Effect of gibberellic acid on the uptake and loss of inorganic phosphate from sweet potato slices

Uptake and loss of irorganic phosphate was determined in sweet potato slices ‘aged’ in the presence or absence of gibberellic acid. Concentrations of uptake medium were varied from 0.1 to 1.2 mM. Slices, aged in gibberellic acid, took up considerably less phosphate, at all concentrations than control, whereas efflux from GA₃ treated discs was greater than control. Whether treated or untreated, kinetics proved to be biphasic hyperbolas. The results suggested an alteration in membrane permeability induced by gibberellic acid.     

Comparison between Aging of Slices and Ethylene Treatment of Whole White Potato Tubers

Cyanide-resistant O(2) consumption can be stimulated by either treating whole white potato tubers (Norchip) with ethylene, in the presence of 100% O(2), or aging slices obtained from untreated potato tubers. A comparison of alternative pathway activity elicited by either treatment was undertaken. The proportion of electrons flowing through the alternative path in the presence of intermediate concentrations of KCN and at various concentrations of salicylhydroxamic acid was identical in both cases. However, the respiration of slices from ethylene-treated tubers was in every case stimulated by KCN, whereas the aged slices never exhibited this phenomenon. Furthermore, the metabolism of d-[U-(14)C]glucose was several hundred times greater in aged slices than in fresh slices from C(2)H(4)-treated tubers. These results, along with the respiratory kinetics of aged slices from ethylene-treated tubers, suggest that aged slices and fresh slices from ethylene-treated tubers are biochemically dissimilar.     

Influence of calcium on sodium and potassium absorption by fresh and aged bean stem slices

The influence of Ca on the aging processes of bean stem (Phaseolus vulgaris) slices and on the absorption of K and Na by fresh and aged slices was investigated. In the presence of Ca, fresh tissue showed a preferential Na uptake. The preference for Na over K resulted from a differential depressive effect of Ca on absorption of these two ions. In aged tissue Na uptake was also depressed, but K absorption was accelerated, with a net result of a much greater absorption of K than Na.     

Investigation of respiratory and ion transport properties of aging bean stem slices

Ion and oxygen uptake were studied on aging bean stem slices. Oxygen uptake was high immediately after slicing, decreased to a minimum at 100 minutes, and then increased again. Ion uptake per unit of O₂ uptake data suggested that metabolic energy was utilized almost exclusively for sodium transport in fresh tissue but was diverted to potassium transport as the slices aged. Oxygen and ion uptake in fresh slices was less sensitive to 2,4-dinitrophenol as compared to the aged slices, indicating major metabolic and physiological changes occurred during aging. This was further substantiated by the tissue response to cyanide and antimycin A. Oxygen uptake was decreased by cyanide (22% by 1 mm) and antimycin A (14% by 1 microgram per milliliter) in fresh slices but not in aged slices. Potassium uptake that developed during aging was sensitive to cyanide and antimycin A. The results are pertinent to understanding the role of the stem in regulating ion transport in plants.     

Effect of Abscisic Acid on Sorbitol Uptake in Growing Apple Fruits

Levels of abscisic acid (ABA) in the fruit flesh of developingapples (cv. Golden Delicious) were measured by electron capturegas chromatography. ABA content of the tissue, calculated ona fresh weight basis, decreased at a constant rate from 200µg g^–1 in young fruit to 10 µg g^–1 inolder fruit and then increased when the ripening process commenced.On a whole fruit basis, the ABA level increased during the initialphase of fruit growth, remained constant during the linear growthphase and increased again when fruits started to ripen. During fruit development the ABA content correlated with therate of sorbitol uptake, when measured after discs of fruittissue were incubated in [¹⁴C]sorbitol. Sorbitol uptake washigh during the initial growth phase and declined at a constantrate during fruit development. ABA present in the incubation medium stimulated sorbitol uptakeinto fruit tissue at concentrations higher than 10^–8 M,whereas indolyl-3-acetic acid had no effect on uptake. When comparing sorbitol uptake in different zones of young fruit,it was found that uptake was higher in discs of outer fruitlayers than in discs from inner fruit zones. Key words: Pyrus malus, Apple, Abscisic acid, Sorbitol     

Jasmonic acid,polyamine, and antioxidant levels in apple seedlings as affected by Ultraviolet-C irradiation

The effect of UV-C irradiation on jasmonates, abscisic acid (ABA), polyamines and antioxidant activities was investigated in apple plants (Malus domestica Borkh.). The EC₅₀ values of O₂ ^–-scavenging activity in the UV-C treated leaves were lower than those in the untreated control. In addition, total ascorbic acid and polyphenolic concentrations in the UV-C treated leaves were generally higher than those in the untreated control. Although the endogenous ABA concentrations were not significantly different between the untreated control and UV-C treatment, the endogenous jasmonic acid, putrescine, and spermine concentrations in the UV-C treated plants were higher than those in the untreated control. However, the application of PDJ before UV treatment inhibited the rise of endogenous jasmonate and putrescine concentrations. The increase of ascorbic acid was also inhibited by PDJ application before UV treatment. These facts suggest that the increase of jasmonates and polyamines may be associated with UV-C stimulation, and UV-C irradiation may be effective for increasing antioxidant activity in apple leaves.     

Influence of Abscisic Acid on Phosphorus Metabolism and Respiration in Potato Tuber Discs. Comparison with the Cycloheximide Effect

Ageing of discs of potato tuber by incubation in an aerated medium, produces an increase in the rates of respiration and of phosphate uptake. The presence of cycloheximide (CHM) or abscisic acid (ABA) in the uptake medium, does not change uptake by fresh tissue over 3–4 h. On the other hand, CHM causes an inhibition of the rate of uptake by aged tissue although ABA does not. The addition of CHM or ABA to the ageing medium, prevents totally (CHM) or only partially (ABA) the increase in phosphate uptake. The analysis of ³²P-incorporation into the various phosphorylated fractions after 24 h of ageing with CHM or ABA show that CHM induces a large inhibition of the rate of uptake with an almost complete inhibition of ³²P-incorporation into the various phosphorylated fractions. By contrast, ABA produces equal inhibition of ³²P labelling of all fractions including all acid-soluble components. CHM prevents the increase in the rate of respiration, whereas ABA causes a slight stimulation. In both cases, no important effect on ATP content was observed. These results are discussed in terms of a comparison of ABA and CHM actions. They lead to the hypothesis of a specific effect of ABA on the development of the uptake mechanism.     

Stimulation by Ethylene of Mitochondrial Development in Wounded Sweet Potato Root Tissue

Ethylene (about 100 µl per liter) markedly stimulatedincreases in respiratory, Cyt c oxidase and succinate dehydrogenaseactivities of the crude mitochondrial fraction as well as mitochondrialmembrane protein during aging of sliced sweet potato root tissue,indicating that it stimulated mitochondrial development in woundedtissue. It had such an effect even when slices were pre-agedin its absence for 1 day and thereafter aged in its presence.The mitochondrial inner membrane from slices aged in ethylene-containingair was denser than that from fresh slices, while the membranefrom slices aged in ethylene-free air was lighter. Chloramphenicolcompletely inhibited the increase in Cyt c oxidase activitywhether slices were aged in the presence or absence of ethylene.Cycloheximide did not inhibit the increase in slices aged inethylene-free air, but did by 50% in those aged in ethylene-containingair. ¹ This work was supported in part by a Grant-in-Aid (No. 411308)for Scientific Research from the Ministry of Education, Scienceand Culture, Japan. (Received April 4, 1981; Accepted July 7, 1981)     

Metabolism of 2-C-(+/-)-abscisic Acid in excised bean axes

Excised embryonic bean axes (Phaseolus vulgaris, var. White Marrowfat) rapidly metabolize 2-¹⁴C-(±)-abscisic acid to two compounds, M-1 and M-2, which have very low growth-inhibitory activity. Chemical tests indicate the M-1 and M-2 are not previously described abscisic acid metabolites. M-2 accumulates in the axes and evidence is presented for the hypothesis that abscisic acid → M-1 → M-2. Zeatin, which partially reverses the abscisic acid-mediated growth inhibition of axes, neither decreases abscisic acid uptake nor causes any major changes in its metabolism. It was observed that axes transferred from abscisic acid-containing solutions to buffer resume control rates of fresh weight increase while still containing considerable quantities of abscisic acid.     

Abscisic Acid Enhances the Ability of the Desiccation-Tolerant Fern Polypodium virginianum to Withstand Drying

Detached fronds of Polypodium virginianum L. survived loss of65–70% of their fresh weight over 10 d of slow-drying.Drying over silica gel resulted in a faster rate of water loss,to a lower fresh weight, which the fronds did not survive. Whenfronds were incubated in abscisic acid for 24 h prior to silica-drying,the amount of water lost was reduced, resulting in survivalof the fronds upon subsequent rehydration. Incubation in abscisicacid for at least 18 h was necessary for survival. Fronds inwhich the final fresh weight after drying was below a criticalamount (i.e. to less than 25% original fresh weight) did notsurvive. A reasonable correlation could be drawn between electrolyteleakage upon rehydration and survival of somedesiccation treatments,although this was not always clear-cut, especially in frondsincubated in abscisic acid for an insufficient time to ensuresurvival. Several polypeptides were synthesized during slow-and silica-drying, and in response to abscisic acid. No novelpolypeptides were identified that were unique to the desiccationregimes which resulted in survival. Nor did ABA induce specificproteins in fronds desiccated after preincubation in this regulatorfor 24 h. Key words: Desiccation-tolerance, abscisic acid, protein synthesis, fern, Polypodium virginianum     

Ricerche sul meccanismo di azione della cocliobolina. A: effetti della tossina su fettine di tubero di patata in riposo

Abstract

STUDIES ON MECHANISM OF COCHLIOBOLIN ACTIVITY. A: EFFECTS OF THE TOXIN ON FRESH POTATO TUBER SLICES. — Cochliobolin, a toxic compound produced by Helminthosporium oryzae, causes inorganic phosphorus leakage from fresh potato tuber slices, higher amounts being found in incubation media with higher toxin concentrations and for longer incubation periods. Organic compounds, also, are released by potato tuber slices, like phenolic products, large amounts of sugars, and some aminoacids, alanine, lysine, tyrosine, valine and γ-aminobutirric acid, which have never been observed in the incubation meida of untreated slices. Cochliobolin uptake by fresh potato tuber slices does not take place actively or, anyhow, at a metabolic level. Indeed, neither 2,4-dinitro-phenol (at concentration, pH, and incubations periods apt to reduce active uptake), nor low temperatures (0–2°C) prevent toxin uptake. Potato tuber slices are not affected by cochliobolin, unless kept at 25°C, and this fact suggests that while toxin uptake does not interest metabolic pathways, it might affect some phase, or structure, essential to cell metabolism. The locus of toxin activity seems therefore to be identified with its influence on cell absortion and retention activities, of products essential for metabolic processes. Thus the impairing of cell permeability, possibly affecting the processes or the structures which regulate the active uptake, seems to be, at the moment, the only evident interpretation of the phenomena observed.     

Influence of Ageing and Abscisic Acid on Potassium Uptake by Potato Tuber Tissues

The potassium uptake by potato tuber discs tissues freshly cut and after 24 h of ageing in the presence or not of abscisic acid was investigated. Uptake kinetics revealed a biphasic dependence on external K⁺ concentrations. At concentration less than 10 mM, uptake was mediated by a saturable component and a linear component became apparent at higher concentrations. At low K⁺ concentrations (lmM), the capacity of K⁺ uptake diminished by 2 times after ageing. Treatment of tissues with ABA increased the rate of K⁺ uptake. In both fresh and aged tissues the uptake was strongly enhanced by fusicoccin and decreased by several metabolic inhibitors and ATPase inhibitors, underlying the active nature of uptake and suggesting the involvement of a plasmalemma H⁺-ATPase in K⁺ transport system. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of Abscisic Acid, Salicylic Acid and Trans-cinnamic Acid on Phosphate Uptake, ATP-level and Oxygen Evolution in Scenedesmus

The effects of abscisic acid, salicylic acid and trans-cinnamic acid were tested on the light-induced phosphorylating reactions and oxygen evolution of the unicellular green alga Scenedesmus. It was found that abscisic acid and cinnamic acid had practically no influence on the total inorganic phosphate uptake, while salicylic acid in the concentration range of 10^-6 to 10^-3M gave a small decrease in the total inorganic phosphate uptake. The ATP level in the cells is in most cases increased when these three acids are given to the algae. The oxygen output is not significantly changed by abscisic acid or salicylic acid. Trans- cinnamic acid inhibits the oxygen evolution at concentrations of 10^-4–10^-3M None of the substances investigated caused such effects on photophosphorylation and oxygen evolution in Scenedesmus as those caused by the inhibitor β-complex from potatoes according to earlier reports. It is suggested that these effects are due to other components in the inhibitor β-complex.     

Insulin prevention of impaired renal calcium transport in alloxan-diabetic rats

The uptake of calcium-45 was measured in slices of kidney cortex from rats with alloxan-diabetes of increasing durations. Slices from untreated diabetic rats had significantly lower slice-to-medium ratios for calcium than did slices from control rats, reflecting decreased uptake of calcium by cortical slices from diabetic rats. Institution of daily insulin therapy immediately upon confirmation of diabetes prevented this depression of calcium uptake, even though the insulin therapy was insufficient to control blood glucose levels. After four weeks of untreated alloxan-diabetes, insulin therapy did not return calcium uptake by kidney cortex slices to control levels.     

The Role of Abscisic Acid in Senescence of Detached Tobacco Leaves

The role of abscisic acid in the regulation of senescence was investigated in detached tobacco leaves (Nicotiana rustica L.). Leaves senesced in darkness showed a sharp rise in abscisic acid level in the early stage of aging, followed by a rapid decline later. The same trend was found when leaves were aged in light, but the rise in abscisic acid occurred four days later than in darkness. Senescence was slower in light than in darkness, while salt stress accelerated the processes. Leaves treated with kinetin which senesced in light and darkness, did not show an increase in abscisic acid. Application of kinetin led to a transformation from free to bound ABA. These results may indicate that ABA and cytokinin are involved in a trigger mechanism which regulates senescence; the stage at which this trigger is activated determines the rate of senescence.     

Variation du taux d'acide abscissique chez deux variétés de poiriers, la Passe-Crassane et la Doyenne du Comice, en fonction de l'induction de la parthénocarpie par l'acide gibbérellique

The content of abscisic acid in the developing fruits of two cultivars of pear (Pyrus communis L.) was determined by UV spectrophotometry and by a colorimetric method based on the possibility of the lactone corresponding to abscisic acid to give a purple colour when sodium hydroxide is added. The young fruits were harvested either from trees treated with gibberellic acid (GA₃), for the purpose of inducing parthenocarpy, or from untreated trees. High levels of abscisic acid were associated with abortion and abscission of young fruits (June drop). There was little difference between fruits of GA₃-treated and untreated trees of the cultivar Passe-Crassane. In the other cultivar, ‘Doyenne du Comice’, the level of abscisic acid in the fruits of GA₃-treated trees was about twice as high as in control fruits and seemed to be directly related to the number of fruits per tree.     

Correlative Bud Inhibition and Abscisic Acid in Acer pseudoplatanus and Syringa vulgaris

Growing shoots of Acer pseudoplatanus and Syringa vulgairs were decapitated and defoliated (“treated”) in June 1972–1975. During the 2–3 weeks after treatment the contents of abscisic acid (ABA) in the out-growing lateral buds at the upper-most node as well as in the petiole stumps and nodes were determined by means of gas-liquid chromatography. The concentration of ABA in lateral buds of intact shoots, calculated on a fresh weight basis, varied greatly from year but was consistently several times higher than in petioles and nodes. Defoliation and decapitation caused out-growth of the lateral buds. This was accompanied by a sharp decrease in the ABA concentration, which finally reached the level of petiole tissue. The concentration of ABA in controls (“untreated”) decreased also, but to a smaller extent, and remained higher than in petioles. In petioles and nodes of treated as well as of untreated shoots, the ABA concentration did not change. The absolute amount of ABA in the buds of treated shoots after 2–3 weeks varied greatly, but was apparently not different from the amount in buds of untreated controls or in buds at the beginning of the experiment. Therefore, the decrease of the ABA concentration was mainly due to fresh weight increase. The results are discussed in relation to a possible role of ABA in correlative bud inhibition.     

Relative Contribution of Cytochrome-mediated and Cyanide-resistant Electron Transport in Fresh and Aged Potato Slices

The respiration of fresh potato (Solanum tuberosum, var. Russet Burbank) slices is predominantly cyanide-sensitive whether in the presence or absence of uncoupler. By contrast, the wound-induced respiration which develops in thin slices with aging is cyanide-resistant, and in the presence of cyanide, sensitive to chlorobenzhydroxamic acid, a selective inhibitor of the cyanide-resistant respiration. Titration of the alternate path in coupled slices with chlorobenzhydroxamic acid, in the presence and absence of cyanide, shows that the contribution of the cyanide-resistant pathway to the wound-induced respiration is zero. Similar titrations with uncoupled slices reveal that the alternate path is engaged and utilized extensively.

The maximal capacity of the cytochrome path (V_cyt) has been estimated in fresh and aged slices in the presence of the uncoupler carbonyl-cyanide m-chlorophenyl hydrazone. It has been found that V_cyt of aged slices is but 30 to 40% higher than that of fresh slices. The results suggest that the bulk of the wound-induced respiration is mediated through the cytochrome pathway which exists in fresh slices in suppressed form, and which is fully expressed by slice aging. The engagement of the alternate path by uncouplers in aged slices is attributed to an increase in substrate mobilization, with the result that the electron transport capacity of the cytochrome chain is exceeded.

     

Ethylene and wound-induced cyanide-resistant respiration climaxes in potato: The synergistic role of CO2 and the selective role of lycorine

When treated with ethylene in O₂, conditioned potato (Solanum tuberosum L. cv. Russet Burbank) tubers – that is, tubers kept at room temperature for 10 days or more – yield slices that are CN^? resistant. Ten % CO₂ in the gas mixture not only synergizes the effect of ethylene, but replaces the need for conditioning as well. The response to CO₂ is more pronounced with increasing time from harvest. By contrast fresh slices from untreated tubers are CN^? sensitive, as are slices from tubers incubated in O₂ or O₂ plus CO₂. The suggestion is made that CN^? resistance is constitutive, and that treatment with ethylene/CO₂ in O₂ confers on potato tuber tissue a resistance to the extensive degradation of membrane phospholipids that normally attends slicing and leads to the loss of CN^? resistance. In this connection respiration inhibition by imidazole, an inhibitor of fatty acid α-oxidation, is extensive in slices of untreated tubers, and sharply diminished in slices of ethylene-treated tubers in proportion to their CN^? resistance. The coextensive rise of respiration rate and CN^? resistance in aged potato slices has led to the presumption that the CN^?-resistant path mediates the respiration climax. Accordingly the alkaloid, lycorine, has been considered to inhibit the development of CN^? resistance in aging potato slices because it curtails the wound-induced respiration. A comparison was carried out on the effect of lycorine on CN^?-sensitive and CN^?-resistant fresh slices – the latter obtained from ethylene/CO₂-treated tubers. Lycorine suppressed the development of the wound-induced respiration without restricting the development of CN^? resistance.     

Induction and development of somatic embryos from spinach (Spinacia oleracea) leaf segments

Somatic embryogenesis was induced in embryogenic calli of Spinacia oleracea L., on a Murashige and Skoog (1962) medium, containing 4.6 μM kinetin as the sole growth regulator. Abscisic acid, gibberellic acid, and indole-3-acetic acid were supplemented to kinetin-containing medium and their effects on the initiation of somatic embryos was studied. Abscisic acid at a particular concentration (4 μM) dramatically increased the number of embryos per g fresh weight of callus, while both gibberellic acid and indole-3-acetic acid suppressed the embryo initiation. It is suggested that the promoting effect of abscisic acid on the embryo initiation may be explained as a stress response of the tissue. The relative number of globular embryos vs. the embryos in heart/torpedo and cotyledonary stages was increased at 4 μM abscisic acid and at all gibberellic acid concentrations (0.3–10 μM). In contrast, the ratio of globular to polar embryos was lower than in controls at 1 μM abscisic acid and indole-3-acetic acid (1 and 10 μM). The effects of growth regulators on the ratio of globular to polar embryos indicate that they interfere with the normal distribution of cell division and cell expansion during early embryogenesis. This revised version was published online in June 2006 with corrections to the Cover Date.