KINETICS OF GLUCOSE TRANSPORT AND GROWTH OF CYCLOTELLA CRYPTICA REIMANN,LEWIN AND GUILLARD1,2

Growth rate as well as rate of glucose uptake of C. cryptica depends on glucose concentration in the medium according to saturation kinetics. The K _g for growth is 1.9 × 10^?5 M, and the K _t, for glucose transport is 5.8 × 10^?5 M. The maximum growth rate in the dark on glucose is considerably slower than the light-saturated growth rate at the same temperature, and does not appear to be determined by the capacity of the cell for glucose uptake. The glucose transport process is highly specific, and depends on energy metabolism. The Q ¹⁰ for the process is 2.2 (15–2.5 C). Glucose taken up by the cells is almost, quantitatively phosphorylated within 10 min, either through the transport process itself or by a high affinity kinase system in the cells.     

Is Cell Elongation Regulated by Extracellular Auxin?

Experiments with isolated epidermal strips of maize coleoptiles, pretreated with auxin and further incubated on sucrose agar containing different concentrations of auxin (indole-3-acetic acid, IAA or naphthalene-1-acetic acid, NAA) and/or naphthylphthalamic acid (NPA), are described. Preincubation for 2h with 2 . 10^?4M IAA or 10^?5M NAA in buffer, followed by 30 min wash in buffer results in measurable cell elongation during a subsequent incubation for 6 h on sucrose agar. Addition of 10^?4M NPA inhibited the response to auxin and this inhibition could be reversed by providing IAA in addition to NPA. Inner tissue fragments (without outer epidermis) did not respond to external IAA. These results lead to the conclusion that auxin secretion at the outer epidermis may be an essential step in auxin-regulated coleoptile growth.     

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.     

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.     

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

Kinetin has a stimulating effect in the Avena straight-growth test. The action of different concentrations of kinetin, 2.5 × 10^?7, 2.5 × 10^?6 and 2.5 × 10^?5M, in combination with different concentrations of IAA was studied in this test. It was shown that the effect of low IAA concentrations, 0.25 × 10^?7 and 1 × 10^?7M, was strongly enhanced by the addition of all the kinetin concentrations investigated. The effect of the highest IAA concentrations, 25 × 10^?7 and 100 × 10^?7M, on the other hand, was inhibited relatively strongly by the highest employed concentration of kinetin. The results are explained as due to a kinetin-produced increase of auxin in the coleoptile segment, which in combination with low IAA concentrations can lead to a growth stimulation and with high IAA concentrations to a growth inhibition. Since kinetin in purification and chromatography of auxin can partly follow IAA, thereby affecting the quantitative yield, it is emphasized that, prior to the test, auxin extracts containing cytokinins should be freed from the latter by, for example, gel filtration or paper electrophoresis.     

The effects of temperature and IAA concentration on the latent period for IAA-induced rapid growth of Avena coleoptile segments

Summary Indoleacetic acid buffered at pH 7.0 induces a high growth rate in Avena coleoptile segments after a latent period, the duration of which is dependent upon both IAA concentration and temperature. A minimum latent period of 7.3 min is observed at 25° C with 10^-3 M IAA in phosphate buffer at pH 7.0.In contrast, 5×10^-3 M IAA made up in 0.01 M KH₂PO₄ alone, promotes elongation almost immediately, regardless of whether the segments have been previously incubated in 0.01 M KH₂PO₄ at pH 4.7, or phosphate buffer at pH 7.0. This immediate response is unaffected by 10^-4 M KCN which abolishes the rapid growth induced by 5×10^-3 M IAA buffered at pH 7.0 but does not affect the immediate appearance of low-pH-induced growth. Since we consistently find solutions of 5×10^-3 M IAA in 0.01 M KH₂PO₄ to have a pH of 3.5, our results indicate that the immediate growth response elicited by this solution is attributable to its low pH rather than to the presence of IAA as previously reported in the literature.     

Die Beurteilung des Einflusses qualitativer Prüffaktoren (Infektionstermine,Sorten) auf den Befall mit Hilfe der Kontingenztafelanalyse

Mycelial growth of some wood‐rotting fungi was studied on a solid modified medium MS (Murashige and Skoog, 1962) with indole‐3‐acetic acid at concentrations of 10^‐6 to 10^‐3 M. The IAA concentrations of 10^‐6 M and 10^‐5 M inhibited mycelial growth of the fungus Phaeolus schweinitzii, Laetiporus sulphureus and Pleurotus ostreatus while the same concentrations stimulated mycelial growth of the fungus Stereum rugosum. The IAA concentrations of 10^‐6 M stimulated mycelial growth in Piptoporus betulinus and temporarily stimulated mycelial growth in Heterobasidion annosum. The IAA concentration of 10^‐4 M appeared critical for wood‐rotting fungi. The IAA concentration of 10^‐3 M inhibited mycelial growth in all the fungi under study.     

SELENIUM STIMULATES GROWTH OF MARINE MACROALGAE IN AXENIC CULTURE1

The marine brown alga Fucus spiralis L. and the red alga Goniotrichum alsidii (Zanard) increase their growth upon the, addition of SeO₃^2- or SeO₄^2- when cultivated axenically in the artificial seawater ASP6 F2. In the concentration range 1 · 10^?10-1 · 10^?7 M there are two optima, one at 3.3 · 10^?10 M and another at 3.3 · 10^?8 M. α-To-copherol, often administered together with selenium to mammals suffering from selenium deficiency, gives no additive effect with selenium, but α-tocopherol in the concentration range 1 × 10^?7-1 × 10^?6 M does influence the morphology of the Fucus plants. Organically bound selenium has no effect.     

The Interaction of Human Serum with the Plant Hormone Indole-3-Acetic Acid

It is well known that human serum inhibits the longitudinal root growth in Lupinus albus L and Triticum sativum Lam. This inhibitory effect has been ascribed to the IAA content in human serum, which unfortunately has never been measured quantitatively. Experiments are presented in which Triticum roots are grown in media with pooled human serum and varying concentrations of IAA. In the presence of 10^?5M p-chlorophenoxy-isobutyric acid (PCIB) and serum, minute IAA additions promoted the growth. This feature hardly could be expected were the serum inhibition in itself an IAA effect. In view of this finding, renewed but unsuccessful attempts were made to demonstrate a similar promotion in media without serum. To explain the observed response curves, it must be further assumed that serum components bind IAA reversibly. In experiments without PCIB in the medium the response curves were similar at a lower level of growth, except that no growth promotion by IAA was discernible. It is concluded, that the inhibiting effect of human serum on the growth of plant roots is not due to free IAA, although IAA in all probability occurs in that fluid.     

Initial phases of indoleacetic acid induced growth in coleoptile segments of Avena sativa L.

The intial phases of auxin-induced growth in coleoptile segments of Avena sativa L. were investigated using a high resolution growth recording technique, based on an angular position sensing transducer. The first response to the hormone is a slight, transient reduction of the growth rate lasting about 5 min. After this phase growth rate increases to a maximum. The duration of the increase and the maximum clearly depend on the concentration of the hormone. With increasing auxin concentration the duration of the growth rate increase is reduced from about 80 min in 10^-9 M indoleacetic acid (IAA) to about 14 min in 10^-4 M IAA. After the maximum the growth rate declines. Looking at the maximum of the growth rate, we obtained a dose-response curve with a sharp increase between 10^-9 M and 10^-6 M IAA and a slight decline between 10^-6 M and 10^-4 M IAA. This result is confirmed by growth rates measured one and two hours after the application of the hormone.Abbreviations IAA indoleacetic acid     

Does indoleacetic acid promote growth via cell wall acidification?

Growth of Triticum aestivum L. cv. Cappelle Desprez coleoptiles is promoted by 5.7×10^–5 M indole acetic acid (IAA) as effectively in pH 3.4 buffer as in water, but IAA is not effective in the presence of buffer at pH 3.0 or 3.2 A combination of 5.7×10^–5 M IAA and pH 3.4 buffer promotes growth to a greater extent than pH 3.2 buffer alone, which is optimal for acid-induced growth. IAA employed at 10^–7 M is still effective at promoting growth in the presence of pH 3.4 buffer, moreover, IAA at 10^–7 M interacts synergistically with the acidic buffer to promote growth. It is concluded that IAA and acid promote growth via separate mechanisms, and that IAA does not promote cell wall loosening by rendering the cell wall more acid.Abbreviation IAA Indoleacetic acid     

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.     

Invertase and auxin-induced elongation in internodal segments of Phaseolus vulgaris

A close positive correlation was observed between segment elongation and the specific activity of soluble acid invertase in stem segments of P. vulgaris incubated for 21 hr in the presence of IAA or of several synthetic auxins and auxin analogues. Optimum concentrations for the stimulation of growth and invertase activity were similar and varied from 10^?6 M (2,4-D) through 10^?5 M (IAA, IBA, α-NAA, β-NAA) to greater than 10^?4 (IPA, PoAA, trans-cinnamic acid). The weak activity of trans-cinnamic acid, a competitive inhibitor of auxin action, may have resulted from cis-trans isomerization during incubation. The concentration of hexose sugars in the segments fell during incubation in the presence of auxin, the greatest decline in hexose concentration occurring in the presence of compounds exhibiting the greatest stimulation of growth.     

NUTRIENT REQUIREMENTS OF THE DINOFLAGELLATE PERIDINIUM GATUNENSE1

Optimum nutrient conditions for growth and photosynthesis of Peridinium gatunense (Nygaard) (Peridinium cinctum fa. westii) were investigated using axenic clones in batch cultures. Selenium (Se) had previously been found to be an indispensable growth factor for P. gatunense. Optimal, suboptimal, and supraoptimal concentrations of HCO₃^?, N, Ca, Cl, Mg, P, K, S, Si, EDTA-Na, Fe, Mo, Zn, Mn, Co, Se, B, Br, I, and various trace element mixtures were determined by measuring biomass development, growth rates, ¹⁴C uptake, and/or oxygen production at various concentration gradients of these elements. The general characteristics of the best formulation, medium-L 16, relative to other media, are its high content of NaHCO₃ (1 meq · L^?1) and Mo (0.2 μM) but low concentrations of NO_3-N (150 μM), PO_4-P (10 μM), and Fe (0.4 μM), in addition to its content of Se. The total content of trace metals, except for Se, may be reduced to one-fourth of that in medium-L 16 without altering the major growth-promoting properties of the medium. Medium-L 16 deviated considerably from Lake Kinneret (Israel) water, being much lower in macroelements except for N and P. The pH (8.1–8.4) was in the same range, but the values of conductivity (140 μS · cm^?1), alkalinity (1 meq · L^?1) and NaCl (200 μM) were > 8, 2, and 30 times higher, respectively, in the lake water. Selenium deficiency may limit the growth of P. gatunense in this lake.     

Effect of IAA and 4-Cl-IAA on growth rate in maize coleoptile segments

The dose-response curves for IAA and 4-Cl-IAA-induced growth of Zea mays L. coleoptile segments were studied as a function of time. Moreover, some characteristic growth parameters for both auxins were compared. The dose-response curve of growth rate measured after IAA or 4-Cl-IAA application was bell-shaped in all experiments. The optimum concentration was 10⁻⁶ M for 4-Cl-IAA and was found not to depend on the time of the growth measurement. However, in the case of IAA the optimum shifted from 10⁻⁶ M at the time of maximal growth rate to 10⁻⁵ M or even 10⁻⁴ M, when growth measured 3–4 hours after auxin application was analysed. The relative activity of 4-Cl-IAA-induced growth rate (as compared to IAA) increased significantly with increasing time from addition of this auxin to the medium. For both auxins the time needed to reach the maximal growth rate was clearly related to their concentrations. These data provided further evidence that 4-Cl-IAA is much more active auxin than IAA and can also suggest that IAA is more rapidly metabolized in comparison to 4-Cl-IAA.     

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.     

INTENSE GRAZING AND PREY‐DEPENDENT GROWTH OF PFIESTERIA PISCICIDA (DINOPHYCEAE)1

Grazing and growth of Pfiesteria piscicida (Pfiest) were investigated using batch and cyclostat cultures with Rhodomonas sp. (Rhod) as prey. Observed maximum growth rates (1.4 d^?1) and population densities (2 × 10⁵ cells·mL^?1) corresponded to values predicted by Monod functions (1.76 d^?1; 1.4 × 10⁵ cells·mL^?1). In batch cultures under a range of prey‐to‐predator ratios (0.1:1 to 180:1) and prey concentrations (1000–71,000 cells·mL^?1), Rhodomonas sp. was always depleted rapidly and P. piscicida concentrations increased briefly. The rate of Rhodomonas sp. depletion and the magnitude of P. piscicida population maxima depended on the prey‐to‐predator ratio and prey concentration. Starvation resulted in cell cycle arrest at G1 and G2+M and ultimately the demise of both P. piscicida and Rhodomonas sp. populations, demonstrating the dependence of P. piscicida on the supply of appropriate prey. The depletion of Rhodomonas sp. populations could be attributed directly to grazing, because P. piscicida did not exert detectable inhibitory effects on the growth of Rhodomonas sp. but grazed intensely, with maximum grazing rates>10 Rhod·Pfiest^?1·d^?1 and with no apparent threshold prey abundance for grazing. The results suggest that 1) the abundance of appropriate prey may be an important factor regulating P. piscicida abundance in nature, 2) P. piscicida may control prey population, and 3) high growth and grazing potentials of P. piscicida along with cell cycle arrest may confer survival advantages.     

Apical callus formation and plant regeneration controlled by plant growth regulators on axenic culture of the red alga Gracilariopsis tenuifrons (Gracilariales, Rhodophyta)

Axenic cultures of Gracilariopsis tenuifrons (Bird et Oliveira) Fredericq et Hommersand (Gracilariales, Rhodophyta) were established in ASP12‐NTA solid medium (0.4% agar and 1.0% sucrose) supplemented with plant growth regulators to evaluate the effects on apical callus formation and plant regeneration. Indole‐3‐acetic acid (IAA), 2,4‐dichlorophenoxyacetic acid (2,4‐D) and 6‐benzylaminopurine (BA) were added individually or in combinations (IAA : BA) over a range of concentrations from 0.5 to 5 mg L^?1. Growth of apical and intercalary segments was stimulated by high concentrations of 2,4‐D (5 mg L^?1) and a high IAA to BA ratio (IAA : BA = 5:1 mg L^?1) respectively. Apical calluses were originated from divisions of apical and cortical cells located at apical regions of thallus segments and lateral branches. Low concentration of IAA (0.5 mg L^?1) or a high IAA to BA ratio (IAA : BA = 5:1 mg L^?1) were the optimal treatments for inducing apical callus formation in apical segments, while high concentration of IAA (5 mg L^?1) stimulated the highest callus induction rate in intercalary segments. Conversely, equal parts IAA and BA (IAA : BA = 1:1 mg L^?1) and low concentration of 2,4‐D (0.5 mg L^?1) stimulated growth of apical calluses from apical and intercalary segments, respectively. Two processes of regeneration were observed: direct regeneration (upright axis originated from cells of proximal region of intercalary segments) and indirect regeneration (adventitious plantlet originated from cells of apical calluses). Direct regeneration was promoted significantly by treatment with a low IAA to BA ratio (IAA : BA= 1:5 mg L^?1), and treatments with IAA (0.5 mgL^?1) or 2,4‐D (0.5 or 5 mg L^?1) significantly stimulated the elongation of upright axis. Plant growth regulators are essential to inducing indirect regeneration, and a high concentration of IAA (5 mg L^?1) and BA (5 mg L^?1) were the optimal treatments for inducing the regeneration of plantlets from apical calluses in apical and intercalary segments, respectively. Regenerating plantlets grew into plants morphologically similar to those formed from germinating spores, and became fertile after 6 weeks. The results suggest that auxins and cytokinins are involved in developmental regulatory processes in G. tenuifrons. The regeneration process from calluses in species of Gracilariales was observed for the first time in the present study. The culture system described for G. tenuifrons could be useful for micropropagation and for biotechnological applications in agarophytic algae.     

Regulation of Sorus Formation by Auxin in Laminariales Sporophyte

Young sporophytes of Laminaria japonica Areshoug were cultured in six indole-acetic acid (IAA) concentrations (0, 10⁻⁸, 10⁻⁷, 10⁻⁶, 10⁻⁵, 10⁻⁴ M) to examine the effect of auxin on growth. The effects of auxin on sorus formation were also examined by using discs taken from the adult sporophyte. The auxin contents and IAA oxidase activities in the thallus and sorus parts of the sporophyte were determined with the blade and sporophyll of other Laminariales plants, Undaria pinnatifida (Harvey) Suringar and Alaria crassifolia Kjellman. The young sporophytes of L. japonica showed highest elongation rate in 10⁻⁵ M IAA. In contrast, the sorus formation on the discs cultured in 10⁻⁵ M IAA was markedly delayed in comparison with other concentrations, indicating that sorus formation was suppressed by IAA. Free and conjugated auxin contents were lower in the reproductive parts than in the vegetative parts. In three Laminariales sporophytes, IAA oxidase activity was about 3–9 times higher in the reproductive parts than in the vegetative parts. Taken together these results suggest that the growth and reproduction of Laminariales sporophytes are regulated by internal auxin levels. Elucidating the regulation mechanism is likely to provide information that is important for the management of plant production and the assessment of the physiological status of plants in the field.     

INTERACTION OF SUGARS AND AUXINS IN PEA EPICOTYL SECTION GROWTH

Purves , W. K., and A. W. Galston . (Yale U., New Haven, Conn.) Interaction of sugars and auxins in pea epicotyl section growth. Amer. Jour. Bot. 47 (8): 665–669. Illus. 1963.—The nature and magnitude of the response of “S₁” etiolated pea-epicotyl sections to auxin are determined by the concentration of sugar in the growth medium. For example, the concentration of IAA inducing maximum elongation shifts through at least 3 orders of magnitude in response to varying sucrose concentrations, from ca. 10^–4 M, with no sucrose, to 10^–7 M, with 2% sucrose. Similarly, the inhibitory action of high levels of IAA on elongation occurs only in the presence of sucrose. By contrast, although sucrose also promotes water uptake, it affects the IAA optimum for this process only slightly. The action of IAA on growth can be detected immediately, but the growth response to sucrose occurs only after a 6–8 hr. lag. If tissues are supplied with sucrose, then 1-hr. exposures to IAA can be as effective on growth as continuous 20-hr. exposures, depending on the time at which such exposures are given. Thus, 10^—4 M IAA applied in the presence of 2% sucrose is markedly inhibitory to elongation in hours 1–3, relatively inactive in hours 4–6, and strongly promotive after hour 7. The change from inhibitory to promotive action thus coincides in time with the length of the lag period for sucrose action.