Ethylene: an urban air pollutant.
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Biomedical subjects
Publications and source records attributed to F B Abeles.
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Ethylene production was induced in sections of dark-grown Sorghum vulgare L. seedlings by treatment with light in the blue and far red regions of the light spectrum. The action spectrum closely resembled the previously reported spectra for high irradiance response; thus, light-induced ethylene production is probably a high irradiance response with phytochrome as the initial photoreceptor.
We observed no exchange between deuterated ethylene (C(2)D(4)) and the hydrogen of pea seedlings (Pisum sativum L. cv. Alaska). This suggests that bonding forces in which exchange could readily occur are not important in the physiological action of ethylene. Deuterated ethylene was just as effective as normal ethylene in inhibiting the growth of pea root sections. These results indicate that splitting carbon to hydrogen bonds did not occur during ethylene action.
Glucanase (endo-beta-1, 3-glucan 3-glucanohydrolase, EC 3.2.1.6, laminarinase, callase) and chitinase (poly-beta-1, 4-[2-acetamido-2-deoxy] -d-glucoside glycanohydrolase, EC 3.2.1.14) were extracted from ethylene-treated bean (Phaseolus vulgaris L. cv. Red Kidney) leaves and purified on hydroxyapatite and carboxymethyl Sephadex columns. The glucanase prepared was homogeneous as judged by analytical centrifugation data, electrophoresis, and antibody-antigen reactions. On the basis of gel filtration, antibody-antigen reactions, and amino acid analysis, the molecular weight was estimated to be between 11,500 and 12,500. However, ultracentrifugation gave a higher estimate of 34,000. The glucanase had an isoelectric point near pH 11 and was specific for beta-1, 3-linkages. The chitinase was only partially purified as judged by electrophoretic behavior.
The role of ethylene in the aging of bean (Phaseolus vulgaris L. cv. Red Kidney) petiole abscission zone explants was examined. The data indicate that ethylene does accelerate aging in addition to inducing changes in break strength. Application of ethylene during the aging stage (stage 1) promoted abscission when followed by a second ethylene treatment during the cell separating stage (stage 2). The half-maximal effective concentration of ethylene to induce aging was around 0.3 microliter per liter; 10 microliters per liter was a saturating dose. CO(2) reversal of ethylene action during stage 1 was incomplete and gave ambiguous results. CO(2) (10%) reversed the effect of 10 microliters per liter ethylene but not 1 microliter per liter ethylene. The possibility that ethylene not only accelerated aging but was also a requirement for it was tested, and experimental evidence in favor of this idea was obtained. It was concluded that ethylene plays a dual role in the abscission of bean petiole explants: a phytogerontological effect and a cellulase-inducing effect.
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The endo-beta-1, 3-glucanase (beta-1, 3-glucan 3-glucanhydrolase, EC 3.2.1.6) extracted from Phaseolus vulgaris L. cv. Red Kidney had a pH optimum of 5 and a temperature optimum of 50 C. Excision of plant tissue resulted in an increase in beta-1, 3-glucanase activity after a 6-hour lag period. The increase could be prevented by indole-3-acetic acid, gibberellic acid, and cytokinins. Ethylene (half-maximal concentration = 0.1 microliter/liter) promoted the synthesis of beta-1, 3-glucanase, and 10% CO(2) overcame some of the ethylene effect. Cycloheximide prevented the induction of beta-1, 3-glucanase, but actinomycin D and chromomycin A(3) had only a partial effect. The amount of callose in sieve tube cells correlated with levels of beta-1, 3-glucanase, suggesting that this enzyme played a role in the degradation of beta-1, 3-glucans.
Ethylene increased the germination of freshly imbibed lettuce (Lactuca sativa L. var. Grand Rapids) seeds. Seeds receiving either red or far-red light or darkness all showed a positive response to the gas. However, ethylene was apparently without effect on dormant seeds, those which failed to germinate after an initial red or far-red treatment. Carbon dioxide, which often acts as a competitive inhibitor of ethylene, failed to clearly reverse ethylene-enhanced seed germination. While light doubled ethylene production from the lettuce seeds, its effect was not mediated by the phytochrome system since both red and far-red light had a similar effect.
Cellulase (beta-1,4-glucan-glucanohydrolase EC 3.2.1.4) activity increased during abscission and was localized in the cell separation layer of Phaseolus vulgaris L. cv. Red Kidney (bean), Gossypium hirsutum L. cv. Acala 4-42 (Cotton) and Coleus blumei Benth. Princeton strain (Coleus) abscission zone explants. Cellulase activity was optimum at pH 7, was reduced by one-half after heating to 55 degrees for 10 min, and was associated with the soluble components of the cell. Explants treated with aging retardants (indoleacetic acid, (6)N-benzyladenine, and coumarin), CO(2), actinomycin D or cycloheximide had less cellulase activity than untreated controls. Ethylene increased cellulase activity of aged explants after a 3-hr lag period but had no effect on cellulase activity of freshly excised explants. It was concluded that 1 of the roles of ethylene in abscission is to regulate the production of cellulase which in turn is required for cell separation.
The construction, operation, and effectiveness of an abscission measuring instrument called an abscissor is described. The device measured the force required for a spring-opposed plunger to shear abscission zone explants and was capable of automatically recording break strength data. Examples of data obtained with the abscissor are presented to demonstrate its capability of rapidly measuring significant changes in explant break strength.
The effect of abscisic acid on cotton (Gossypium hirsutum L. cv. Acala 4-42) and bean (Phaseolus vulgaris L. cv. Red Kidney) explants was 2-fold. It increased ethylene production from the explants, which was found to account for some of its ability to accelerate abscission. Absci is acid also increased the activity of cellulase. Increased synthesis of cellulase was not du to an increase in aging of the explants but rather was an effect of abscisic acid on the processes that lead to cellulase synthesis or activity.
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Ethylene was the most effective abscission accelerant examined, with decreasing activity shown by propene, carbon monoxide, acetylene, vinyl fluoride, 1-butene, and 1,3-butadiene. Carbon dioxide inhibited abscission, but its effect was overcome by ethylene. Oxygen was required for abscission as an electron acceptor for respiration and not as a potentiator or activator of the ethylene attachment site. The molecular requirements for abscission were similar to those shown by other workers for other biological processes under the influence of ethylene.
The cell separation aspect of abscission is thought to involve the action of specific cell wall degrading enzymes. Enzymes represent synthesis which in turn is preceded by the synthesis of specific RNA molecules, and it follows that inhibition of either of these processes would also block abscission. Since abscission is a localized phenomenon usually involving 2 or 3 cell layers, RNA and protein synthesis should also be localized. Manipulations of plant material which either accelerate or retard abscission may be due to the regulation of RNA and protein synthesis. This paper is a review of literature concerned with these and related questions.
Excision of Phaseolus vulgaris L. c.v. Red Kidney abscission zone explants results in senescence, mobilization, and abscission. Because these processes take place at about the same time, there has been some question as to whether they are causally related or are occurring in an independent but simultaneous fashion. Data presented here suggest that the latter interpretation is correct. After abscission zone explants are isolated from the leaf an aging process is set into motion and a degradation of metabolites in the pulvinus takes place. During the aging process the explants also become increasingly sensitive to ethylene which in turn promotes cell separation. Indoleacetic acid, cytokinins, and coumarin appear to retard aging since both degradative processes and abscission are inhibited. However, ethylene increased abscission without increasing degradative processes indicating that abscission and senescence are independent processes occurring at the same time.
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Ethylene stimulated the incorporation of (32)P into RNA in the abscission zone of bean explants (Phaseolus vulgaris L. var. Red Kidney). The enhancement was observed in all fractions separated by methylated albumin kieselguhr column chromatography, although the magnitude of the increase was not the same for each fraction. Differential extraction of the nucleic acids indicated that the ethylene stimulation was confined to the fraction extracted with sodium lauryl sulfate, with the increase mainly in Fraction III (Ribosomal RNA) and Fraction IV (Messenger RNA). Actinomycin D, which blocks ethylene-stimulated abscission, inhibited (32)P incorporation into all column fractions. 5-Fluorouracil, which blocked 50% of the ethylene-enhanced (32)P incorporation, did not inhibit ethylene-enhanced abscission. The results indicate that ethylene may regulate abscission through control of specific RNA's.
The stimulation of ethylene production from seedling tissue of Phascolus vulgaris, Helianthus annuus and Zea mays by growth regulators was inhibited by actinomycin D and puromycin and to a lesser extent by 2-thiouracil and p-fluorophenylalanine. It is concluded that the mechanism of action of growth regulators on the enhancement of ethylene production is the formation of enzymes involved in ethylene biogenesis.