Analysis of the Growth Response of Air-Grown Rice Coleoptiles to Submergence

The effect of submergence of air-grown rice seedlings (Oryza sativa L. var. Sasanishiki) on coleoptile growth and ultrastructure, extensibility and chemical composition of the cell walls was investigated. The lag-time between start of submergence and the onset of the enhancement of growth was less than 4 h. The growth response was associated with a drastic thinning of the cell walls and an increase in wall extensibility. At the outer epidermal wall of both air-grown and submerged coleoptiles electron-dense (osmiophilic) particles were detected. During submergence, the net accumulation of cellulose and hemicellulose was reduced, but the increase in pectic substances was unaffected. Submergence caused an 80% inhibition of the net accumulation of wall-bound phenolics (ferulic- and diferulic acid) compared with air-grown controls. The osmotic concentration of the tissue saps was not affected by submergence. Our results support the hypothesis that rapid coleoptile elongation under water is caused by an inhibition of the formation of phenolic cross-links between matrix polysaccharides via diferulate, which results in a mechanical stiffening of the cell walls in the air-grown coleoptile.     

Osmotic Stress-Induced Growth Suppression of Dark-Grown Wheat (Triticum aestivum L.) Coleoptiles

Application of 60 mM polyethylene glycol (PEG) to dark-grownwheat (Triticum aestivum L.) roots substantially reduced growthof coleoptiles. However, when PEG was removed, the growth rateof these coleoptiles greatly increased. Cell walls of stressedcoleoptiles remained loosened as compared with those of unstressedones. The osmotic potential of the stressed coleoptiles decreasedto that of the 60 mM PEG solution. On the other hand, the extentof decrease in the osmotic potential of stressed roots was smallerthan that of stressed coleoptiles. The osmotic potential difference() between the cell sap and the incubation medium of stressedroots was substantially higher than that of unstressed ones.The amount of ink moved from roots, where it was applied, tothe apical region of coleoptiles was significantly reduced underosmotic stress conditions. When water was exogenously appliedto abraded coleoptiles, the growth of these stressed coleoptileswas greatly promoted. These results suggest that inhibitionof coleop-tile growth under osmotic stress conditions is notdirectly related to a decrease in cell wall extensibility orto loss of the capacity to maintain osmotic potential gradients,but is caused by the reduction of the water supply from theroots to the coleoptiles. (Received August 26, 1996; Accepted December 18, 1996)     

Geotropic Response of Coleoptiles under Anaerobic Conditions

Coleoptiles of Avena sativa and Zea mays do not develop a geotropic response under anaerobic conditions.     

Effects of an Epidermal Growth Factor on the Growth of Zea Coleoptiles

Sections of the elongation zones of coleoptiles of Zea maysL. were incubated in buffers containing various concentrationsof human epidermal growth factor (EGF). The growth rate of sectionsincubated with 10, 1.0 or 0.1 fig liter¹ EGF was higher thanthat of control. EGF at 10 fig liter ^–1 increased thegrowth rate by 13% of the control rate. (Received September 25, 1990; Accepted June 19, 1991)     

Effect of Peeling on IAA-induced Growth in Avena Coleoptiles

The act of peeling removes the epidermis exclusively from Avenacoleoptiles. Peeling inhibits IAA-induced growth, by inhibitingthe growth of segments incubated in the presence of IAA, andpromoting that of those incubated in water. The magnitude ofthe inhibition of IAA-induced growth is proportional to theamount of epidermis removed. It is shown that neither lateralswelling, wounding, anaerobiosis, nor exposure to supraoptimalconcentrations of IAA cause the inhibition. It is concludedthat in Avena coleoptiles the epidermis regulates the rate ofexpansion of the underlying parenchyma cells and is the principaltarget of IAA-action. Avena sativa L., oat, coleoptile, indol-3-ylacetic acid, auxin, extension growth     

Geotropic Response of Wheat Coleoptiles in Absence of Amyloplast Starch

Young coleoptiles of wheat (Triticum durum var. Henry), depleted of amyloplast starch by incubation at 30°C with gibberellin plus kinetin, retained their geotropic responsiveness. Depleted coleoptiles curved upward more slowly than controls, but this was commensurate with their slower growth. The ratio of curvature to growth was about 50° per mm of elongation in both cases. Newly excised coleoptiles, though containing much more starch than incubated controls, curved only about 25° per mm. The tissue treated in gibberellin plus kinetin appeared to contain no starch when examined (a) freshly squashed, (b) as fixed material sectioned thin and stained by the PAS procedure, and (c) as electron micrographs. Shrunken, starch-free amyloplasts could be identified in certain regions, but these did not show evidence of asymmetric distribution under the influence of gravity. The possibilities that other organelles function as statoliths are considered, and it is concluded not only that georeception is independent of starch grains, but further that it may not be due to statoliths at all in an ordinary sense.     

The Response of Triticum aestivum Treated with Plant Growth Regulators to Acute Day/Night Temperature Rise

The fluctuation in temperature adversely affects grain development when the climate changes intermittently. This study investigated the effect of high day/night temperatures (34/30 °C, 38/34 °C and 42/38 °C) for two stress durations (24 h and 48 h) on Triticum aestivum. To ascertain the role of plant growth regulator (PGR) in alleviating the deleterious effects of high temperature stress, the combination of various PGRs (e.g., methyl jasmonate, salicylic acid, ascorbic acid, calcium chloride and indole acetic acid) were foliar sprayed twice; one week prior to commencement of anthesis stage and immediately after the exposure to high temperature stress. In general, the high temperature reduces plant growth, grain setting, and 100-grain weight. High temperature stress causes deterioration of plant photosynthetic machinery through a significant decline in energy dissipation, linear electron flow (LEF) and quantum yield of photosystem II (Phi2) which led to plant death. An increase in the antioxidant enzymes activity (SOD, APX, and CAT) was observed at 38/34 °C, while their activity declined sharply at 42/38 °C. Grain setting and filling were completely inhibited in plants exposed to 42/38 °C even when treated with different combinations of PGRs. Salicylic acid along with methyl jasmonate was the most effective PGR combination resulting in significant improvements in Phi2, NPQt, SOD, grain filling and grain protein content under high temperature stress. A strong correlation was observed between LEF and chlorophyll contents against the number of grains per spike and 100-grain weight. In summary, acute day and night temperature stress adversely affected wheat morphological, physiological, and yield traits, while foliar application of PGRs was partially effective in mitigating these harmful changes.

     

Effects of Morphactin on Indol-3yl-acetic Acid Transport, Growth, and Geotropic Response in Cereal Coleoptiles

The effects of the morphactin 2-ehloro-9-hydroxyfluorene-9-carboxylicacid methyl ester [CFM] on growth, geotropic curvature and transportand metabolism of indol-3yl-acetic acid [IAA-5-³H] in the coleoptilesof Zea mays and A vena saliva have been investigated. A strongcorrelation has been found to exist between the inhibition ofthe geotropic response and the inhibition of auxin transport.CFM supplied at concentrations sufficient to abolish auxin transporthas been shown to promote the elongation of Zea, but not ofAvena, coleoptile segments. CFM does not change the patternof metabolism of IAA in Zea coleoptile segments. In these segmentsIAA is metabolized when its concentration is high, but the radioactivitytransported basipetally, or laterally in geotropically stimulatedcoleoptiles, is virtually confined to the IAA molecule. Radioactivityexported into the basal receiver blocks is wholly confined toIAA. It is concluded that CFM inhibits the geotropic responsein coleoptiles by suppression of the longitudinal and lateralauxin transport mechanisms. The growth-promoting propertiesof this substance cannot be linked with its effects on eitherauxin metabolism or transport.     

Blue-Light-Induced Shrinking of Protoplasts from Maize Coleoptiles and Its Relationship to Coleoptile Growth

Protoplasts isolated from red-light-grown maize (Zea mays L.) coleoptiles shrank transiently upon brief exposure (e.g. 30 s) to blue light under background irradiation with red light. The maximal volume reduction (about 4% at a saturating fluence) occurred about 5 min after blue-light stimulation. The response was prevented by the anion-channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoic acid. Red light and far-red light did not induce any comparable response. Protoplasts of different sizes and those isolated from different coleoptile positions showed similar responses. After treatment with a saturating blue-light pulse, the protoplasts became responsive to a second pulse and gained full responsiveness within 5 min, suggesting that the photoreceptor system involves a dark-reversible component. The response to continuous blue light was also found to be transient. The protoplast volume was reduced during about 6 to 9 min of irradiation and returned within the next 30 min to the control level. The response to continuous blue light was saturated at 30 [mu]mol m-2 s-1. However, when the fluence rate was enhanced 10-fold after a period of irradiation at 30 [mu]mol m-2 s-1, the protoplasts showed another shrinking response. These and other kinetic results indicate that the photoreceptor system undergoes a photosensory adaptation. Growth in different zones of the coleoptile was inhibited by blue light transiently after pulse stimulation, as well as during continuous stimulation. It was concluded that the observed protoplast shrinking is related to the blue-light-induced inhibition of coleoptile growth.     

Growth and Geotropic Responses of Avena Coleoptiles to 4-Amino-3,5,6-Trichloropicolinic Acid

The elongation and geotropic responses of coleoptile sections of Avena sativa L. to various concentrations of 4-amino-3,5,6-trichloropicolinic acid (Tordon) proved to be qualitatively similar to those previously reported for 2,3,6-trichlorobenzoic acid (TCBA). Tordon stimulated growth in a range of concentrations from 1 × 10^?6 to 1 × 10^?4M but higher concentrations were inhibitory. Geotropic curvature was extensively depressed by 1 × 10^?5 and 1 × 10^?4M Tordon, concentrations which accelerated elongation. A similar differential effect has been reported for TCBA and other auxins. Several other picolinic acids and related compounds were tested, but only very slight responses were noted.     

Osmotic Stress in Coleoptiles and Primary Leaves of Wheat. II. Main Phosphorylated Fractions

Seedlings of wheat (Triticum durum, cv. Balcarceno-INTA) werewater-stressed in darkness with 20% polyethylene glycol (PEG)6000 or 0.3 M mannitol added to the root medium. At differenttimes and up to a total of 36 h of treatment the coleoptilesand primary leaves were cut and analysed for acid soluble-P,lipid-P, protein-P, alkali-stable organic-P, and nucleic acid-P.All the phosphorylated fractions were expressed on 100 mg oftotal coleoptile and primary leaves dry wt. Acid soluble-P (14.6µmol) accounted for most of the phosphorus. The lipid-Pfraction remained at an unchanged level (6.2 µmol/100mg dry wt) in control or in mildly osmotic stressed shoots ofseedlings over the period of treatment. The protein-P representedonly 2.4% (0.8 µmol) of the total phosphorus found incoleoptiles and primary leaves. The alkali-stable organic-Pand nucleic acid-P fractions represented 22.2% (7.2µ mol)and 11.1% (3.6 µmol) of the total phosphorus content ofshoots, respectively. All the phosphorylated fractions, exceptingthe lipid-P fraction, underwent a significant (P < 0.025)fall over periods of up to 36 h of wheat seedlings growth indarkness. A common characteristic found in all the phosphorylatedfractions was the fact that there were not significant differencesbetween control and 20% PEG 6000 or 0.3 M mannitol treated seedlingsduring 36 h of treatment and growth in darkness. However, whenseedlings were pulse-labelled with 32P during imbibition, someeffects of the osmotic stressants on several fractions couldbe seen. Specific radioactivity fell in acid soluble-P of controlshoots, but increased in 20% PEG 6000 and in 0.3 M mannitoltreated seedlings. Radioactive phosphorus was not found in thealkali-stable organic-P fraction. Lipid-P, nucleic acid-P, andprotein-P fractions increased their specific activities during36 h of shoot growth in control plants. Such 20% PEG 6000 and0.3 M mannitol restricted this increase during the same periodof time, the former being more active. Possible implicationsof phospholipid and phosphoprotein turnover in relation to waterstress are discussed.     

Physiological and Biochemical Mechanisms Involved in the Response to Abscisic Acid in Maize Coleoptiles

The role of abscisic acid (ABA) in controlling growth and developmenthas been studied in maize (Zea mays L.) coleoptile segments.Application of ABA reduces the elongation rate by about 50%and affects ion fluxes. In particular, proton extrusion is decreasedwhile potassium efflux is greatly enhanced. Apparently, ABAdoes not: seem to influence calcium influx from the apoplastinto the cytosol, but more likely it influences its efflux.Alteration of cytosolic calcium concentration may also be obtainedby increasing its release from internal stores. This possibilitymight be sustained by the increased hydrolysis of phosphatidylinositolupon ABA application. Change in the balance of ion fluxes shouldresult from regulation of transport mechanisms at the membranelevel and should produce changes in the transmembrane electricalpotential. The H⁺- ATPase and the ATP-dependent calcium transportactivities are both influenced by the treatment with ABA, –55%and –40%, respectively. Under these conditions [Ca²⁺]_cytand pH_cyt can be modified and, as a consequence of their regulation,they may play an important role in mediating the physiologicaland biochemical effects of ABA, acting as second intracellularmessengers. ¹Research supported by National Research Council of Italy, SpecialProject RAISA, Sub-Project N. 2, Paper n. 2782.     

Short-Term Stimulation of Growth Induced by the Apical Application of IAA to Intact Maize Coleoptiles

Indole-3-acetic acid (IAA), mixed with lanolin, was appliedto the surface of defined parts of the intact coleoptile ofmaize (Zea mays L.) seedlings, and the elongation growth ofthe coleoptile zone was monitored for a period of 8 h. WhenIAA was applied to a subapical region, the growth of zones locatedbelow was stimulated only temporarily, although stimulationwas continuous when IAA was applied directly to the zone thatwas monitored for growth. This short-term stimulation was observedat all concentrations tested (0.03–30 mg of IAA per gof lanolin). The duration of stimulated growth, which variedfrom 1 to 5 h, was longer as the distance from the site of IAAapplication was reduced and the concentration of IAA was increased.Growth during the post-stimulation phase after application ofa high dose of IAA could not be enhanced again by applying IAAnear the site of the first application, but it was enhancedwhen IAA was applied directly to the zone used to measure growth.These results suggest that the supply of IAA from the site ofapplication to lower zones is controlled internally such thatthe elevated concentration of IAA in these zones returns tothe concentration before IAA application. Either the feedbackregulation of auxin transport or the feedforward regulationof auxin metabolism may account for the suggested control ofIAA supply. (Received May 18, 1995; Accepted October 20, 1995)     

The Phototropic Response of Avena Coleoptiles following Localized Continuous Unilateral Illumination

When only part of the elongating zone on an Avena coleoptileis subject to continuous unilateral illumination, the majorphototropic response is restricted to the illuminated area.It is argued that previous studies have greatly exaggeratedthe contribution of any differential growth in non-illuminatedzones to the development of second positive phototropic curvature. Key words: Phototropism, Avena, Blue light, Growth rate     

Oxygen Dependence of Elongation Growth and Oxygen Uptake in Maize Coleoptiles

Auxin-mediated elongation growth of maize coleoptile segments is inhibited by reducing the O₂ concentration in the incubation medium to GT 100 μmol . 1^?1. The half-maximal elongation rate is reached at 40 μmol . 1^?1 O₂, i.e. about two orders of magnitude higher than with mitochondrial respiration. O₂ uptake of the segments measured under similar conditions with an O₂ electrode shows a very similar dependence on O₂ concentration. Auxin increases O₂ uptake by 5–10% when it induces growth. About 40% of the O₂ uptake is insensitive to inhibition by KCN. Auxin has no effect on O₂ uptake in the presence of KCN. The possibility that auxin-mediated elongation growth depends on a KCN-sensitive oxidative process, other than cytochrome c oxidase-catalyzed respiration, is discussed.     

Role of the Plasmalemma in Controlling the Gravitropic Response of Wheat Coleoptiles

The changes in the specific binding of ³H-IAA to the plasmalemma from segments of wheat (Triticum aestivumL.) coleoptiles and the physiological activity of the IAA–protein complexes thus formed in dependence on the duration of gravitational stimulation (GS) (1 g) were studied. The overall inhibition of the formation of IAA–protein complexes was accompanied by a transverse polarization of their functional activity occurring as early as within two minutes after the onset of GS. The pretreatment of plasmalemmal vesicles with 0.1 M CaCl₂prevented the in vitroIAA–protein complex formation in the plasmalemma. It is suggested that the GS results in an increase in the plasmalemma permeability for Ca²⁺, which reduces the capacity of the plasmalemma to bind IAA at the early stages of the gravitropic response.     

Some Responses of Avena Coleoptiles to Ethylene

Ethylene pretreatment of intact Avena seedlings or of excisedcoleoptile sections results in an increased response of thecells to auxin. It is suggested that ethylene brings about theacceleration of hydrolytic reactions controlling the physicalproperties of cell walls and hence increases their capacityfor growth. Coleoptile elongation of intact seedlings is inhibitedby ethylene; this inhibition is concurrent with a lateral expansionof the entire coleoptile. It is suggested that under a givenset of conditions coleoptile cells are capable of attaininga finite volume and that the preferential lateral expansioninduced by ethylene is accomplished at the expense of longitudinalextension. Experiments with intact and deseeded plants indicatethat lateral expansion depends on the supply of some factorfrom the endosperm.     

Timing of the Auxin Response in Coleoptiles and Its Implications Regarding Auxin Action

The timing of the auxin response was followed in oat and corn coleoptile tissue by a sensitive optical method in which the elongation of about a dozen coleoptile segments was recorded automatically. The response possesses a latent period of about 10 min at 23°C, which is extended by low concentrations of KCN or by reducing the temperature, but is not extended by pretreatments with actinomycin D, puromycin, or cycloheximide at concentrations that partially inhibit the elongation response. Analysis of the data indicates that auxin probably does not act on the elongation of these tissues by promoting the synthesis of informational RNA or of enzymatic protein. Not excluded is the possibility that auxin acts at the translational level to induce synthesis of a structural protein, such as cell wall protein or membrane protein. While the data do not provide direct support for this hypothesis, the speed with which cycloheximide inhibits elongation suggests that continual protein synthesis may be important in the mechanism of cell wall expansion.