Biochemical differences between carrot inbreds differing in plant regeneration potential.

Cultures derived from domestic carrot (Daucus carota L.) inbreds were found to vary with respect to regeneration potential as measured by the production of somatic embryos in suspension cultures. A number of biochemical parameters previously reported to distinguish embryogenic from non-embryogenic cultures of other species were measured in these carrot cell lines. Ethylene production was found to be inversely related to regeneration potential. The cell line producing the greatest number of somatic embryos exhibited the lowest rate of ethylene biosynthesis, even when grown on 2, 4-D-containing maintenance medium. A specific isozyme of acid phosphatase was associated with embryogenic calli. Proteins visualized by SDS-PAGE did not discriminate between embryo-forming and proliferating calli in all inbreds.

Triglycerides in embryogenic conifer calli: a comparison with zygotic embryos.

Triglycerides in developing zygotic embryos of Norway spruce and loblolly pine were found to accumulate continuously during the course of development, comprising nearly 50% of the fresh weight of a mature embryo. Embryogenic calli of these two species contained dramatically lower levels of triglycerides. Abscisic acid treatments promoted both embryo production and triglyceride accumulation in Norway spruce cultures. A method used to determine triglyceride levels in human serum, commercially available in kit form, was adapted for use with plant tissues. Low levels of triglycerides in the cultured tissues may be related to difficulties in the development and germination of conifer somatic embryos.

DL-alpha-difluoromethyl[3,4-3H]arginine metabolism in tobacco and mammalian cells. Inhibition of ornithine decarboxylase activity after arginase-mediated hydrolysis of DL-alpha-difluoromethylarginine to DL-alpha-difluoromethylornithine.

DL-alpha-Difluoromethylarginine (DFMA) is an enzyme-activated irreversible inhibitor of arginine decarboxylase (ADC) in vitro. DFMA has also been shown to inhibit ADC activities in a variety of plants and bacteria in vivo. However, we questioned the specificity of this inhibitor for ADC in tobacco ovary tissues, since ornithine decarboxylase (ODC) activity was strongly inhibited as well. We now show that [3,4-3H]DFMA is metabolized to DL-alpha-difluoromethyl[3,4-3H]ornithine [( 3,4-3H]DFMO), the analogous mechanism-based inhibitor of ODC, by tobacco tissues in vivo. Both tobacco and mammalian (mouse, bovine) arginases (EC 3.5.3.1) hydrolyse DFMA to DFMO in vitro, suggesting a role for this enzyme in mediating the indirect inhibition of ODC by DFMA in tobacco. These results suggest that DFMA may have other effects, in addition to the inhibition of ADC, in tissues containing high arginase activities. The recent development of potent agmatine-based ADC inhibitors should permit selective inhibition of ADC, rather than ODC, in such tissues, since agmatine is not a substrate for arginase.

Arginine decarboxylase and polyamines required for embryogenesis in the wild carrot.

Embryogenic cultures of Daucus carota treated with 1 millimolar alpha-difluoromethylarginine, a specific inhibitor of arginine decarboxylase, exhibited nearly a 50 percent reduction in embryo formation compared with controls. Putrescine and spermidine concentrations in the treated cells were greatly reduced. Addition of putrescine, spermidine, or spermine to the culture medium restored embryogenesis in the treated cultures. Embryogenesis was not significantly affected by alpha-difluoromethylornithine, an inhibitor of ornithine decarboxylase. These results suggest that polyamines have a major function in plant embryo development and that the wild carrot synthesizes polyamines through the biosynthetic pathway involving arginine decarboxylase rather than ornithine decarboxylase.

Relative role of caloric restriction and exercise training upon susceptibility to isoproterenol-induced myocardial infarction in male rats.

To determine the relative effect of body weight and physical inactivity on susceptibility to drug-induced myocardial infarction, randomly selected groups of 100-day-old male Sprague-Dawley rats were subjected to a 10-wk program of exercise training and (or) to caloric restriction followed by two subcutaneous injections of L-isoproterenol. Two groups of rats were fed a restricted diet consisting of minimum calories to maintain body weight and were either exercised (R-Ex) or remained sedentary (R-C), one group was fed ad libitum and subjected to exercise (Ex), and one group remained sedentary (C), but was provided only enough food to maintain body weight in a range similar to Ex rats. Initially, there was no difference between group body weights, but Ex and C rats exhibited a significantly greater final body weight. All Ex, R-Ex, and R-C rats survived the isoproterenol injections, but 50% of C rats died. Group C rats exhibited significantly greater activity of total plasma lactate dehydrogenase (LDH), whereas R-Ex rats had the lowest total LDH activity (p less than 0.05). R-Ex and R-C rats had a significantly lower activity of plasma LDH-1, the heart isozyme, than did the heavier Ex and C rats. More specifically, R-C rats exhibited a significantly decreased amount of plasma LDH-1 activity when compared with Ex rats, indicating that smaller, untrained rats had less myocardial damage than the heavier, exercise-trained rats. These data suggest that either exercise or maintenance of body weight is beneficial toward prevention of the drug-induced myocardial infarction, but when weight maintenance is combined with exercise additional protection is provided.