Morphological analysis historically has been used to determine parentage of unknown hybrids. This can be difficult when potential parents have similar appearance, as in the case of three azaleodendron cultivars, Rhododendron L. ‘Fragrans’, ‘Fragrans Affinity’, and ‘Fragrant Affinity’. These cultivars are similar in name and appearance, and all are purported hybrids of R. catawbiense Michx. or R. ponticum L. and R. viscosum (L.) Torr. Amplified fragment length polymorphism (AFLP) analysis was conducted to determine whether the cultivars are synonyms or distinct clones and to elucidate the parental species. The three cultivars, suspected to be hybrids between taxa in subgenera Hymenanthes (Blume) K.Koch (evergreen rhododendrons) and Pentanthera (G.Don) Pojarkova (deciduous azaleas), and related taxa from each subgenus were evaluated using 31 AFLP primer combinations. Genetic similarity, calculated using Jaccard's coefficient, among the hybrids ranged from 53% to 71%, indicating that they are distinct cultivars and not a single clone. Genetic similarity was highest between the hybrids and R. ponticum among the evergreen rhododendrons, and R. viscosum among the deciduous azaleas. A dendrogram generated using the genetic similarity matrix grouped taxa into their respective subgenera, with the three cultivars nested intermediately between subgenera but more closely with subgenus Hymenanthes and particularly R. ponticum, suggesting it is the evergreen rhododendron parent. Furthermore, principle components grouped R. ponticum more closely with the hybrids and there were 18 AFLP fragments unique to R. ponticum and the hybrids. However, no unique AFLP bands were shared exclusively among the hybrids and the purported deciduous azalea parent, R. viscosum, suggesting that the original azalea parents may have been hybrids.
Ryan N. Contreras, Thomas G. Ranney, Susana R. Milla-Lewis, and G. Craig Yencho
Jennifer A. Kimball, Thomas G. Isleib, William C. Reynolds, Maria C. Zuleta, and Susana R. Milla-Lewis
New elite st. augustinegrass cultivars with improved cold tolerance and desirable turf quality are needed for the turf industry, especially in the transitional climatic region of the United States. To efficiently use sources of cold tolerance in a breeding program, an understanding of the genetic control of this trait and its relationship to important turf quality traits is required. Therefore, the objective of this study was to estimate general and specific combining abilities for cold response and turf quality traits. Six diploid genotypes of st. augustinegrass were selected as parents for a diallel mating design without reciprocals and evaluated over 3 years at two locations. The true hybridity of crosses was confirmed using simple sequence repeats (SSR). Combining ability analysis revealed that both general and specific combining abilities were significant across years and locations. Specific combining ability (SCA) was the largest source of genetic variation for winter survival, genetic color, turf density, and end-of-season cover indicating that nonadditive gene effects play a key role in the inheritance of these traits. The parental genotype ‘GF2’ was identified as a promising parent for future breeding efforts as it provided positive general combining ability (GCA) effects for both cold tolerance and turf quality traits, which were not significantly correlated with one another. Lines identified as parental selfs generally showed lower cold response and inferior turf quality than the original parental lines indicating that inbreeding depression can occur in st. augustinegrass. This study provides information regarding the combining ability of cold response and turf quality traits in st. augustinegrass, which will ultimately aid in parental selection for our future breeding efforts.
Jennifer A. Kimball, M. Carolina Zuleta, Matthew C. Martin, Kevin E. Kenworthy, Ambika Chandra, and Susana R. Milla-Lewis
St. augustinegrass [Stenotaphrum secundatum (Walt.) Kuntze] is a popular turfgrass in the southern United States as a result of its superior shade tolerance and relatively low input requirements. However, it is the least cold-tolerant of commonly used warm-season turfgrass species. ‘Raleigh’, released in 1980, has superior cold tolerance and is adapted and widely used in U.S. Department of Agriculture hardiness zones 8 to 9. More than 25 years after its release, ‘Raleigh’ is still the industry’s standard in terms of cold tolerance. However, the original foundation and breeder stock fields of the cultivar have been lost, placing the integrity of the cultivar at risk. The objectives of this study were to investigate whether current ‘Raleigh’ production fields across the southern United States are true to the original source. In this study, 15 amplified fragment length polymorphism (AFLP) primer combinations were used to assess levels of genetic variability among three original stocks of ‘Raleigh’ and 46 samples obtained from sod farms and universities in six states. Genetic similarities among the original stocks were Sij = 1, whereas similarities between this group and all other samples ranged from 0.24 to 1.0. Results based on cluster analysis, principal coordinate analysis, and analysis of molecular variance (AMOVA) revealed separation between original stocks of ‘Raleigh’ and some commercial samples. Results from this study offer further evidence that molecular markers provide a useful and powerful technique for identity preservation of clonally propagated cultivars and the detection of genetic variants in sod production fields and turfgrass breeding programs.
Karen R. Harris-Shultz, Susana Milla-Lewis, Aaron J. Patton, Kevin Kenworthy, Ambika Chandra, F. Clint Waltz, George L. Hodnett, and David M. Stelly
Zoysiagrass (Zoysia sp.) is used as a warm-season turfgrass for lawns, parks, and golf courses in the warm, humid and transitional climatic regions of the United States. Zoysiagrass is an allotetraploid species (2n = 4x = 40) and some cultivars are known to easily self- and cross-pollinate. Previous studies showed that genetic variability in the clonal cultivars Emerald and Diamond was likely the result of contamination (seed production or mechanical transfer) or mislabeling. To determine the extent of genetic variability of vegetatively propagated zoysiagrass cultivars, samples were collected from six commercially available zoysiagrass cultivars (Diamond, Emerald, Empire, JaMur, Meyer, Zeon) from five states (Arkansas, Florida, Georgia, North Carolina, Texas). Two of the newest cultivar releases (Geo and Atlantic) were to serve as outgroups. Where available, one sample from university research plots and two samples from sod farms were collected for each cultivar per state. Forty zoysiagrass simple sequence repeat (SSR) markers and flow cytometry were used to compare genetic and ploidy variation of each collected sample to a reference sample. Seventy-five samples were genotyped and an unweighted pair group method with arithmetic mean clustering revealed four groups. Group I (Z. japonica) included samples of ‘Meyer’ and Empire11 (‘Empire’ sample at location #11), Group II (Z. japonica × Z. pacifica) included samples of ‘Emerald’ and ‘Geo’, Group III (Z. matrella) included samples of ‘Diamond’ and ‘Zeon’, and Group IV (Z. japonica) consisted of samples from ‘Empire’, ‘JaMur’, ‘Atlantic’, and Meyer3 (‘Meyer’ at sample location #3). Samples of ‘Empire’, ‘Atlantic’, and ‘JaMur’ were indistinguishable with the markers used. Four samples were found to have alleles different from the respective reference cultivar, including two samples of ‘Meyer’, one sample of ‘Empire’, and one sample of ‘Emerald’. Three of these samples were from Texas and one of these samples was from Florida. Three of the four samples that were different from the reference cultivar were university samples. In addition, one sample, Empire11, was found to be an octoploid (2n = 8x = 80). For those samples that had a fingerprint different from the reference cultivar, contamination, selfing, and/or hybridization with other zoysiagrasses may have occurred.
Jeffrey C. Dunne, W. Casey Reynolds, Grady L. Miller, Consuelo Arellano, Rick L. Brandenburg, A. Schoeman, Fred H. Yelverton, and Susana R. Milla-Lewis
Bermudagrass, Cynodon spp. is one of the most commonly grown turfgrass genera in the southern United States having excellent drought tolerance, but poor tolerance to shade. Developing cultivars tolerant to shade would allow bermudagrass to become more prevalent in home lawns or other recreational areas in the southeast, where trees dominate the landscape. In this field study, nine accessions collected from Pretoria, South Africa were evaluated for their ability to grow under shade with varying fertility treatments. These accessions and cultivars ‘Celebration’, ‘TifGrand’, and ‘Tifway’ were evaluated under 0%, 63%, and 80% continuous shade during 2011–12. For both years, significant differences among shade levels, genotypes, and the interaction of the two were observed. As expected, the progression from 0% to 63% to 80% shade reduced normalized difference vegetation index (NDVI), percent turfgrass cover (TC), and turf quality (TQ) readings for all accessions. Some genotypes, however, were able to maintain adequate quality and aggressiveness under 63% shade. ‘Celebration’, WIN10F, and STIL03 performed better than ‘Tifway’ (P ≤ 0.05), the susceptible control. Overall, our results indicate that there are promising genotypes among the bermudagrass materials collected from South Africa. These accessions represent additional sources of shade hardiness to be used in bermudagrass breeding. Furthermore, higher nitrogen fertility provided increased NDVI and TQ in some instances suggesting an added benefit of fertility under low-light conditions. However, the increased economic value attributed to the added inputs associated with these increases is outweighed by the low impacts offered.