Activity of Various Growth Substances in the Avena First Internode Test

The elongation growth of the Avena first internode segments was studied in the presence of one or several of the following growth substances: indoleacetic acid (IAA), 6-fur-furylamino purine (FAP, kinetin), 6-benzylamino purine (BAP), gibberellin A₃ (GA₃) and A₄₊₇ (GA₄₊₇), and abscisic acid (ABA). The cytokinins at concentrations of 10^?7 to 10^?6M stimulated growth with 4 to 6 per cent but this effect was not statistically significant. Concentrations higher than 5 × 10^?6M inhibited growth. FAP and BAP (from 10^?8M to 10^?6M) had no significant interaction with any other growth substance used. The two-factor interactions of IAA × ABA, IAA × GA₃, and GA₃× ABA, as well as the three-factor interaction IAA × ABA × GA₃ were significant. However, the IAA × ABA interaction was significant only when high concentration (10^?6M) of ABA was used. The growth inhibition produced by 10^?7 and 10^?6M ABA was overcome by about equimolar concentrations of IAA. The stimulation of growth by GA₃ and GA₄₊₇ (10^?9 to 10^?7M) was prevented by simultaneous application of ABA, and it was reduced significantly by application of IAA (10^?7 to 10^?8M). GA3 at 10^?8M combined with different concentrations of IAA gave slightly higher elongation than IAA alone but the observed values were significantly lower than expected assuming independent additive action.     

Effects of Organic Nutrients and Hormones on Growth and Development of Tissue Explants from Coconut (Cocos nucifera) and Date (Phoenix dactylifera) Palms Cultured in vitro

The growth response (increase in weight) of cultured explants from seedling date (Phoenix dactylifera L.) and mature coconut (Cocos nucifera L. cv. Malayan Dwarf) palms to source and concentration of organic nitrogen. carbohydrate, auxins, cytokinins and gibberellins was examined. Growth was strongly stimulated by the presence of auxins (10^?7 to 10^?6M), cytokinins (10^?6 to 10^?5M), high concentrations of sucrose (0.2 M), and in the absence of NH₄Cl, by organic sources of reduced nitrogen. Higher concentrations of auxin (2,4-D or NAA at 10^?6 to 10^?5M) which still stimulated growth of Phoenix tissue, proved inhibitory to growth of freshly excised Cocos tissues. Explants from both palms initiated roots when subcultured on a medium with increased levels of auxin (NAA, 2.5 × 10^?6 to 2.5 × 10^?5M) and reduced levels of cytokinin (6-BAP, 5 × 10^?8M). Isolated roots excised from these explants continued growth and produced new laterals when subcultured on media with GA₃ (5 × 10^?7M) and reduced levels of auxin, cytokinin, and either minerals or sucrose.     

The Effect of Exogenous IAA and Kinetin on Nitrate Reductase,Nitrite Reductase and Glutamate Dehydrogenase Activities in Excised Pea Roots

Nitrate reductase (NO₃R) activity, nitrite reductase (NO₂R) activity and NADH₂ dependent glutamate dehydrogenase (GDH) activity were followed in extracts from excised pea roots incubated under aseptic conditions for 9 and 24 h in nitrate containing nutrient medium to which IAA was added in concentrations promoting lateral root formation (1 × 10^?5; 3 × 10^?5; 5 × 10^?5 M) and kinetin in concentrations which reduce lateral root formation (0.1; 1; 5 mg 1^?1, that is 4.65 × 10^?7;4.65 × 10^?6 and 2.3 × 10^?5 M). NO₃R activity was not influenced by IAA, NO₂R activity was slightly depressed by IAA after 24 h incubation and GDH activity was slightly increased after 24 h incubation in the presence of IAA. Kinetin decreased NO₃R activity significantly both after 9 h and 24 h incubation, slightly increased NO₂R activity after 9 h incubation but slightly decreased it after 24 h incubation, and did not affect GDH activity after 24 h incubation. However, when applied together with IAA, kinetin abolished the promoting effect of IAA on GDH activity. IAA neither reversed nor accentuated the effect of kinetin on NO₂R activity. Nevertheless the depressing effect of kinetin on NO₃R activity was emphasized by the presence of IAA after 9 h incubation. The results obtained indicate that reduced nitrate assimilation due to the depression of nitrate reductase activity caused by kinetin probably contributes to the negative growth effect of kinetin in pea root segments grown in nitrate medium.     

Root Growth Activity of Barban in Relation to Auxin and Other Growth Factors

The effect of barban (4-Cl-2-butynyl-N-3-Cl-phenylcarbamate) on the growth of roots of wheat seedlings has been studied. In concentrations of 10^?7 to 5 · 10^?7M barban causes rapid inhibitions of cytokineses and cell elongation, the effects of which are spontaneously reversible. The reversion of the meristem inhibition is enhanced by thymidylic acid and indole-3-acetic acid (IAA). Initiation of cell elongation is slowed down or ceases during cytostasis; its reversal, on the other hand, is promoted by IAA and kinetin but inhibited by Fe. The final cell elongation attained is strongly reduced by barban and reversed under transient aberrant elongation. This inhibition and the recovery appear both to be additive to cell elongation actions of auxin and antiauxin but reversed by nucleic acid components. The inhibition of elongation is increased by Fe. The following explanation for this phenomenon is suggested: the primary effect of barban is known to be the blocking of metaphases under anaesthesis; this blocking then leads to reduced activation of IAA, kinetin and other metabolites. Auxin is required for cell divisions and initiation of elongation: the apical root growth equals in this respect that of shoot apices and lateral meristems. Initiation of cell elongation is closely dependent upon metabolites produced in dividing meristematic cells, whereas the limitation of cell stretching is independent of the meristem activity. No explanation is offered for the role of Fe.     

Cytokinin Reversal of Abscisic Acid Inhibition of Growth and α-Amylase Synthesis in Barley Seed

The effect of cytokinin, kinetin, on abscisic acid (dormin) inhibition of α-amylase synthesis and growth in intact barley seed was investigated. Abscisic acid at 5 × 10^?5M nearly completely inhibited growth response and α-amylase synthesis in barley seed. Kinetin reversed to a large extent abscisic acid inhibition of α-aniylase synthesis and coleoptile growth. The response curves of α-amylase synthesis and coleoptile growth in presence of a fixed amount of abscisic acid (6 × l0^?6M) and increasing concentrations of kinetin (from 5 × l0^?7M to 5 × 10^?5 M) showed remarkable similarity. Kinetin and abscisic acid caused synergistic inhibition of root growth. Gibberellic acid was far less effective than kinetin in reversing abscisic acid inhibition of α-amylase synthesis and coleoptile growth. A combination of kinetin and gibberellic acid caused nearly complete reversal of abscisic acid inhibition of α-amylase synthesis but not the abscisic acid inhibition of growth. The results suggest that factors controlling α-amylase synthesis may not have a dominant role in all growth responses of the seed. Kinetin possibly acts by removing the abscisic acid inhibition of enzyme specific sites thereby allowing gibberellic acid to function to produce α-amylase.     

Stimulation of Growth of Peppermint (Mentha piperita) by Phosfon, a Growth Retardant

The shoot growth and fresh weight of Mentha piperita grown in soil were stimulated at concentrations of 1.26 × 10^?5M to 7.77 × 10^?4M phosfon (2,4-dichlorobenzyl tributyl phosphonium chloride) while higher concentrations resulted in retardation of growth. Concentrations of 6.30 × 10^?7M to 3.78 × 10^?5M caused retardation of growth in mineral nutrient solution, and even death at the highest concentrations. However, when the M. piperita plants were grown in mineral nutrient solutions at concentrations of phosfon which had been sequentially lowered from 2.52 × 10^?8M to 2.52 × 10^?12M, the shoot growth and fresh weight were stimulated as in the case of plants grown in phosfon treated soil.     

Induction of apogamy inEquisetum arvense

InEquisetum arvense, apogamous sporophytes were produced on medium containing 5×10^?6–5×10^?8 g/ml kinetin. NAA, IAA, GA₃, glucose and saccharose were ineffective for the induction of apogamy. On medium containing 5×10^?7–5×10^?8 g/ml kinetin, the gametophytes passed into sporophytic structures directly. On medium containing 5×10^?6 g/ml kinetin, some gametophytes passed into sporophytic structures directly, and others became a callus-like cell mass from which an apogamoun shoot arose. The results of the morphological observations on them were reported and compared with the sexually produced sporophyes. The apogamous sporophytes induced by 5×10^?7 g/ml kinetin were haploid in their nuclear phase and some of those induced by 5×10^?6 g/ml kinetin had a tendency to become diploid.     

The Uptake of Cytokinins by Protoplasts and Root Sections of Brassica campestris

The effect of cytokinins was studied on the incorporation of ¹⁴C-labelled precursors into the nucleic acid fraction of protoplasts isolated from callus or roots of Brassica campestris. Protoplasts from callus and roots took up ¹⁴C-uridine from the incubation medium and incorporated this precursor into the ribonucleic acid fraction during the experimental period of 16 h. Low concentrations of kinetin (10^?8-5 × 10^?6M) did not stimulate the incorporation, and kinetin inhibited this process at higher concentrations (5 × 10^?5M). This result led to an investigation on the uptake of cytokinins by protoplasts of roots. In contrast to a rapid uptake of radio-actively labelled adenine and uridine. protoplasts from roots took up only small amounts of labelled kinetin. zeatin, zeatin riboside and zeatin nucleotides from the incubation medium. Root sections took up far more adenine and kinetin than protoplasts from roots. The ratio between the amount of kinetin taken up and applied was much higher for the sections than for protoplasts, indicating that intact root cells took up kinetin far more rapidly than protoplasts. It is suggested that the plasmalemma and cell wall play an essential role in the uptake of cytokinins or that the differences in the uptake rates are related to differences between the rates of metabolism of cytokinins in root sections and in protoplasts.     

A Requirement for DNA Synthesis during Auxin Induction of Cell Enlargement in Tobacco Pith Tissue

IAA (indoleacetic acid) is known to induce cell enlargement without cell division in tobacco pith explants grown on an agar medium without added cytokinin. The very long lag period before IAA (2 × 10^?5M) stimulates growth, about 3 days, can be useful to study the metabolic changes which lead to the promotion of growth. When the disks are transferred to a medium without IAA after 2 days or less of treatment with IAA, the IAA does not stimulate growth. Disks transferred after 3 days, subsequently show an auxin response, almost as great as those given IAA continuously. At 5 × 10^?4M, 5-fluorodeoxyuridine (FUDR), which inhibits DNA synthesis by blocking formation of thymidylate, completely suppresses the lAA-induced growth if it is added together with the IAA or 1 day later. When the FUDR is given 2 days after the IAA, there is a small increment of auxin-induced growth, and an even greater amount if added after 3 days. The period when exogenous auxin must be present to stimulate growth corresponds to the period of FUDR sensitivity. The FUDR inhibition is prevented by thymidine but not by uridine. Other inhibitors of DNA synthesis, hydroxyurea and fluorouracil, also inhibit auxin-induced growth. Thus DNA synthesis seems to be required for auxin induction of cell enlargement in tobacco pith explants. In contrast, FUDR does not inhibit auxin-induced growth in corn coleoptile and artichoke tuber sections.     

Seasonal Effects in the Hormonal Control of Growth in the Submerged Aquatic Macrophyte Ceratophyllum demersum

The season dependent changes in growth response to treatment with auxin or gibberellin were studied in the aquatic macrophyte Ceratophyllum demersum. Control plants show, under experimental conditions, a maximum growth in length in February. In the same period most of the lateral buds appear. Growth of the lateral buds occurs later. IAA causes a stimulation of growth in length from late November or December until February, in concentrations of 10^?9M and 10^?6M. There is almost no stimulation of lateral bud formation by IAA, only a slight increase from late November until December occurs by the lowest concentrations used. The highest concentration used, 10^?4M, is in most cases supraoptimal for lateral bud formation; only when plants become dormant (August), this high dose may stimulate the process. GA₃, in concentrations of 10^?9, 10^?6 or 10^?4M, exhibits a dose dependent increase of the response with respect to growth in length and lateral bud formation. The response occurs earlier than that for IAA: already early in November, or December, until February. Growth of the lateral buds may show only a slight stimulation by IAA as well as GA in winter. From February until April all GA concentrations used could cause a small increase of the growth of sprouts. In the case of IAA, however, only the lowest concentration could cause a small increase.     

The Interrelation of RNA, Auxin and Auxin-oxidases in Lentil Roots

The correlation between auxin and RNA metabolism was investigated in lentil roots. IAA and NAA both cause a considerable rise in the RNA level of germinating lentil roots, though no effect of IAA was found on the DNA level. In untreated germinating roots various sections were isolated and a direct relation found between RNA and auxin content, and an indirect relation between RNA content and auxin oxidase activity. In excised roots, incubated for 24 hours, the loss of RNA is paralleled by a loss of endogenous auxin. Excised roots treated with 10^?4M IAA or M 10^?4 NAA loose little RNA. The findings suggest that in lentil roots the RNA levels may be controlled by auxin levels, which in turn may be controlled by the levels of auxin oxidase.     

Involvement of cytokinin in the stimulation of nodulation by low concentrations of ammonium in Pisum sativum

Nodulation in pea (Pisum sativum L.) grown in hydroponic and sand culture systems is stimulated by low concentrations (<1.0 mM) of ammonium, but the physiological mechanisms underlying this stimulation are unknown. The current study involves a series of experiments, which investigate if the ammonium‐induced stimulation of nodulation involves changes in endogenous hormone (auxin and cytokinin) levels. P. sativum L. cv. Express was grown in growth pouches for 1 week with mineral N (0.5 and 2.0 mM NH₄⁺ or NO₃^–) or for 3 weeks exposed to exogenous indole‐3‐acetic acid (IAA) or 6‐benzylaminopurine (BAP) at a range of concentrations (10^‐9?10^‐5 M). Ammonium enhanced nodulation on the basis of both early whole plant (nodules plant^?1) and specific nodulation (nodules g^?1 root DW), especially in 0.5 mM treatment in which nodulation was approximately 4‐fold of the mineral‐N‐free control 1 week after inoculation. Correspondingly, the roots treated with ammonium contained much higher levels of t‐zeatin (Z) and lower t‐zeatin riboside (ZR) than that the control or nitrate‐treated plants. There was no significant difference in IAA levels between the control and ammonium treatments. Exogenous application of BAP for 3 weeks at concentrations of 10^‐7?10^‐5 M strongly inhibited nodulation. However, 10^?9 M BAP, but not IAA, significantly enhanced nodulation. These data support the theory that a relatively high ratio of cytokinin:auxin in roots is favourable for nodule initiation, but that an excessively high level of cytokinin inhibits nodulation. Based on these results we propose that stimulation of nodulation by low concentrations of ammonium may be mediated through increasing Z level in roots, which alters the balance of cytokinin and auxin, which in turn induces cortical cell divisions leading to nodule initiation.     

Inhibition of Lettuce Seed Germination and Root Elongation by Derivatives of Auxin and Reversal by Derivatives of Cycocel

Lettuce seed germination or lettuce root elongation after germination in water was inhibited by 5.7 × 10^-6M indoleacetic acid (IAA) and designated other IAA derivatives. These IAA-inhibited growth responses were reversed by 10^-5 to 10^-6M Cycocel, (2-chloroethyl) trimethylammonium chloride, which alone was without effect. Only those Cycocel analogs, which have previously been shown to be active as plant growth retardants, were effective in reversing IAA inhibition of germination or root elongation. These results are consistent with the concept that Cycocel at low concentrations acts as an auxin antagonist. However. Cycocel did not reverse the inhibitory effects from indole-3-propionic acid or indole-3-butyric acid.     

Differential IAA dose response relations of the axr1 and axr2 mutants of Arabidopsis

In comparison to wild type Arabidopsis thaliana, the auxin resistant mutants axr1 and axr2 exhibit reduced inhibition of root elongation in response to auxins. Several auxin-regulated physiological processes are also altered in the mutant plants. When wild-type, axr1 and axr2 seedlings were grown in darkness on media containing indoleacetic acid (IAA), promotion of root growth was observed at low concentrations of IAA (10^?11 to 10^?7M) in 5-day-old axr2 seedlings, but not in axr1 or wild-type seedlings. In axr1 there was little or no measurable root growth response over the same concentration range. In wild type, root growth was inhibited at concentrations greater than 10^?10M and no detectable root growth response was observed at lower concentrations. In addition, production of lateral roots in response to IAA increased in axr2 seedlings and decreased in axr1 seedlings relative to wild type. Promotion of root elongation and initiation of lateral roots in axr2 seedlings in response to auxin indicate that axr2 seedlings are able to perceive and respond to IAA.     

The Influence of pH on the Cadmium-Repressed Growth of the Alga Coelastrum proboscideum

The inhibition of growth by different concentrations of CdCl₂ in the range 4,5 × 10^?7 to 5.6 × 10^?7M was studied in the green alga Coelastrum proboscideum Bohlin in inorganic media at pH 4.3, 5.3 and 6.2. The factorial destgn of the experiments was evaluated as an analysis of 2² factors. Below pH 4.0 and above pH 6.5 growth was depressed without adding Cd. Cd concentrations exceeding 5.6 × 10^?8M reduced algal growth significantly with a 50% inhibition at 5.6 × 10^?7M Cd. The Cd concentration of 5.6 × 10^?7M was less toxic at pH 6.2 than at pH 5.3 and 4.3, thus revealing a negative interaction between protons and Cd.     

The Role of Auxin Metabolism in Root Meristem Regeneration by Pinus lambertiana Embryo Cuttings

Since the existence of root promoting substances that consist of a complex between auxin and another molecule has been suggested, we have examined the role of auxin conversion products in root regeneration by Pinus lambertiana embryo cuttings. Auxin conversion products were detected using radioactive forms of the auxins IAA (indoIe-3-acetic acid), NAA (a-napthaleneacetic acid) and 2,4-D (2,4-dichlorophenoxyacetic acid). 10^?7M NAA was more effective than 10^?6M IAA at promoting rooting, yet it formed conversion products much less rapidly. Also continuous exposure to IAA was necessary for optimum root formation. Based on these and other findings, we conclude that free auxin, and not the conversion products we detected, is essential to root meristem formation.     

Rapid Effects of Abscisic Acid on Ion Uptake in Sunflower Roots

Short-term effects of ABA, ABA + kinetin and kinetin on ion (⁸⁶Rb-potassium and phosphate) and water uptake in sunflower plants (Helianthus annuus var. californicus) were examined with a continuous-recording technique. Ion uptake in the roots and transport to the shoots were also investigated by conventional tracer uptake experiments and by sap bleeding experiments with excised roots. After addition of 5 × 10^?6-4 × 10^?5M ABA to the root medium there was an immediate decrease (30–70%) in the rate of ion uptake which lasted 30–70 min. The rate of water uptake was not significantly affected as measured with this method. Ion transport to the shoots and to the bleeding sap of excised roots was decreased by ABA. ABA-induced inhibition of ion uptake was abolished by the presence of kinetin, and uptake was slightly stimulated by 2 × 10^?5M kinetin alone. We suggest that concentration gradients of ABA or rapid changes in the ABA-kinetin balance in the roots affect ion uptake and transport.     

Interaction of Kinetin and Abscisic Acid in the Avena Straight-Growth Test

Different amounts of abscisic acid, 2.7 × 10^?9? 5 × 10^?8 moles, were chromatographed in isopropanol: ammonia: water (100:14:6), firstly alone and secondly together with 5 × 10^?8 moles kinetin. The same amount of kinetin was also chromatographed alone. The chromatograms were tested biologically with the Avena straight-growth test. Whereas a large part of the chromatograms of kinetin gives growth stimulation, the Rf region 0.4–0.6 of abscisic acid chromatograms is strongly growth-inhibiting. The inhibition within this Rf region does not become less if abscisic acid and kinetin are chromatographed together.     

Callus and Cell Suspension Cultures of Carnation

Callus cultures of carnation, Dianthus caryophyllus L. ev. G. J. Sim, were grown on a synthetic medium of half strength Murashige and Skoog salts, 3 % sucrose, 100 mg/l of myo-inositol, 0.5 mg/l each of thiamin, HCl, pyridoxin, HCl and nicotinic acid and 10 g/l agar. Optimal concentrations of growth regulators were observed to be 3 × 10^?6M indoleacetic acid (JAA) combined with 3 × 10^?6M benzylaminopurin (BAP) or 10^?6M 2,4-dichlorophenoxy acetic acid (2,4-D) alone. IAA + BAP caused a 100 fold increase in fresh weight over 4 weeks at 25°C. Addition of casein hydrolysate increased growth further. Cell suspension cultures worked best in media containing 2,4-D in which they had a doubling time of about 2 days. Filtered suspensions were successfully plated on agar in petri dishes, but division was never observed in single cells. The cultures initiated roots at higher concentrations of IAA or NAA, but all attempts to induce formation of shoots or em-bryoids gave negative results.     

Simple Determination of Trace Amounts of Anionic Surfactants in River Water by Spectrophotometry Combined with Solid-phase Extraction

A simple and sensitive specrophotometric method combined with solid-phase extraction (SPE) for the simultaneous determination of sodium linear-dodecylbenzenesulfonate (DBS) and sodium dodecyl sulfate (SDS) is described. The C2 (ethyl group bonded silicagel) cartridge could be repeatedly used more than 500 times for SPE, and it enabled the anionic surfactants to be concentrated by 50-fold. The calibration graph for DBS was linear in the range from 1.6×10^?8 M to 5.0×10^?7 M and for SDS from 2.0×10^?9 M to 3.0×10^?7 M. The relative standard deviation (n=5) for 5.0×10^?7 M DBS was 3.1% and for 2.5×10^?7 M SDS was 1.7%. The proposed method was applied to the simultaneous determination of DBS and SDS in river-water samples.