Low levels of oxygen promote internodal elongation in floating rice independently of enhanced ethylene production

Submergence induces rapid elongation of internodes in floating rice(Oryza sativa L. cv. Habiganj Aman II). The initial signalfor such internodal elongation has been considered to be the reduced partialpressure of oxygen in submerged internodal cavities, which promotes theelongation of internodes through the enhancement of ethylene synthesis. Weexamined the relationship between low oxygen pressure and ethylene production inthe rapid elongation of floating rice internodes using ethylene biosynthesisinhibitors, aminooxyacetic acid (AOA) and CoCl₂. When floating ricestem segments were incubated in an atmosphere of low O₂, internodalelongation accelerated and ethylene production increased. However, in stemsegments treated with AOA or CoCl₂, low O₂ stillstimulated the elongation of internodes although the ethylene production by theinternodes was less than by those in control stem segments where internodalelongation was not promoted. These results indicate that low O₂ iscapable of causing rapid elongation of internodes of floating rice independentlyof enhanced production of ethylene. In addition to low O₂,submergence, ethylene and gibberellic acid each enhanced the production ofethylene by internodal tissues, suggesting that enhanced ethylene production isa common phenomenon accompanied by the acceleration of internodal elongation infloating rice.     

Thigmomorphogenesis: Ethylene evolution and its role in the changes observed in mechanically perturbed bean plants

Mechanical perturbations, in the form of either rubbing or wounding, cause ethylene evolution from bean internodes ( Phaseolus vulgaris L. cv. Cherokee Wax). This evolution begins 45 to 60 min after perturbation or wounding and peaks about 2 h later. Maximal thigmomorphogenesis occurs if internodes are perturbed when they are 10 mm or less in length. Maximal ethylene evolution, however, occurs in longer internodes. When one internode is perturbed, ethylene evolution is not observed from other internodes even though they respond thigmomorphogenetically by exhibiting decreased elongation. Ethylene evolution is apparently a result of increased 1-amino-cyclopropane-1-carboxylic acid (ACC) production after perturbation. Inhibitors of ACC and ethylene synthesis block increased radial growth but not reduced elongation. Ethylene may therefore be only one of several factors causing thigmomorphogenesis.     

Mechanical Stress and Gibberellin: Regulation of Hollowing Induction in the Stem of a Bean Plant, Phaseolus vulgaris L.

In pole bean plants, mechanical stress (MS) inhibited stem elongationand induced radial thickening of the stem. Application of uniconazole,an inhibitor of gibberellin biosynthesis, also reduced stemgrowth but had no effect on stem diameter. Both MS and uniconazolesignificantly reduced hollowing of the first internodes, butonly the former increased ethylene evolution from the firstinternode. Application of GA₃ increased the length of the firstinternode and decreased its diameter in bush bean plants; thiswas accompanied by a significant promotion of stem hollowing.Aminooxyacetic acid (AOA) decreased ethylene evolution fromthe GA₃-treated internodes, though it did not reduce the GA₃-inducedhollowing of the first internodes. Application of GA₃ affectedneither ethylene evolution nor cellulase activity in the firstinternodes of bush bean plants. Application of GA₃ stimulatedmuch greater cell elongation in the center of pith tissue thanin the outer surrounding tissues, suggesting a possible physicalbreakage of the inner cells, which leads the hollowing of beanstems. These results suggest that gibberellin is a factor responsiblefor stem hollowing in bean plants. Because MS is known to reducegibberellin content in bean plants [Suge (1978) Plant Cell Physiol.21: 303] MS may inhibit stem hollowing by reducing the amountof endogenous gibberellin. (Received July 1, 1994; Accepted November 8, 1994)     

Development of Nitrogen Assimilation Enzymes during Photoautotrophic Growth of Chenopodium rubrum Suspension Cultures

We have shown previously that ethylene, which accumulates in the air spaces of submerged stem sections of rice (Oryza sativa L. cv “Habiganj Aman II”), is involved in regulating the growth response caused by submergence. The role of gibberellins in the submergence response was studied using tetcyclacis (TCY), a new plant growth retardant, which inhibits gibberellin biosynthesis. Stem sections excised from plants that had been watered with a solution of 1 micromolar TCY for 7 to 10 days did not elongate when submerged in the same solution or when exposed to 1 microliter per liter ethylene in air. Gibberellic acid (GA₃) at 0.3 micromolar overcame the effect of TCY and restored the rapid internodal elongation in submerged and ethylene-treated sections to the levels observed in control sections that had not been treated with TCY. The effect of 0.01 to 0.2 micromolar GA₃ on internodal elongation was enhanced two- to eight-fold when 1 microliter per liter ethylene was added to the air passing through the chamber in which the sections were incubated. GA₃ and ethylene caused a similar increase in cell division and cell elongation in rice internodes. Thus, ethylene may cause internodal elongation in rice by increasing the activity of endogenous GAs. In internodes from which the leaf sheath had been peeled off, growth in response to submergence, ethylene and GA₃ was severely inhibited by light.     

Gibberellin-enhanced elongation of inverted Pharbitis nil shoot prevents the release of apical dominance

Ethylene evolution resulting from the gravity stress of shoot inversion appears to induce the release of apical dominance in Pharbitis nil (L.) by inhibiting elongation of the inverted shoot. It has been previously demonstrated that this shoot inversion release of apical dominance can be prevented by promoting elongation in the inverted shoot via interference with ethylene synthesis or action. In the present study it was shown that apical dominance release can also be prevented by promoting elongation of the inverted shoot via treatment with gibberellic acid (GA₃). A synergistic effect was observed when AgNO₃, the ethylene action inhibitor, was applied with GA₃. Both GA₃ and AgNO₃ increased ethylene production in the inverted shoot. These results are consistent with the view that it is ethylene-induced inhibition of elongation and not any direct effect of ethylene per se which is responsible for the outgrowth of the highest lateral bud.     

The biophysical basis of elongation growth in internodes of deepwater rice

Partial submergence induces rapid internodal elongation in deepwater rice (Oryza sativa L., cv Habiganj Aman II). We measured in vivo extensibility, tissue tension, hydraulic conductance and osmotic potential in the region of cell elongation in the uppermost internode. The in vivo extensibility of the internode, measured by stretching of living tissue with a custom-made constant stress extensiometer, rose rapidly following submergence of the plant. Both the elastic (E_el) and plastic (E_pl) extensibility increased when growth of the internode was induced. The submerged internode displayed tissue tension (elastic outward bending of longitudinally split internode sections); in air-grown control internodes, no such bending occurred. The hydraulic conductance, estimated from the kinetics of tissue shrinkage in 0.5 molar mannitol and subsequent swelling in distilled water, was not changed by submergence. The osmotic potential, measured with a dew-point hygrometer using frozen-thawed tissue, was only 18% less negative in the submerged internode than in the air-grown control. This indicates that osmoregulation takes place in rapidly elongating rice internodes. We suggest that the rapid expansion of the newly formed internodal cells of submerged plants is controlled by the yielding properties (E_pl) of the cell walls. Experiments with excised stem sections indicate that gibberellin is involved in increasing the E_pl of the elongating cell walls.     

The Never ripe Mutant Provides Evidence That Tumor-Induced Ethylene Controls the Morphogenesis of Agrobacterium tumefaciens-Induced Crown Galls on Tomato Stems

We confirm the hypothesis that Agrobacterium tumefaciens-induced galls produce ethylene that controls vessel differentiation in the host stem of tomato (Lycopersicon esculentum Mill.). Using an ethylene-insensitive mutant, Never ripe (Nr), and its isogenic wild-type parent we show that infection by A. tumefaciens results in high rates of ethylene evolution from the developing crown galls. Ethylene evolution from isolated internodes carrying galls was up to 50-fold greater than from isolated internodes of control plants when measured 21 and 28 d after infection. Tumor-induced ethylene substantially decreased vessel diameter in the host tissues beside the tumor in wild-type stems but had a very limited effect in the Nr stems. Ethylene promoted the typical unorganized callus shape of the gall, which maximized the tumor surface in wild-type stems, whereas the galls on the Nr stems had a smooth surface. The combination of decreased vessel diameter in the host and increased tumor surface ensured water-supply priority to the growing gall over the host shoot. These results indicate that in addition to the well-defined roles of auxin and cytokinin, there is a critical role for ethylene in determining crown-gall morphogenesis.     

Pea Xyloglucan and Cellulose : III. Metabolism during Lateral Expansion of Pea Epicotyl Cells

Lateral expansion of the third internodes of pea epicotyls was evoked by treatment with either 2,4-dichlorophenoxyacetic acid (2,4-D) or ethylene gas. During growth, 2,4-D enhanced and ethylene inhibited the deposition of xyloglucan and cellulose in the cell wall, with the result that the wall framework (ghost) from ethylene-treated swollen tissue was much thinner than that from 2,4-D-treated. The level of activity of xyloglucan synthase, alkali-insoluble β-glucan synthases, and endo-1,4-β-glucanases were all enhanced by 2,4-D treatment but not by ethylene. Both 2,4-D and ethylene treatments led to increased osmotic potential in the swelling tissues. Accordingly, swelling after 2,4-D treatment was accompanied by xyloglucan degradation, concomitant with substantial net synthesis, but swollen tissue as a result of ethylene treatment was characterized by walls whose integrity was weakened by relatively low levels of newly deposited polysaccharides rather than by the degradation.     

Ethylene Evolution from Maize (Zea mays L.) Seedling Roots and Shoots in Response to Mechanical Impedance

The effect of mechanical impedance on ethylene evolution and growth of preemergent maize (Zea mays L.) seedlings was investigated by pressurizing the growth medium in triaxial cells in a controlled environment. Pressure increased the bulk density of the medium and thus the resistance to growth. The elongation of maize primary roots and preemergent shoots was severely hindered by applied pressures as low as 10 kilopascals. Following a steep decline in elongation at low pressures, both shoots and roots responded to additional pressure in a linear manner, but shoots were more severely affected than roots at higher pressures. Radial expansion was promoted in both organs by mechanical impedance. Primary roots typically became thinner during the experimental period when grown unimpeded. In contrast, pressures as low as 25 kilopascals caused a 25% increase in root tip diameter. Shoots showed a slight enhancement of radial expansion; however, in contrast to roots, the shoots increased in diameter even when growing unimpeded. Such morphological changes were not evident until at least 3 hours after initiation of treatment. All levels of applied pressure promoted ethylene evolution as early as 1 hour after application of pressure. After 1 hour, ethylene evolution rates had increased 10, 32, 70, and 255% at 25, 50, 75, and 100 kilopascals respectively, and continued to increase linearly for at least 10 hours. When intact corn seedlings were subjected to a series of hourly cycles of pressure, followed by relaxation, ethylene production rates increased or decreased rapidly, illustrating tight coupling between mechanical impedance and tissue response. Seedlings exposed to 1 microliter of ethylene per liter showed symptoms similar to those shown by plants grown under mechanical impedance. Root diameter increased 5 times as much as the shoot diameter. Pretreatment with 10 micromolar aminoethoxyvinyl glycine plus 1 micromolar silver thiosulfate maintained ethylene production rates of impeded seedlings at basal levels and restored shoot and root extension to 84 and 90% of unimpeded values, respectively. Our results support the hypothesis that ethylene plays a pivotal role in the regulation of plant tissue response to mechanical impedance.     

On the role of abscisic Acid and gibberellin in the regulation of growth in rice

Submergence induces rapid elongation of rice coleoptiles (Oryza sativa L.) and of deepwater rice internodes. This adaptive feature helps rice to grow out of the water and to survive flooding. Earlier, we found that the growth response of submerged deepwater rice plants is mediated by ethylene and gibberellin (GA). Ethylene promotes growth, at least in part, by increasing the responsiveness of the internodal tissue to GA. In the present work, we examined the possibility that increased responsiveness to GA was based on a reduction in endogenous abscisic acid (ABA) levels. Submergence and treatment with ethylene led, within 3 hours, to a 75% reduction in the level of ABA in the intercalary meristem and the growing zone of deepwater rice internodes. The level of GA₁ increased fourfold during the same time period. An interaction between GA and ABA could also be shown by application of the hormones. ABA inhibited growth of submerged internodes, and GA counteracted this inhibition. Our results indicate that the growth rate of deepwater rice internodes is determined by the ratio of an endogenous growth promoter (GA) and a growth inhibitor (ABA). We also investigated whether ABA is involved in regulating the growth of rice coleoptiles. Rice seedlings were grown on solutions containing fluridone, an inhibitor of carotenoid and, indirectly, of ABA biosynthesis. Treatment with fluridone reduced the level of ABA in coleoptiles and first leaves by more than 75% and promoted coleoptile growth by more than 60%. Little or no enhancement of growth by fluridone was observed in barley, oat, or wheat. The involvement of ABA in determining the growth rate of rice coleoptiles and deepwater rice internodes may be related to the semiaquatic growth habit of this plant.     

Interactions between abscisic acid, ethylene and gibberellin in internodal elongation in floating rice: the promotive effect of abscisic acid at low humidity

The enhancement of internodal elongation in floating or deepwater rice (Oryza sativa L. cv. Habiganj Aman II) by treatment with ethylene or gibberellic acid (GA₃) at high relative humidity (RH) is inhibited by abscisic acid (ABA). Here, we examined the interactive effects of ethylene, gibberellin (GA) and ABA at low RH on internodal elongation of deepwater rice stem segments. Although ethylene alone hardly promoted internodal elongation of stem sections at 30% RH, it enhanced the internodal elongation induced by GA₃. Application of ABA alone to stem segments had no effect on internodal elongation. However, in the presence of ethylene and GA₃ at 30% RH, ABA further promoted internodal elongation. This promotive effect of ABA was not found in the internodes of stem segments treated either with ethylene or with GA₃ at 30% RH or in the internodes of stem segments treated with ethylene and/or GA₃ at 100% RH.     

Author index volume 38 (1985)

Mechanical perturbation (MP, rubbing) of internodes of Pharbitis nil shoots initiates release of lateral buds (LB) from apical dominance within 48 h. Evidence is presented which suggests that MP promotion of LB outgrowth is mediated by ethylene-induced restriction of main shoot growth. Ethylene production in the internodes is stimulated by MP within 2 h. Effects of MP are mimicked by treatments with 1-aminocyclopropane-1-carboxylic acid (ACC) and are negated by the inhibitors of ethylene production or action, aminoethoxy vinylglycine (AVG) and AgNO₃. The fact that effects of MP, ACC and ethylene inhibitors are observed to occur on main shoot growth at least 24 h before they are observed to occur on LB growth suggests a possible cause and effect relationship. MP also causes an increase in internode diameter. MP stimulation of ethylene production appears to be mediated by ACC synthase. The results of this study and our previous studies suggest that apical dominance may be released by any mechanism which induces ethylene restriction of main shoot growth.     

Effect of atmospheric pressure on maize root growth and ethylene production

Maize (Zea mays) seedlings were exposed to elevated atmospheric pressures while growing in moist sand in open plastic envelopes to evaluate the effects of directly applied atmospheric pressure on ethylene production and root growth. Effects were evaluated after 24 h. The threshold pressures necessary to promote ethylene production and decrease root elongation were about 600 and 400 kPa, respectively. Direct atmospheric pressure, at levels up to 300 kPa, mimicked the control decrease in root diameter and increased diameter only slightly at 500 to 1200 kPa. In contrast, in previous work it was shown that physical impedance resulting from compression of the growth medium by external application of 100 kPa increased the ethylene production rate 4-fold and the root diameter 7-fold while reducing elongation 75% in 10 h. The relative insensitivity of roots to direct atmospheric pressure suggests that they perceive physical impedance, achieved experimentally by compressing the growth medium, via a surface mechanism rather than via a pressure-sensing mechanism.     

Turnover of cell wall components during internode growth in <Emphasis Type="Italic">Merremia emarginata</Emphasis>

The internodes of Merremia emarginata plant showed exceptionally high stretchability throughout the development period. Therefore, it provides excellent material to study the changes undergoing during cell elongation. In this study, the influence of the hormone treatments (GA₃, PAA and NAA) on the wall component synthesis was analyzed in relation to elongation growth during internode development. A clear increasing trend of wall components was observed with increase in internode length. The non-esterified pectic substances were markedly correlated with internode length while esterified pectic substances showed correlation only in hormone treated internodes. Low molecular weight xyloglucans content showed correlation only in GA₃ and NAA treated internodes, while high molecular weight xyloglucans were significantly correlated with length of internodes treated with PAA and NAA.     

Particle-based production of antibiotic rebeccamycin with Lechevalieria aerocolonigenes

Rebeccamycin is a promising antibiotic synthesized by the filamentous bacterium Lechevalieria aerocolonigenes. To date, the main investigations were focused on the biological effect or the structure elucidation of rebeccamycin. The aim of this study was to develop an efficient cultivation process based on shake flask experiments. The relationship between morphology and productivity and the positive effects of micro- and macroparticle enhanced cultivation on rebeccamycin production with this strain in the pellet-like morphology was investigated. The addition of talc microparticles increased the rebeccamycin concentration to 120 mg L⁻¹ and 3-fold compared to the control without microparticles. An up to 9-fold increase in rebeccamycin concentration was achieved using surface modified talc particles. Through the addition of glass beads as macroparticles, the highest rebeccamycin concentration of approximately 120 mg L⁻¹ was achieved by the induction of mechanical stress. Cultivation with glass beads enhanced the rebeccamycin concentration 22-fold compared to the control without macroparticles. By further increasing the diameter of the glass beads and increasing mechanical stress, the morphology was driven toward mycelial growth and the productivity was decreased. An adjusted mechanical stress, through the addition of macro-shaped glass bead particles, led to the desired increase in productivity.     

Effects of gibberellic Acid, calcium, kinetic, and ethylene on growth and cell wall composition of pea epicotyls

The influence of gibberellic acid (GA), calcium, kinetin, and ethylene on growth and cell-wall composition of decapitated pea epicotyls (Pisum sativum L. var. Alaska) was investigated. Calcium, kinetin, and ethylene each caused an inhibition of GA-induced elongation of pea stems. Gibberellic acid did not reverse the induction of swelling by Ca²⁺, kinetin, or ethylene. Both Ca²⁺ and ethylene significantly inhibited the stimulatory effects of GA on the formation of residual wall material. Although GA promoted the development of walls relatively low in pectic substances and pectic uronic acid, Ca²⁺, kinetin, and ethylene favored the formation of walls rich in these constituents. Calcium, kinetin, and GA, alone or in combination, had no effect on the production of ethylene by pea epicotyls.     

Influence of cobalt on soybean hypocotyl growth and its ethylene evolution

Development of dark-grown “Clark” soybean (Glycine max [L.] Merr.) seedlings is abnormal at 25 C but normal at 20 and 30 C. At 25 C, hypocotyls swell and fail to elongate normally; lateral root formation and seedling ethylene evolution are enhanced.

Co²⁺ promoted hypocotyl elongation of etiolated “Clark” soybean seedlings by 28% when grown at 25 C. The same growth-promoting concentration reduced hypocotyl thickness and primary root elongation by 28 and 43%, respectively. Co²⁺ inhibited ethylene production both of intact seedlings and of apical 1-centimeter hypocotyl segments with attached epicotyls and cotyledons by 65 and 60%, respectively. These results suggest that Co²⁺ exerts its effects on the hypocotyl growth by inhibiting ethylene production, and also confirm our previous conclusion that abnormal ethylene production at 25 C is responsible for the inhibition of hypocotyl elongation and for its swelling.

     

Influence of the Volume Fraction,Size and Surface Coating of Hard Spheres on the Microstructure and Rheological Properties of Model Mozzarella Cheese

This study investigated the influence of model filler particles (glass beads) on the microstructure and rheological properties of Mozzarella cheese. Model Mozzarella cheese composites with increasing volume fractions of glass beads of various sizes and surface properties were processed in a Rapid Visco Analyser (RVA). Confocal laser scanning microscope images showed that all the hard spheres were dispersed in the protein phase, rather than in the fat phase. Dynamic oscillatory rheology revealed that the volume fraction of the glass beads had a major influence on the complex modulus (G*) of the cheese composites, whereas the size and the coating of the glass beads had no influence. However, the zero shear viscosity (η ₀), measured using the creep-compliance test, was affected by both the size and the volume fraction of the glass beads. This indicated that there were some interactions between the glass beads and the cheese matrix. Filler–matrix interactions played a major role in the fracture properties of the cheese composites. The fracture stress (σ _f ) was highly dependent on the coating and the size of the glass beads. Simple equations for filled gels from the literature fitted well with the experimental results and could be successfully applied for future predictions. According to this study, the transfer of knowledge from filled polymer composites to model cheese appears relevant. This can provide a good basis for designing new dairy product structures.