The objective was to distinguish between cultivars and evaluate genetic relatedness of poinsettia (Euphorbia pulcherrima) using two methods of DNA fingerprinting—DNA Amplification Fingerprinting (DAF) and Arbitrary Signatures from Amplification Profiles (ASAP). Eleven red poinsettia cultivars were studied, including `Celebrate 2', `Darlyne', `Freedom Red', `Lilo', `Nutcracker Red', `Peterstar Red', `Petoy', `Red Sails', `Supjibi', `V-14 Glory', and `V-17 Angelika'. Amplification was with 10 octamer primers. Gels were visually scored for presence or absence of bands. The 10 primers generated 336 bands. The average number of bands (≈1000 bp) per primer was 34 ranging from 19 to 43. Thirty-one percent of bands were polymorphic and distinguished between each cultivar. The number of unique profiles varied from two to nine. Genetic relationships were evaluated by SAHN cluster analysis based on the distance estimator of Jaccard using the NTSYS-pc program (Numerical taxonomy and multivariate analysis system, version 1.8). The resulting dendrogram closely agreed with known pedigree data. ASAP analysis was used to further assess cultivar identification of two cultivars that were genetically and morphologically similar. Markers were found that separated `Nutcracker Red' and `Peterstar Red'. ASAP analysis separated cultivars within the Freedom series that DAF failed to distinguish. Two cultivars in the Freedom series, `Jingle Bells' and `Marble', were characterized from other cultivars in the series with ASAP.
Terri Woods Starman and Shane Abbitt
Terri Woods Starman and Shane Abbitt
The objective was to distinguish between series of cultivars of Pelargonium xhortorum (zonal geranium), Pelargonium hybrids (seed geranium), and Pelargonium peltatum (ivy leaf geranium) using DNA amplification fingerprinting (DAF) demonstrating the utility of DAF for patent protection to prevent infringement of inventor's rights. Leaf tissue of 10 plants of each cultivar of seedling geranium was bulked for DNA extraction, and cutting and ivy geranium cultivars were bulks of five plants of each cultivar. Isolated DNA from different cultivars of a series were bulked together in their respective series. Seedling geranium series included Dynamo, Glamour, Multibloom, Orbit, Pinto, and Ringo 2000. Cutting geranium series included Designer and Showcase. Ivy geraniums were from the Guillou group. Amplification was with one of two octamer primers, followed by reamplifying with one of four different mini hairpin primers. Gels were visually scored for presence or absence of bands. The four primers generated 336 bands. The average number of bands (_1000 bp) per primer was 40. Twenty percent of bands were polymorphic and distinguished between each series of cultivars. Genetic relationships were evaluated by SAHN cluster analysis based on the distance estimator of Dice using the NTSYS-pc program (Numerical taxonomy and multivariate analysis system, version 1.8). Series were grouped according to species. Seedling geraniums were in one large group, the two cutting geraniums were grouped together and the ivy leaf geraniums were a separate branch.
Terri Woods Starman and Shane Abbitt
Our objective was to distinguish between eight cultivars of two geranium species, Pelargonium ×hortorum L.H. Bailey (cutting and seed geranium) and Pelargonium peltatum (L.) L'Hér. ex Ait. (ivy geranium), and evaluate their genetic relationships using the nucleic acid scanning techniques of DNA amplification fingerprinting (DAF) and/or arbitrary signatures from amplification profiles (ASAP). Cultivars used in the study represented three commercial types: cutting, seed, and ivy geranium. Two seed geranium cultivars from each of the Dynamo and Orbit series were included. Cutting geranium cultivars were `Designer Lilac Chiffon' and `Starburst Red' and the ivy geraniums were `Bernardo Guiber' and `Vinco Guivin'. The ASAP amplification protocol used one of two arbitrary octamer primers, followed by reamplification with one of four different minihairpin primers. ASAP profiles were complex, with 66% of bands being polymorphic and useful in distinguishing between cultivars. Genetic relationships were evaluated by principal coordinate analysis and cluster analysis based on the Jaccard distance estimator. This analysis grouped cultivars by species according to commercial type, i.e., seed geraniums were in one large group, the cutting geraniums were grouped together, and the ivy geraniums were a separate branch.
Terri Woods Starman, Xiangrong Duan, and Shane Abbitt
DNA amplification fingerprinting (DAF) was used to evaluate the genetic relationships among 11 cultivars of poinsettia (Euphorbia pulcherrima Willd.). Amplification was with 10 octamer oligonucleotide primers that generated 336 DNA bands. Thirty-one percent of the bands were polymorphic and distinguished among cultivars. Genetic relationships were evaluated by cluster analysis, and the resulting dendrogram closely agreed with published cultivar relationships. Arbitrary signatures from amplification profiles (ASAP) were further used to characterize two cultivars, `Nutcracker Red' and `Peterstar Red', that were previously found to be genetically and morphologically similar, as well as five cultivars in the “Freedom” series. The DAF products generated with arbitrary octamer primers were reamplified with mini-hairpin decamer primers in these experiments. The ASAP profiles were complex and yielded a total of 231 bands, 38% of which were polymorphic and capable of distinguishing each Freedom cultivar. Five of the eight primer combinations distinguished `Nutcracker Red' from `Peterstar Red'. Thus, closely related cultivars of poinsettia can be separated using new and improved molecular fingerprinting protocols.
Elizabeth Will, Terri W. Starman, James E. Faust, and Shane Abbitt
The objective was to study the flowering response of garden cultivars of Dendranthemum × grandiflorum (Ramat.) Kitamura to temperature and photoperiod. Fifteen garden mum cultivars were grown in ten temperature (18 and 24°C constant day and night greenhouse temperatures) and photoperiod (8, 10, 12, 14, and 16 h) combinations. Rooted cuttings were pinched above the fifth node and placed in the temperature/photoperiod treatments. When axillary shoots developed, all but one shoot was removed to produce a single stemmed plant. Photoperiods were provided by delivering 8 h sunlight, then pulling black cloth and providing daylength extension with incandescent bulbs. Days to visible bud, days to first bud color, days to flower, node number, and stem length were measured. By 11 weeks after the start of photoperiod treatments, no difference was measured in days to flower in the 8-, 10-, and 12-h photoperiods at 18°C. Days to flower increased as photoperiod increased from 12 to 14 h. At 18°C, five cultivars flowered in the 16-h photoperiod, while 10 cultivars developed crown buds, i.e., flower buds that initiated but had not developed. At 24°C, there was no difference in days to flower in the 8and 10-h photoperiod, while days to flower increased as photoperiod increased from 10to 12-h treatment. Cultivars formed crown buds but had not reached flowering in the 14and 16-h photoperiods at 24°C. Regardless of temperature, stem length increased as photoperiod increased above 10 h.