Some Physical-kinetic Considerations in Penetration of Naphthaleneacetic Acid through Isolated Pear Leaf Cuticle

The penetration of naphthaleneacetic acid (NAA) through enzymatically isolated pear leaf cuticle (Pyrus communis L. cv. Bartlett) is reported herein. Penetration of NAA increased with increasing lime and attained a steady state in approximately 20 minutes. The quantity of NAA penetrating was directly related to the concentration of the donor solution. NAA that penetrated the cuticle was shown to he unaltered. The Penetration of NAA from inside to outside is similar to that from outside to inside. Isolated stomatous lower cuticle permitted approximately 10-foId greater penetration of NAA than the astomatous upper cuticle. The penetration of NAA through isolated pear leaf cuticle is highly temperature dependent, exhibiting a temperature coefficient (Q₁₀) of about 5.6 between 15° and 25 C. The low quantities of chemicals penetrating through the isolated cuticle reported herein and elsewhere are considered to he a characteristic of the technique and not an absolute limitation of the cuticle. Cuticular penetration could account for physiological quantities of NAA entering the plant.     

Cation Penetration through Isolated Leaf Cuticles

The rates of penetration of various cations through isolated apricot Prunus armeniaca L. leaf cuticles were determined. Steady state rates were measured by using a specially constructed flow-through diffusion cell. The penetration rates of the monovalent cations in group IA followed a normal lyotropic series, i.e., CS⁺ ≥ Rb⁺ > K⁺ > Na⁺ > Li⁺. The divalent cations all penetrated through the cuticle more slowly than the monovalent cations. Comparison of the relative values of k (permeability coefficient) and D (diffusion coefficient) indicates that the penetration of ions through isolated cuticles took place by diffusion and was impeded by charge interactions between the solute and charge sites in the penetration pathway. Cuticular penetration rates of K⁺ and H₂O at pH above 9 were of similar magnitude. At pH 5.5 H₂O penetration was not affected but that of K⁺ was greatly reduced. From this observation and from data on cuticle titration and ion adsorption studies, we hypothesize that cuticular pores are lined with a substance (perhaps a protein) which has exposed positively charged sites.     

Rheological Properties of Enzymatically Isolated Tomato Fruit Cuticle

Rheological properties were determined for cuticular membranes (CMs) enzymatically isolated from mature tomato (Lycopersicon esculentum Mill. cv Pik Red) fruit. The cuticle responded as a viscoelastic polymer in stress-strain studies. Both CM and dewaxed CM expanded and became more elastic and susceptible to fracture when hydrated, suggesting that water plasticized the cuticle. Dewaxing of the CM caused similar changes in elasticity and fracturing, indicating that wax may serve as a supporting filler in the cutin matrix. Exposure of the cuticle to the surfactant Triton X-100 did not significantly affect its rheological properties.     

Effects of Gibberellic Acid and Naphthaleneacetic Acid on Petiole Senescence and Subtended Peduncle Growth of Cyclamen persicum Mill

Removal of the blade from the leaf subtending the first flowerbud on Cyclamen persicum ‘Swan Lake’ plants causedthe petiole of that leaf to senesce, but had no effect on thegrowth of the flower peduncle in the debladed petiole's axil.A 10 mg NAA l^–1 application generally had no effect onpetiole senescence, peduncle elongation or flowering date whenapplied to the cut end of the petiole after blade removal. A25 mg GA₃ l^–1 application or a combination of 25 mg GA₃l^–1 application or a combination of 25 mg GA₃ l^–1plus 10 mg NAA l^–1 delayed petiole senescence and enhancedpeduncle elongation and subsequent flowering. No treatment significantlyaltered peduncle length at the time of flowering. Cyclamen persicum Mill, ‘Swan Lake’, tissue receptivity, flowering, GA₃, NAA     

Metabolism of Auxin in Pine Tissues: Naphthaleneacetic Acid Conjugation

Incubation of sections of various tissues of Pinus pinea L. and Pinus halepensis Mill, with α-naphthaleneacetie acid-1-¹⁴C (NAA) resulted in two metabolites which proved to be l-O-(α-naphthylacetyl)-β-D-glucose (NAGlu) and α-naphthylacetylaspartic acid (NAAsp). NAGlu was purified by means of insoluble polyvinyl-polypyrrolidone (Polyclar AT) column chromatography and preparative thin layer chromatography and identified by its ‘H-nuclear magnetic resonance and mass spectra. NAAsp was partially purified by means of preparative thin-layer chromatography and identified by co-chromatography with a synthetic standard and hydrolysis to the parent compounds. NAA and L-aspartic acid. Needles and shoot bark rapidly converted NAA mainly to NAGlu. In contrast, woody roots and shoot wood showed a much slower rate of conjugation with the formation of both NAGlu and NAAsp. Preincubation of wood sections in NAA increased the formation of NAGlu, whereas the formation of NAAsp was almost unaffected.     

Observations on the Three-dimensional Structure of the Leaf Cuticle in Certain Plants

The three-dimensional structure of the leaf cuticle has beenstudied microscopically on cuticular preparations from a numberof plants. The cuticle is one- to several-layered, the latterincluding, besides the epidermis, sub-epidermal layers suchas hypodermis, palisade, and spongy mesophyll.     

脱落酸对玉米胚芽和大豆子叶线粒体呼吸的影响

在提取玉米和大豆线粒体后,以ABA处理线粒体,发现在不同底物存在下,ABA均可增加线粒体的耗氧速率,呼吸速率随ABA浓度变化的结合常数（Kd）值为1．43μmol／L。ABA对4态呼吸的促进作用更为显著,因而导致呼吸控制下降,P／O比降低。蛋白质合成抑制剂环己酰亚胺不影响ABA的效应。以ABA预处理大豆子叶,虽然也促进了呼吸作用,但不改变呼吸控制和P／O.     

Effect of Auxins (2,4-Dichlorophenoxyacetic acid, Indole-3-Acetic Acid and Naphthaleneacetic Acid) on the Accumulation of {gamma}-Aminobutyric Acid in Excised Rice Root Tips

-Aminobutyric acid (GABA) accumulation occurs in cultured ricecells when ammonium is added to the medium [Kishinami and Ojima(1980) Plant Cell Physiol. 21: 581–589]. Whether thisphenomenon occurs in rice plant tissues was examined with respectto exogenously supplied auxins: 2,4-dichlorophenoxyacetic acid(2,4-D), indole-3-acetic acid (IAA) and naphthalene-acetic acid(NAA). In intact rice plants grown in medium containing ammonium withoutauxin, glutamine first increased, then asparagine graduallyincreased. In both shoots and roots, the asparagine contentbecame the highest among four amino acids after 4 days of cultureperiod. GABA did not increase at all, its level remaining lowin both shoots and roots throughout the culture period. GABA accumulation was observed in excised rice root tips whenthey were incubated in the medium containing ammonium in thepresence of 2,4-D, IAA or NAA. In the absence of auxin, however,excised rice root tips accumulated asparagine and glutamine,but not GABA. Rice root segments obtained from a region in whichroot cells had already developed to maturity did not accumulateGABA but asparagine and glutamine in the presence of both ammoniumand 2,4-D. These results suggest that GABA accumulation occurs in rapidlygrowing and dividing tissue, such as the apical meristem ofrice root in the presence of auxin during ammonium assimilation. (Received June 15, 1987; Accepted March 14, 1988)     

Uptake and Release of Abscisic Acid by Isolated Photoautotrophic Mesophyll Cells, Depending on pH Gradients

Uptake and release of abscisic acid (AbA) by isolated mesophyll cells of Papaver somniferum is characterized by the following observations: (a) Uptake rate is a linear function of the external AbA concentration in the range from 10⁻⁶ to 5 × 10⁻⁵ molar, and decreases with increasing pH. At any pH, uptake rate is linearly related to the concentration of undissociated abscisic acid, calculated from the pK = 4.7 according to the Henderson-Hasselbalch equation. At low external pH (5.0), AbA accumulation in the cells is about 10-fold. (b) Uptake of AbA is completely inhibited by salts such as KNO₂ or sodium acetate, which decrease the pH gradient between medium and cells. KCN or m-chlorocarbonylcyanide phenylhydrazone inhibits AbA uptake only after longer incubation periods (20-40 minutes). (c) Uptake rate as well as equilibrium concentration is significantly higher in light than in darkness. (d) At low external pH, release of AbA from preloaded cells is strongly stimulated by KNO₂. It is concluded that AbA is distributed between leaf cells and free space according to pH gradients, with the undissociated abscisic acid being the main penetrating species. Uptake and release occur via diffusion, without participation of a carrier.     

Effect of Growth Regulators and Herbicides on Photosynthetic Partial Reactions in Isolated Leaf Cells

The effect of indoleacetic acid, gibberellic acid, 2,4-dichloro-phenoxyacetic acid and amitrole at various concentration levels ranging from 0.05 to 1.0 mM on the photosynthetic evolution of O₂ and ¹⁴CO₂ fixation by isolated leaf cells was studied. The plant growth regulators enhanced O₂ evolution and ¹⁴CO₂ fixation at low concentrations and were inhibitory beyond a critical level. The amitrole had an inhibitory effect at all the concentration levels used. All the substances exhibited similar patterns of effect on the ferricyanide reduction by isolated chloroplasts and on the electron transport rates of sub-chloro-plast particles containing PS-I and PS-II independently, under non-phosphorylating conditions. As was seen from the response in all the three electron transport systems of the chloroplast studied, the electron transport chain connecting PS-II and PS-I could be considered as a possible site of action at least for the growth regulating substances as it is the only part that is common to all the three reactions. The phosphorylation associated with this part of the electron transport was “inhibited” by the substances even at the lowest concentration used. The stimulation of non-phosphorylating electron flow, with a simultaneous reduction in the rate of ATP synthesis, at low concentration levels indicated that these substances played a possible uncoupling role. The amitrole on the other hand appeared to have a generalized non-specific inhibitory action on all the partial reactions of photosynthesis.     

Effect of Intraluminal pH on the Absorption of Pteroylmonoglutamic Acid

The administration of pteroylmonoglutamic acid and either sodium bicarbonate or phenytoin sodium in normal subjects and epileptics results in lower serum levels of folic acid than when administered alone. Bicarbonate administration was shown to produce a more alkaline pH in the jejunum, while an alkaline pH was found in some epileptics on phenytoin therapy. The physicochemical mechanisms whereby folic acid absorption is reduced in an alkaline medium are considered, and the implications on folate absorption in other disease states are discussed. The mechanism by which phenytoin sodium alters the intrajejunal milieu is obscure.     

Effect of Salicylic Acid on Leaf Anatomy and Chloroplast Ultrastructure of Barley Plants

Light and electron microscopy were used to relate histological and ultrastructural differences of barley leaves treated with different concentrations of salicylic acid (SA, 100 µM-1 mM). Light microscopy revealed that the thickness of all leaf tissue components decreased in SA-treated plants. The effect was most pronounced on the width of the adaxial epidermis and on the size of the bulliform cells. The chloroplast ultrastructure was also affected by SA treatment. Swelling of grana thylakoids in various degrees, coagulation of the stroma, and increase in chloroplast volume were observed. 1 mM SA caused a vast destruction of the whole plastid structure.     

Effect of Cations on Effective Permeability of Leaf Cuticles to Sulfuric Acid

Many plants are exposed to prolonged episodes of anthropogenic acid precipitation with pH values of 4 or less, but there is little evidence of widespread direct damage to the plant cells. Acids appear to permeate leaf cuticle via charged pores, which act as a fixed buffer that delays but does not stop acid movement. We investigated the effect of cations on the movement of protons through astomatous isolated leaf cuticles of pear (Pyrus communis L.) and rough lemon (Citrus limon [L.] Burm. fils cv Ponderosa). Chloride salt solutions of Na, K, Ca, Cd, Mg, Gd, or Y in a diffusion apparatus were applied to the morphological inner surface of the cuticle, while the outer surface faced a large volume of pH 3 or 4 sulfuric acid. Effective permeability was calculated from the change in the pH of the inner solution as measured with a pH microelectrode. Monovalent cations caused either no change (pear) or promotion (rough lemon) of proton movement. Divalent cations reduced proton movement in a concentration-dependent manner (both species), whereas trivalent cations (rough lemon only) caused the effective permeability to decrease to near zero. Inhibition by 10 mM CaCl2 was reversed with water. The effects of these cations on the permeability of cuticles to protons was used to elucidate mechanisms by which cations can protect leaves from acid precipitation in nature.     

Effect of Abrading the Cuticle Using Emery

Abrading the cuticle is sometimes required in order to measureauxin-stimulated acidification. We have determined, however,that the process should be used with caution. The diameter ofthe Edmund Scientific emery No. 305 particles was found to benear 40 times greater than cuticle thickness (0 –14 µm).Thus it was not surprising to find that even gentle abrasionmight cause damage to the epidermal cells. The resultant woundingmay complicate interpretastion of experimental results becausethe healing process shown to follow the wounding is likely tohave an auxin – regulated component. Experiments shouldbe designed to differentiate wound – related auxin effectsfrom elongation-related auxin effects.