The role of auxin in cell separation in the dehiscence zone of oilseed rape pods

Concentrations of both free and conjugated indole-3-acetic acid(IAA) were studied during development of pod wall, dehiscencezone and seeds of Brassica napus pods. A decrease in auxin contentprior to moisture loss in the pods was observed specificallyin the dehiscence zone, which was correlated with a tissue specificincrease in ß-1,4-glucanase activity. Furthermore,treatment of the pods with the auxin mimic 2-methyl-4-chlorophenoxyaceticacid resulted in a delay of 10 d of ß-1,4-glucanaseactivity and con-comitant cell separation in the dehiscencezone. This indicates that the activity of hydrolytic enzymesinvolved in cell separation in the dehiscence zone is regulatedby auxin activity. Comparison of parthenocarpic pods with seeded pods pointed tothe seeds as the source of IAA. Levels in the dehiscence zoneof these pods were low over the entire sampling period, whilecell separation in the dehiscence zone was delayed by about4 d. These results indicate that a low level of auxins in thedehiscence zone is necessary for dehiscence to take place, butother factors may also be important. Key words: Brassica napus, pod dehiscence, auxin, cellulase     

Hormonal Regulation of Pedicel Abscission in Begonia Flower Buds

In order to analyse the hormonal regulation of flower bud shedding in Begonia, levels of indoleacetic acid (IAA), abscisic acid (ABA) and ethylene were determined in buds and pedicels. The translocation and metabolism of ¹⁴C-labeled IAA in pedicel segments were also studied. In a monoecious Begonia fuchsioides hybrid, abscising male flower buds contain about 1% of the IAA present in non-abscising female flowers. In a male Begonia davisii hybrid, the seasonal variation in bud drop coincides with changes in the IAA content of the buds, while also the release of IAA from the bud to the pedicel is hampered. Abscission zones of these pedicels always contain abscission promoting ethylene concentrations. The tissue is prevented from responding with abscission by IAA from the flower buds. The buds also contain ABA but without influencing abscission considerably. Pretreatment with ethylene or ABA does not affect IAA transport in pedicel segments. The rate of this transport is 4–6 mm × h^–1:; the capacity increases with the transverse area. In young segments, IAA is decarboxylated and also otherwise metabolized.     

Auxin Physiology of Decapitated Stems of Phaseolus vulgaris L. Treated with Indol-3yl-acetic Acid

Physiological amounts of indol-3yl-acetic acid (IAA) were accumulatedby decapitated stems of Phaseolus vulgaris L. seedlings fromlanolin pastes, containing 0.1 per cent IAA, applied to thecut surfaces of the stumps. Both the levels and gradients ofextractable and diffusable IAA detected in the treated stumpscompared favourably with those reported for the whole plants.A considerable portion of IAA that entered the tissue was metabolizedto a compound that had chromatographic properties similar toindol-3yl-acetylaspartic acid. Two other metabolities were tentativelyidentified as indol-3yl-acetylaspartic acid indol-3yl-acetylglucose.The accumulated IAA appears to be transported as indol-3yl-aceticacid at an apparent velocity of 28 mm/h down the decapitatedinternodes.     

Apical Dominance in Phaseolus vulgaris L.

Although determinations of the ABA content of lateral buds ofPhaseolus vulgaris revealed no difference between decapitatedand intact control plants in the first 12 h following decapitation,a relative decrease in the ABA content of lateral buds of decapitatedplants was detectable 24 h following decapitation. Shoot decapitationwas also observed to result in a decrease in the ABA contentof stem tissue. The application of IAA to the stem of decapitatedplants prevented these changes and increased the ABA contentof stem tissue relative to that of intact plants. The levelsof IAA and ABA were also determined in the stem tissue fromthe nodes of intact bean plants. The possible interdependenceof these two plant hormones was further investigated by a studyof [2_–14ClABA metabolism. The results are discussed inrelation to the possible role of these hormones in apical dominance. Key words: Apical dominance, Abscisic acid, Indole-3-acetic acid     

Morphogenic Potentialities of Flower Buds of Kalanchoe pinnata Pers. grown in vitro

Excised flower buds of Kalanchoe pinnata Pers. representingtwo early stages of development (designated sets I and II),were cultured on modified White's medium (WB). They failed toattain full development on WB or on WB containing any of thefollowing supplements: indole-3yl-acetic acid (IAA), 2,4-dichlorophenoxyaceticacid (2,4-D), naphthaleneacetic acid (NAA), kinetin, or coconutmilk (CM). A slight stimulation of the growth of corolla wascaused by kinetin (10 ppm). IAA (I ppm) and NAA (I ppm) inducedrooting from the cut end of the pedicel and from the proliferatedtorus tissue situated between the sepals and petals. 2,4-D (Ippm) either singly or in concert with CM (10 per cent) stimulatedthe formation of shoot buds and root growth. Addition of kinetin(I and 10 ppm) to WB favoured shoot formation, but suppressedrooting. Flower buds of set II developed shoot buds more readilythan those of set I. Thus, the primordia floral organs presentin the immature buds lose their ability for normal morphogenesisunder culture conditions. Buds destined to form flowers canbe made to revert to vegetative growth.     

The Effect of Ultraviolet Radiation and Sucrose on IAA Levels in Spirodela oligorhiza

U.v. radiation significantly lowers the IAA level in frondsof Spirodela oligorhiza. Fronds grown in the light on mediumsupplemented with sucrose had much higher IAA levels than frondsgrown on medium lacking sucrose. The results suggest that u.v.-inducedabscission in Spirodela and the reduction of abscission by sucroseare both related to their effect on endogenous IAA levels.     

Tracheary Element Differentiation Induced in Isolated Cylinders of Lettuce Pith: a Bipolar Gradient Technique

To study the influence of morphogenetic gradients on vasculardifferentiation patterns, a new technique was developed whichallows different substances to be applied at opposite ends ofa tissue block. It yielded information on the mobility of particularmorphogens and on the dependence of callus formation and trachearyelement differentiation on their presence. Application of indol-3ylacetic acid (1AA) (10 mg l^–1), zeatin (0.1 mg l^–1)and sucrose (3 per cent, w/v) in various combinations to theends of cylindrical explants of lettuce pith (Lactuca sativaL.) showed that (a) callus formation was stimulated by IAA,whereas induction of tracheary elements required both IAA andzeatin; (b) callus was confined to a few millimetres at theends of the explants, and tracheary elements occurred mainlywithin the callus; (c) sucrose or its metabolic products diffusedthe 10 mm length of the explants, while IAA and zeatin wereeffective only close to the application site; and (d) some callusand tracheary elements formed when no sucrose was applied, butboth increased with sucrose application, though inhibition oftracheary elements formation occurred with high sucrose concentrations. differentiation, pith explant, tissue culture, xylogenesis, indol-3yl acetic acid, sucrose, zeatin, lettuce, Lactuca sativa     

Apical Dominance in the Tomato: Some Further Observations on Isogenic Lines Showing Varying Degrees of Side-shoot Development

A study has been made of the distribution of substances resemblingindol-3-ylacetic acid (IAA), abscisic acid (ABA) and cytokinin-likesubstances in the stem tissue of Craigella tomato plants ascompared with that found in two isogenic lines of this variety,Craigella Blind (blbl) and Craigella Lateral Suppressor (lsls),in both of which side shoot growth is suppressed to varyingdegrees. There was no evidence to suggest that the distributionof these hormones in the stem had any association with the differentpatterns of side shoot development of the three types, thoughsome of the lateral suppressor plants which exhibited only partialbud inhibition did show a relation between high auxin and abscisicacid levels and lack of side shoot development from the centralnodes of the shoot. Decapitation led to a stimulation of bud outgrowth from allnodes of the Craigella plants but the lateral suppressor plantsremained inhibited. The blind plants were found to initiatebud primordia at the cotyledonary nodes only when the severedapex was replaced by exogenous IAA. The results are discussed in relation to our knowledge of themechanisms controlling apical dominance in the tomato. Lycopersicon esculentumL, tomato, apical dominance, growth regulation, indol-3-ylacetic acid, abscisic acid, cytokinins     

Factors Affecting the Abscission of Reproductive Organs in Yellow Lupins (Lupinus luteus L.): III. ENDOGENOUS GROWTH SUBSTANCES IN VIRUS-INFECTED AND HEALTHY PLANTS AND THEIR EFFECT ON ABSCISSION

Extracts of small and mature-size lupin pods yielded four substancesaffecting the growth of wheat-coleoptile sections: one acidpromotor (A), two acid inhibitors(B and X), and one neutralinhibitor(Y). Inhibitor B was extremely active, however, coleoptile sectionsshowed no signs of toxic effects; they resumed growth at a rapidrate after rinsing them and adding ß-indolylaceticand (IAA) to the medium. 1 µg of IAA was required to counteractthe effect of ‘B’ extracted from 230 mg. Of tissue.On an equal fresh weight basis the inhibiting action of ‘B’in lupin pods was 500–1,500 times more potent than thatof ‘inhibitor ß’ in etiolated pea seedlings. Small pods of plants infected with pea-mosaic virus yielded3 times the amount of ‘A’ of healthy plants (equivalentto 1 µg. IAA 0.3 µg. IAA per 25 g. of tissue respectively),and approximately the amount of ‘B’. Mature podsof virus-infected plants again yielded more‘A’,but also 2? times more ‘B’ than pods of healthyplants. Healthy pods yielded more ‘A’ than virus-infectedpods, and there was no difference in ‘X’. A lupin abscission test was developed and the effects of proximaland distal application of -naphthyl acetic acid (NAA) are presented,and discussed with respect to results of other abscission tests. ‘A’ accelerated abscission when applied proximally,and delayed or prevented it when applied distally. ‘B’strongly accelerated abscission when applied in either way.A possible mechanism explaining the abscission-inducing effectof developing pods on later flowers is discussed in terms ofthe substances ‘A’ and ‘B’. The partlyprevented abscission observed on virus-infected plants was foundto agree well with the proposed mechanism.     

Optimal and Spatial Analysis of Hormones,Degrading Enzymes and Isozyme Profiles in Tomato Pedicel Explants During Ethylene-Induced Abscission

Activities of degrading enzymes, hormones concentration and zymogram patterns were investigated during control and ethylene-induced abscission of tomato pedicel explants. Exogenous ethylene accelerated abscission of pedicel explants. It was showed that IAA concentration in abscission zone tended to decline at first and then was reduced before separation in control and ethylene-treatment. Moreover, IAA (indole acetic acid) and ABA (abscise acid) concentrations were elevated in each segment when exposing to ethylene, but GA_{1 + 3} (gibberellin1 + gibberellin3) concentration was decreased in abscission zone and the proximal side. Activities of cellulase, polygalacturonase and pectinesterase in the explants were induced in the separating process and strengthened by ethylene. However, comparing with the proximal side, cellulase and polygalacturonase activities in abscission zone and distal side were higher. Electrophoresis of isozymes revealed that at least three peroxidase and three superoxidase isozymes appeared in the explants, respectively. One peroxidase isozyme exhibited differentially among the three positions in control and ethylene-treatment. One esterase isozyme weakened or disappeared in the following hours, but three novel esterase isozymes were detectable from beginning of the process. The data presented support the hypothesis that the distal side, together with abscission zone of explants plays a more important role in separation than does the proximal side. The possible roles of degrading enzymes, hormones and isozymes in three segments during ethylene-induced abscission of tomato pedicel explants are discussed.     

The Anther Smut of Sea Campion: A Study of the Role of Growth Regulators in the Dwarfing Symptom

The sympton of dwarfing in the sea campion, Silene vulgaris(Moench) Garcke sub sp. maritima(With.) A. & D. Löveinfected with the anther smut fungus Ustilago violacea (Pers.)Fuckel, a systemic, perennial parasite, has been investigated. Extracts of both healthy and diseased plants contain IAA andGA₃, but diseased plants contain less gibberellins than healthyplants. Neither IAA nor gibberellins were detected in significantquantities in the medium when U. violacea was grown in pureculture, but IAN was present. IAN was also found m extractsof diseased plants and it is tentatively suggested that it isformed by the fungus and may accumulate in the host owing tothe inability of the plant to convert IAN to IAA The exogenousapplication of IAN to healthy plants does not produce any diseasesymptoms. The dwarfing symptom of the diseased plant may be due to thelower levels of gibberellins which it contains compared withhealthy plants since the exogenous application of GA₃, restoresthe diseased plant to normal growth     

AUXIN OF TOMATO FRUIT AT DIFFERENT STAGES OF ITS DEVELOPMENT WITH A SPECIAL REFERENCE TO HIGH TEMPERATURE INJURIES

Acidic auxin in tomato fruit at various developmental stageswas examined by paper chromatography and Avena curvature test. Auxin activity of the fruit extracts was detected only in thesame R_f zone as that of authentic IAA with four solvent systems,i.e., isopropanol-ammonium hydroxide-water 10:1:1 v/v, n-butanol-ammoniumhydroxide-water 25:1:5 v/v, ethanol-water 7:3 v/v and n-butanol-aceticacid-water 4:1:1 v/v. Auxin content of fruit was the highest on the seventh day afteranthesis, when the two-celled proembryo began to grow rapidlyand the endosperm consisted of multi-cells. The second peakof auxin activity occurred on the 30th day, when the embryo,which consisted of the cotyledons, hypocotyl, and radicle, finishedits most rapid growth. No auxin activity was detected in theripe fruit. Auxin contents in fruit on the fifth, seventh, and tenth daysafter anthesis were lowered by a 4 hr high temperature treatmentat 40 given 0–3 days after anthesis. The nature of tomato auxin and its possible role in the fruitdevelopment were discussed. (Received June 30, 1966; )     

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.     

In vitro Propagation of Red Cabbage (Brassica oleracea L. var. capitata)

By manipulation of various growth regulators and physical conditions,plants have been regenerated from excised roots, stem segments,cotyledons, leaves, and callus cultures of red cabbage (Brassicaoleracea var. capitata) grown under in vitro conditions. Shootbuds were induced on isolated root segments (1 cm long) culturedon Murashige and Skoog's medium and the frequency of bud formationwas greatly enhanced by the addition of kinetin (0.5 part 10^–6).Callus obtained from the seeds, cotyledons, and hypocotyl segmentscultured on a medium fortified with 2,4-D (1 part 10^–6),kinetin (0.1 part 10^–6), and coconut milk (10%, v/v) hasbeen repeatedly subcultured. The callus is slow growing, andon transference to a kinetin (2 parts 10^–6) and IAA (2parts 10^–6) medium underwent morphogenesis to give riseto plants. The significance of the propagation of red cabbageby in vitro culture is pointed out.     

Effects of Gibberellin on Auxin-Induced Hyperpolarization of Cell Membranes in Hypocotyls of Vigna unguiculata

Auxin activates pumping of protons from the symplast to theapoplast and causes hyperpolarization of the symplast membranein the elongation zone of Vigna stems prior to the accelerationof growth. This auxin-induced hyperpolarization has been studiedin most cases in hypocotyl segments excised from the elongationzone. In the present study, mature-zone segments were perfusedwith IAA by the xylem perfusion technique in an effort to determinewhether or not IAA has any effects in the mature zone. Althoughno hyperpolarization of the symplast membrane was observed uponthe perfusion with auxin alone, auxin-induced hyperpolarizationwas observed when mature-zone segments had been pretreated withGA₃, in the absence of an increase in the growth rate. Theseresults suggest that cells in the mature zone have lost theability to activate the proton-pumping machinery in responseto auxin but that this ability can be restored by treatmentwith GA₃. This effect of GA₃ suggests the possibility that theconcentration of gibberellin in a tissue controls one of thecell's responses to auxin, namely, activation of the protonpump. (Received January 10, 1994; Accepted June 11, 1994)     

INVESTIGATIONS ON AUXIN-GIBBERELLIN INTERACTION ON THE PHOTOTROPISM OF HELIANTHUS ANNUUS HYPOCOTYL

On applying IAA-GB and IAA-NAA mixtures to the hypocotyls ofHelianthus annuus, the plants showed an increase and a decrease,respectively, in their phototropic response compared with thecontrol treated with IAA-H₂O only. In the in vitro experimentsit was observed that the illumination (with 1,500 lux light,lasting for 24 hr) caused photoinactivation of IAA in the mixturesof IAA-H₂O, IAA-GB and IAA-NAA by 50%, 24% and 0%, respectively.In the presence of 0.01% riboflavin as a photosensitizer inthe same mixtures, the degree of photodestruction of IAA causedby the same illumination was 92%, 97% and 86%, respectively. The stronger positive phototropism in the IAA-GB treated hypocotylwas accounted for by the situation that on the illuminated sidethe photo-inactivation of IAA was accelerated by GB, while onthe shaded side IAA and GB acted synergistically. On the otherhand, the weaker phototropism in the IAA-NAA treated hypocotylwas explained as due to a partial inhibition of photoinactivationof IAA by NAA. (Received October 6, 1962; )     

Polar transport and content of indole-3-acetic acid in wounded sweet potato root tissues

In roots of sweet potato (Ipomoea batatas Lam. cv. Kokei 14),the metabolic response to wounding was remarkable in the proximalside and developed in the acropetal direction. We assumed thatthe polarity resulted from the increase in polar movement ofindoleacetic acid (IAA) (1977, Plant Physiol. 60: 563–566).Transport of IAA and change of the IAA level in the woundedtissue of sweet potato roots were investigated. Transport ofthe label from ¹⁴C-IAA was obviously polarized in the acropetaldirection. ¹⁴C-IAA administered to the wounded tissue was mainlymetabolized into two conjugates of IAA. The amount of IAA inthe wounded tissue, determined by the spectrofluorometric method,increased about 3-fold after 18 hr of incubation prior to thedevelopment of activities of some enzymes. The increase in IAAcontent was not affected with aseptic incubation, therefore,the possibility of IAA production by microorganisms on the woundedtissue was excluded. The results obtained strongly support ourhypothesis that IAA plays an important role in the metabolicresponse to wounding. (Received May 2, 1979; )