A microcomputer-based image processing system was used to simplify the large number of visual comparisons required to identify various Corylus spp., cultivars, and clonal accessions using polyacrylamide gel electrophoresis isozyme patterns. Photographs of gels stained for peroxidase, acid phosphatase, and phenol oxidase were digitally captured and selected lanes were enhanced and scanned. The scan data were analyzed to locate bands and normalize their position to that of standards. Such data were plotted and a computer-generated isozyme pattern was displayed. Compressed image data were then stored in a database for subsequent automated isozyme pattern comparisons. Photographic records that were previously used in published reports were reevaluated with the computerized system. Species, cultivars, or clones that were characterized in visual evaluations were similarly characterized using the computer method. Computer evaluations usually identified more bands. Band positions were only rarely different and probably resulted from better normalization relative to standard bands when using the computerized procedure.
A.R. Dixon, R.B. Boone, A. Gardea, L.S. Daley and T.L. Righetti
A.M. Shirazi, Y.M. Moreno, L.H Fuchigami and L.S. Daley
Previously, we reported recovery of plants from “Near-Lethal” (NL) (Sub-Lethal) stresses was dependent on stage of development and post-stress environment Dormant plants exposed to NL-heat, freezing, and hydrogen cyanamide either died or were severely injured when stored at 0°C or recovered at 23°C and natural condition. This study reports on the changes in the evolution of metabolic heat in dormant red-osier dogwood (Cornus sericea L.) stem tissues after beat stress. Heat stress (51°C for half an hour) was followed by one of two post-stress environment (PSE) (0° or 23°C dark condition). Isothermal measurements of the heat of metabolism of the tissues were taken after 0, 1, 2, 5, 7 and 11 days of PSE. A significant reduction of metabolic heat generation occured in heat stressed plants at 0°C PSE from one to 11 days of incubation as compared to the non-stressed tissues. At 23°C PSE, no significant differences of heat generation between stressed and non stressed tissues were found within 7 days of incubation. The rate of metabolic. heat measured by decreasing temperature scanning microcalorimetry (21° to 1°C) were lower in beat stressed tissues. Arrhenius plots of metabolic heat rate gave a linear slope for non-stressed tissues and a complex slop for NL-stressed tissues at lower temperatures. Energy of activation (Ea) between 1°-8°C were 15.45 and 83.882 KJ mol-1 for NL-heat and non-stressed tissues, respectively.
A.A. Gardea, Y.M. Moreno, A.N. Azarenko, P.B. Lombard, L.S. Daley and R.S. Criddle
The pattern of respiratory changes during dormancy and development of `Pinot noir' primary grape (Vitis vinifera L.) buds through the broken-bud stage was determined using differential scanning calorimetry. Metabolic heat rates [μJ·s-1·mg-1 dry weight (dw)] decreased during endodormancy and increased during ecodormancy and bud development. Respiration of ecodormant buds was at its minimum (≈1 μJ·s-1·mg-1 dw) in January. Respiration steadily increased, beginning in February and continuing as buds passed through the four developmental stages: ecodormant, initial swelling, fully swollen, and broken bud. Heat of metabolism increased from 5 to 18, 28, and 29 μJ·s-1·mg-1 dw as the buds developed from ecodormant to broken-bud stage. Carbon dioxide evolution from respiring buds increased from 15.9 to 22.2, 57.9, and 68.3 × 10-6 μmol·s-1·mg-1 dw as bud development advanced. Metabolic inefficiency, as quantified by heat (kJ) released per CO2 (mol) produced, was lowest in ecodormant buds, then increased at initial bud swelling and declined again in fully swollen and broken buds. The temperature coefficient of metabolism (μ) [apparent activation energy (Ea)/gas constant (R)] was lower at low temperatures (5 to 15C) in ecodormant buds than at the three more advanced stages. At higher temperatures (20 to 35C), buds at the ecodormant and initial swelling stages had a higher μ than the fully swollen and broken buds. These results suggest there may be a major change in metabolic pathways between the ecodormant bud stage and bud swelling.
A. A. Gardea, Y. M. Moreno, A. N. Azarenko, P. B. Lombard, L. S. Daley and R. S. Criddle
The increasing respiration of breaking `Pinot Noir' buds was measured by Differential Scanning Calorimetry. Bud development was classified into ecodormant, initial swelling, fully swollen, and breaking buds. Metabolic and CO2 evolution heat rates increased as the buds developed. Activation energy decreased steadily as development proceeded, which implied that less energy was required for metabolism to continue at later bud stages. A decrease in metabolic efficiency noted by a low calorespirometric ratio was observed during the transition from ecodormant to the initial swelling stage. From the second stage on, metabolic efficiency increased. The responsive nature of grape buds to warm temperatures was explained by increasing Q10 (10-20C) values from 2.8 to 3.8, 3.2, and 3.6 for the four developmental stages described above.
A. A. Gardea, P. B. Lombard, R. L. Kohnert, A. N. Azarenko, Y. M. Moreno and L. S. Daley
Changes in water content of `Pinot Noir' endo- and ecodormant primary buds were gravimetrically partitioned into extracellular (ECW) and intracellular water (ICW). During endodormancy, water status remained unchanged with values of 0.6 and 0.1 mg/mg dw for ICW and ECW, respectively. Ecodormant buds, prior to budbreak, increased in ICW from 0.5 to 4.4 mg/mg dry weight for Jan. and Apr., respectively. Liquid water in the buds was determined by H-NMR. The spin-spin relaxation time (T2) at -30C represented the bound fraction, which peaked in Jan. at 0.3 mg/mg dw followed by a decrease to 0.2 mg/mg dw in March. During the dormant season the free fraction was always larger than the bound fraction. No vascular connection between bud and stem was observed by mid Jan. Changes in bound water indicated that there is a transient fraction changing to the free form. These changes were not strictly related to the bud's dormancy status.