Regional differences in the sensitivity of the metabolism of coelenterate larvae to external influences]

The regional differences in the sensitivity of protein synthesis and free radical processes to temperature, trypsin, urea and LiCl were studied in Obelia flexuosa by means of autoradiography. The regional differences were also determined with respect to the rate of incorporation and excretion of a labelled aminoacid under the normal conditions. The metabolic reactions of larvae can be divided in primary (the first 30 min following the effect) and subsequent adaptive ones. The primary reactions are characterized by the greater sensitivity of the anterior larval regions to all factors under study. The subsequent reactions are characterized by synchronous and unidirectional metabolic starts in both the anterior and posterior larval regions, the starts being bigger in the anterior regions. The restoration of the normal ratios of metabolic activities of the opposite larval regions does not always correlate with the restoration of the normal absolute level of metabolism. The adaptive reactions are better expressed for protein synthesis, rather than for free radical processes. The anterior larval region has the greatest metabolic non-stability by a series of indices.     

The quantitative response to cytokinin in the moss Funaria hygrometrica does not reflect differential sensitivity of initial target cells

Exposure to sufficient cytokinin induces the formation of buds from responsive cells in the protonema of Funaria hygrometrica. Initial perception of the phytohormone results in a Ca+2 cascade within minutes. A second cytokinin-mediated event occurs some days later, and converts incipient buds into stably committed buds. The concentration of exogenous cytokinin also regulates the total number of buds produced from a protonemal colony. This concentration-dependent production of buds has been thought to reflect differential sensitivity of target cells. Under that hypothesis, the regulation of bud number occurs during initial perception of hormone. This paper presents direct experimental evidence to the contrary and supports the alternate hypothesis that bud formation involves the gating of large numbers of responding cells by later events. Experiments transferring protonema between media with different levels of cytokinin show that the cytokinin concentration during the initial perception of cytokinin is unimportant in controlling bud number. Instead, bud number is found to be regulated by the concentration of exogenous cytokinin as incipient buds or bud initials become stably committed buds.     

Trehalose breakdown in germinating spores of Mucor rouxii

Germinating spores of Mucor rouxii rapidly broke down their large (23% of the dry weight) trehalose reserve. More than 50% of this trehalose was broken down to ethanol. About one-third of the trehalose was converted to glycerol, which started to leak out of the spores after some 20 min germination. The synthesis of glycerol was not associated with any major change in glycerol 3-phosphatase activity in the spores. Since its rate of leaking was much smaller and the internal concentration reached was much higher in spores subjected to osmotic stress, glycerol might play a role in the initial water uptake and swelling of the germinating spores.     

Periods of sensitivity to chilling in germinating cotton

Cotton seedlings were subjected to a 96 hour chilling treatment (5° or 10°) after periods of germination at 31° ranging from 0 to 48 hours. Inhibition of subsequent growth at a favorable temperature by chilling was dependent on level of low temperature and stage of seedling development when chilled. Two periods of chilling hypersensitivity were observed during germination: 1) coincident with subjection of seed to a germination environment; and 2) after 18 to 30 hours of germination at 31°. Subsequent growth of seedlings chilled after 12 to 18 hours or 48 hours of germination at 31° was relatively unaffected. It is suggested that chilling alters specifically timed events that occur at the initiation of germination and after 18 to 30 hours of germination, and that alteration of these germination processes is visited on long term subsequent growth of the plant.     

Osmotic sensitivity in relation to salt sensitivity in germinating barley seeds

Abstract Cultivars of barley (Hordeum vulgare L.) were tested for germination sensitivity to progressively higher concentrations of salt, mannitol, and betaine. The three solutes were equally inhibitory at equal osmotic potential, but there was a consistent difference in osmotic sensitivity between two cultivars, CM-67 and Briggs (Briggs was the most sensitive). There was no difference between the two cultivars in salt or water uptake from salt solutions during imbibition. Brief presoaking in water did not improve salt resistance, indicating that a hydration-dependent decrease in membrane permeability is not involved in salt tolerance. The calcium content of Briggs was higher than CM-67. These results suggest that salt inhibits barley germination primarily by osmotic effects, and that salt influx during imbibition does not play a role in this inhibition. A hypothesis regarding salt effects on germination is discussed.     

Gravity-directed calcium current in germinating spores of Ceratopteris richardii

 Gravity directs the early polar development in single cells of Ceratopteris richardii Brogn. It acts over a limited period of time during which it irreversibly determines the axis of the spore cell's development. A self-referencing calcium selective electrode was utilized to record the net movement of calcium across the cell membrane at different positions around the periphery of the spore during the period in which gravity orients the polarity of the spore. A movement of calcium into the cell along the bottom and out of the cell along the top was detected. This movement was specific, polarized, and strongest in a direction that opposed the vector of gravity. Treatment with nifedipine, a calcium-channel blocker, diminished the calcium current and caused the cell to lose its responsiveness to the orienting influence of gravity. Results shown suggest that calcium plays a crucial role in the ability of a single cell to respond to gravity and in the subsequent establishment of its polarity. Received: 13 June 1999 / Accepted: 1 September 1999     

Dry mass changes in germinating spores of Diplodia maydis

Summary The dry mass of two-celled Diplodia maydis spores was measured both before and after germination by quantitative interference microscopy. The dry mass of spores declined approximately 50% during germination. However, the dry mass of germinating spores plus the dry mass of their germ tubes was greater than the dry mass of spores before germination. We conclude that the germinating spores absorbed nutrients released from non-germinating spores.The dry mass of fungal spores can be estimated by weighing large numbers of spores and determining the mean from sample spore counts. Mumford and Pappelis(4) determined the total dry mass of individual spores of Fusarium roseum and the contained lipid bodies before and after spores germinated using quantitative interference microscopy. The mean spore dry mass before germination was 57 pg. Lipid bodies accounted for about 61% of that mass and decreased as spores germinated. The total dry mass of the spore and germ tube 24 hr later greatly exceeded that of the spore before germination. Quantitative interference microscopy has been used to measure the dry mass of various types of cells. Kulfinski and Pappelis (3) recently reviewed how this technique has been applied to plant cells. Technical aspects of interference microscopy have been described by Ross (6).The purpose of this study was to examine the dry mass changes in Diplodia maydis (Berk.) Sacc. with and without germ tubes through the use of interference microscopy.     

Resistance of germinating Phycomyces spores to desiccation, freezing and acids

Abstract Phycomyces spores are remarkably resistant to desiccation, freezing and mineral acids, but this resistance gradually disappears during germination. Dehydration resistance decreases much faster in acetate-activated spores than in heat-activated spores. This may be related to the more extensive breakdown of trehalose in the former case. The water content of the spores doubles 10–40 min after activation in both cases. This may be related to the rapid loss of frost resistance in both heat- activated and acetate-activated spores. Resistance to HCl, on the other hand, decreases only after 1 h germination, and is not related to any change in water or trehalose content of the spores.     

Growth initiation in the geotropic response of the wheat node

Summary The geotropic response in stems of Triticum aestivum occurs at the leaf sheath bases. Growth is initiated in the lower halves of these organs after orientation into the horizontal position. Lateral transport of growth hormone is not involved in the development of curvature. Continuous stimulation is necessary to maintain the response.Growth can be initiated in small pieces of tissue excised from the leaf sheath base but this is entirely dependent upon the precise orientation of the tissue in the gravitational field. Growth occurs only when the segment is orientated horizontally with the outer epidermis facing downwards. IAA is able to initiate growth in nongeo-induced tissue segments of the leaf sheath base but this effect appears to differ from the geo-induced growth.IAA-Indole-3-acetic acid; ABA-RS(±) abscisic acid; CFM-chlorofluorenol methyl ester; GA₃-gibberellic acid.     

Spectral dependence of the light-induced geotropic response in Zea roots

The induction by light of geotropic responsiveness in the primary roots of Zea mays L. (cv. Golden Cross Bantam 70) was found to be governed by the all-or-none law. The response was induced by light energies above a threshold value, but the maximal curvature of geo-stimulated roots was constand irrespective of the light energy above that threshold. The action spectrum for this light effect showed a large peak at 650, a small peak at 410, and a shoulder at 663 nm. The effect of red light was not reversed by far-red light. Thus, the geotropic response in Zea roots may not be controlled by phytochrome.     

Acid-induced growth and the geotropic response of the wheat node

Summary Growth is initiated in segments of the leaf sheath base of Triticum aestivum by gravitational stimulation and by incubation in buffer solutions of pH 3. Both responses involve only an increase in segment length, are rapidly terminated on removal of the stimulus, have a similar Q₁₀ and are dependent upon cell turgor. They differ, however, in that the response to acid solutions is rapid and unaffected by anoxia. Acid-induced growth can be stopped and started repeatedly by changing the pH, an increase in pH from 3 to 5 or 7 being sufficient to terminate the response. The maximum growth induced by low pH is not increased by simultaneous stimulation by gravity.     

Cell elongation in the grass pulvinus in response to geotropic stimulation and auxin application

Summary Horizontally-placed segments of Avena sativa L. shoots show a negative geotropic response after a period of 30 min. This response is based on cell elongation on the lower side of the leaf-sheath base (pulvinus). Triticum aestivum L., Hordeum vulgare L. and Secale cereale L. also show geotropic responses that are similar to those in Avena shoots. The pulvinus is a highly specialized organ with radial symmetry and is made up of epidermal, vascular, parenchymatous and collenchymatous tissues. Statoliths, which are confined to parenchyma cells around the vascular bundles, sediment towards the gravitational field within 10–15 min of geotropic stimulation. Collenchymatous cells occur as prominent bundle caps, and in Avena, they occupy about 30% of the volume of the pulvinus. Geotropic stimulation causes a 3- to 5-fold increase in the length of the cells on the side nearest to the center of the gravitational field. Growth can also be initiated in vertically-held pulvini by the application of indole-3-acetic acid, 1-naphthaleneacetic acid or 2.4-dichlorophenoxyacetic acid. 2.3.5.-triiodobenzoic acid interferes with growth response produced by geotropic stimulation as well as with the response caused by auxin application. Gibberellic acid and kinetin have no visible effect on the growth of the pulvinus. Polarization microscopy shows a unique, non-uniform stretching of the elongating collenchymatous cells. Nonelongated collenchymatous cells appear uniformally anisotropic. After geotropic stimulation or auxin application, they appear alternately anisotropic and almost isotropic. Such a pattern of cell elongation is also observed in collenchyma cells of geotropically-stimulated shoots of Rumex acetosa L., a dicotyledon.Abbreviations 2.4-D 2.4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - NAA l-naphthaleneacetic acid - TIBA 2.3.5-triiodobenzoic acid     

Limited period of graviresponsiveness in germinating spores of Ceratopteris richardii

Rhizoids of the fern Ceratopteris richardii Brogn. usually emerge 40 h after germination is initiated by light, and more than 90% of them emerge growing in a downward direction. However, when the spores are germinated on a clinostat, the emerging rhizoids show no preferential orientation. This indicates that under normal 1 · g conditions the initial growth direction of rhizoids can be oriented by gravity. If the orientation of the spores is changed 3 h or less after the start of germination, the growth direction of most emerging rhizoids becomes downward relative to the new orientation. However, if the orientation of the spores is changed by 180° 8 h or more after germination is initiated by light, most rhizoids emerge growing upward; i.e., the same direction as if there had been no orientation change. Emerged rhizoids also do not change their direction of growth if their orientation is changed. These results indicate that the growth direction of emerging rhizoids is set by gravity prior to actual emergence, and that the time of full orientation responsiveness is limited to a period ranging from the initiation of germination to about 3–4 h after the start of germination. There is a gravity-oriented nuclear movement beginning at about 13 h after germination, and this movement appears to predict the initial growth direction of rhizoids.These studies were made possible by grant NAGW 1519 to S.J.R. and grant NGT-51065 to E.S.E., both from the National Aeronautics and Space Administration.     

Isocitrate lyase in germinating spores of the fern Anemia phyllitidis

Isocitrate lyase was partially purified from germinating spores of the fern Anemia phyllitidis. The enzyme requires Mg²⁺ and thiol compounds for maximal activity and has a pH optimum between 6.5 and 7.5. The K_m of the enzyme for threo-Δ_s-isocitrate is 0.5 mM. Succinate inhibits the enzyme non-competitively (K_i. 1.8 mM). The increase of isocitrate lyase activity is closely correlated with the induction of the germination process. The fall of enzyme activity during germination is associated with the decline in triglyceride reserves.     

Sensitivities of germinating spores and carvacrol-adapted vegetative cells and spores of Bacillus cereus to nisin and pulsed-electric-field treatment

Treatment of Bacillus cereus spores with nisin and/or pulsed-electric-field (PEF) treatment did not lead to direct inactivation of the spores or increased heat sensitivity as a result of sublethal damage. In contrast, germinating spores were found to be sensitive to PEF treatment. Nisin treatment was more efficient than PEF treatment for inactivating germinating spores. PEF resistance was lost after 50 min of germination, and not all germinated spores could be inactivated. Nisin, however, was able to inactivate the germinating spores to the same extent as heat treatment. Resistance to nisin was lost immediately when the germination process started. A decrease in the membrane fluidity of vegetative cells caused by incubation in the presence of carvacrol resulted in a dramatic increase in the sensitivity to nisin. On the other hand, inactivation by PEF treatment or by a combination of nisin and PEF treatments did not change after adaptation to carvacrol. Spores grown in the presence of carvacrol were not susceptible to nisin and/or PEF treatment in any way.     

Ornithine and arginine decarboxylase activities and effect of some polyamine biosynthesis inhibitors on Gigaspora rosea germinating spores

The pathways for putrescine biosynthesis and the effects of polyamine biosynthesis inhibitors on the germination and hyphal development of Gigaspora rosea spores were investigated. Incubation of spores with different radioactive substrates demonstrated that both arginine and ornithine decarboxylase pathways participate in putrescine biosynthesis in G. rosea. Spermidine and spermine were the most abundant polyamines in this fungus. The putrescine biosynthesis inhibitors alpha-difluoromethylarginine and alpha-difluoromethylornithine, as well as the spermidine synthase inhibitor cyclohexylamine, slightly decreased polyamine levels. However, only the latter interfered with spore germination. The consequences of the use of putrescine biosynthesis inhibitors for the control of plant pathogenic fungi on the viability of G. rosea spores in soil are discussed.     

Cyclic AMP regulation of glucose transport in germinating Pilobolus longipes spores

Pilobolus longipes spores were activated by either glucose or 6-deoxyglucose. Glucose-induced spore activation was previously shown to follow an increase in intracellular cyclic AMP. Concurrent with glucose-induced spore activation, were shifts in 6-deoxyglucose transport kinetics towards higher V _max and K _m values. Cyclic AMP derivatives also caused spore activation and similar changes in the kinetic parameters of 6-deoxyglucose transport. The time course of activation was paralleled by changes in transport activity. Inhibition of phosphodiesterase alone did not cause activation or induce changes in transport activity, but in combination with sub-optimal levels of either 6-deoxyglucose or cAMP derivatives, it amplified the germination signals to produce large increases in both spore activation and 6-deoxyglucose transport activity. These results support the conclusion that glucose transport in germinating spores is regulated by cAMP.Abbreviations IBMX 3-isobutyl-1-methylxanthine; monobutyryl cyclic AMP - N⁶ monobutyryladenosine 3:5-cyclic monophosphate - 8-bromo cyclic AMP 8-bromoadenosine 3:5-cyclic monophosphate     

Inhibitors of the geotropic response in plants: A correlation of molecular structures

Selected compounds from several groups of chemicals known to affect the root geotropic response in plants have been assessed for their effects on both stem and root geotropism in cress seedlings in order to compare their relative activities. It is suggested that a class of compound which affects geotropism can be distinguished, and that the molecular requirements for its activity can be tentatively defined. It is suggested that an ortho carboxylic acid function attached to an aromatic ring, which is separated by a conjugated system of atoms from a second aromatic ring, is required for high activity. Substituents on the second aromatic ring increase activity if they assist in the achievement of a minimum molecular size.