Peroxidase Activity in the Abscission Zone of Bean Leaves during Abscission

Peroxidase activity and localization in the abscission zone of bean leaves were studied histochemically and by gel electrophoresis. Deblading of bean leaves resulted in an increase in peroxidase activity in the abscission zone 2 to 4 days after deblading with highest activity just prior to separation. In debladed plants, the cell division in six to eight layers of cells preceded separation. An ethylene treatment (8 microliters per liter) induced separation of debladed petioles in approximately 24 hours and of intact plants in 36 to 48 hours. Ethylene treatment produced similar results in both debladed and intact plants. In ethylene-treated plants, whether debladed or not, enzyme localization was restricted to only two to three layers of cells with no cell division apparent prior to separation. Infrequent cell divisions were observed after treatment with 2-chloroethylphosphonic acid (1000 micrograms per liter) (Ethephon); however, other changes were similar to those observed with ethylene. Deblading and ethylene treatment resulted in changes in the six peroxidase isozymes observed in the abscission zone. Only four were observed in samples collected 2 centimeters below the abscission zone. Peroxidase bands IV and V increased significantly in debladed and ethylene-treated plants and peroxidase VI decreased only in debladed plants. The changes in peroxidase activity were invariably observed prior to separation in all treatments.     

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     

Influence of Reproduction on the Abscission Process of Cotton (Gossypium barbadense L.) Leaves

Abscission responses of debladed petioles of young and olderleaves were analysed during flowering, fruiting and post fruitingstages of development of G. barbadense plants. Identical abscissionexperiments were performed with materials collected from plantsmaintained in a vegetative condition by removal of flower buds. Inhibition of the abscission of debladed petioles by NAA wasgreater in debudded plants as compared to normal plants andthe extent of inhibition gradually declined during growth. Promotiveeffects of ethrel and abscisic acid were higher in normal plantsthan in debudded plants. The duration of auxin-inhibitablc stage-I of abscission wasextended in debudded plants and it gradually declined with theprogress of development. Debudded plants were characterizedby higher abscission inhibition during stage-I and lower abscissionpromotion during stage-II as a result of application of auxincompounds to the debladed petioles. Laminar tissues of debudded plants contained higher amountsof endogenous IAA and lesser amount of abscisions than did thoseof normal plants and in both cases the levels of these compoundschanged markedly during plant development. Decrease of total RNA content in the distal tissues of the abscissionzones was accompanied by increase in proximal tissues duringabscission in both normal and debudded plants. This tendencywas more pronounced in normally grown plants as compared todebudded plants.     

Categories of Petal Senescence and Abscission: A Re-evaluation

In a previous paper (Woltering and van Doorn, 1988, Journalof Experimental Botany39: 1605–1616) we identified threetypes of flower life cessation: by petal wilting or withering,which was either ethylene-sensitive or insensitive, and by abscissionof turgid petals, which was ethylene-sensitive. These categoriestended to be consistent within families. Here we re-examinethese relationships by testing a further 200 species, and anumber of other families. As previously, flowering shoots wereexposed to 3 ppm ethylene for 24 h at 20 °C, in darkness.Most monocotyledonous species tested showed ethylene-insensitivepetal wilting, although ethylene-sensitive wilting occurredin the Alismataceae and Commelinaceae. Petals of the dicotyledonousspecies tested were generally sensitive to ethylene, exceptfor a few groups showing wilting (Crassulaceae, Gentianaceaeand Fumariaceae, and one subfamily in both the Ericaceae andSaxifragaceae). Petal abscission was generally ethylene-sensitive,but ethylene insensitivity was found in some Tulipa cultivarsand three Saxifraga species. In most tulip cultivars tested,the petals wilted and then fell. It is concluded that (a) theresponse to ethylene is often consistent within either familiesor subfamilies; and (b) a fourth category, ethylene-insensitivepetal abscission, exists both in monocotyledons and dicotyledons.Copyright 2001 Annals of Botany Company Ethylene sensitivity, flower longevity, petal abscission, petal wilting, petal withering, petal senescence, taxonomic categories     

Effect of Treatments with Growth Retardant Chemicals on Petiole Abscission in Coleus

A bscission of debladed petioles of Coleus was observed following spray applications of growth retardant chemicals and particularly of Phosfon D to the foliage. Sprays were applied to some branches, which were left intact (inducing branches), or to adjacent branches the leaves of which were later debladed (induced branches). In all experiments two applications of growth retardant chemicals were made, after which the induced branches were debladed. Treatments on induced branches accelerated the petiole abscission relative to the controls. Treatments on inducing branches, instead, decreased abscission speed of debladed petioles. The evidence suggests that phosfon D affects abscission by interfering with the indoleacetic acid mechanism.     

A Senescence Factor and Foliar Abscission in Catharanthus roseus

Abscission-accelerating activity was found in naturally senescentpetioles, and senescent leaves of Catharanthus indicating thepresence of a senescence factor (SF) in these tissues. No SFactivity was observed in non-senescent petioles, or non-senescentleaves at various defined developmental stages. The resultsare discussed in relation to the auxin-auxin balance theoryfor the control of leaf abscission and the auxin-SF balancetheory.     

Abscission: potentiating action of auxin transport inhibitors

Reduction in petiolar auxin transport has been proposed as one of the functional actions of endogenous or exogenous ethylene as it regulates intact leaf abscission. If this hypothesis is correct, auxin-transport inhibitors should hasten the rate or amount of abscission achieved with a given level of ethylene. Evidence presented here indicates that the hypothesis is correct. Three auxin transport inhibitors promoted ethylene-induced intact leaf abscission when applied to specific petioles or the entire cotton plant (Gossypium hirsutum L., cv. Stoneville 213). In addition, the transport inhibitors caused rapid abscission of leaves which usually do not abscise under the conditions employed. No stimulation of abscission occurred during the initial 3 to 5 days after plants were treated with transport inhibitors unless such treatments were coupled with exogenous ethylene or that derived from 2-chloroethylphosphonic acid. However, vegetative cotton plants did abscise some of their youngest true leaves during the 2nd and 3rd weeks of exposure to transport inhibitor alone. Taken as a whole, the results indicate that reducing the auxin supply to the abscission zone materially increases sensitivity to ethylene, a condition which favors a role of endogenous ethylene in abscission regulation. Such a role of ethylene indicates the importance of auxin-ethylene interactions in the over-all hormone balance of plants and specific tissues.     

Assessing susceptibility of Hevea clones to Microcyclus ulei

Leaf disks of 7-day-old Hevea leaves floating on water produced lesions of varying sizes following inoculation with conidia of Microcyclus ulei, the cause of South American leaf blight (SALB) of Hevea. The resistance ratings of 188 Hevea clones classified according to lesion size on leaf disks and to leaf area infected in the field were correlated. Lesion size varied little with small differences in leaf age or inoculum level. Leaves which had been treated with sodium hypochlorite and stored for 3 days could still be infected by desiccated conidia, suggesting that Hevea leaves from South East Asia and conidia of M. ulei from South America could be sent to a central laboratory for rapid screening for resistance to SALB.     

An Induced Mutant of Corchorus olitorius L. with Enhanced Abscission Rate

A mutant of Corchorus olitorius L. (cultigen, JRO-632) isolatedfrom R₂ generation of 90 kR X-rayed progeny is characterizedby excessive leaf-fall. Although the initial rate of leaf-fallis comparable with that of the mother strain, after 65 daysof growth, leaf-fall is much more rapid as revealed from comparativecounts of the number of leaves and nodes. Formation of a separation layer has been observed at the abscissionzone of the sixth leaf above the last abscissed leaf of themutant, whereas, in the mother strain it is initiated just abovethe last abscissed leaf under the same climatic conditions inthe field. A crude extract from senescent leaves of the mutant acceleratesseparation layer formation in the abscission zone of the motherstrain. The possibility of indole-3yl acetic acid repressionand abscissin-like activity accelerating abscission is suggested.     

Fine Structure and Cytochemistry of the Abscission Zone Cells of Phaseolus Leaves: II. Localization of Peroxidase and Acid Phosphatase in the Separation Zone Cells

The sub-cellular localization of acid phosphatase and peroxidaseactivities has been studied by cytochemical procedures in cellsat the surface of the separation layer during the abscissionof leaves of Phaseolus vulgaris. Intense staining for both enzymeswas found in the cell walls, Golgi bodies and endoplasmic reticulum.The wall staining for acid phosphatase was chiefly associatedwith the middle lamellar region while staining for peroxidasewas found throughout the wall. These observations are discussedin relation to the possible involvement of these enzymes inthe changes occurring in the wall during abscission and to therole of the Golgi bodies in the separation process.     

Mechanism of Action of Abscission Accelerators

Abscission zone explants of Gossypium hirsutum L., Cassia fistula L., and Coleus blumei Benth. were used to investigate correlations between endogenous rates of ethylene evolution and time of abscission. Additions of 0.1 nl/ml ethylene to the explants markedly accelerated abscission; continuous aeration of the explants, to prevent accumulation of small amounts of endogenously produced ethylene, inhibited abscission compared with that of sealed controls. Substances that stimulated abscission simultaneously accelerated ethylene evolution on all three species and at any position of application. The positional effects of auxin are explained as being due to differences in transport in the explant. Thus, distally applied auxin inhibits abscission, regardless of the accelerated rate of ethylene evolution, by being rapidly transported to the abscission zone. Auxin applied proximally stimulates abscission because it is unable to move as rapidly to the abscission zone and the ethylene effect becomes dominant. Ethylene was found to be most effective on aged tissues, and it is concluded that abscission rates are determined by an increase in sensitivity of the tissue to the ethylene that is already being produced.     

Abscission in Coleus: Light and Phytohormone Control

Light control of leaf abscission in Coleus (Coleus blumei Benthcv. Ball 2719 Red) appears to be regulated by the quantity ofendogenous auxin transported from the leaf blade to the abscissionzone. Gas chromatographic—mass spectrophotometric analysisindicated that diffusate collected from leaf tissue treatedwith red light contained significantly higher levels of auxinthan dark and far-red light-treated leaf tissue. In addition,diffusate from red light-treated tissue inhibited abscissionof leafless petioles while diffusate from far-red light-treatedtissue promoted abcission when compared with diffusate fromdark-treated tissue. The effect of red light on abscission couldbe mimicked by IAA, but not by other phytohormones. An auxintransport inhibitor, 2, 3, 5-triiodobenzoic acid (TIBA), appliedeither as a lanolin ring around the petiole or vacuum infiltratedinto tissue, could completely eliminate any red light effecton abscission. The data are consistent with a phytochrome-mediatedlight regulation of endogenous auxin level in the leaf whichthen controls abscission. Key words: Abscission, Coleus, IAA, plant hormones, red (far-red) light, TIBA     

Perianth Abscission in Montbretia (Crocosmia x crocosmiiflora)

The anatomy, histochemistry, kinetics and hormonal regulationof perianth abscission in Crocosmia x crocosmiiflora (Montbretia)has been investigated. The abscission zone is anatomically welldefined, with cell divisions occurring in this region at anthesis.Abscission is first detectable 3 d after perianth opening, whenthe walls of a group of cells beneath the adaxial epidermisshow reduced staining with polyanion-specific stains, and adecline in penanth break strength also occurs. Abscission isachieved by cell wall breakage in thc abscission zone, whichprogresses eccentrically from the adaxial epidermis throughthe abscission zone, rather than the separation of intact cellsas occurs in flowers of dicotyledons. Experiments on detachedflowers suggest similarities in the hormonal regulation of abscissionin Crocosmia to that of dicotyledons, in that an ethylene promotion,and possibly an auxin inhibition, mechanism may exist in Crocosmia.Ovary expansion occurs throughout the development and senescenceof unpollinated flowers, but does not appear to be the solecause of wall breakage in the abscission zone. It is suggestedthat hormonally regulated wall hydrolases weaken the cell wallsin the abscission zone, and allow wall breakage and subsequentabscission to occur. Cdrocosmia x crocosmiiflora, Montbretia, anatomy, breakstrength, cell wall changes, histochemistry, flowers, monocotyledons, perianth, senescence, ethylene, 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.     

C]Ethylene Metabolism during Leaf Abscission in Cotton

Changes in ¹⁴C₂H₄ metabolism in the abscission zone were monitored during cotton (cv. Deltapine 16) leaf abscission. Rates of ¹⁴C₂H₄ oxidation to ¹⁴CO₂ and tissue incorporation in abscission zone segments cut from the second true leaf of nonabscising leaves of intact plants were similar (about 200 disintegrations per minute per 0.1 gram dry weight per 5.5 hours) and relatively constant over a 5-day period. Deblading to induce abscission caused a dramatic rise in ¹⁴C₂H₄ oxidation, but tissue incorporation was not markedly affected. This rise occurred well before abscission, reaching a peak of 1,375 disintegrations per minute per 0.1 gram dry weight per 5.5 hours 2 days after deblading when abscission was 40%. The rate then gradually declined, but on day 5 when abscission reached completion, it was still nearly three times higher than in segments from nonabscising leaves. Application of 0.1 millimolar abscisic acid in lanolin to the debladed petiole ends increased the per cent abscission slightly and initially stimulated ¹⁴C₂H₄ oxidation. In contrast, naphthaleneacetic acid applied in a similar manner delayed and markedly inhibited both abscission and ¹⁴C₂H₄ oxidation.     

Leaf Abscission in Soybean: Cytochemical and Ultrastructural Changes following Benzylaminopurine Treatment

6-benzylaminopurine (BAP) delays leaf abscission of soybeanGlycine max (L.) Merr. Abscission of the distal pulvinus ofprimary leaves was induced in 12-d-old seedlings or explantsby removal of the leaf blade. BAP applied to the cut end ofthe pulvinus following leaf blade removal delayed abscission.Discoloration of the pulvinus occurred before abscission commencedand the number of grana in chloroplasts within cortical parenchymacells of the pulvinus decreased over time following leaf bladeremoval. BAP prevented discoloration of pulvinus tissues anda decrease in grana number. Starch grains within amyloplastsof cells of the starch sheath in the pulvinus disappeared followingleaf blade removal, whereas starch accumulated within the abscissionzone prior to abscission. BAP prevented this apparent redistributionof starch and instead promoted an increase in starch withinplastids of cortical parenchyma cells of the pulvinus. Duringthe abscission process, cells within the separation layer enlargedand their nuclei and nucleoli became more evident prior to theirseparation from one another. Cell separation resulted from breakdownof middle lamellae and partial degradation of primary cell walls.Cycloheximide applied directly to the external surface of theabscission zone inhibited abscission in a similar way to theBAP treatment. These results suggest that BAP prevents abscissionby altering patterns of starch distribution in the pulvinusand abscission zone and by inhibiting the synthesis of proteinsthat typically appear de novo in induced abscission zone tissues. Key words: Benzylaminopurine, BAP, Soybean, Pulvinus, Abscission, amyloplast.     

Changes in Polygalacturonase Activity and Solubility of Polyuronides during Ethylene-stimulated Leaf Abscission in Sambucus nigra

Leaflet abscission in Sambucus nigra is precipitated by cellwall degradation which is restricted to the site of cell separation.Accompanying wall breakdown is an increase in the activity ofthe enzyme polygalacturonase (PG) (E.C. 3.2.1.15 [EC] ) and this riseis primarily confined to the abscission zone tissue. The polygalacturonasehas a pH optimum of 4·4 and has the characteristics ofan endo-acting enzyme. The elevation in enzyme activity is theresult of an increase in at least two isoforms of PG as revealedby polyacrylamide gel electrophoresis of the natured protein.Leaflet abscission in S. nigra is associated with an increasein the solubility and depolymerization of polyuronides fromthe cell wall. These observations are discussed in relationto the mechanism of cell separation during ethylene-stimulatedleaf abscission. Key words: Elder, Sambucus nigra, abscission, polygalacturonase, polyuronides, ethylene     

Abscisic Acid, Auxin, and Ethylene in Explant Abscission

Experiments with explants of Phaseolus vulgaris L., cv. CanadianWonder, show that abscission and the associated rise in oarboxymethyl-cellulaseactivity in the separation zone are initiated by a peak in ethyleneproduction during senescence of pulvinar tissue distal to thezone. Distal applications of abscisic acid (ABA) induce an earlierpeak in ethylene production, increase cellulase activity, andpromote abscission. ABA is more effective in these ways if treatmentis delayed from 0 to 24 h after excision. With increasing concentrations of ABA the maximum rate of ethylene production is achievedsooner. Indol-3yl-acetic acid (IAA) and ABA are antagonisticin this system and have opposing effects. IAA retards the timeof peak ethylene-production and delays abscission. Explantsmay be retained for long periods without abscinding if incubatedin an ethylene-free atmosphere: the addition of ethylene forany one 24-h period (except the first 24 h after excision) willinduce abscission. The initial period of insensitivity to ethyleneis extended by distal applications of IAA. Ethylene-inducedabscission can be inhibited by IAA applied up to 72 h afterexcision provided the ethylene is not applied first. It is proposedthat abscission in the explant is controlled at two levels:(1) an auxin-dependent stage determining the duration of insensitivityto ethylene; (2) the timing of a rise in ethylene productionin senescing tissue distal to the separation zone. An auxin-ethylenebalance-mechanism at the separation zone is discussed.     

Effect of calcium, (2-chloroethyl) phosphonic acid and ethylene on bean leaf abscission

Summary The effects of CaCl₂, (2-chloroethyl) phosphonic acid (Ethephon) and ethylene on leaf abscission of debladed and intact bean plants (Phaseolus vulgaris L.) were studied. Ethephon (1000 g/l) and ethylene (8 l/l) induced abscission in debladed and intact plants in 24–72 h whereas IAA (10^-5M), cycloheximide (10^-5M) and CaCl₂ (0.068M) delayed abscission in debladed plants. CaCl₂ completely inhibited the abscission-enhancing effect of Ethephon in intact bean leaves. When CaCl₂ and Ethephon were applied simultaneously to separate halves of the leaf blade, leaves with Ethephon applied closest to the pulvinus abscised rapidly; when CaCl₂ was applied closest to the pulvinus, abscission was prevented. Calcium pre-treatment prior to ethylene (8 l/l) treatment of debladed plants delayed abscission as compared to those treated with ethylene alone.Michigan Agricultural Experiment Station Journal Article No. 6299.     

Leaf Abscission Zones in Molinia caerulea (L.) Moench, the Purple Moor Grass

Leaf separation and loss in the grass Molinia caerulea (L.)Moench was investigated using scanning electron microscopy.Leaf senescence and subsequent shedding of leaves was precededby the formation of a leaf abscission zone. The zone containscells which have more than doubled their wall thickness to greaterthan 0-4 µm. The line of fracture associated with thezone principally followed the middle lamellae, leaving intactcells on the fracture faces. Molinia should prove an interestingmodel in which to study abscission processes in the Gramineae. Molinia caerulea, leaf abscission zone, electron microscopy