Kenneth H. Quesenberry

Genetic diversity and DNA content of three South American and three Eurasiatic Trifolium species

Six species of Trifolium (T. polymorphum Poir., T. riograndense Burkart, T. argentinense Speg., T. medium L., T. pratense L. and T. repens L.) were analyzed using inter-simple sequence repeats (ISSR) markers. Six selected primers generated 186 polymerase chain reaction (PCR) products exploring 112 loci in 34 genotypes analyzed with molecular sizes ranging from 200 to 1300 bp. These primers were able to discriminate among and within species, with the PCR products being on average 41.6% species-specific and 59.9% polymorphic at the within species level. Nuclear DNA content was determined by flow cytometry and revealed variation among species. The 1Cx genome size values were calculated and were found to range from 0.46 pg (T. pratense) to 0.96 pg (T. polymorphum). Genome size values of South American species were higher than those of Eurasiatic origin. The analyses of the molecular data grouped the six species in agreement with their geographical origin and clearly differentiate T. polymorphum from T. argentinense. The Eurasiatic group showed the highest average of species-specific bands (45.3%) and the South American group exhibited the highest amount of total bands (59.7). The highest level of intra-species polymorphisms was detected in T. argentinense (92.9%), followed by T. medium (89.5%).

Tetraploid bahiagrass hybrids: breeding technique, genetic variability and proportion of heterotic hybrids

Manipulating apomixis for fixing tetraploid bahiagrass (Paspalum notatum Flüggé) hybrids exhibiting superior agronomic characteristics would be a valuable tool for the genetic improvement of this species. The objectives were to create a second generation of hybrids by crossing sexual first-generation tetraploid hybrids (FGTH) and unrelated apomictic FGTH or ecotypes, determine the segregation for mode of reproduction, and estimate the resulting genetic variation and heterosis for several agronomic characteristics. The segregation for mode of reproduction was analyzed using mature embryo sac observations. Field measurements and visual ratings were used to evaluate the growth habit, production of inflorescences, cool-season growth and freeze resistance of hybrids. A ratio of 4.6:1 between sexual and apomictic hybrids was observed. Only 3% of the progeny was classified as highly apomictic. The low proportion of highly apomictic progeny restricts the feasibility of this breeding approach. Large genetic variation was observed among hybrids for growth habit, cool-season growth and freeze resistance. The proportion of hybrids exhibiting heterosis was high for plant height, cool-season growth, and freeze resistance. The tetraploid bahiagrass germplasm contains a large diversity for traits of agronomic importance that can be exploited for the genetic improvement of this species.

Growth and nitrogen fixation ofAeschynomene under water stressed conditions

SummaryStudies on the tolerance ofAeschynomene americana L. to periods of flooding or soil moisture deficit were conducted in an attempt to elucidate nitrogen fixation as affected by soil moisture. Nitrogenase activity was not reduced significantly in pot-grown Aeschynomene plants subjected to flooding in greenhouse conditions. After 20 days of withholding water from the soil, nitrogenase activities of the drought-stressed plants were much lower than those of either the well-watered or flooded plants. Leaf water potentials were similar in flooded and control plants; however, the droughted plants had leaf water potentials that were 4 bars lower than those of the control plants. Aeschynomene plants were tolerant to long-term periods of flooding, but exhibited a reduction in nitrogenase activity and leaf water status when subjected to soil moisture deficits.

Potential root depth development and nitrogen uptake by tetraploid bahiagrass hybrids

The objectives were to develop a screening technique to evaluate the tetraploid germplasm of Paspalum notatum Flüggé (bahiagrass) for potential rate of root depth development (RRDD), estimate the variability for RRDD among hybrids, and analyze the association between RRDD and nitrogen (N) uptake. First, a screening technique was developed based on the evaluation of two clones grown in clear acrylic columns of different sizes, filled with mineral or organic substrates. The RRDD response was determined to be constant across substrates, and tube sizes. Second, differences in RRDD among thirteen clones were compared by growing plants in acrylic columns filled with mineral substrate. Genetic variability was identified among the bahiagrass germplasm for RRDD. Greater RRDD values resulted in greater root and shoot mass, indicating that RRDD was related to early vigor. Finally, to evaluate the relation between RRDD and N uptake, labeled N (15N) was injected either at the depth of the deepest root of each clone or at the maximum depth of all clones. Rapid root depth development was a determining factor for rapid access and uptake of nitrogen present in deep soil layers. This relationship might explain why clones exhibiting rapid root penetration also showed greater early vigor.

Selection in Setaria sphacelata for winter survival

Abstract Choice of warm‐season grasses for use in North Florida, United States, is limited because of the cold winters, which are characterised by both radiation frosts and freezes. With the objectives of increasing the adaptation of the forage Setaria sphacelata to these conditions, a breeding programme was undertaken at the University of Florida from 1997 to 2000. A recurrent selection procedure was used for three cycles to select for winter survival and early spring growth after frost and freezes in Gainesville (29.6°N latitude) and for one cycle in Quincy (31 °N latitude). Heritability estimates for the first two cycles in Gainesville and the cycle in Quincy ranged from 0.35 to 0.50 on a family basis. Predicted gains ranged from 10.4 to 17.4% in winter survival over the preceding cycle. These populations were compared in Gainesville to the original population and to cultivars ‘Solander’ and ‘Splenda’ in two experiments, one wide‐ and one close‐spacing. Results showed that selection for winter survival in Gainesville reduced canopy height in the second and third cycles and the reproductive height in the third cycle. Leaf dry matter yield was decreased in the first and second cycles and leaf percentage in the first cycle. These effects were reversed in the remaining cycles. Selection did not have a consistent effect on the width of leaves, length or number of inflorescences, or on total and stem dry matter yields. However, selection for winter survival in Quincy increased canopy height and the number of inflorescences. Selection for winter survival after frosts and freezes in Gainesville was successful because all cycles of selection had a higher survival rate after the winter of 1999–2000 than the original population. This winter, which killed many of the Setaria plants, was effective in providing additional screening for winter survival after frosts and freezes. Further research with this species for Florida conditions appears warranted.

Selection of morphological traits to improve forage characteristics of Setaria sphacelata grown in Florida

Abstract A genetically diverse population of Setaria sphacelata was developed at Gainesville, Florida by intercrossing the cultivars ‘Narok’, ‘Ka‐zungula’ and ‘Solander’ and a local persistent population. To determine heritability of traits that may improve setaria as a forage plant in Florida, progeny of the intercrossed population underwent two cycles of selection for inflorescence number and one cycle for leaf width. The selected populations, together with cultivars and introductions, were evaluated for morphological characters, yield and forage quality in field experiments with close‐ and wide‐spaced plants. Increase of inflorescence number was not obtained by selection but resulted in decreased plant height, inflorescence length, yield, leaf width, and leafiness. Leaf width was increased by selection and was associated with increased leafiness and whole‐plant crude protein content. The results suggest good prospects for selection of setaria cultivars suited to Florida with improved yield and quality providing they tolerate winter cold and reliably produce seed.

Cytological and geographical characterizations of Hemarthria

SummaryChromosme numbers were determined for 56 Hemarthria altissima(Poir.) Stapf et C. E. Hubb. introduction, forty of these were diploid (2n=18) and 16 were tetraploid (2n=36). These results, combined with previous findings, show that all but three of the tetraploids in the USDA Hemarthria altissima collection originated north of 20 S latitude. All other introductions collected throughout southern Africa were diploid. Meiosis was regular in a hexaploid, 6 tetraploid and 32 diploid introductions but a low frequency of quadrivalents was found in the teraploids. H. uncinataR. Br. and H. uncinata var. spathacea (Domin) Vickery were found to have 2n=36 chromosomes H. compressa (L.f.) R. Br., introduced from Japan, had 2n=54 chromosomes. No major cytological abnormalities exist in these Hemarthria species. Significant variation exists among the ploidy levels for cold tolerance and in vitro organic matter digestion to provide the basis for a forage improvement program.

Heritability estimates for callus growth and regeneration in desmodium

The F2 and F3 generations of two crosses (6123×13083 and 6123×144, with 6123 the regenerating parent) were evaluated for callus growth and regeneration capacity. Based on joint scaling tests and variance partitioning, neither callus growth nor regeneration fitted a simple additive-dominant genetic model. Heritability estimates obtained from parent-offspring regression analyses ranged from 0.65 to 0.77 for callus growth and from 0.19 to 0.46 for regeneration, with the range in both influenced by the cross and numerical scale employed. Members of two F3 families exhibited much more vigorous and prolific regeneration than the regenerating parental genotype. Because many individuals in the segregating generations showed no evidence of regeneration, population distributions for this trait were severely truncated, or censored. Regression-order analysis was used to estimate means and variances of these censored populations. The association between poor callus growth and high regeneration capacity observed in the parental lines was absent from the F2 and F3 generations, indicating that no association between callus growth and regeneration was present.

Clover, red (Trifolium pratense).

Genetic modification of plants by the insertion of transgenes can be a powerful experimental approach to answer basic questions about gene product function. This technology can also be used to make improved crop varieties for use in the field. To apply this powerful tool to red clover, an important forage legume, a population of red clover with high potential for regeneration in tissue culture has been developed. Here we provide a detailed procedure for Agrobacterium-mediated transformation of genotypes derived from this regenerable population. We have successfully used this methodology to express β-glucuronidase (GUS) reporter genes as well as for hairpin RNA-mediated silencing of endogenous genes for polyphenol oxidase and a transferase crucial in phaselic acid accumulation.

Comparative assessment of variation in the USA Arachis pintoi (Krap. and Greg.) germplasm collection using RAPD profiling and tissue culture regeneration ability.

Arachis pintoi accessions were used to study genetic diversity using RAPD markers. Concurrently, two tissue culture protocols were evaluated for organogenesis and the capacity to generate somaclonal variation. Data were collected on callus growth, callus weight gain, and number of regenerated plants. Robust RAPD profiles were obtained and eight primers amplified 100 different bands with 98% polymorphisms. The proportion of polymorphic RAPD loci was 89%. Average genetic distance was 0.36 and indicated that a large amount of genetic diversity exists within the germplasm evaluated. Genetic distances were used to prepare a dendogram for the A. pintoi accessions that separated them into four groups. A large degree of variability for callus induction and callus weight gain was observed among the accessions. Shoot regeneration was achieved for several accessions on both media with no structures indicative of somatic embryogenesis detected. Root induction was difficult to obtain, and many shoots died during this process. RAPD band profiles of regenerated tissue culture plants were similar to their parent plants, and therefore no somaclonal variation was evident using these methods.