Ethephon induced oxidative stress in the olive leaf abscission zone enables development of a selective abscission compound.

BACKGROUND: Table olives (Olea europaea L.), despite their widespread production, are still harvested manually. The low efficiency of manual harvesting and the rising costs of labor have reduced the profitability of this crop. A selective abscission treatment, inducing abscission of fruits but not leaves, is crucial for the adoption of mechanical harvesting of table olives. In the present work we studied the anatomical and molecular differences between the three abscission zones (AZs) of olive fruits and leaves. RESULTS: The fruit abscission zone 3 (FAZ3), located between the fruit and the pedicel, was found to be the active AZ in mature fruits and is sensitive to ethephon, whereas FAZ2, between the pedicel and the rachis, is the flower active AZ as well as functioning as the most ethephon induced fruit AZ. We found anatomical differences between the leaf AZ (LAZ) and the two FAZs. Unlike the FAZs, the LAZ is characterized by small cells with less pectin compared to neighboring cells. In an attempt to differentiate between the fruit and leaf AZs, we examined the effect of treating olive-bearing trees with ethephon, an ethylene-releasing compound, with or without antioxidants, on the detachment force (DF) of fruits and leaves 5 days after the treatment. Ethephon treatment enhanced pectinase activity and reduced DF in all the three olive AZs. A transcriptomic analysis of the three olive AZs after ethephon treatment revealed induction of several genes encoding for hormones (ethylene, auxin and ABA), as well as for several cell wall degrading enzymes. However, up-regulation of cellulase genes was found only in the LAZ. Many genes involved in oxidative stress were induced by the ethephon treatment in the LAZ alone. In addition, we found that reactive oxygen species (ROS) mediated abscission in response to ethephon only in leaves. Thus, adding antioxidants such as ascorbic acid or butyric acid to the ethephon inhibited leaf abscission but enhanced fruit abscission. CONCLUSION: Our findings suggest that treating olive-bearing trees with a combination of ethephon and antioxidants reduces the detachment force (DF) of fruit without weakening that of the leaves. Hence, this selective abscission treatment may be used in turn to promote mechanized harvest of olives.

Transplacental effects of maternal feeding with high fat diets on lipid exchange and response of the splenic lymphoid system in mice offspring exposed to low doses of carcinogen.

We studied whether feeding pregnant female mice with different fats affects lipid exchange and activity of the splenic lymphoid system in offspring exposed to low doses of carcinogen. Female mice were fed diets with either 7% or 15% corn oil or olive oil. The 4-week-old offspring of these mice were transferred to a chow diet, and exposed to a low dose of the carcinogen, dimethylbenz(a)antracene (2 mg/kg). Results of experiments were studied 5 months later. Concentrations of polyunsaturated linoleic and oleic acids were determined in the blood and liver of mothers and offspring. The activity of the splenic immune system in offspring was studied using immunohistochemical methods for evaluating the number of different types of lymphocytes (B and T cells), mitotic and apoptotic indexes and the number of Fas-positive lymphocytes. Serum concentrations of the fatty acids examined were unchanged in the blood of the mothers and their offspring. Concentration of both linoleic and oleic acids was significantly higher in the liver of mothers fed the 15% olive-oil or corn-oil diets. This high level was maintained in linoleic acid in offspring of mothers fed the 15% olive-oil diet. Spleen weight was higher in offspring of mothers fed a 15% corn-oil diet compared to those fed the 7% corn-oil diet. The 15% olive-oil diet slightly decreased the weight of the spleen compared to counterparts fed the 15% corn-oil diet. Immunohistochemical studies showed that the olive diet, partially of 15%, significantly stimulated B-cell blast transformation. The finding reflects the reaction of B lymphocyte-producing splenic zones to the carcinogenic effect, though to a weak extent. T lymphocyte-producing zones did not respond to the diets studied, probably due to the weak carcinogenic effect and lack of tumor appearance. The Fas activity of both B and T cells in the spleen was stimulated by the carcinogen and enhanced by feeding the mothers on the olive-oil diet. Maternal feeding with a diet rich in olive oil before pregnancy results in stimulation of morphological and functional attributes of the splenic immune system of the offspring, particularly related to producing of B lymphocytes.

Prunin identification, biological activity and quantitative change in comparison to naringenin in dormant peach buds.

Prunin (naringenin 7-glucoside), which was identified in dormant peach buds, was found to act as a growth inhibitor of wheat coleoptile elongation.Quantitative estimation on a fresh weight basis of the flavonoid in peach buds revealed a high level in late summer, a steep decrease in autumn that became more moderate during the winter, with lowest values near bud swell in spring. Two periods of reaccumulation of prunin during October and January fitted 2 periods of arrested growth of the resting bud. A negative correlation was demonstrated between prunin and naringenin levels during August through October. It is proposed that prunin might act as a precursor for naringenin that accumulates in resting buds in autumn. It is suggested that the system glucoside-aglucone-beta glucosidase might have regulatory properties in the dormant bud.

Endogenous cytokinins in maturing manzanillo olive fruits.

Cytokinin activity was found in mature black olive fruits while in green fruits the activity was markedly lower. The substances with cytokinin activity showed chromatographic properties similar to those of nucleotides. Acid hydrolysis caused a shift of the active zone in the organic phase of 1-butanol-acetic acid-water (4:1:5) from R(F) 0.1 to R(F) 0.8. The possible relation of these endogenously appearing cytokinins to anthocyanin accumulation and aging of the fruit is discussed.

Ethylene Evolution following Treatment with 1-Aminocyclopropane-1-carboxylic Acid and Ethephon in an in Vitro Olive Shoot System in Relation to Leaf Abscission.

1-Aminocyclopropane-1-carboxylic acid (ACC) supplied via the cut base of detached olive shoots caused a burst of ethylene from leaves, but other cyclopropanes tested did not exhibit this effect. Ethephon (ET) and another ethylene-releasing compound caused a prolonged increase in ethylene evolution. ACC had only a very limited effect on leaf abscission regardless of concentration, whereas shoots placed with cut bases in ET for 60 to 80 minutes exhibited 100% leaf abscission within 90 hours. Shoots with inflorescences treated with ET just prior to anthesis began to wilt in vitro within 20 to 30 hours and failed to exhibit leaf abscission. At earlier stages of development, ET induced more leaf abscission on reproductive shoots than on vegetative shoots. It is suggested that the duration of ethylene evolution from the leaves governs their potential for abscission and that bursts of ethylene evolution even though large in amount may not induce abscission.

Structural physiological, and biochemical gradients in tobacco pith tissue.

Explants of tobacco pith taken at various distances from the apex of a mature stem show a sharp gradient in growth potential in vitro; growth is highest in the extreme apical and basal explants, and is minimal in explants removed ca. 75 cm from the apex. Calluses produced by the vigorously growing basal explants are harder and more compact than those produced from more apical explants. The gradient in growth potential is directly correlated with gradients in RNA, protein of cell sap and soluble N per unit fresh weight, but is inversely correlated with peroxidase activity. Cell size increases from apex to base of plants.The peroxidase activity of pith explants is electrophoretically resolvable into 2 isoperoxidases, moving anodically at pH 9.0. During in vitro culture, this activity rises, due to the formation of several new isozymes moving toward the cathode. The appearance of these isozymes occurs most rapidly in apical and extreme basal explants.

Changes in the Activity of Catalase (EC 1.11.1.6) in Relation to the Dormancy of Grapevine (Vitis vinifera L.) Buds.

Catalase activity in grapevine (Vitis vinifera L.) buds cv. ;Perlette.' increased to a maximum in October and thereafter decreased within 3 months to less than half its maximal rate. The decrease in catalase activity coincided with the decline in temperature during winter. The rate of sprouting of buds forced at 23 degrees C was negatively related to the activity of catalase. Artificial chilling of grapevine canes at 5 degrees C resulted in a 25% decrease of catalase activity in the buds after 3 days and 31% after 17 days. The activity of catalase increased to the control level only 96 hours after removing canes from 5 degrees C to room temperature. Efficient buddormancy breaking agents, such as thiourea and cyanamide decreased catalase activity to 64 and 50% of the controls respectively, while the activity of peroxidase remained the same under those conditions. A less efficient dormancy breaking agent dinitro-ortho-cresol, did not decrease catalase activity.

Effect of water vapor pressure on the thermal decomposition of 2-chloroethylphosphonic Acid.

Decomposition of 2-chloroethylphosphonic acid (Ethephon) was studied in dried films at various water vapor pressures (0.6 to 86.9 millimeters Hg, 3.2 to 93.9% relative humidity) and temperatures (20, 30, 40, and 50 C) at pH 6.3 and 7.0. The rate of decomposition could be determined equally well by [(14)C]Ethephon or ethylene measurements. The rate increases at increasing water vapor pressures at a constant temperature and pH, up to an optimum. The optimum vapor pressure for decomposition approximately doubles for each 10 C increase. The activation energy for the decomposition reaction in water vapor pressures of 3.2 to 12 millimeters Hg is 8.7 and 14.3 kilocalories per mole at pH 6.3 and 7.0, respectively.Decomposition of Ethephon is inhibited above an optimum vapor pressure. The inhibition is stronger at lower temperatures and at pH 6.3 than at pH 7.0. The rate of decomposition and the inhibition observed at a low temperature (20 C) was found to be similar on various surfaces, including olive leaves.Failure to induce olive (Olea europaea L.) fruit abscission under certain environmental conditions can be readily attributed to rapid breakdown of Ethephon at elevated temperatures and low relative humidities.