Effect of colchicine on the carbohydrate metabolism of mycelial felts of cunninghamella Sp.

Summary An experiment is reported in which 5 days old Cunninghamella spec. mats were incubated at 25°C over Richard's medium alone or together with colchicine. The results show that, up to 20 p.p.m., colchicine had no effect on dry weight and soluble sugar content of the fungal mats but caused an acceleration in the rate of sugar absorption and utilisation and polysaccharide accumulation especially the glucosans. 10 p.p.m. concentration further induced an increase in CO₂ production and synthesis of mononucleotides and nucleoproteins as indicated by excessive accumulation of conjugated pentoses and the pentosan fraction of polysaccharides.     

Colchicine after-effects on the absorption and utilisation of phosphates and nitrates by mycelial felts of Cunninghamella sp.

Summary Colchicine at 5 and 10 p.p.m. increased both phosphorus uptake and incorporation into organic forms (nucleoproteins or other simpler organophosphorus compounds). Continuous supply of colchicine at 20 p.p.m., almost checked phosphorus uptake during the second 24 hours of the experiment.The absorbed nitrates were utilised through the classical reduction steps. It appears also that colchicine had an inhibitory effect on the nitrite reductase that increased by increasing the concentration of the drug. Continuous supply of colchicine at its highest concentration (20 p.p.m.) checked completely the protein building during the second 24 hours of the experiment, though nitrate absorption continued; a phenomenon that caused the accumulation, in the tissue medium systems, of large amounts of peptide nitrogen.The mechanism of nitrate reduction as affected by colchicine treatment was fully discussed.     

Effects of colchicine and 2-Br-alpha-ergocryptine-methane-sulfonate (CB 154) on the release of prolactin and growth hormone by functional pituitary tumor cells in culture

The effects of colchicine and 2-Br-α-ergocryptine-methane-sulfonate (CB 154) on the release of prolactin and growth hormone have been studied in a clonal strain of rat pituitary tumor cells (GH₃) in monolayer culture. These cultures produce both prolactin and growth hormone and release both proteins spontaneously into the medium without storing them in large amounts. Immunological methods were used to measure both intracellular and extracellular concentrations of the hormones. Colchicine (5 × 10^?6 M for 3 hours) caused a 2- to 3-fold increase in intracellular concentrations of prolactin and growth hormone but, under basal conditions, had little or no measurable effect on the amounts of hormone accumulated in the medium during the course of the standard three hour treatment period. This latter finding evidently is due to a lag in the onset of drug action. Colchicine had little or no effect on accumulation of extracellular prolactin during the first two hours of treatment whereas such accumulation was depressed by over 60% during the third hour of treatment. Previous studies have shown that treatment of GH₃ cells with thyrotropin releasing hormone (TRH) and hydrocortisone (HC) increases both intra and extracellular levels of prolactin and growth hormone, respectively. In cultures treated with TRH (5 × 10^?8 M), colchicine (5 × 10^?6 M for 3 hours) increased intracellular prolactin by about 70% and decreased extracellular hormone by 10%. In cultures treated with HC (3 × 1O^?6 M), colchicine increased intracellular growth hormone by more than 100% and decreased medium concentrations of the hormone by 15%. Colchicine did not significantly alter total hormone (intracellular + extracellular) accumulation, cellular uptake of ³H-amino acids, or total cell protein synthesis. The synthetic ergot alkaloid, CB 154, (3.3 × 10^?6 M for 3 hours) caused an 80% increase in intracellular, and a nearly 50% decrease in extracellular, prolactin without affecting the accumulation of growth hormone, the uptake of ³H-labeled amino acids, or overall protein synthesis in the cultures. Elevation of medium potassium concentration from a basal value of 5.3 mM to 3–5 × 10^?2 M (by addition of KCl) decreased intracellular levels of prolactin by 85% and growth hormone by 55%. These effects of high potassium were blocked by colchicine and by CB 154. We conclude that colchicine, after a lag period of two hours, acts to inhibit the release of prolactin and growth hormone from GH₃ cells. By the end of three hours of treatment, this inhibition is over 60% complete in the case of prolactin. The qualitatively different effects of colchicine and CB 154 on prolactin and growth hormone release suggest that these two secretory blocking agents probably act on GH₃ cells by different mechanisms.     

Effects of anti-microtubule drugs onin vitro polymerization of tubulin from mung bean

Effects of anti-microbule drugs on tubulin polymerizationin vitro were investigated using purified mung bean (Vigna radiata) tubuli. Colchicine induced the formation of macrotubules at the relatively low concentration of 10 μM. and the appearance of corkscrew-like filaments from the ends of the macrotubules at concentrations of more than 100 μM. Vinblastine substantially inhibited polymerization at 1 μM and caused the formation of paracrystals at concentrations greater than 10 μM. Oryzalin inhibited polymerization at 1 μM partially and at 10 μM completely. Paracrystal formation was also induced by cremart at 10 μM, but these paracrystals appeared to be more rigid than those induced by vinblastine. Amiprophos methyl (APM), with a chemical configuration similar to cremart, substantially inhibited polymerization at 1 μM, but the formation of paracrystals was weak. Griseofulvin at 10 μMalso inhibited the polymerization of tubulin while at higher concentrations aggregates of helices were formed. Inhibition of polymerization by phenylcarbamate herbicides was more effective than that caused by benzimidazoylcarbamate fungicides. The effects of drugs onin vitro preformed (MTs) were also investigated. Colchicine and vinblastine showed identical effects to those on the polymerization process. Griseofulvin, cremart and APM induced only macrotubule formation while the other drugs tested had no major effects     

Potential role of ATP‐binding cassette transporters against acaricides in the brown dog tick Rhipicephalus sanguineus sensu lato

ATP‐binding cassette (ABC) transporters have been shown to be involved in pesticide detoxification in arthropod vectors and are thought to contribute to the development of drug resistance. Little is currently known about the role they play in ticks, which are among the more important vectors of human and animal pathogens. Here, the role of ABC transporters in the transport of fipronil and ivermectin acaricides in the tick Rhipicephalus sanguineus (Ixodida: Ixodidae) was investigated. Larvae were treated with acaricide alone and acaricide in combination with a sub‐lethal dose of the ABC transporter inhibitor cyclosporine A. The LC₅₀ doses and 95% confidence intervals (CIs) estimated by mortality data using probit analysis were 67.930 p.p.m. (95% CI 53.780–90.861) for fipronil and 3741 p.p.m. (95% CI 2857–4647) for ivermectin. The pre‐exposure of larvae to a sub‐lethal dose of cyclosporine A reduced the LC₅₀ dose of fipronil to 4.808 p.p.m. (95% CI 0.715–9.527) and that of ivermectin to 167 p.p.m. (95% CI 15–449), which increased toxicity by about 14‐ and 22‐fold, respectively. The comparison of mortality data for each separate acaricide concentration showed the synergic effect of cyclosporine A to be reduced at higher concentrations of acaricide. These results show for the first time a strong association between ABC transporters and acaricide detoxification in R.sanguineus s.l.     

Effects of indole‐3‐butyric acid on growth,pigments and UV‐screening compounds in Nostoc linckia

In the present research, the effect of indole‐3‐butyric acid (IBA) on the growth, and the production of some primary and secondary metabolites was studied in Nostoc linckia. In this respect, algae cultures were supplied with 0, 0.01, 0.1, 1, 10, and 100 μM IBA for 14 days. IBA at concentrations of 10 and 100 μM induced algal growth expressed as fresh weight in N. linckia. Treatment with IBA at all concentrations stimulated heterocyst formation. In addition, low concentrations of IBA (0.01, 0.1, and 1 μM) had a stimulatory effect on chlorophyll a and carotenoids accumulation. In contrast, higher concentrations of IBA induced the accumulation of phycocyanin, allophycocyanin, and phycoerythrin in the treated algae. In this case, IBA at the concentration of 10 μM was more effective. A significant decrease in protein content was observed in the algae treated by 0.01 μM IBA. All concentrations of IBA caused a decrease in sugar content, but lower concentrations were more effective. IBA application in all of the concentrations except 100 μM increased oligosaccharide‐linked mycosporine‐like amino acids (OS‐MAAs) content. Lower concentrations had a more significant effect on increasing OS‐MAAs content. However the concentrations of 10 and 100 μM IBA decreased scytonemin content. These results indicated the stimulatory impact of IBA on weight, heterocyst formation, and photosynthetic pigments in N. linckia.     

Impact of auxin-compounds produced by the antagonistic fungus Pythium oligandrum or the minor pathogen Pythium group F on plant growth

Plant growth promotion induced by the antagonistic fungus, Pythium oligandrum, is the result of a complex interaction which includes an indirect effect through control of pathogens in the rhizosphere and/or a direct one mediated by plant-induced resistance. The present study shows an increased plant growth associated with direct interaction between P. oligandrum and roots, which is mediated by a fungus-produced auxin compound, tryptamine (TNH₂). In vitro experiments provided evidence that P. oligandrum metabolised specifically indole derivatives, such as tryptophan and indole-3-acetaldehyde, to produce THN₂ through the tryptamine pathway. When P. oligandrum grew in sterile root exudates, it also produced an auxin-like compound. Additional experiments on P. oligandrum–root interaction showed that, in amended nutrient solution of plants, the antagonist metabolised Trp into TNH₂ and that root absorption of this newly formed auxin-compound in appropriate concentrations was associated with enhancement of plant growth. This phenomenon was observed only when nutrient solution was amended with low tryptophan (Trp) concentrations, i.e. 0.05 and 0.1 mM; higher concentration (0.5 and 1 mM Trp) induced abnormal root development. Similar experiments were performed with Pythium group F, a minor pathogen known for its ability to produce auxin-compounds through the tryptamine pathway. In this case, irregular root development was always noticed with all Trp concentrations added to the nutrient solution of plants. Moreover, Pythium group F colonization of roots was associated with leakage of auxin-compounds in the nutrient solution. Our results, therefore, highlight that the production of similar auxin-compounds by two Pythium species has contrary effects on plant development.     

Antioxidant enzyme activities and hormonal status in response to Cd stress in the wetland halophyte Kosteletzkya virginica under saline conditions

Salt marshes constitute major sinks for heavy metal accumulation but the precise impact of salinity on heavy metal toxicity for halophyte plant species remains largely unknown. Young seedlings of Kosteletzkya virginica were exposed during 3 weeks in nutrient solution to Cd 5 µM in the presence or absence of 50 mM NaCl. Cadmium (Cd) reduced growth and shoot water content and had major detrimental effect on maximum quantum efficiency (F_v/F_m), effective quantum yield of photosystem II (Y(II)) and electron transport rates (ETRs). Cd induced an oxidative stress in relation to an increase in O₂^?? and H₂O₂ concentration and lead to a decrease in endogenous glutathione (GSH) and α‐tocopherol in the leaves. Cd not only increased leaf zeatin and zeatin riboside concentration but also increased the senescing compounds 1‐aminocyclopropane‐1‐carboxylic acid (ACC) and abscisic acid (ABA). Salinity reduced Cd accumulation already after 1 week of stress but was unable to restore shoot growth and thus did not induce any dilution effect. Salinity delayed the Cd‐induced leaf senescence: NaCl reduced the deleterious impact of Cd on photosynthesis apparatus through an improvement of F_v/F_m, Y(II) and ETR. Salt reduced oxidative stress in Cd‐treated plants through an increase in GSH, α‐tocopherol and ascorbic acid synthesis and an increase in glutathione reductase (EC 1.6.4.2) activity. Additional salt reduced ACC and ABA accumulation in Cd+NaCl‐treated leaves comparing to Cd alone. It is concluded that salinity affords efficient protection against Cd to the halophyte species K. virginica, in relation to an improved management of oxidative stress and hormonal status.     

Effect of zinc and cadmium on physiological and production characteristics in <Emphasis Type="Italic">Matricaria recutita</Emphasis>

Effects of zinc (12–180 μM) alone and in mixtures with 12 μM Cd on metal accumulation, dry masses of roots and shoots, root respiration rate, variable to maximum fluorescence ratio (F_V/F_M), and content of photosynthetic pigments were studied in hydroponically cultivated chamomile (Matricaria recutita) plants. The content of Zn in roots and shoots increased with the increasing external Zn concentration and its accumulation in the roots was higher than that in the shoots. While at lower Zn concentrations (12 and 60 μM) the presence of 12 μM Cd decreased Zn accumulation in the roots, treatment with 120 and 180 μM Zn together with 12 μM Cd caused enhancement of Zn content in the root. Presence of Zn (12–120 μM) decreased Cd accumulation in roots. On the other hand, Cd content in the shoots of plants treated with Zn + Cd exceeded that in the plants treated only with 12 μM Cd. Only higher Zn concentrations (120 and 180 μM) and Zn + Cd mixtures negatively influenced dry mass, chlorophyll (Chl) and carotenoid content, F_V/F_M and root respiration rate. Chl b was reduced to a higher extent than Chl a.     

Interactions in the Physiological Effects of Growth Substances on Plant Development

The interrelated effects of 3-indolylacetic acid and sodium2 : 4-dichlorophenoxyacetate on the growth and development ofLemna minor and Helianthus annuus have been investigated bymeans of multifactorial experiments, involving three to fivelevels of each compound. A clonal population of L. minor wasmaintained under constant conditions of light (550 foot-candles)and temperature (25 C.) and the growth regulators were addedto a phosphate-buffered culture solution (pH 51), which waschanged every 2 days. In the H. annuus experiments the compounds,in aqueous solution, were placed by means of a micro-pipetteon the leaves of young plants grown in a greenhouse or in theopen. Indolylacetic acid, at concentrations up to 25 p.p.m., increasesthe relative growth rate of L. minor on either a dry-weightor frond-area basis. Sodium dichlorophenoxyacetate at 0025p.p.m. has little effect on growth, but at and above concentrationsof 02 p.p.m. growth is depressed. The ratio of frond area tototal dry weight is increased by indolylacetic acid but is depressedby the phenoxyacetate. In the presence of indolylacetic acid,more particularly at the higher levels, the relative inhibitionof growth caused by sodium dichlorophenoxyacetate is reduced. When sunflowers are treated daily with indolylacetic acid (0,6, 30, 150 µg./ plant) or initially with sodium dichlorophenoxyacetate(0, 20, 100 µ.g./plant) in all possible combinations,then the relative growth rates of the treated leaves, the leavesabove the treated leaves, the first internode, and the rootsare depressed when each compound is applied alone. On the otherhand, the growth rate of the hypocotyl is increased by bothgrowth regulators, while for the second internode indolylaceticacid decreases the growth rate but the phenoxyacetate at thelower level accelerates it. The combined action of the two compoundson the different parts tends to be cumulative, save for thehypocotyl, where the effect of indolylacetic acid is dependenton the level of the phenoxyacetate. It is concluded that these results cannot be explained on theconcept of wholly additive effects. There is evidence that thecompounds can mutually interfere in their actions and it wouldseem that more than one mechanism is involved. Many of the resultscan be satisfactorily interpreted on the basis that the twocompounds act on a common system and that they compete withone another, but the results are not compatible with the postulatethat the dichlorophenoxyacetic acid directly inactivates indolylaceticacid.     

Fungal Inoculation and Elevated CO2 Mediate Growth of Lolium Mutiforum and Phytolacca Americana,Metal Uptake,and Metal Bioavailability in Metal-Contaminated Soil: Evidence from DGT Measurement

Fungal inoculation and elevated CO₂ may mediate plant growth and uptake of heavy metals, but little evidence from Diffusive Gradients in Thin-films (DGT) measurement has been obtained to characterize the process. Lolium mutiforum and Phytolacca americana were grown at ambient and elevated CO₂ on naturally Cd and Pb contaminated soils inoculated with and without Trichoderma asperellum strain C3 or Penicillium chrysogenum strain D4, to investigate plant growth, metal uptake, and metal bioavailability responses. Fungal inoculation increased plant biomass and shoot/root Cd and Pb concentrations. Elevated CO₂ significantly increased plants biomass, but decreased Cd and Pb concentrations in shoot/root to various extents, leading to a metal dilution phenomenon. Total Cd and Pb uptake by plants, and DGT-measured Cd and Pb concentrations in rhizosphere soils, were higher in all fungal inoculation and elevated CO₂ treatments than control treatments, with the combined treatments having more influence than either treatment alone. Metal dilution phenomenon occurred because the increase in DGT-measured bioavailable metal pools in plant rhizosphere due to elevated CO₂ was unable to match the increase in requirement for plant uptake of metals due to plant biomass increase.     

Investigations on Cytokinins. I. Effect of 6-Benzylaminopurine on Growth and Starch Content of Lemna minor

An excessive starch accumulation in the fronds of Lemna minor L. was observed microscopically and confirmed by direct starch determinations after application of growth inhibiting 6-benzylaminopurine concentrations. As a high growth rate was coupled with a low starch content and vice versa, the influence of the cytokinin on growth and starch content was compared. It appeared that apart from the expected influence of the growth rate on the starch content, 6-benzylaminopurine favoured starch accumulation also by itself.     

Effect of lanthanide chloride on photosynthesis and dry matter accumulation in tobacco seedlings

To investigate the physiological effects of rare earth ions, we have studied the effect of LaCl₃ on the photosynthetic light reactions in tobacco (Nicotiana tobacum). When treated with 5–20 mg/L LaCl₃ in Hoagland solution by water culture, the dry matter accumulation of different parts in tobacco, the content of chlorophyll increased gradually, but decreased when the concentration of LaCl₃ was ≥ 50 mg/L. The optimum concentration for growth appeared to be about 20 mg/L of LaCl₃ in nutrient solution. La³⁺ promoted the activities of the Hill reaction, Mg²⁺-ATPase, and stimulated the rate of photophosphorylation in chloroplast at low concentrations, but inhibited them at high concentrations. It is concluded that La³⁺ stimulated the growth of tobacco seedlings and accelerated the photosynthetic light reactions at suitable concentration in vivo.     

The Relative Toxicity of Manganese and Cobalt to the Tomato Plant

1. At equivalent concentrations, cobalt was more effective thanmanganese in inducing chlorosis and necrosis in tomato plants. 2. Chlorotic plants suffering from either cobalt or manganesetoxicity alone, or from the two conditions together, containedas much or more iron than normal plants, and chlorotic leavesresponded positively to painting with a solution of ferroussulphate. 3. When cultures were supplied with 5 p.p.m. Mn, distinct injuryof the lower leaves occurred regardless of the cobalt supply,but the addition of cobalt up to 0·01 p.p.m. broughtabout a gradual decrease in the chlorosis of the top leavesand an increase in dry weight. 4. With cobalt at the 0·1 p.p.m. level, severe chlorosisand necrosis occurred and growth was stunted, but by raisingthe manganese level to 5·0 p.p.m. a substantial antidotingeffect on the toxicity was obtained. 5. Cobalt and manganese had an antidoting effect on each other,whereby at high manganese supplies small quantities of cobaltalleviated manganese toxicity, while toxic concentrations ofcobalt were antidoted by high levels of manganese.     

High frequency in vitro propagation of <Emphasis Type="Italic">Trichopus zeylanicus</Emphasis> subsp. <Emphasis Type="Italic">travancoricus</Emphasis> using branch–petiole explants

Trichopus zeylanicus subsp. travancoricus (known as Arogyapacha), an endangered ethnomedicinal plant of the Western Ghats of South India, serves as the major source of the commercial drug Jeevani. The present study established a long-term high frequency in vitro propagation protocol for Arogyapacha. Callus obtained from the branch–petiole explants cultured on Murashige and Skoog (MS) medium with 4.5 μM 2,4-dichlorophenoxyacetic acid upon subculture to medium with different concentrations of 6-benzyladenine (BA) either alone or in combination with an auxin favoured shoot morphogenesis. Medium with 13.3 μM BA alone facilitated high frequency shoot bud (mean of 93.2) formation. Medium with lower concentrations of BA (4.4, 6.6 and 8.8 μM) alone or in combination with lower concentration of α-naphthaleneacetic acid (NAA) or indole-3-butyric acid (IBA) favoured better shoot growth than 13.3 μM BA containing medium, but with reduced number of shoot buds. Subsequent cultures on medium with lower concentrations of BA and also on MS basal media facilitated shoot formation as well as growth of shoots. The shoot regeneration potential showed no decline up to 5 years. Culture of the in vitro-derived whole branch–leaf explants on MS basal medium developed shoots directly from the node. On medium with 19.6 μM IBA, the whole branch–leaf explants induced nodular callus from the node, which developed shoots later. Subsequent cultures on medium with BA exhibited high frequency shoot formation. The transfer of shoots after 10–15 days culture on half-strength MS medium containing 2.7 μM NAA to half-strength basal medium induced a mean of 11.3 roots. Field survival of plantlets relied on the soil mix: a 1:4 ratio of sand and red-soil exhibited the highest plantlets survival (86.6%). RAPD profile of the source plant and plants regenerated from calli after 4 years showed no polymorphism. The established plantlets with morpho-floral features similar to that of the source plants flowered normally and set fruits.     

Biphasic Effect of Transforming Growth Factor-β1 on in Vitro Angiogenesis

Although the existence of an increasing number of angiogenesis-regulating cytokines is well documented, the response elicited by combinations of these cytokines is largely unknown. Using an in vitro model in which microvascular endothelial cells can be induced to form capillary-like tubes within three-dimensional collagen or fibrin gels, we have investigated the effect of transforming growth factor-β₁ (TGF-β₁) on basic fibroblast growth factor (bFGF)-induced and vascular endothelial growth factor (VEGF)-induced angiogenesis. Endothelial cell invasion and capillary lumen formation were inhibited by TGF-β1 at relatively high concentrations (5-10 ng/ml), while lower concentrations (100 pg/ml-1 ng/ml) of TGF-β1 potentiated the effect of bFGF- and VEGF-induced invasion. The optimal potentiating effect was observed at 200-500 pg/ml TGF-β1. At invasion-potentiating doses of TGF-beta;1, lumen size in fibrin gels was markedly reduced compared to that in cultures treated with bFGF alone. These results show that TGF-β1 exerts a biphasic effect on bFGF- and VEGF-induced angiogenesis in vitro. Our studies support the notion that the nature of the angiogenic response elicited by a specific cytokine is contextual, i.e., depends on the presence and concentration of other cytokines in the pericellular environment of the responding endothelial cell.     

Hg and Cd Induced Changes in Proline Content and Activities of Proline Biosynthesizing Enzymes in Phaseolus Aureus and Triticum Aestivum

The effect of mercury and cadmium, in the form of HgCl₂ and CdCl₂ respectively, on proline accumulation and two key proline biosynthesizing enzymes, ¹-pyrroline-5-carboxylate synthetase (P5CS) and ¹-pyrroline-5-carboxylate reductase (P5CR), was investigated in Phaseolus aureus Roxb. and Triticum aestivum L. The 5-d-old seedlings were exposed to 0.05, 0.1, 0.2 or 0.4 mM concentrations of the metals in Hoagland solution for 12 and 36 h. T. aestivum exhibited considerably greater accumulation of proline than P. aureus in response to the metal treatment. Among the two metals, Hg induced greater accumulation of proline than Cd. The activity of P5CS increased significantly in response to the metal treatment, particularly in T. aestivum in which the activity of the enzyme in the control was much higher than that was in P. aureus. The activity of P5CR on the other hand mostly decreased in response to the metal treatment. The study indicated a strong dependence of the metal induced proline accumulation on the constitutive P5CS content of the plants.     

The effect of plant growth regulators on growth, morphology and condensed tannin accumulation in transformed root cultures of Lotus corniculatus

This study concerns the effects of four different classes of plant growth regulators on root morphology, patterns of growth and condensed tannin accumulation in transgenic root cultures of Lotus corniculatus L. (Bird's-foot trefoil). Growth of transformed roots in 2,4-dichlorophenoxyacetic acid (2,4-D) resulted in decreased tannin levels relative to controls at concentrations of 10^-6 M and above, while gibberellic acid (GA₃) inhibited tannin accumulation at concentrations of 10^-7 M and above. Benzyladenine (BA) had little effect at low concentrations (10^-7 M and below) but resulted in an increase in tannin levels at 10^-6 M. Abscisic acid had little effect on levels of condensed tannins at any of the concentrations used. Experiments involving growth regulator addition and medium transfer demonstrated that 2,4-D inhibition of tannin accumulation could be reversed by GA₃ and BA, while GA₃ downregulation could only be reversed by the addition of 2,4-D. Although 2,4-D inhibited tannin accumulation, addition of 2,4-D to root cultures grown for 14 or 28 days in the absence of plant growth regulators stimulated both growth and tannin biosynthesis. Characteristic alterations in root morphologies accompanied growth regulator-mediated modulation of tannin biosynthesis. Growth in 2,4-D resulted in partially de-differentiated root cultures while growth in GA₃ produced roots with an elongated phenotype. Restoration of tannin biosynthesis in 2,4-D-treated roots was accompanied by root re-differentiation and the production of new lateral roots.Abbreviations ABA abscisic acid - BA benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid 3 - FW fresh weight