgrass improvement could readily be distinguished based on SRAP.

Impacts
The development of new seeded and vegtative cultivars of buffalograss that will use less water, fertilizer, pesticides and require less mowing. Management systems will be developed that will encourage society to use this environmently friendly species. Use of fine fescues for overseeding buffalograss can extend turfgrass quality and color by as much as three months each growing season. Use of PCR-based markers such as SRAP is an effective tool for estimating genetic diversity, identifying unique genotypes as new sources of alleles for enhancing turf characteristics, and analyzing the evolutionary and historical development of cultivars at the genomic level in a buffalograss breeding program.

Publications

• Budak, H., R.C. Shearman, I. Parmaksiz, R.E. Gaussoin, T.P. Riordan, and I. Dweikat. 2003. Molecular characterization of buffalograss germplasm using sequence related amplified polymorphism (SRAP) markers. Theor. Appl. Genet. DOI 10.1007/s00122-003-1428-4.

Progress 10/01/01 to 09/30/02

Outputs
This project is part of a long-term, team effort to develop buffalograss cultivars that use less water and have fewer negative environmental impacts than other commercially available turfgrasses. As this five-year project nears its end, the goals of this project have all been met except one. New northern and southern cultivars have been released, patented and commercialized. 'Legacy' sod is available in the northern U. S. and plugs are available nationwide by mail order in areas where adapted. Discussions are also taking place that would allow an excellent warm season cultivar to be made available in the south by mail order. Although progress has been made toward the effort of making buffalograss resistant to Round-Up, we are still unable to regenerate a plant that will reach maturity. Efforts are being made to solve this problem. The responsibility of this research project has been tranferered to a different scientist, Dr. Robert C. Shearman; who will continue many of the research thrusts, but also bring some new ideas to the project.

Impacts
'Legacy' buffalograss continues to have significant impact as a drought tolerant turfgrass in arid regions of the United States. Utilization is increasing and performance is excellent. A new seeded cultivar, 'Bowie', is doing well during its initial year of introduction.

Publications

• Fei, S., Riordan, T. P. and Read, P. E. 2001. Stepwise Decrease of 2, 4-D and Addition of BA in Subculture Medium Stimulated Shoot Regeneration and Somatic Embryogenesis in Buffalograss. Journal of Plant Cell, Tissue and Organ Culture 60: 197-203.
• Heng-Moss, T.M., F.P. Baxendale, and T.P. Riordan. 2001. Interactions between the parasitoid, Rhopus nigroclavatus (Hymenoptera: Encyrtidae) and its mealybug hosts. Biological Control. Vol 22(3), p. 201-206.
• Erusha,K. S., R. C. Shearman, T. P. Riordan and L. A. Wit. 2002. Kentucky Bluegrass Cultivar Root and Top Growth Responses When Grown in Hydroponics. Crop Sci. 42:570-571.
• `Bowie' Buffalograss, PVP 200100201. 2002

Progress 10/01/00 to 09/30/01

Outputs
This project is part of a long-term, team effort to develop cultivars that use less water and have fewer negative environmental impacts than other commercially available turfgrasses. A new cultivar, 'Legacy', was available nationally as sod and plugs for the first time in 2001. Legacy has performed well and has been well received in the marketplace. It has a darker color, improved density and better quality than other buffalograss cultivars. It can be ordered and delivered nationally in two days as plugs. It was selected as the mail order green plant or the year in 2001. The major effort of or project in 2001 was transform buffalograss using biolistic bombardment with the goal making buffalograss Round-Up resistant. Three genotypes were selected for this effort, Legacy, '609' and experimental variety 91-118. Each of these genotypes is a female and there is no pollen shed; therefore, reducing the chance of outcrossing by a great degree. Also, these genotypes would be planted, propagated and marketed vegetatively, further reducing the potential of gene movement. This three year project has allowed us to develop procedures for transforming callus lines and for regenerating plants. However, to date we have been unable to regenerate a live transformed buffalograss genotype. After the plants begin to develop, approximately seedling size, they turn dark brown and die. It is thought that there is either a genetic problem or the medium is lacking something that is required for the seedling to mature. Major efforts are being made to solve this problem. We are developing new, younger callus lines and exploring new media for regeneration. If we are successful in developing a Round-Up resistant buffalograss, it would increase the potential for utilization of a species that would use less water and require less inputs than other commercially available turfgrass species. This would occur because the turfgrass manager would be better able to control weeds, especially bermudagrass, in this somewhat noncompetitive species. Ideally, the consumer would be able to have an environmentally friendly turfgrass with relatively good turfgrass quality.

Impacts
The impact of our newest buffalograss, Legacy, is significant throughout the northern United States. This cultivar is performing well and is being grown by eight sod producers and is also available by mail order. Homeowners who use this cultivar re duce environmental impact while maintaining turfgrass quality. The impact of Round-Up resistance would allow and encourage use of this species in a larger geographical region and with improved success.

Publications

• Fei, S., Riordan, T. P. and Read, P. E. 2001. Stepwise Decrease of 2, 4-D and Addition of BA in Subculture Medium Stimulated Shoot Regeneration and Somatic Embryogenesis in Buffalograss. Journal of Plant Cell, Tissue and Organ Culture 60: 197-203.
• Fei, S., Bishnoi, U. S., Riordan, T., and Read, P. 2001. Embryogenic callus induction and plant regeneration of buffalograss through leaf base and seedling segment culture. ITS Jour. Vol. 9, Part 1, p. 162-4.
• Fei, S., Yu, T., clemente, T., and Riordan, T. 2001. Nodal segment explant as a potential target for the genetic engineering of buffalograss. ITS Jour. Vol. 9, Part 1, p. 165-168.
• Johnson, P.G., and Riordan, T. P. 2001. Unbalanced chromosome number and inbreeding effects on fertility and plant vigor in buffalograss. ITS Jour. Vol. 9, Part 1, p. 176-179.
• Frank, K. W., Gaussoin, R. E., Riordan, T. P., Stroup, W. W., and Bloom, M. H. 2001. Nitrogen allocation of turfgrasses: I. Recovery of 15N-labeled ammonium nitrate applied to buffalograss [Buchloe dactyloides (Nutt.) Engelm]. ITS Jour. Vol. 9, Part 1, p. 268-276.
• Frank, K. W., Gaussoin, R.E., Riordan, T. P., Stroup, W. W., and Bloom, M. H. 2001. Nitrogen allocation of turfgrasses: II. Recovery of 15 N-labeled ammonium nitrate applied to Kentucky bluegrass (poa pratensis L.) and tall fescue (Festuca arundinacea Schreb). ITS Jour. Vol. 9, Part 1, p. 277-286.

Progress 10/01/99 to 09/30/00

Outputs
This project is part of a long-term effort to develop buffalograsses (Buchloe dactyloides) that use less water and require fewer inputs than other turfgrasses that are used in the Central Great Plains. An additional requirement that was considered, because of the requirements of our granting agency, the United States Golf Association, was that the buffalograsses have potential to be used on golf courses. In 1998 three new cultivars were released, in 1999 they received plant patents, and in 2000 the first cultivar, Legacy (86-61), was marketed. Legacy has an excellent turfgrass quality, dark green color, density, sod strength, and it has excellent low mowing tolerance that makes it well adapted to golf course use. Initial adoption has been very good and potential use in the Northern U.S could be quite good over the next few years. Currently it is being marketed as sod and vegetative plugs that are being mail ordered in the U.S. Another cultivar 91-118 is number 1 in the National Turfgrass Evaluation Program national trial; Legacy is number 2. This cultivar 91-118 is adapted to the Southern U.S., but so far has not been produced by our southern cooperator. Our cooperator is being encouraged to initiate production in order to have a turfgrass that well perform well under the drought conditions that our occurring in their marketing area in Texas. A weakness of buffalograss is establishment; this is due to its non-competitiveness and problems with weed pressure. To address this concern a major effort has been made to introduce the gene for Round-Up resistance into buffalograss. This has been accomplished this year and progress is being made to regenerate resistant calli into plants which would replace the current cultivars on the market. Buffalograss continues to perform well and have the potential to reduce inputs that are used on turf areas.

Impacts
The impact of a Round-Up resistant buffalograss could be significant if a transgenic plant is accepted by society and there continue to be droughts and shortages of water in the U.S. As a proprietary turfgrass, there could be a significant financial return to Nebraska. However, the most important impact will be the positive effect on the environment through the development and use of a species that uses less water, fertilizer, pesticides and requires less mowing.

Publications

• Kyoung-Nam K., R.C. Shearman, and T.P. Riordan. 1999. Top growth and rooting responses of tall fescue cultivars grown in hydroponics. Crop Science 39:1431-1434. (J. Series No. 9476).
• Shuizhang Fei, P.E. Read, and T.P. Riordan. 2000. Improvement of embryogenic callus induction and shoot regeneration of buffalograss (Buchlbe dactyloides nutt. engelm) by AGN03. J. of Plant, Cell, Tissue and Organ Culture (J. Series No. 948378).
• Johnson-Cicalese, J.M., P.G. Johnson, T.P. Riordan, F.P. Baxendale, R.E. Gaussoin, J.E. Watkins, and R. V. Klucas. 2000. Registration of '120' Buffalograss. Crop Sci. 40(2): 571-572. (J. Series 12362).
• Johnson, P.G., T.P. Riordan, J.M. Johnson-Cicalese, F.P. Baxendale, R.E. Gaussoin, R.C. Shearman, and R. V. Klucas. 2000. Registration of '61' Buffalograss. Crop Sci. 40(2): 569-570. (J. Series 12361). Riordan, T.P., P.G. Johnson, J.M. Johnson-Cicalese, R.E. Gaussoin, F.P. Baxendale, R.C. Shearman and R. V. Klucas. 2000. Registration of '118' Buffalograss. Crop Sci. 40(2): 570-571. (J. Series 12363).

Progress 10/01/98 to 09/30/99

Outputs
NTG-5, NTG-7 and FW-3 are being evaluated at the John Seaton Anderson Turfgrass and Ornamental Research Facility. Seeds West has indicated that they would still like to market NTG-5. Patents have been approved for releases NE 86-61, NE 86-120 and NE 91-118. Publication of the crop registration should occur this winter in the Crop Science. NE 86-61 has been named 'Legacy' and 35 acres are being produced by Todd Valley Farms at Mead, Ne. Legacy will be available in the northern United States next year. Crenshaw Turf is now part of Turfgrass America, Inc. along with Thomas Bros. Sod and Milberger Turf. Their goal is to be the premier supplier of proprietary southern turfgrasses, i.e. bermudagrass, zoysiagrass, St. Augustinegrass, and buffalograss. This would include genetically enhanced turfgrasses. Sales of buffalograss for 1999 are down, but they have indicated that this is due more to reorganization of the companies than lack of interest in buffalograss. Performance levels continued to improve with the establishment of a new breeding nursery in 1999. Numerous accessions in this nursery have exhibited increased establishment rate over commercially available cultivars. Newly released cultivars continue to show their superiority over older varieties with improved sod strength, color, turfgrass quality, and density. The establishment of six new crossing blocks in 1999 with selections exhibiting fairway type characteristics should provide germplasm with higher levels of turf quality and adaptation to golf course management systems. Buffalograss seed production has received major attention in 1999. To insure the successful use of buffalograss, seed production characteristics must be a major factor in the selection process. The buffalograss project has initiated a three-phase approach to provide high turf quality varieties with high seed yields. Phase one involves breeding of high yielding female lines with advanced male accessions that contribute to seed yield, seedling vigor, and turf performance characteristics. The second phase is the use of flow cytometry to identify crossing accessions of similar ploidy levels. The third phase is to explore chemical applications of plant hormones to enhance seed production. Establishment of buffalograss has always been a major objective of the project. Great strides have been made with buffalograss establishment since the project was first initiated. Initial studies in the 1980's indicated that pre-rooted plugs had an advantage in establishment over seeded varieties. However, present studies have shown that seeded varieties now establish as rapidly as vegetative plugs. These improvements are due to improved selections that exhibit quicker germination and improved seedling vigor, and because of better production of seed by producers. Sprigging of buffalograss has also shown potential to provide very rapid establishment rates. The research project will focus on improving sprigging characteristics in addition to seed production.

Impacts
Since 1984, this project has generated over 1.5 million dollars in grants and royalties. In addition, eight cultivars have been released. These cultivars are being used to reduce water use, fertilizer use and pesticide use in the urban landscape. A new cultivar 'Legacy' and the possibility of a Round-Up ready buffalograss will have a very positive impact on the use of buffalograss over the next few years.

Publications

• Johnson-Cicalese, J.M., F.P. Baxendale T.P. Riordan and T.M. Heng-Moss. 1998. Identification of mealybug (Hornoptera: Pseudococcidae) resistant turf-type buffalograss germplasm. J. Econ. Entomol. 91: pp 340-346.
• Johnson, P.G., T.P. Riordan, and K. Arumuganathan. 1998. Ploidy level determination in buffalograss clones and populations. Crop Science 38. p478-482.
• Heng-Moss, T.M., EP. Baxendale and T.P. Riordan. 1999. Influence of Rhopus nigroclavatus (Ashmead) (Hymenoptera: Encyrtidae) on the buffalograss mealybugs, Trl&scus sporoboli (Cockerell) and Trionymus sp. Environ. Entomol. 28(l). pp 123-127..
• Baxendale, ER, T.M. Heng-Moss, and T.P. Riordan. 1999. Blissus occiduus Barber (Hemiptera: Lygaeidae): A chinch bug pest new to buffalograss turf. J. Econ. Entomol. vol. 92(5). pp 1172-1176.
• Johnson, P.G. , and T.P. Riordan. 1999. A Review of Issues Pertaining to Transgenic Turfgrasses. J. of HortScience. Vol. 34(4). pp 594-498.

Progress 10/01/97 to 09/30/98

Outputs
Level of buffalograss performance continues to improve with each generation of selection. Newly release cultivars continue to show their superiority over older varieties with improved sod strength, color, quality, and density. Accessions from fairway maintained areas look very promising and show continuing improvements towards a high quality, low maintenance fairway turf. The top performers in the Nebraska National Buffalograss trial were 91-118 and 86-61, which are being commercialized. The seeded varieties Cody and Tatanka showed little differentiation during the first year of this study. However, in 1998 the advance-seeded types began to show better performance than the common types like Texoka. Under low mowing and no wear the female clone 92-135, performed very well again in 1998 along with female clone 92-31. However, two male clones, 92-141 and 92-116, had the best overall performance in 1998. All seed established experimentals exhibited average color and quality characteristics. Under wear results indicated that male and monoecious clones exhibited the most damage and wear tolerance of female cultivars was significantly better than males, but not as good as for mixed seeded types. In a management study, NE 91-118 and 378 had the highest quality ratings at the 2.5 cm mowing heights for years 1996-1998. Cody and Texoka had poor quality ratings at the 2.5 cm mowing height for all years. In 1998, NE 91-118, 378, and Cody had the highest quality ratings at the 5.1 cm mowing height. At the 7.6 cm mowing height Cody and Texoka had the highest quality rating in 1997 but Cody and 378 had the highest quality ratings in 1998. Management recommendations for 378 and NE 91-118 are 2.5 or 5.1 cm mowing heights and a nitrogen rate of 10 g N m2 year. Recommendations for Cody and Texoka are 5.1 or 7.6 cm mowing heights and a nitrogen rate of 10 g N m2 year. Eleven buffalograss cultivars/selections (`Cody', `Bonnie Brae', `Tatanka', `Texoka', NE 91-118, NE 86-120, NE 86-61, `315', `378', `609', and NE 84-45-3) were screened for resistance to B. occiduus in 2 greenhouse trials. Using chinch bug numbers and plant damage ratings to assess levels of resistance, the 11 buffalograss cultivars/selections were separated into categories of resistance. `Cody' and `Tatanka' consistently exhibited high levels of resistance to chinch bug feeding, while `Bonnie Brae' and NE 91-118 showed high to moderate levels of resistance. Other cultivars/selections, including `378', `315', NE 84-45-3, and NE 86-61, were moderately to highly susceptible. `Cody' and `Tatanka' maintained acceptable turf quality although both became heavily infested with chinch bugs. This suggests tolerance may be a mechanism of the resistance

Impacts
(N/A)

Publications

• Heng-Moss, T.M., F.P. Baxendale and T.P. Riordan. 1998. Beneficial arthropods associated with buffalograss. J. Econ. Entomol. 91:5 pp1167-1172.
• Heng-Moss, T.M., F.P. Baxendale and T.P. Riordan. 1998. Rhopus nigroclavatus (Ashmead) and Psuedaphycus sp. (Hymenoptera: Encyrtidae): two parasitoids of the buffalograss mealybugs, Tridiscus sporoboli (Cockerell) and Trionymus sp. J. Kansas Entomol. Soc. 71: pp85-86.
• Johnson, P.G., T.P. Riordan, and K. Arumuganathan. 1998. Ploidy level determination in buffalograss clones and populations. Crop Science 38:(p478-482).
• Johnson-Cicalese, J.M., F.P. Baxendale T.P. Riordan and T.M. Heng-Moss. 1998. Identification of mealybug (Homoptera: Pseudococcidae) resistant turf-type buffalograss germplasm. J. Econ. Entomol. Journal Series No. 11560 (Vol 91 pp 340-346).
• Frank, K.W., R.E. Gaussoin, T.P. Riordan and E.D. Miltner 1998. Date of Planting Effects on Seeded Turf-Type Buffalograss. Crop Science 38: 1210-1213.