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Richard T. Olsen and Thomas G. Ranney

Inheritance of two mutant foliage types (purple and mottled variegated) was investigated for diploid, triploid, and tetraploid tutsan (Hypericum androsaemum). Segregation ratios were determined for diploid crosses in reciprocal dihybrid F1 and F2, BC1P1, and BC1P2 families. F2 tetraploids were derived from autotetraploid F1s. Triploid segregation ratios were determined from crosses between autotetraploid F1s and diploid F1s. Diploid di-hybrid crosses fit the expected 9:3:3:1 ratio for a single, simple recessive gene for both traits, with no evidence of linkage between each trait. Data from backcross and triploid crosses generally supported this model. In tetraploid crosses we observed twice as many variegated phenotypes as predicted which was not explained by random chromosome or chromatid assortment. Inheritance of purple foliage did not deviate from random chromosome assortment at the tetraploid level.

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Richard T. Olsen and Thomas G. Ranney

A diverse collection of germplasm representing 24 taxa from Catalpa sect. Catalpa Paclt and sect. Macrocatalpa Grisebach, Chilopsis D. Don, and ×Chitalpa Elias & Wisura were screened for susceptibility to powdery mildew (PM), Erysiphe elevata (Burr.) U. Braun & S. Takam, and Catalpa sphinx larvae (CSL), Ceratomia catalpae (Boisduval), feeding. The PM screening was conducted in 2004–05, with plants grown in a lathhouse (50% shade) in 2004, and a gravel pad (100% full sun) in 2005. The PM causal organism was identified as Erysiphe elevata both years. Disease incidence and severity were recorded at 2-week intervals for 6 weeks and used to calculate area under the disease progress curves (AUDPC) for each taxon for each year. North American Catalpa in sect. Catalpa, Chilopsis, and ×Chitalpa taxa were all moderate to highly susceptible to PM. Chinese Catalpa in sect. Catalpa and West Indian species in sect. Macrocatalpa were resistant to PM. Hybrids between North American and Chinese Catalpa in sect. Catalpa varied in susceptibility, indicating inheritance of partial resistance to PM. A no-choice feeding study conducted with CSL in 2005 found no differences in survival or growth of larvae reared on taxa from Chilopsis, ×Chitalpa, or either section of Catalpa. Future breeding of ×Chitalpa can utilize two different sources of PM resistance, but a source for resistance to CSL was not identified.

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Richard E. Bir, T. G. Ranney and R. K. Jones

Twelve shrub rose cultivars were evaluated for pest resistance in the southern Blue Ridge mountains under high humidity and rainfall (1.34 inches per week average during the 1994 and 1995 growing seasons). `Albo plena', `Blanc Double de Coubert', `Fru Dagmar Hastrup', `Roseraie de l'Hay', R. rugosa `Alba', `Sarah van Fleet', and `Topaz Jewel' were highly resistant to black spot and Cercospora sp. leaf spot. `Alba Meidiland', `Linda Campbell', `Pink Meidiland', and `Scarlet Meidiland' were susceptible, while `Bonica' displayed intermediate resistance to both diseases. `Sarah van Fleet' foliage and the flowers of `Albo plena', `Blanc Double de Coubert', and R. rugosa `Alba' were damaged by Japanese beetle feeding. No other cultivars were damaged by Japanese beetles.

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Richard T. Olsen, Thomas G. Ranney and Zenaida Viloria

A series of studies were conducted to determine medium components necessary for ovule and embryo culture of ×Chitalpatashkentensis Elias & Wisura hybrids in order to improve recovery of interploid crosses. Ovules were collected at 2, 3, 4, 5, and 6 weeks after pollination (WAP) from selfed tetraploid × Chitalpa (S) and tetraploid × Chitalp × diploid Catalpabignonioides Walt. (3×) hybrids. Excised ovules were placed in petri dishes with Schenk and Hildebrandt (SH) medium and 0.7% agar, with or without coconut-water (2%) and three sucrose concentrations (20, 40, or 80 g·L-1). No ovules germinated for either cross in any treatment at 2, 3, and 4 WAP. Selfed ovules germinated at 5 WAP, in both 20 and 40 g·L-1 sucrose. At 6 WAP, 3× ovules germinated in 20 g·L-1 sucrose. Coconut water provided no apparent benefit. Embryos were apparent at 6 WAP, so a new study was initiated to compare ovule vs. embryo culture at this sample date. Excised embryos germinated in greater percentages than ovules, in all treatment combinations at 6 WAP. Germination in 80 g·L-1 sucrose was observed only for S embryos without coconut water. Greatest 3× germination (16.7%) was observed for embryos in 20 g·L-1 sucrose without coconut water. A final study was conducted to investigate the effect of gibberellic acid (GA3) on embryo germination. Embryos were harvested at 7 WAP for both crosses and grown in SH medium supplemented with 20 g·L-1 sucrose and 0, 1, 2, or 4 μm GA3. The addition of GA3, regardless of concentration, increased germination from 30.6% to 99.1% for S embryos and from 11.1% to 99.1% for 3× embryos.

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Richard T. Olsen, Thomas G. Ranney and Zenaida Viloria

×Chitalpa tashkentensis Elias & Wisura is a sterile intergeneric hybrid [Catalpa bignonioides Walt. × Chilopsis linearis (Cav.) Sweet]. To restore fertility in ×Chitalpa the following were evaluated: 1) oryzalin as a polyploidization agent, 2) fertility of induced polyploids, and 3) in vitro culture methods for embryo rescue of interploid crosses. Meristems of ×Chitalpa `Pink Dawn' were submerged in an aqueous solution of 150 μm oryzalin for 0, 6, 12, or 24 hours and ploidy analyzed via flow cytometry. As treatment duration increased, recovery of diploids decreased as mixoploids and shoot mortality increased. Two tetraploid shoots occurred in the 24-hour treatment. Four tetraploids and two cytochimeras were stabilized in total. Tetraploids flowered sparsely; however, cytochimeras flowered profusely and these were used to study fertility at the tetraploid level. Diploid ×Chitalpa `Pink Dawn' pollen was essentially nonviable, but cytochimera pollen stained and germinated equal to or greater than pollen of C. bignonioides and C. linearis `Bubba'. Cytochimera ×Chitalpa were selfed yielding tetraploid seedlings, crossed with C. bignonioides to yield triploids, but failed in reciprocal crosses with C. linearis `Bubba' and `Burgundy Lace'. To increase recovery of triploids, germination of triploid and tetraploid embryos was investigated, as either intact ovules or excised embryos, on Schenk and Hildebrandt (SH) basal salts supplemented with sucrose at 20, 40, and 80 g·L-1, presence or absence of 2% coconut-water, and gibberellic acid (GA3) at 0, 1, 2, or 4 μm, and harvested weekly beginning 2 weeks after pollination (WAP). Germination of triploids (cytochimera ×Chitalpa × diploid C. bignonioides) and tetraploids (selfed cytochimera ×Chitalpa) were greatest with excised embryos at 7 WAP on SH supplemented with sucrose at 20 g·L-1 and ≥1 μm GA3. Germination of triploids (diploid C. linearis × cytochimera ×Chitalpa) was <5% at 4, 5, or 6 WAP on the same medium as above. Oryzalin effectively induced polyploidy and restored fertility in ×Chitalpa `Pink Dawn'. Successful crosses between hybrid and parental taxa of different ploidy levels, coupled with embryo culture will facilitate a ×Chitalpa breeding program. Chemical names used: 4(dipropylamino)-3,5-dinitrobenzenesulfonamide (oryzalin).

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Richard T. Olsen, Thomas G. Ranney and Dennis J. Werner

Inheritance of two mutant foliage types, variegated and purple, was investigated for diploid, triploid, and tetraploid tutsan (Hypericum androsaemum). The fertility of progeny was evaluated by pollen viability tests and reciprocal crosses with diploids, triploids, and tetraploids and germinative capacity of seeds from successful crosses. Segregation ratios were determined for diploid crosses in reciprocal di-hybrid F1, F2, BCP1, and BCP2 families and selfed F2s with the parental phenotypes. F2 tetraploids were derived from induced autotetraploid F1s. Triploid segregation ratios were determined for crosses between tetraploid F2s and diploid F1s. Diploid di-hybrid crosses fit the expected 9: 3: 3: 1 ratio for a single, simple recessive gene for both traits, with no evidence of linkage. A novel phenotype representing a combination of parental phenotypes was recovered. Data from backcrosses and selfing support the recessive model. Both traits behaved as expected at the triploid level; however, at the tetraploid level the number of variegated progeny increased, with segregation ratios falling between random chromosome and random chromatid assortment models. We propose the gene symbol var (variegated) and pl (purple leaf) for the variegated and purple genes, respectively. Triploid pollen stained moderately well (41%), but pollen germination was low (6%). Triploid plants were highly infertile, demonstrating extremely low male fertility and no measurable female fertility (no viable seed production). The present research demonstrates the feasibility of breeding simultaneously for ornamental traits and non-invasiveness.

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Richard T. Olsen, Thomas G. Ranney and Charles S. Hodges

A diverse collection of germplasm representing 24 taxa from Catalpa sect. Catalpa Paclt and sect. Macrocatalpa Grisebach, Chilopsis D. Don, and ×Chitalpa Elias & Wisura were screened for susceptibility to powdery mildew (PM) incited by Erysiphe elevata (Burr.) U. Braun & S. Takam and feeding by catalpa sphinx larvae (CSL), Ceratomia catalpae (Boisduval). PM screening was conducted on plants grown in a lathhouse (50% shade) in 2004 and a gravel pad (100% sun) in 2005. The PM causal organism was identified as E. elevata both years. Disease incidence and severity were recorded at 2-week intervals for 6 weeks and used to calculate area under the disease progress curves (AUDPC) for each year. North American Catalpa in sect. Catalpa, Chilopsis, and ×Chitalpa taxa were all moderately to highly susceptible to PM. Chinese Catalpa in sect. Catalpa and West Indian species in sect. Macrocatalpa were resistant to PM. Hybrids between North American and Chinese Catalpa in sect. Catalpa varied in susceptibility, indicating transmission of partial resistance to PM. No differences in survival or growth were found in a no-choice feeding study with CSL reared on taxa from Chilopsis, ×Chitalpa, or either section of Catalpa. Future breeding of ×Chitalpa can use sources of PM resistance identified in this study, but a source of resistance to CSL was not found.

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Clara E. Trueblood, Thomas G. Ranney, Nathan P. Lynch, Joseph C. Neal and Richard T. Olsen

Although Hypericum androsaemum L. is a valuable landscape plant, the species can be weedy and potentially invasive in certain locations. Infertile, non-invasive cultivars of H. androsaemum with desirable ornamental features would be ecologically beneficial and valuable for the horticultural industry. The male and female fertility of 10 triploid H. androsaemum, developed with a combination of variegation and foliage colors, was investigated under greenhouse (controlled pollination) and field conditions (natural pollination). Male fertility was evaluated based on pollen viability tests (pollen staining and pollen germination). Female fertility was based on fruit set, seed set, germinative capacity of seeds, and number of seedlings produced for each flower. Although values for different measures of fertility varied among triploid clones, pollen germination was significantly reduced for all triploids and nine of the 10 triploids produced no viable seed. These results represent 100% failure of ≈171,000 potential fertilization events based on fertility levels of diploid controls. The remaining triploid clone produced two seedlings per flower compared with 260 seedlings per flower for the controls. However, the seedlings produced by the triploid clone died shortly after germination. This research documented that the triploid H. androsaemum tested are highly infertile with no measurable female fertility. These clones will provide ideal alternatives to fertile forms of H. androsaemum where invasiveness is a concern. These methods also provide a useful protocol for evaluating fertility of other taxa that are selected or developed as non-invasive cultivars of potentially weedy species.

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J. Kevin Parris, Thomas G. Ranney, Halina T. Knap and W. Vance Baird

The genus Magnolia includes over 250 species that range in ploidy level from diploid to hexaploid. Although there is basic information on ploidy levels of various species, sampling has been limited and little information on specific cultivars and hybrids is available. The objective of this research was to determine relative genome sizes and relationships to ploidy levels among a diverse collection of species, hybrids, and cultivars using flow cytometry. Nuclei were extracted, stained with 4′, 6-diamidino-2-phenylindole (DAPI), and analyzed using a flow cytometer. Relative genome sizes were determined using Pisum sativum as the reference genome. Genome size was calibrated with ploidy level for species with documented chromosome numbers. Relative genome size for a given ploidy level varied significantly among most taxonomic sections indicating these groups have undergone considerable genomic divergence. These data also indicate it is desirable to calibrate ploidy level with relative genome size for each section separately. Within a section, relative 2C genome sizes, for a given ploidy level, had narrow ranges and could be used to clearly distinguish between euploid levels. Genome size estimates, determined with DAPI or propidium iodide fluorochromes, varied (by 0% to 14%) as a function of species and base pair (bp) composition. Both methods were suitable for determining euploid level. Base pair composition of representative Magnolia species ranged from 61.6% to 63.91% AT. Genome sizes and ploidy levels are presented for a broad range of species and hybrids within genus Magnolia. This information also provides further insight into reproductive biology, substantiation of numerous hybrids and induced polyploids, and comparison of methods for determining genome size that will help facilitate the development of improved hybrids in the future.

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W.C. Dunwell, D. Fare, M.A. Arnold, K. Tilt, G. Knox, W. Witte, P. Knight, M. Pooler, W. Klingeman, A. Niemiera, J. Ruter, T. Yeager, T. Ranney, R. Beeson, J. Lindstrom, E. Bush, A. Owings and M. Schnelle

The Southern Extension and Research Activities/Information Exchange Group-27 (SERA/IEG-27) is sponsored by the Southern Association of Agricultural Experiment Station Directors. Thirteen universities and the U.S. National Arboretum cooperate with official representatives from extension and research programs. The objective of the group is to identify, evaluate, select, and disseminate information on superior, environmentally sustainable, landscape plants for nursery crop production and landscape systems in the southeastern U.S. Plants are distributed to members responding to a request from cooperators for plant evaluation. Those who agree to cooperate are expected to grow the selected liner to landscape size, then transplant it in a landscape setting. The plant is rated for insect, disease, and cold damage, heat stress, growth rate, ornamental flowering and fruiting, fall color, commercial production potential, landscape potential, invasiveness potential, and insect disease transmission potential. Growth rate is evaluated annually by recording plant height and width. Initial bloom date is reported followed by bloom duration in days. Following evaluation, the group collectively and individually disseminates information gained from the plant evaluation system to a wide variety of audiences.