The release of the cotyledonary shoot ofpisum sativum from inhibition in relation to changes in the levels of endogenous auxins,cytokinins, and gibberellins

Both axillary buds belonging to the cotyledons (cotyledonary buds) start to grow on decapitated pea seedlings, but one of them (the dominant shoot) prevails in growth over the other (the inhibited shoot). If the dominant' cotyledonary shoot is removed, the inhibited shoot is released from inhibition and starts to grow. This release from inhibition of the inhibited cotyledonary shoot is accompanied within two hours from the removal of the dominant cotyledonary shoot by a marked increase in the level of endogenous cytokinin-like substances and by a decrease in the level of endogenous IAA. By contrast, a significant increase in IAA level and a decreasing trend in the level of cytokinin-like substances occur in the originally inhibited cotyledonary shoot between hour 4 and hour 48 after the release from inhibition of the inhibited cotyledonary shoot. The level of gibberellin-like substances in the cotyledonary shoot released from inhibition steadily increases from the beginning of the release.     

Effect of auxins and cytokinins on plant regeneration from hypocotyls and cotyledons in niger (Guizotia abyssinica Cass.)

Tissue cultures were established from hypocotyl and cotyledonary leaf segments ofGuizotia abyssinica Cass. on MS medium supplemented with various concentrations of auxins (IAA, NAA, IBA or 2,4-D) and cytokinins (KN or BA). Expiants cultured on media with cytokinins or in combination with auxins produced shoot buds. Maximum number of shoot buds (20–25 per culture) were differentiated from cotyledonary leaf segments on medium with 2 mg 1^-1 each of KN and IBA. Rooting of regenerated shoot buds was acheived on medium with NAA. The obtained plantlets were successfully transferred to soil.     

Further studies on the effects of different auxins and cytokinins on flower formation in thin cell layers of Nicotiana tabacum: The effects of zeatin riboside, dihydrozeatin and dihydrozeatin riboside

Organogenesis in thin cell layers of Nicotiana tabacum L. was studied in relation to the effects of natural and synthetic auxins in combination with various cytokinins. All cytokinins tested, benzyladenine (BA), kinetin, zeatin (Z), zeatin riboside (ZR), N⁶-Δ²-isopentenyl) adenine (IPA), dihydrozeatin [(diH)Z] and dihydrozeatin riboside [(diH)ZR], seem to be active in flower bud formation. In addition to the initiation of flower buds, vegetative buds or roots were also formed on the explants in the presence of BA, Z or IPA as exogenous cytokinins. Only dihydrozeatin and its riboside stimulated the initation of flower buds alone (as is known for kinetin), especially if supplemented with indole-3-acetic acid (IAA) as exogenous auxin. A high number of explants with flower buds was also found with high cytokinin/2,4-D ratios. In these conditions the presence of (diH)Z yielded the higest number of flower buds per explant.     

Mutual interaction of auxin and cytokinins in regulating correlative dominance

Correlative dominance requires correlative signals from a dominant to a dominated organ. Auxins, particularly IAA, and cytokinins are obviously important components of this correlative system. Using a vegetative pea shoot and a generative apple and tomato fruit system it can be demonstrated that dominant organs always export more IAA and have a higher ³H-IAA transport capacity and velocity compared to dominated organs. In both systems the dominant organ can be replaced by the application of auxin, e.g. NAA, which maintains the differences in IAA export. This is an indication that similar regulatory mechanisms control dominance in both of these diverse systems. The possibility of replacing a dominant organ by auxin also makes it unlikely that growth of that organ or allocation of nutrients regulates the correlative inhibition of the dominated organ.It is suggested that differences in IAA export from, and transport capacities of, dominant and dominated shoots, may be explained by a mechanism of auxin transport autoinhibition (ATA), whereby the earlier and stronger export of IAA from the dominant shoot inhibits auxin export from the dominated shoot at the point where the two auxin streams converge. This hypothesis was tested with explants of pea, apple and tomato. It was shown that the basal application of cold IAA significantly reduced endogenous as well as exogenous IAA transport through these explants.Since the reduced IAA transport of dominated organs was not followed by an accumulation of IAA in the auxin producing subtending organ, it was concluded that IAA biosynthesis was possibly reduced and/or IAA conjugation stimulated. This could have been one of the determinants of their growth inhibition. ATA might also explain how the unidirectional IAA signal may affect the growth rate of organs even lateral or acropetal to its transport pathway and thus polar IAA-transport becomes a ``multidirectional' signal. From the experiments demonstrated it seems that ATA is a sufficient mechanism to impose growth inhibition in the dominated organ, without the need of other regulators.However, to release dominated organs from dominance cessation of ATA may not be sufficient and cytokinins are obviously a powerful antagonist to auxins. Their repeated exogenous application turns dominated lateral buds into strongly growing organs which ultimately may even dominate the previously dominant apex. These lateral shoots finally gain a strong IAA export capacity and inhibit, by ATA, IAA export from the hitherto dominant apex.In other experiments it was shown that interruption of polar IAA transport leads to a strong increase in root derived cytokinins. This can largely be prevented, in a concentration dependent manner, by the application of auxin, indicating that basipolar auxin may control cytokinin production in the roots and its possible delivery to lateral buds. In turn, the increased delivery of cytokinins to the lateral buds promotes a strong increase in IAA production and export. Thus there is a strong mutual interaction between auxin production in the shoots and cytokinin production in the roots, which may be important in regulating the balance between root and shoot growth.     

Effect of apex excision and replacement by 1-naphthylacetic acid on cytokinin concentration and apical dominance in pea plants

As known from literature lateral buds from pea ( Pisum sativum ) plants are released from apical dominance when repeatedly treated with exogenous cytokinins. Little is known, however, about the endogenous role of cytokinins in this process and whether they interact with basipolar transported IAA, generally regarded as the main signal controlling apical dominance. This paper presents evidence that such an interaction exists.
The excision of the apex of pea plants resulted in the release of inhibited lateral buds from apical dominance (AD). This could be entirely prevented by applying 1-naphthylacetic acid (NAA) to the cut end of the shoot. Removal of the apex also resulted in a rapid and rather large increase in the endogenous concentrations of zeatin riboside (ZR), isopentenyladenosine (iAdo) and an as yet unidentified polar zeatin derivative in the node and internode below the point of decapitation. This accumulation of ZR and iAdo, was strongly reduced by the application of NAA. The observed increase in cytokinin concentration preceded the elongation of the lateral buds, suggesting that endogenous cytokinins play a significant role in the release of lateral buds from AD. However, the effect of NAA on the concentration of cytokinins clearly demonstrated the dominant role of the polar basipetally transported auxin in AD. The results suggest a mutual interaction between the basipolar IAA transport system and cytokinins obviously produced in the roots and transported via the xylem into the stem of the pea plants.     

The character of endogenous growth regulators in the course of development in the fungus Lentinus tigrinus

The character of endogenous growth regulators in different stages of growth and development ofLentinus tigrinus was followed by means of bioassays. The methanolic extract of differently old mycelium and fruiting body was chromatographically separated and tested for auxins by means of Avena coleoptile segments, for gibberellins by using lettuce seedlings and for cytokinins by measuring the content of chlorophyl in leaf segments of barley. Auxins were found only in young vegetative mycelium and in the growing young stalk. The level of gibberellins is more significant and it reaches the highest values in the vegetative mycelium, in the growing young stalk and in the differentiated cap. The content of auxin and gibberellin substances did not increase even in the medium after cultivation of mycelium. Bioassays of cytokinins show a slight stimulation in the vegetative mycelium and in the primordia of caps. Significant inhibition was ascertained in the fully developed stalk. The high quantities of cytokinins were found in caps when basidiospores were formed. The results show a close connection of endogenous growth substances with the growth and the development of the fungus and indicate their participation in differentiation processes.     

Regeneration of calabrian pine from juvenile needles

Cytokinins applied in an agar medium induced adventitious buds on cultured needles from seedling of Pinus brutia Ten. Cytokinins applied as pulses to the explants prior to culture were less effective. Irrespective of the mode of cytokinin application, 8 weeks was the time required to bring about bud formation. Organogenetic potential of the cultured needles decreased with chronological age of the explanted seedlings. The induced buds grew into elongated shoots on culture medium without cytokinins, but the inclusion of activated charcoal (1%) doubled the elongation rate. There was an indication that mixtures of cytokinins were more effective than separate cytokinins in producing buds on explants, but the difference between treatments did not achieve statistical significance. Parenchyma cells in mesophyll layers were evidently the target cells responding to the culture conditions. After a period of activity and division in these cells, meristematic zones developed which later led to formation of bud primordia, and subsequently these primordia developed into well-formed adventitious buds. Subsequent rooting (64%) of shoots was achieved using a combination of two auxins and a low level of cytokinin.     

THE ROLE OF AUXINS AND CYTOKININS IN THE RELEASE OF BUDS FROM DOMINANCE

The paper deals with the general problem of the physiological basis of branching, and the roles of known and unexplored factors in sensitivity to apical dominance. It is shown that when pea seedling shoots are completely or partially inhibited by other shoots on the same plant auxin can promote their elongation, even though it does not have this effect on inhibited buds. This influence of auxin is only exerted on internodal elongation and not on apical growth. When kinetin in a solution of alcohol and carbowax is applied directly to the lateral buds of pea seedlings, it releases them from inhibition by the growing apex. It is shown that the role of alcohol in this solution is to act as a surfactant, permitting good contact with the buds, while that of carbowax, being hygroscopic, is to maintain a thin film of solution over the buds. Buds thus released from apical dominance by kinetin do not elongate as much as do uninhibited control buds. Such kinetin-treated buds can, however, be made to elongate normally by the application of auxin locally to their apices. It is concluded that growing shoots are relatively insensitive to correlative inhibition because they synthesize two types of growth substances, namely, auxin, which antagonizes the inhibitory effect on internodal elongation, and cytokinins, which permit the apex itself to develop. In the discussion it is brought out that many cases of branching, which appear at first to bear little relation to one another, can be understood on the basis of two principles, namely: (1) Any reduction in the growth rate of a dominant apex reduces its inhibitory effect on other apices, and (2) once an apex starts growing it becomes less sensitive to inhibition by other apices These generalizations and the experimental results are tentatively interpreted in terms of an interaction between the syntheses of auxin and of cytoldnin.     

The Influence of Different Cytokinins and Auxins on Flower Neoformation in Thin Cell Layers of Nicotiana tabacum L.

Neoformation of flower buds was studied in thin cell layersderived from floral branches of Nicotiana tabacum L. Organogenesiswas found to be expressed in different ways according to thekind of auxin and cytokinin present in the culture medium. Inthe presence of synthetic auxins (NAA, CTA), more flower budswere generated than with the natural auxins (IAA, IBA), if theculture medium was also supplemented with a cytokinin. Theseconditions also stimulated the development of the buds to flowersor inflorescences. The most active cytokinin was BA. Naturalcytokinins (zeatin, IPA) appeared much less active but led toremarkable development of roots together with flower buds. Anorganogenetic effect of 2,4-D was observed in this experimentalsystem, leading to the conclusion that organogenetic activityof cytokinins and auxins in thin cell layer cultures stronglydepends on the molecular structure of the growth regulators. (Received September 30, 1987; Accepted March 23, 1988)     

Effects of light quality on apical dominance in Xanthium strumarium and the associated changes in endogenous levels of abscisic acid and cytokinins

Summary Apical dominance in Xanthium strumarium was influenced by the quality of illumination received at the end of the photoperiod. The involvement of the red/far-red regions of the spectrum was apparent. The persistence of the effects was partially dependent on the age of the individual buds concerned. Plants receiving 30 minutes of illumination from tungsten lamps after a 16-hour photoperiod from fluorescent tubes failed to branch, whereas plants given an identical photoperiod, both in terms of day-length and photosynthetically available light energy, but lacking the far-red from tungsten lamps, branched profusely.The influence of the spectral distribution of illumination on the levels of cytokinins and abseisic acid in the plant, and the correlation with the degree of branching, is presented and discussed. The cytokinin content was much higher in inhibited than released buds. The cytokinins present were probably not able to particinate in bud growth because of an accumulation of inhibitors resembling abscisic acid. The concentration of the inhibitors in inhibited buds was 50 to 250 times that occurring in all other plant parts examined.     

Periodicity of bud bursting in willow (Salix viminalis) as affected by growth regulators

Saunders, P. F. and Barros, R. S. 1987. Periodicity of bud bursting in willow ( Salix viminalis ) as affected by growth regulators.
Lateral vegetative buds of willow ( Salix viminalis L.) were only innately dormant for 3–5 weeks in October; during this time their apices were correlatively inhibited by the bud leaflets. Exogenous gibberellins stimulated the opening of cultured buds when the plants were dormant or entering dormancy. As dormancy was being released, however, cultured buds became more responsive to exogenous cytokinins. Thus the demand for gibberellins and cytokinins for bud opening seemed to be sequential rather than simultaneous. Dormant buds cultured in the presence of abscisic acid remained unopened, but they opened after a chilling treatment. Subsequent growth of such buds as measured by dry matter accumulation, was observed only if a cytokinin was added to the medium.     

Hormone Levels and Apical Dominance in the Aquatic Fern Marsilea drummondii A. Br

Terminal buds and successively subjacent lateral buds of the water fern, Marsilea drummondii, were examined to determine the pattern of hormone distribution in relation to apical dominance. Quantitative levels of indole-3-acetic acid (IAA), abscisic acid (ABA), zeatin and zeatin riboside (Z and ZR), and isopentenyladenosine (iPA) were determined by a solid-phase immunoassay using polycional antihormone antibodies. Enzyme-linked immunosorbent assay was used following a one-step HPLC purification procedure to obtain the free hormones. Active shoot apices contained the most IAA and Z-type cytokinins and inhibited buds the least. No significant differences in ABA levels were found leading to the conclusion that ABA did not play any role in apical dominance. The normal precedence of the most rapid outgrowth of the youngest inhibited bud as observed previously in decapitated plants was well correlated with its very high level of iPA observed in this study. The same phenomenon was observed in the median buds but with a weaker amplitude. The presence of this storage form could indicate that a bud at its entry into quiescence eventually looses the ability to hydroxylate iPA to Z-type cytokinins when it is fully inhibited. IAA and Z + ZR are concluded to be essential for lateral bud growth.     

Endogenous plant growth regulators and rooting capacity of different walnut tissues

Rooting ability of three different materials, namely cotyledonar linking areas, cotyledonary portions and embryonic microshoots from walnut seeds was determined. It was shown that while cotyledonar linking areas may be defined as a espontaneous rooting system, the other two tissue types correspond to inducible rooting systems, needing exogenous application of auxins for root induction. Endogenous zeatin and isopentenyl type cytokinins, indole-3-acetic acid (IAA), and abscisic acid (ABA) contents were determined in the three kinds of plant material prior to their culture. Our results show that the most active rooting systems has high levels of cytokinins, particularly dihydrozeatin (diH)Z and its riboside, and low levels of IAA and ABA. This situation could indicate that oxidative metabolism was taking place in cotyledonar linking areas, which is generally associated with the initiation phase of the rooting process. So that, we can assume that the induction phase is already overpassed in these tissues. On the contrary, the other two systems in which high levels of auxins and reduced levels of cytokinins were found, need to undergo the induction phase to be able to show root manifestation. In this case, the exogenous auxins are responsible for triggering this process, by enhancement of oxidative conditions. Therefore, it can be conclude that dynamics of growth regulators levels during the different phases of rooting are more important than specific single levels at fixed time.     

Response to strigolactone treatment in chrysanthemum axillary buds is influenced by auxin transport inhibition and sucrose availability

Axillary bud outgrowth is regulated by both environmental cues and internal plant hormone signaling. Central to this regulation is the balance between auxins, cytokinins, and strigolactones. Auxins are transported basipetally and inhibit the axillary bud outgrowth indirectly by either restricting auxin export from the axillary buds to the stem (canalization model) or inducing strigolactone biosynthesis and limiting cytokinin levels (second messenger model). Both models have supporting evidence and are not mutually exclusive. In this study, we used a modified split-plate bioassay to apply different plant growth regulators to isolated stem segments of chrysanthemum and measure their effect on axillary bud growth. Results showed axillary bud outgrowth in the bioassay within 5 days after nodal stem excision. Treatments with apical auxin (IAA) inhibited bud outgrowth which was counteracted by treatments with basal cytokinins (TDZ, zeatin, 2-ip). Treatments with basal strigolactone (GR24) could inhibit axillary bud growth without an apical auxin treatment. GR24 inhibition of axillary buds could be counteracted with auxin transport inhibitors (TIBA and NPA). Treatments with sucrose in the medium resulted in stronger axillary bud growth, which could be inhibited with apical auxin treatment but not with basal strigolactone treatment. These observations provide support for both the canalization model and the second messenger model with, on the one hand, the influence of auxin transport on strigolactone inhibition of axillary buds and, on the other hand, the inhibition of axillary bud growth by strigolactone without an apical auxin source. The inability of GR24 to inhibit bud growth in a sucrose treatment raises an interesting question about the role of strigolactone and sucrose in axillary bud outgrowth and calls for further investigation.     

Changes in the content of endogenous auxins in apical buds ofchenopodium rubrum L. Induced with respect to the endogenous rhythm in capacity to flower

The content of endogenous auxins was examined in apical buds ofChenopodium rubrum plants induced by a photoperiodic cycle of 16h darkness and 8h light followed by a dark period of various duration so as to correspond with either maximal or minimal flowering response in the endogenous rhythm in capacity to flower initiated by the photoperiodic treatment. Apical buds of potentially generative plants contained less auxins than apical buds of plants which remained in the vegetative state. Apical buds from plants treated with kinetin (1. 10^-3 M) and therefore remaining in the vegetative state showed an auxin level comparable to that of untreated plants exhibiting minimal flowering response irrespective of the duration of the second dark period. Plants cultivated on a sucrose solution (0.6 M) during the second dark period became generative even at the normal minimum of flowering. The auxin content of the apical buds was low, similarly as in untreated plants induced for a period leading to maximal flowering response. On the other hand, apical buds from plants grown on sucrose solution during a dark period leading to the manifestation of maximal flowering response showed a relatively high auxin content comparable to that found in untreated plants which had obtained a more extended induction by three photoperiodic cycles. The results are discussed with respect to the possible role of endogenous auxins in the regulation of the changes in growth correlations occurring in the shoot apex during photoperiodic induction and in the expression of the competence to flower.     

The role of endogenous cytokinins in correlation between cotyledon and its axillary bud and in hypocotyl regeneration of flax

When flax seedlings are decapitated above cotyledons and three days later one of the two cotyledons is removed then the remaining cotyledon stimulates in four to five days growth of its axillary bud. It has been found that content of endogenous cytokinins was higher in the stimulated bud as compared with the other one already 12 h after the cotyledon removal. Flax seedlings decapitated under cotyledons regenerate adventitious buds on thy hypocotyl stump during 5–6 days. The endogenous fytohormonal preparation of this regeneration was investigated in the 20 mm apical part of the hypocotyl stump. Decrease in auxin and increase in gibberellins was already found during the first day after decapitation while the level of cytokinins increased as late as three days after the apex removal.     

Release of cotyledonary shoots of pea (<Emphasis Type="Italic">Pisum sativum</Emphasis>) seedlings from inhibition as related to endogenous zeatin and ethylene and exogenous IAA and benzyladenine

This paper deals with apical dominance using a dicotylar model obtained after decapitation of pea seedlings with two shoots — one dominant and the other inhibited. When the dominant shoot was decapitated the inhibited one is released from inhibition and after 24 to 72 h begins to grow. However, the levels of trans-zeatin and production of ethylene increase within 4 and 6 hours respectively after release from inhibition, and within an interval of 72 h the levels of both phytohormones begin gradually to decrease. This indicates that also in this model, the release from apical dominance is associated with an increase in the level of cytokinin zeatin and, thereafter, also with an increased production of ethylene. If indolyl-3-acetic acid (IAA) is applied on the decapitated main stem after decapitation of the dominant shoot, the growth of the initially inhibited one is very strongly retarded; if, however, IAA is applied on the decapitated dominant shoot, this inhibition is significantly weaker. This means that the inhibiting effect of IAA on the inhibited shoot originates to a greater degree from the main stem rather than from the dominant shoot. The effect of benzyladenine (BA) is transferred equally from the decapitated main stem and from the decapitated dominant shoot because the initially inhibited shoot begins to grow as well as also other shoots from serial cotyledonary buds.     

Indole auxins in spinach plants grown in long and short days

The endogenous auxin-like substances were analyzed in the shoot extracts of young spinach seedlings, exposed to photoperiodic induction. At least eight indole auxins were found. One of them was identified as tryptophan, the other one is most probably IAA. The plants grown in long days had a higher level of ether soluble auxins than the controls in short days. Separate extractions of plants after each of the eight inductive days showed that the auxin content was not constant, but subjected to irregular oscillations. However, parallel oscillations were also found in control plants grown in short days. Staminate plants were found to contain more endogenous auxins than the pistillate ones. It is concluded that the quantitative changes in auxins during the photoperiodic induction are probably not related to flowering, but to some other growth process, common to all plants in that phase of growth. The higher level of auxins in staminate plants may be the cause of their faster elongation before the onset of flowering.     

The role of endogenous growth regulators in the differentiation processes of walnut (Juglans regia L.)

The character of endogenous growth substances was investigated in developing buds, young fruits and mature walnut leaves. The relatively high content of auxins and gibberellin-like substances was found by means of bioassays in the youngest primordia of vegetative buds. The level of auxins drops in the further course of primordia transformation into the staminate catkins. The development of leaf-buds is characterized by the accumulation of inhibitory activity as revealed by theAvena bioassay, whereas the data obtained from the lettuce bioassay indicate a pronounced stimulation. The onset of terminal bud development is also accompanied by inhibitions and it is only with pistillate flower differentiation that the temporary rise in auxin level is observed. An inhibitory activity was found in these extracts using lettuce bioassay. There is a relatively high auxin level in young fruits, mature leaves and resting buds during the mid-summer period whereas the accumulation of clearcut inhibitions is signalled by the results of lettuce bioassay. The regulatory role of growth substances in differentiation may be better understood during the second year as many leaf-abnormities appear only with the outgrowing of the bud. Abnormal catkins differ in the number of florets and stamens and some even bear pistillate flowers. Fruit development is liable to deviations in the early stages of differentiation. Abnormal fruits enable us to elucidate many structural peculiarities.     

Role of mineral nutrients and growth regulators in the apical dominance in Solanum sisymbrifolium

Summary The lower axillary buds of intact plants of Solanum sisymbrifolium are released from complete inhibition by supplying high doses of potassium to the soil while complete apical dominance is shown by plants grown at low, but not deficiency levels of K. Nitrate and phosphate supplied alone or together are ineffective but when either of them is supplied with K, the effect of the latter on the growth of axillary buds is somewhat enhanced. The buds thus released from inhibition elongate further only when supplied with gibberellin A₃ (GA₃). Indoleacetic acid (IAA) alone has a considerably weaker effect but when supplied with GA₃ the stimulatory effect is greater than is caused by the latter alone. The completely inhibited buds of low-K plants can be released from inhibition by supplying kinetin. In such buds IAA promotes further extension but GA₃ does not. The partially inhibited buds of the high-K plants, on the other hand, are not affected by exogenous kinetin.Dedicated to Professor H. D. Wulff on the occasion of his 60th birthday.