STUDIES ON THE PHYSIOLOGY OF ABSCISSION: EFFECT OF TREATMENTS WITH GIBBERELLIC ACID

Abscission of debladed petioles of Coleus was observed following spray applications of gibberellic acid (GA) to the foliage. Sprays were applied to some branches which were left intact (inducing branches), or to adjacent branches whose leaves were later debladed (induced branches). In all experiments three applications of GA were made after which the induced branches were debladed, but in one series deblading was delayed for a week after the last spray application. All treatments resulted in accelerated petiole abscission relative to the controls. Differences between the results of these experiments and the results of similar, earlier experiments with indoleacetic acid (IAA) are discussed. The evidence suggests that GA accelerates abscission by a different mechanism than does IAA.     

Auxin transport and the regulation of petiole abscission in cotton (Gossypium hirsutum L.): A comparison of indol-3yl-acetic acid and phenylacetic acid

Distal applications of indol-3yl-acetic acid (IAA) to debladed cotyledonary petioles of cotton (Gossypium hirsutum L.) seedlings greatly delayed petiole abscission, but similar applications of phenylacetic acid (PAA) slightly accelerated abscission compared with untreated controls. Both compounds prevented abscission for at least 91 h when applied directly to the abscission zone at the base of the petiole. The contrasting effects of distal IAA and PAA on abscission were correlated with their polar transport behaviour-[1-¹⁴C]IAA underwent typical polar (basipetal) transport through isolated 30 mm petiole segments, but only a weak diffusive movement of [1-¹⁴C]PAA occurred.Removal of the shoot tip substantially delayed abscission of subtending debladed cotyledonary petioles. The promotive effect of the shoot tip on petiole abscission could be replaced in decapitated shoots by applications of either IAA or PAA to the cut surface of the stem. Following the application of [1-¹⁴C]IAA or [1-¹⁴C]PAA to the cut surface of decapitated shoots, only IAA was transported basipetally through the stem. Proximal applications of either compound stimulated the acropetal transport of [¹⁴C]sucrose applied to a subtending intact cotyledonary leaf and caused label to accumulate at the shoot tip. However, PAA was considerably less active than IAA in this response.It is concluded that whilst the inhibition of petiole abscission by distal auxin is mediated by effects of auxin in cells of the abscission zone itself, the promotion of abscission by the shoot tip (or by proximal exogenous auxin) is a remote effect which does not require basipetal auxin transport to the abscission zone. Possible mechanisms to explain this indirect effect of proximal auxin on abscission are discussed.     

LOCALIZATION OF DEHYDROGENASE AND ACID PHOSPHATASE IN THE ABSCISSION ZONE OF BEAN LEAVES

Leaf abscission in Phaseolus vulgaris L. cv. ‘Contender’ is associated with enzymatic changes during and prior to separation. Deblading resulted in a localized increase in dehydrogenase and acid phosphatase in the abscission zone. Increased enzyme activities were observed 24–48 hr after deblading. In debladed plants separation was complete in 6–8 days. At separation, dehydrogenase activity appeared to decrease and localization was specific to the protective layer, while the petiole side had no activity. In contrast, acid phosphatase activity was observed in some layers of cells on the petiole side after separation. Ethylene treatment promoted abscission and separation occurred in 24–48 hr in both debladed and intact plants. No protective layer was formed during ethylene-induced abscission. Enzymatic changes similar to those observed in debladed control plants were observed with ethylene treatment. Ethylene induced an additional abscission layer between the pulvinus and petiole, where an abscission layer normally does not form. In this ethylene-induced abscission layer, similar enzyme activities were detected.     

Retardation of abscission of citrus leaf and fruitlet explants by brassinolide

Brassinolide (BR), a novel plant growth-regulating steroidal lactone, markedly retarded the abscission of leaf explants of Calamondin (Citrus madurensis Lour.), when dissolved in water and fed through the petiole. BR was effective at concentrations as low as 0.021 M, and showed a stronger effect than IAA which also retarded abscission. Trifluoperazine (TFP), an inhibitor of the calmodulin-calcium complex, accelerated abscission, and this acceleration could be counteracted by a simultaneous addition of IAA or BR, the effect of IAA being stronger. BR in lanolin applied to the cut surface of the leaf blade of the explant showed a weaker abscission-retarding effect than that applied in water via the petiole. BR and IAA also markedly retarded the abscission of fruitlet explants of Calamondin.     

GIBBERELLIN AND AUXIN RELATIONSHIPS IN ABSCISSION

Gibberellic acid (GA) has no effect on abscission when applied proximally or distally to the abscission zones of debladed petioles of Coleus. Application of GA to the stem apex increases the rate of abscission of debladed petioles. The effect on abscission is accompanied by an increase in the level of endogenous auxin in the stem. Correspondingly proximal applications of indoleacetic acid (IAA) accelerate abscission, whereas the longevity of the debladed petiole approaches that of the intact leaf only in the presence of a continuous distal supply of IAA. No correlation is found between petiole elongation and its longevity. The experimental data support the view that auxin acts at the abscission zone in regulating separation processes and that the effect of GA is through its effect on the level of endogenous auxin.     

Methyl jasmonate induces the formation of secondary abscission zone in stem of Bryophyllum calycinum Salisb

Methyl jasmonate (JA-Me) at a concentration of 0.5 % induced the formation of secondary abscission zone and senescence in several types of stem explants (only internode segment, internode segment with nodes and without leaves, internode segment with nodes and debladed petioles) of Bryophyllum calycinum when it was applied in various places of the stem or the debladed petiole as lanolin paste. In the presence of small leaves in stem explants methyl jasmonate also induced the formation of secondary abscission zone and senescence but the presence of larger leaves completely inhibited methyl jasmonate-induced processes. Auxin, (indole-3-acetic acid, IAA), at a concentration of 0.1 % extremely prevented the formation of secondary abscission zones and senescence in the stem tissues induced by methyl jasmonate. Similar relationship between auxin and methyl jasmonate to induce the formation of secondary abscission zone and senescence was found in decapitated shoot of the intact plant. Mechanisms of the formation of secondary abscission zone are also discussed in terms of the interaction of methyl jasmonate with auxin.     

An explanation for the light requirement of AgNO3 to inhibit ethylene-induced abscission

The loss of the antiethylene activity of Ag⁺ on leaf abscission by incubation in the dark was investigated. When primary leaves were removed from cuttings of Vigna radiata previously sprayed with AgNO₃, dark-induced abscission of the petioles was inhibited, compared to untreated leafless controls, in the presence or absence of ethephon, an ethylene-releasing compound. Malformin did not negate inhibition of petiole abscission induced by Ag⁺. Although leaf removal restored the antiethylene activity of Ag⁺ in the dark, macerates of leaves from dark-aged cuttings did not negate the ability of Ag⁺ to inhibit petiole abscission in the dark. Abscisic acid completely abolished the ability of Ag⁺ to counteract ethephon-induced leaf abscission in the light, and almost completely abolished the Ag⁺-induced inhibition of petiole abscission from explants in the dark. It is proposed that the phytochrome requirement for the antiethylene activity of Ag⁺ on ethephon-induced leaf abscission involves prevention of the formation, accumulation, or transport of a substance in leaves in the dark which negates Ag⁺ activity. This substance may be abscisic acid or another substance with similar biological activity.     

Meristem differentiation in Riella helicophylla (Bory et Mont.) Mont. under the influence of auxin or antiauxin

During the development of the unistratose gemmae of Riella helicophylla, the single intercalary meristem of the very young gemmae is subdivided into two lateral meristems. The duration of the cell reproduction cycle increases from the margin to the median part of the gemmae. This polarization within the meristem disappears after addition of the antiauxin PCIB to the culture medium. PCIB leads to a retardation or blockage of the cell cycle during the light period of the culture. Under the influence of PCIB the amount of starch in the chloroplasts is strikingly increased, probably because of a reduction of starch degradation. Addition of sugars compensates the effect of PCIB on the cell cycle. The effects of PCIB are counteracted by auxin. The results are taken as evidence that auxin plays a role in directing the transport of substances needed for the continuation of the cell reproduction cycle between adjacent cells of the meristem.Abbreviations IAA indole-3-acetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - PCIB p-chlorophenoxyisobutyric acid - G-l-P glucose-l-phosphat - EDTA ethylenediaminetetraacetic acid     

Inhibition of Abscission of Bean Petiole Explants by Lepidimoide

The effect of lepidimoide on the process leading to abscission was studied in bean (Phaseolus vulgaris L. cv. Masterpiece) petiole explants. The assays, involving observations on the junction of the petiole of primary leaves and the pulvinus, were conducted in the light. Lepidimoide, at concentrations of 1 μm or higher, delayed the abscission process; however, the progression of abscission proceeded at normal rates, and complete abscission resulted. On the other hand indoleacetic acid inhibited the normal senescence resulting in greatly decreased abscission during the observation period. These observations show that lepidimoide only delays abscission, and the kinetics seem to indicate that lepidimoide and indoleacetic acid affect abscission through different mechanisms. Received March 1, 1996; accepted November 4, 1996     

Effect of Protein Synthesis Inhibitors, Auxin, and Gibberellic Acid on Abscission

Chloramphenicol, actinomycin D, and other inhibitors of protein synthesis promote abscission in several plant genera. Abscission is accelerated in species where an abscission layer is present, as well as in tissue where no abscission layer develops prior to abscission. The inhibitors promote abscission in species where cell division is reported to precede the separation processes as well as in tissues where no cell division is associated with the initiation of abscission. Indoleacetic acid (IAA) or auxin precursors, when applied with chloramphenicol and aclinomycin D, overcome the promotive effects of the inhibitors on abscission. These inhibitors apparently do not promote abscission through their effects on auxin precursor conversion, IAA transport, and IAA destruction in the petiole. IAA increases the incorporation of leucine-1-¹⁴C into a trichloroacetic acid precipitable fraction of the abscission zone under conditions where abscission is retarded. A low concentration of IAA which accelerates abscission, decreases incorporation of leucine into protein. Other promoters of abscission — chloramphenicol, d-aspartic acid, and gibberellic acid —also decrease the incorporation of leucine into the protein of the abscission zone. The data indicate that enzymes required for the degradative processes associated with abscission are already present in the abscission zone whereas a continuous synthesis of protein is required for the retention of the leaf.     

Die hormonale Regulation des Blattfalls bei Coleus rehneltianus Berger

Summary In extracts of Coleus rehneltianus shoots abscisic acid was identified by thin layer chromatography and by spectropolarimetry.Diffusates from the petiole of isolated leaves were tested in an abscission test. According to previous results (Part I) these experiments allow conclusions concerning the hormonal flux from the lamina.The next step was to separate the diffusates by paper chromatography. The relative amounts of IAA and ABA were estimated on the basis of a biological assay. Young leaves yield ten times as much IAA to an agar receptor as green adult leaves and twenty times as much as yellow senescent leaves. On the other hand young leaves do not transmit detectable amounts of ABA to the receptor. Diffusates from adult leaves contain only a small amount of ABA, whereas those of senescent leaves contain a considerable amount.Evidently the induction of leaf abscission is not effected by a strong decrease of the IAA content during the development of a young leaf into an adult leaf. One is lead to assume that the rapid increase of the ABA flux during senescence is responsible for the induction of the abscission process.

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Zweiter Teil einer Dissertation des Fachbereiches Biologie der Universität Hamburg     

Ethylene-induced leaf abscission in cotton seedlings : the physiological bases for age-dependent differences in sensitivity

The speed of ethylene-induced leaf abscission in cotton (Gossypium hirsutum L. cv LG-102) seedlings is dependent on leaf position (i.e. physiological age). Fumigation of intact seedlings for 18 hours with 10 microliters per liter of ethylene resulted in 40% abscission of the still-expanding third true (3°) leaves but had no effect on the fully expanded first true (1°) leaves. After 42 hours of fumigation with 50 microliters per liter of ethylene, total abscission of the 3° leaves occurred while <50% abscission of the 1° leaves was observed. On a leaf basis, endogenous levels of free IAA in 1° leaves were approximately twice those of 3° leaves. Free IAA levels were reduced equally (approximately 55%) in both leaf types after 18 hours of ethylene (10 microliters per liter) treatment. Ethylene treatment of intact seedlings inhibited the basipetal movement of [¹⁴C]IAA in petiole segments isolated from both leaf types in a dose-dependent manner. The auxin transport inhibitor N-1-naphthylphthalamic acid increased the rate and extent of ethylene-induced leaf abscission at both leaf positions but did not alter the relative pattern of abscission. Abscission-zone explants prepared from 3° leaves abscised faster than 1° leaf explants when exposed to ethylene. Ethyleneinduced abscission of 3° explants was not appreciably inhibited by exogenous IAA while 1° explants exhibited a pronounced and protracted inhibition. The synthetic auxins 2,4-D and 1-naphthaleneacetic acid completely inhibited ethylene-induced abscission of both 1° and 3° explants for 40 hours. It is proposed that the differential abscission response of cotton seedling leaves is primarily a result of the limited abscission-inhibiting effects of IAA in the abscission zone of the younger leaves.     

FORMATION OF CALLOSE AND LIGNIN DURING LEAF ABSCISSION

Deblading of bean leaves promoted the formation of callose and lignin in the abscission zone. The abscission layer became evident three days after deblading. The greatest increase in callose occurred in about two layers of cells during the development of the abscission layer. Four days after deblading, only a few layers of cells on the distal side of the abscission layer showed an increase in lignin. Lignification continued to expand to 8–10 layers of cells at the time of separation. After separation, the lignified cells remained with the petiole. Sieve elements in the abscission zone were covered with callose plugs and the vessels were occluded with tyloses.     

DOES AUXIN INHIBIT THE ABSCISSION OF COLEUS LEAVES BY ACTING AS A GROWTH HORMONE?

The hypothesis that auxin prevents abscission, in Coleus blumei, by causing growth has been confirmed in a number of different ways: (1) in the intact plant, petioles grow until just before abscission; (2) excising the blades causes uniformly fast abscission, which is correlated with uniform absence of elongation; (3) if one stimulates the debladed petioles to renewed growth by substituting IAA for the leaf-blades, one can restore their longevity to that of the intact leaves; (4) increasing the concentration of IAA added to debladed petioles increases both the elongation and the longevity. However, the parallel between elongation and longevity was not exact: IAA concentrations giving full replacement of the blades in preventing abscission gave less than full replacement of elongation in petioles 2 and 3 and more than full replacement in petioles 5–8. Following the time-course revealed that if an IAA-treated debladed petiole elongates as much or more than normal during the first week after deblading, then it will have normal longevity.     

Jasmonates promote abscission in bean petiole expiants: Its relationship to the metabolism of cell wall polysaccharides and cellulase activity

Jasmonic acid (JA) and its methyl ester (JA-Me) promoted the abscission of bean petiole expiants in the dark and light, and the activity of these compounds was almost same. JA and JA-Me did not enhance ethylene production in bean petiole expiants in the light, indicating that the abscission-promoting effects of these compounds are not the result of ethylene. Cells in the petiole adjacent to the abscission zone expanded during abscission but not in the pulvinus, and JA-Me promoted cell expansion in the petiole and the pulvinus. JA-Me had no effect on the total amounts of pectic and hemicellulosic polysaccharides in 2-mm segments of the abscission region, which included 1 mm of pulvinus and 1 mm of petiole from the abscission zone. On the other hand, the total amounts of cellulosic polysaccharides in this region were reduced significantly by the addition of JA-Me in the light. JA-Me had no effect on the neutral sugar composition of hemicellulosic polysaccharides during abscission. The decrease in the endogenous levels of UDP-sugars in the petiole adjacent to the abscission zone was accelerated during abscission by the addition of JA-Me in the light. Cellulase activities of pulvinus and petiole in 10-day-old seedlings were enhanced by the addition of JA. These results suggest that the promoting effect of JA or JA-Me on the abscission of bean petiole explants is due to the change of sugar metabolism in the abscission zone, in which the increase in cellulase activity involves the degradation of cell wall polysaccharides. Jasmonic acid (JA) and its methyl ester (JA-Me) are considered to be putative plant hormones for a number of reasons, including their wide occurrence in the plant kingdom, biologic, activities in multiple aspects at low concentrations, and their interaction with other plant hormones (for reviews see Parthier 1991, Hamberg and Gardner 1992, Sembdner and Parthier 1993, Ueda et al. 1994a). We have already reported that JA and JA-Me and C₁₈-unsaturated fatty acids, which are considered to be the substrates of the biosynthesis of jasmonates, are powerful senescence-promoting substances (Ueda et al. 1982b, 1991a). Senescence symptoms induced by these compounds are identical to those of natural senescence. Recently we have also found that JA inhibited indole-3-acetic acid (IAA)-induced elongation of oat (Avena sativa L. cv. Victory) coleoptile segments by inhibiting the synthesis of cell wall polysaccharides (Ueda et al. 1994b, 1995). These facts led us to study the mode of actions of JA and JA-Me on promoting abscission, which is considered the last dramatic phenomenon of senescence. In this paper we report that JA and JA-Me promote abscission in bean (Phaseolus vulgaris L. cv. Masterpiece) petiole expiants and that the changes in the metabolism of cell wall polysaccharides in the petiole and the pulvinus adjacent to the abscission zone are involved in the promotive effects of these compounds.Abbreviations ABA abscisic acid - ACC 1-aminocyclopropane-1-carboxylic acid - DCB 2,6-dichlorobenzonitrile - HPLC high performance liquid chromatography - IAA indole-3-acetic acid - JA jasmonic acid - JA-Me methyl jasmonate - MES 2-(N-morpholino)ethane-sulfonic acid, monohydrate - TCA trichloroacetic acid - Tris 2-amino-2-hydroxymethy-1,3-propanediole     

Involvement of Abscisic Acid in Ethylene-Induced Cotyledon Abscission in Cotton Seedlings

Cotton (Gossypium hirsutum L. cv LG102) seedlings raised from seeds exposed to 100 [mu]M norflurazon (NFZ) during imbibition contained reduced levels of free abscisic acid (ABA) and were visibly achlorophyllous. Exposure of untreated cotton seedlings to ethylene concentrations >1 [mu]L/L for 24 h resulted in cotyledon abscission. In contrast, exposure of NFZ-treated seedlings to concentrations of ethylene [less than or equal to]50 [mu]L/L elicited no cotyledon abscission. Application of ABA, an ABA analog, or jasmonic acid to NFZ-treated seedlings restored ethylene-induced abscission. Isolated cotyledonary node explants prepared from NFZ-treated seedlings exhibited an altered dose-response pattern of ethylene-induced petiole abscission. Endogenous levels of free IAA were unaltered in NFZ-treated seedlings. Ethylene treatment (50 [mu]L/L, 24 h) had no effect on free indoleacetic acid (IAA) levels in either control or NFZ-treated seedlings. Levels of conjugated (ester plus amide) IAA were substantially increased in NFZ-treated seedlings regardless of ethylene treatment. These results indicate that endogenous ABA plays an essential, but physiologically undefined, role in ethylene-induced cotyledon abscission in cotton.     

THE REGULATION OF ABSCISSION AND IAA BY SENESCENCE FACTOR AND ABSCISIC ACID

The effects of the endogenous “senescence factor” (SF) and synthetic (±)-abscisic acid (ABA) on basipetal transport of indoleacetic acid (IAA) in Coleus blumei Benth. were studied. When used to pretreat explant petioles, both SF and ABA accelerated abscission and decreased basipetal movement of IAA through petiolar sections but had no effect on acropetal movement, which was negligible in all experiments. SF decreased the intensity but not the velocity of the basipetal movement of IAA. Pretreatment with either SF or ABA decreased the free IAA and increased the IAA-aspartate extractable from the sections. The results support the hypothesis that SF and ABA hasten abscission by lowering the amounts of IAA transport in the tissue. Known properties of SF are similar to those of ABA. Efforts are under way to elucidate the chemical identity of SF.     

Stimulation of abscission of Petioles of Phaseolusavulg ris by Sucrose and Gibberellic acid

The influence of Grand sucrose, alone and in combination, on the abscissionarate of petioles of the common be n plant was investigated. application of either Gaorasucrose to the distalatip of one of apair of opposite petiolesa celebrated the abscission of the treated petiole. When the two compounds were applied simult neously less than additive effect was observed. The abscission rate of untreated petioles remain ed practically unchanged by treatment of their opposite partners with G a or sucrose or combinations of these two compounds.     

ABSCISSION OF LEAVES AND LEAFLETS IN ACER NEGUNDO AND FRAXINUS AMERICANA

A comparative study of leaf and leaflet abscission in Acer negundo and Fraxinus americana was undertaken with special emphasis on leaflet abscission. Leaf fall in both species is accomplished by orderly, fragmentary abscission of leaflets followed by petiole abscission. Leaflet fall was presaged by differentiation of a separation layer at leaflet bases 10–15 days prior to leaflet fall, without an accompanying protective layer. Anatomical studies of petiole abscission revealed early differentiation of a protective layer followed by differentiation of a separation layer at petiole bases just prior to petiolar fall. Abscission at both sites was facilitated by cell division and dissolution of cell walls within separation layers.     

The Role of Auxin in the Apical Regulation of Leaf Abscission in Cotton (Gossypium hirsutum L.)

The petiole abscission induced by deblading cotyledonary leavesof cotton (Gossypium hirsutum L. cv. Delta Pine) was acceleratedby the presence of the intact shoot apex or, in decapitatedplants and explants, by application to the stem (proximal application)of indol-3yl-acetic acid (IAA) or 1-aminocyclopropane-l-carboxylicacid (ACC). IAA and ACC accelerated the abscission of debladedpetioles whether applied above or below the cotyledonary node.Transport of IAA to the node was not required for the responseto proximal IAA. [2,3-¹⁴C]ACC was readily transported to thenodal region whether applied to the stem above or below thenode. Application of IAA or ACC to the stem did not induce theabscission of intact leaves or of debladed petioles treateddistally with IAA The acceleration of abscission by proximal IAA, but not thatcaused by ACC, was prevented if explants were treated with a-aminooxyaceticacid (AOA), an inhibitor of ACC-synthase. AOA also preventedthe acceleration of abscission caused by the shoot apex. Theprogress of abscission in debladed explants was greatly delayedby silver thiosulphate (STS—an inhibitor of ethylene action),whether or not the explants were treated with IAA or ACC. Itis suggested that the speeding effects of the shoot apex andof proximal auxin on the abscission of debladed petioles requiresauxin-induced ACC synthesis. The possibility is discussed thatACC may function as a mobile abscission promoter Key words: Abscission, ACC, ACC-synthase, cotton (Gossypium), proximal auxin