Abscission and thinning of young fruit and thier regulation by plant hormones and bioregulators

Natural abscission of young fruit and its regulation by plant hormones isconsidered and compared to the generally accepted model of senescencetriggered abscission of, for example, leaves or mature fruit. It isconcluded that abscission of young fruit cannot be explained by this model.Alternatively, it is suggested that the senescence triggered initial step inthe classical abscission model should be replaced by a correlativelytriggered step. Polar basipetal IAA transport with its autostimulation andautoinhibition components is the main regulating signal in this correlativeacting system and replaces ethylene as the initial driving force from thesenescence triggered model.Results supporting this model are presented and tested against existingresults from the literature. Finally, this hypothesis is tested as a possibleexplanation of the mode of action of some thinning chemicals orbioregulators. It is speculated how a thinning chemical should be designedto function in a more reliable way, at least as far as its interference with the endogenous hormone system is concerned.     

Use of hydrogen cyanamide for apple and plum thinning

Effects of various concentrations of Dormex (a.i. 49% hydrogen cyanamide) on fruit thinning of Rome Beauty apple (Malus domestica Borkh.), Friar and Simka plums (Prunus salicina Lindley) were studied. A full bloom application of Dormex at all tested concentrations decreased Rome Beauty apple fruit set and yield, and increased fruit weight. Dormex at 0.25% (v/v) resulted in adequate apple thinning, indicated by production of an optimum fruit weight (320 g). Prebloom and full bloom applications of Dormex at greater than 0.75% reduced plum fruit set and yield in Friar. Full bloom application of Dormex at 0.50% showed a satisfactory fruit set, yield, and fruit weight in Friar plum. Prebloom Dormex application had no significant effect on `Simka' plum fruit set or yield, but full bloom application decreased fruit set and yield.     

Cytokinins in cut carnation flowers. IV effects of benzyladenine on flower longevity and the role of different longevity treatments on its transport following application to the petals

A low concentration of benzyladenine (4.44 × 10^-5 M) accelerated the senescense of cut carnation flowers. This effect could be reversed by STS-treatment but not by keeping the flowers in a holding solution containing 4% ethanol. This appears to be the first report indicating that cytokinins at a specific level may actually enhance flower senescense. The higher levels tested (1.11 × 10^-4 and 2.22 × 10^-4 M) retarded senescense, being in agreement with published results. The applied cytokynin was metabolized slowly in the petals to a compound(s) which co-chromatographed with ribosylbenzyladenine when separated by TLC and when fractionated by HPLC. In the experiments applying (¹⁴C) benzyladenine to the petals, a small degree of transport of the ¹⁴C was detected in naturally senescing (control) cut flowers and when treated with ethrel. The transported ¹⁴C was detected in both the ovaries and in the stems and co-chromatographed with benzyladenine. Where flower senescense was delayed (ethanol or STS) no movement from the petals was detected. This suggests that the cytokinin moved within the assimilate stream, along with sugars.     

Cell division and cell enlargement in fruit of Lagenaria leucantha as influenced by pollination and plant growth substances

The effects of NAA (naphthaleneacetic acid), GA₃ (gibberellic acid), CPPU (N-(2-chloro-4-pyridyl)-N'-phenylurea) and pollination on fruit set, cell division and enlargement were studied in Lagenaria leucantha, an important vegetable. NAA and GA₃ were ineffective in inducing parthenocarpy, whereas CPPU induced parthenocarpic fruit significantly larger than fruit that resulted from pollination. Cell division, which occurred during the first 4 days after pollination was not reactivated by NAA or GA₃, but was effectively reactivated by CPPU. The cell number of the total cross-section of CPPU-treated fruit was 117.4% of that of pollinated fruit and 154.4% of that of unpollinated at 12 DAA (days after anthesis) respectively. The CPPU-induced parthenocarpic fruit had the largest cell cross-sectional area followed, successively, by pollinated fruit, NAA-treated fruit, GA₃-treated fruit and unpollinated fruit. These results indicate that CPPU induced parthenocarpic fruit growth by directly reactivating cell division and expansion.     

<Emphasis Type="Italic">In vitro</Emphasis> nitrogen nutrition and hormonal pattern in bromeliads

Summary Four cytokinins (CKs), indole-3-acetic acid (IAA) and its ester and amino conjugates, and abscisic acid (ABA) levels of two bromeliads, Ananas comosus (L.) Merril and Vriesea gigantea Gaudich., grown in 5 mM (NH₄)₂SO₄ or urea as the sole nitrogen (N) form, were investigated. In both bromeliads, zeatin (Z) and zeatin riboside ([9R]Z) were the most abundant CKs. In A. comosus, CKs levels decreased drastically (≊ 12 times) after 7 and 30 d in media with ammonium and urea, respectively. After 3 d in media with N, V. gigantea CK levels decreased 30 and 20 times in the presence of ammonium and urea, respectively. N-starved A. comosus and V. gigantea exhibited similar ABA levels, but ABA decreased faster in V. gigantea when plants were transferred to media with N. Free IAA levels decreased until the 15th and 30th day when A. comosus was transferred to a medium with ammonium and urea, respectively. N-starved A. comosus amide, ester, and free IAA amounted to 81%, 14%, and 4%, respectively. There was a transient increase in the proportion of amide IAA and a corresponding decrease of the ester and the free IAA proportion when N-starved plants were transferred to media with N. The relationship between the internal hormonal patterns and the different ecological adaptations of the two bromeliads are discussed.     

Influence of ethylene on apex development and mobilization of soluble saccharides in the corm of Liatris

We studied the influence of ethephon (2-chloroethylphosphonic acid) on the first stages of development of the main apex of Liatris spicata cv. Callilepis and its relation to the mobilization of soluble saccharides in the corm. Ethephon enhanced cell expansion during the first weeks of cultivation but did not affect the timing of flower initiation. Soluble saccharide concentration in the corm reached a maximum after sprouting, probably due to rapid growth of the shoot. There was no change in the concentrations of soluble saccharides immediately prior to flower initiation, which suggested that quantitative changes in these saccharides were not responsible for activating flower initiation. When corms were treated with ethephon there was a greater degree of soluble saccharide mobilization during the first weeks of cultivation than in untreated corms. However, this did not seem to be the direct result of the addition of this phytohormone and was more probably the consequence of cell growth being encouraged. This revised version was published online in July 2006 with corrections to the Cover Date.     

The Roles of Phytohormones in Development as Studied in Transgenic Plants

The study of transgenic plants has greatly advanced our understanding of the control of development and metabolism. The ability to isolate and modify genes greatly extends the range of what is technically feasible. In the area of hormone biology, transgenic plants have helped to elucidate the pathways of synthesis, the metabolic control points, and the biological functions of the various phytohormones. This review covers the available genes that modulate the metabolism and perception of the phytohormones. One of the most significant conclusions coming out of transgenic plant work is the complex interaction among the different classes of phytohormones. For example, increasing the level of the auxin indole-3-acetic acid (IAA) in a plant has the secondary effect of inducing ethylene biosynthesis. This complication can be circumvented by combining transgenic plants modulating multiple hormones or through the use of available mutants. In this manner, transgenic plants have been utilized to unambiguously define the roles of auxin, cytokinin, and ethylene in the control of apical dominance. The power of transgenic plants as tools in hormone biology is perhaps best illustrated by work on ethylene. In this case, the modular characterization of genes led to elucidation of the biosynthetic pathway. Availability of the biosynthetic genes has permitted detailed analysis of the regulation of synthesis, definition of the role of ethylene in the control of several developmental processes, and the application of that knowledge for agricultural improvement.     

Benzyladenine-induced increase in DNA content in the nucleus of bean leaf cells

Primary leaves of bean plants (Phaseolus vulgaris L.) were treated with benzyladenine (BA) from 7 days after sowing. Nuclei were isolated and the DNA content per nucleus was determined. BA increased the amount of DNA per nucleus of untreated controls by about 50% without nuclear division.     

Tissue culture and wetland establishment of the freshwater monocots <Emphasis Type="Italic">Carex,Juncus, Scirpus</Emphasis>, and <Emphasis Type="Italic">Typha</Emphasis>

Summary Cell cultures of freshwater wetland monocots were regenerated, plants were grown in the greenhouse, and then established and evaluated in wetlands. Typha (cattail), Juncus (rushes), Scirpus (bulrushes), and Carex (sedges) were studied because they are common, dominant, high biomass wetland-adapted plants, tolerant of chemically diverse ecosystems. The goal was to define micropropagation and wetland establishment protocols. Tissue culture systems defined for numerous monocot crop species can be readily applied to wetland plants, with a few modifications. Issues addressed were selection of explant material, shoot and root regeneration conditions, culture age verses regenerability, greenhouse acclimatization needs, plant uniformity and requirements for wetland establishment. In vitro-germinated seedlings were an excellent source of pathogen-free regenerable tissue. T. latifolia, T. angustifolia, and J. accuminatus were regenerated from callus induced in the dark with picloram, then transferred to medium with benzyladenine in the light to promote shoot organogenesis. J. effusus, S. polyphyllus, and C. lurida could not be regenerated from callus, which turned black. They could be regenerated directly by culturing intact seedlings directly on cytokinin media in the light. Shoots rooted with little or no auxin. J. effusus rooting was promoted by the addition of charcoal to the medium. Covering plants for the first 2 wk with plastic facilitated greenhouse establishment. There were high rates of greenhouse and wetland survival. No abnormal plants were observed. These regeneration systems could be utilized for the production of wetland plants for potential application in habitat restoration and wetland creation, and would provide an alternative to field collection.     

Ethylene synthesis and growth in embryogenic tissue of Norway spruce: Effects of oxygen, 1-aminocyclopropane-1-carboxylic acid, benzyladenine and 2,4-dichlorophenoxyacetic acid

Ethylene production from an embryogenic culture of Norway spruce ( Picea abies L.) was generally low. ca 2.5 nl g⁻¹ h⁻¹, whereas 1-aminocyclopropane-1 -carboxylic acid (ACC) concentration was high, fluctuating between 50 and 500 nmol g⁻¹ during the 11-day incubation period. Hypoxia (2.5 and 5 kPa O₂) rapidly inhibited ethylene production without subsequent accumulation of ACC. Exogenous ACC (1, 10 and 100 μ M ) did not increase ethylene production, but the highest concentrations inhibited tissue growth. Ethylene (7 μl I⁻¹) did not inhibit growth either when supplied as ethephon in the medium or in a continuous flow system. Benzyladenine (BA) had little effect on ethylene production, although it was necessary for sustaining the ACC level. Omission of 2.4-dichloro-phenoxyacetic acid (2.4-D) from the medium caused ethylene production to increase from about 2.5 to 7 nl g⁻¹ h⁻¹ within the 11-day incubation period. Although 2.4-D did not specifically alter the endogenous level of ACC, the lowest ACC level, 33 nmol g⁻¹, was observed in tissue treated with 2.4-D (22.5 μ M ) and no BA for 11 days. Data from this treatment were used to estimate the kinetic constants for ACC oxidase, the apparent K_m was 50 μ M and V_max 2.7 nl g⁻¹ h⁻¹. Growth of the tissue was strongly inhibited by 2.4-D in the absence of BA, but weakly in the presence of BA (4.4 μ M ). The results suggest that ethylene or ACC may be involved in the induction of embryogenic tissue and in the early stages of embryo maturation.     

A comparison of the effects of mechanically-induced stress,ethephon and silver thiosulphate on the growth of cauliflower seedlings

Mechanically-induced stress (MIS) was applied to cauliflower seedlings by brushing with paper for 1.5 minutes each day. With the possible exception of the effect on leaf 1 thickness, none of the growth responses induced by MIS were significantly nullified by spraying the seedlings with the ethylene inhibitor, silver thiosulphate (STS). The ethylene-releasing compound, ethephon, induced some changes in cauliflower growth similar to those caused by MIS, such as reduced shoot and leaf 1 weight, a reduction in cotyledon and leaf 1 area, and an increase in the thickness of leaves 1 and 2. However, other effects of ethephon were different from those of brushing. Petiole length and diameter and the weight and area of leaf 2 were reduced by brushing but generally increased by ethephon. STS reversed, at least partially, most of the ethephon effects on growth, thus demonstrating that it acts as an inhibitor of ethylene action in cauliflower.The results are discussed in relation to the possible endogenous control of MIS growth responses by endogenous ethylene and auxin.     

Inhibition of the growth of apple pollen tubes by EDTA, surfactants and fungicides

The effects of EDTA (ethylene diamine tetraacetic acid), Kresoxim‐methyl, Wettable Sulphur 80 WP and the surfactants Triton X‐100, Triton X‐114, Dehydol TA 5 and Dehydol TA 29 on pollen tube growth of Malus domestica, Borkh. cvs ‘Golden Delicious’ and ‘Mondial Gala’ were determined by image analysis. The ED₅₀ ‐ and ED₉₀ ‐ values (effective doses giving 50% and 90% reduction of growth) of the chemicals, except for the surfactants, were determined. The selected test substances showed an inhibiting effect on pollen tube growth, with Kresoxim‐methyl being the most active. Surfactants are components of pesticides and they also caused inhibition of pollen tube growth but only at application rates more then 10‐fold higher than typically recommended. Pollen of both apple cultivars were almost equally sensitive to the chemicals.     

Experimental control of flowering and vivipary in timothy (Phleum pratense)

Environmental and hormonal control of flowering and vivipary in four Norwegian timothy ( Phleum pratense L.) cultivars has been studied in phytotron and by aseptic culture of inflorescence explants. The critical photoperiod for flowering increased with increasing temperature (12–18°C) and it was 13 to 15 h for the southern and 14 to 16.5 h for the northern cultivars. Diurnal temperature fluctuation significantly stimulated flower formation compared to the corresponding constant temperature treatment. Plants grown in 16-h photoperiod contained normal sexual flowers, but a high percentage of spikes developed in 12- or 14-h photoperiod contained viviparous plantlets. One- to four-weeks in continuous light before treatment with 12-h photoperiod increased the number of spikes per plant, but did not enhance the frequency of vivipary. Experiments with aseptic cultures showed that generative versus vegetative development of timothy inflorescence was affected by plant hormones. Kinetin stimulated the vegetative development and induced proliferation both in inflorescence initials and in spikelets isolated at heading time.     

Synergistic action of auxin and cytokinin mediates aluminum‐induced root growth inhibition in Arabidopsis

Auxin acts synergistically with cytokinin to control the shoot stem‐cell niche, while both hormones act antagonistically to maintain the root meristem. In aluminum (Al) stress‐induced root growth inhibition, auxin plays an important role. However, the role of cytokinin in this process is not well understood. In this study, we show that cytokinin enhances root growth inhibition under stress by mediating Al‐induced auxin signaling. Al stress triggers a local cytokinin response in the root‐apex transition zone (TZ) that depends on IPTs, which encode adenosine phosphate isopentenyltransferases and regulate cytokinin biosynthesis. IPTs are up‐regulated specifically in the root‐apex TZ in response to Al stress and promote local cytokinin biosynthesis and inhibition of root growth. The process of root growth inhibition is also controlled by ethylene signaling which acts upstream of auxin. In summary, different from the situation in the root meristem, auxin acts with cytokinin in a synergistic way to mediate aluminum‐induced root growth inhibition in Arabidopsis.     

Cytokinin metabolism in tomato plants. II. Metabolites of kinetin and benzyladenine in decapitated roots

Decapitated tomato plants were supplied via the roots with [8-¹⁴C]-kinetin or [8-¹⁴C]-benzyladenine in a nutrient solution for a period of 24 h. After this time the root material, the root sap produced during the 24 h period and the nutrient solution remaining at the end of the experiments were analysed for cytokinins. HPLC techniques and chemical treatments were used to tentatively identify radioactive metabolites formed. Uptake of kinetin and benzyladenine by the roots was found to be limited but once within the root tissues metabolism was both rapid and extensive.At least 14 metabolites of kinetin were recovered from root tissue and root sap. Many of these appeared to be degradation products. There was, however, some evidence of formation of zeatin-like derivatives. Side-chain cleavage of the original kinetin which occurs rapidly is suggested as a possible route for the eventual production of these endogenous cytokinin forms.The benzyladenine taken up by the roots was apparently both ribosylated and glucosylated. No unmetabolized benzyladenine was detected in the root tissues after 24 h. Only very low levels of radioactivity were associated with the retention time of adenine, suggesting that in the case of benzyladenine side-chain cleavage is of limited importance.The significance of these reactions in relation to the potential use of cytokinins in the regulation of plant growth is discussed.     

Effects of fruit thinning agents on apple tree canopy photosynthesis and dark respiration

Effects on photosynthesis of the fruit thinning agents naphthaleneaceticacid (NAA) and three commercial plant growth regulator formulations,naphthaleneacetic acid ('Rhodofix') and naphthaleneacetamide('Amidthin') and 2-chloroethylphosphonic acid('Ethrel')were evaluated with respect to the stress they impose on the fruit tree, usingthe alternate-bearing sensitive apple cv. 'Elstar'. This work wasbased on the hypothesis that plant stress in the form of large reductions inleaf photosynthesis are a pre-requisite for successful fruit thinning. A newtechnology was employed for continuous recording of tree canopyphotosynthesis, dark respiration and carbon balance of apple trees. This wasbased on six canopy chambers, which enclosed apple trees under naturalconditions in the field, with on-line measurements and continuous analysis ofCO₂ exchange and automated data acquisition. All employed thinningagents reduced whole tree canopy photosynthesis consistently by3–34% on the five days following their application, withphotosynthesis still declining thereafter in the case of the NAA and'Amid-thin' application. The reduction after application of either'Rhodofix' or 'Ethrel', declined within five days, suchthat most of the original photosynthetic potential was restored, indicatingacceptable phytotoxicity of these three plant growth regulators at theconcentrations used. The effects on dark respiration differed markedly. NAA and'Ethrel' increased dark respirationover-proportionally by up to 106%, whereas 'Amid-thin' and'Rhodofix' decreased it by up to 46%inthe first night after application, thereby drastically affecting the carbonbalance of the tree in opposite ways. These results are integrated into ahypothesis linking basipetal auxin transport, phloem loading, translocation anddeficiency of photoassimilates.     

The Effects of Phytohormones and 5-Azacytidine on Apoptosis in Etiolated Wheat Seedlings

The development of etiolated wheat (Triticum aestivum L.) seedlings is necessarily accompanied by apoptosis in their coleoptiles and first leaves. Internucleosome DNA fragmentation, which is characteristic of apoptosis, was detected in the coleoptile as soon as six days after germination. After eight days of germination, DNA fragmentation was clearly expressed in the coleoptile and was noticeable in the apical part of the first-leaf blade. Growing of intact seedlings or incubation of their shoots in the presence of such phytohormones as benzyladenine, gibberellin A₃, fusicoccin C, and 2,4-D at the concentration of 10^–5 M did not essentially affect DNA fragmentation in the coleoptile. As distinct from antioxidants, none of the phytohormones used prevented apoptosis in wheat seedlings. In contrast, ABA (10^–5 M) and an ethylene producer, ethrel (2-chloroethylphosphonic acid, 10^–2–10^–3 M), stimulated sharply DNA fragmentation in the coleoptile. An inhibitor of DNA methylation, 5-azacytidine, was very efficient in the stimulation of DNA fragmentation in the coleoptiles of eight-day-old seedlings at its concentration of 100 g/ml. Thus, some phytohormones can regulate apoptosis, and DNA methylation is involved in this process. Our results indicate that apoptosis activation by some phytohormones may be mediated by their regulation of DNA methylation/demethylation, which is responsible for the induction of genes encoding apoptogenic proteins and/or the repression of antiapoptotic genes.     

Effect of indole-3-acetic acid and kinetin on tuberisation parameters of different cultivars and transgenic lines of potato in vitro

The effect of indole-3-acetic acid or kinetin on the weight and numberof microtubers formed was studied on single node cuttings of sevendifferent potato (Solanum tuberosum L.) cultivars as well astransgenic lines harbouring rolB or rolC genes undercontrol of the patatin class I (B33) promoter. Plants were cultivatedin vitro in the dark on solidified MS medium containing 1 to8% sucrose with or without phytohormones. Most of thenontransformed potato cultivars and transgenic lines responded tohormone application by an increase in tuber yield. Auxin and cytokininacted differently: IAA increased predominantly the tuber size whilekinetin increased the number of tubers. RolC transformantsdisplayed an altered response to sucrose and especially to auxin. Thedegree of phytohormone effect on tuberisation parameters depended onsucrose content of the medium and potato genotype.