Rhythmic and Blue Light-Induced Turgor Movements and Electrical Potential in the Laminar Pulvinus of Phaseolus vulgaris L.

The well-known circadian movement in Phaseolus leaves is usuallysuperimposed with small and somewhat irregular rhythmic changesof pulvinar movement. This study examines the relationship betweenthese small pulvinar movements and the membrane potential ofthe motor cell as well as the effect of light on them. Bluelight affected both the movements and potential while red andgreen lights of the same photon flux had little effect. Also,the difference in the membrane potential between the extensorand flexor cells was found to be closely related to the rhythmicturgor movement of the pulvinus. Changes in the potential differencealways preceded the movement. Sequential changes of the potentialdifference and turgor pressure in the motor cells, includingthe light-induced ones, are discussed in relation to the leafmovement. (Received August 8, 1985; Accepted November 11, 1985)     

The Phototropic Pulvinus of Bean Phaseolus vulgaris L. - Functional Features

Abstract: The laminar pulvinus of bean ( Phaseolus vulgaris L.) is a highly specialized organ for reorienting the lamina, and exhibits positive phototropic curvature. Structural and ultrastructural features of the pulvinus were studied to determine their possible role in its phototropic response. The vascular tissue forms a flexible, relatively inextensible central core enclosed by a starch sheath and surrounded by a multi-layered motor tissue. Phototropic curvature is a result of opposite anisotropic changes in volume of the cells in the exposed and opposite sectors of the motor tissue. Radial inflexibility of the epidermis and axial plasticity constrain these changes to the pulvinar axis. Anisotropic changes in volume of motor cells reduce the osmotic work involved. Motor cells exhibit features that are associated with high synthetic activity: thick cytoplasm with numerous ribosomes, polysomes, RER and SER, well-developed mitochondria and a large nucleus. Numerous, well-developed chloroplasts, with little starch, are increasingly abundant toward the periphery. The intercellular system is limited and partially filled with matrix. Stomata are absent and the motor tissue lacks vascularization. These features support the suggestion that the primary role of the chloroplasts in the photonastic response is photophosphorylation and photosynthetic electron transport (Koller et al., 1995^[339]).     

Diurnal Rb+ Transport from Roots to the Laminar Pulvinus in Phaseolus vulgaris L.

The primary leaves of kidney bean (Phaseolus vulgaris L.) openunder light and close in the dark by the deformation of thepulvinus resulting from diurnal distribution changes of K⁺,Cl^–, organic acid (or H⁺) and NO₃^–. When Rb⁺ was added as a tracer of K⁺ to the seedlings throughtheir roots, it was transported to the pulvinus cells duringthe light period but not during the dark period. Transpirationoccurred vigorously in the light but almost stopped in the dark.We concluded that Rb⁺ absorbed by the roots was carried to thepulvinus by the transpiration stream. Phaseolus vulgaris L., pulvinus, Rb⁺, diurnal transport transpiration stream     

The Rhythmic Leaf Movements after Regeneration of Partially Excised Pulvinus in Phaseolus vulgaris L.

Unlike the petiole or stem, the laminar pulvinus of the primaryleaves of Phaseolus vulgaris L. regenerated after a partialexcision. The histological and physiological aspects of theseregeneration processes have been studied. On the third day afterthe excision of the flexor (or extensor) region, the pulvinuswas regenerated. When the major part of the extensor was cutaway, the period and phase of the circadian leaf movements wereunchanged whereas the amplitude was greatly reduced. When theflexor region was excised, period, phase and amplitude weremaintained. Some changes could be seen in the ultradian movementsafter excision of flexor as well extensor regions. (Received August 31, 1988; Accepted March 30, 1989)     

Effects of Anoxia and Red Light on Changes Induced by Blue Light in the Membrane Potential of Pulvinar Motor Cells and Leaf Movement in Phaseolus vulgaris L.

In the dark, the membranes of the pulvinar motor cells of Phaseolusvulgaris L. were rapidly depolarized under anoxic conditionsand repolarized with re-aeration. This change in potential mayhave been due to suppression by anoxia of a respiration-dependent,electrogenic ion pump in the motor cells. When the pulvinuswas irradiated with blue light (BL) in the depolarized stateunder anoxic conditions, no marked depolarization occurred.Furthermore, a short pulse of BL did not induce transient depolarization.On continuous irradiation with red light (RL), the motor cellunder anoxic conditions showed slow recovery of the depolarizedmembrane potential. When a pulse of BL was superimposed on theRL after the recovery, transient depolarization occurred again. The leaf showed a small downward movement under anoxic conditionsbut recovered with re-aeration. Upward movement of the leafcaused by continuous application of BL to flexor cells changedto a downward movement under anoxic conditions, and re-aerationled to a return to upward movement. Unidirectional irradiationby BL of the flexor side did not cause upward movement of theleaf under anoxic conditions. However, unidirectional irradiationof RL to the flexor side caused downward movement of the leafunder anoxic conditions, which could be changed to upward movementby superimposition of irradiation with BL. The experimentalresults clearly show that BL acts mainly to inhibit (depolarize)an electrogenic component of the membrane potential in pulvinarmotor cells which is dependent on a supply of energy from respirationor photophsphorylation. (Received November 1, 1989; Accepted April 12, 1990)     

Cytokinin Effects on Leaf Architecture in Phaseolus vulgaris L.

Treatment of expanding primary leaves of bean plants (Phaseolnsvulgaris L. cv. Limburgse vroege) with benzyladenine (BA) orkinetin at 0.5 mM for five consecutive days resulted in thickerleaves showing a significant decrease in intercellular air spacevolume. Compared with control plants, exposed mesophyll cellsurface area was lower per unit tissue volume, but unchangedwhen expressed per unit leaf surface area. Stomata of treatedplants were not fully closed in the dark and they did not openas wide as controls in the middle of the light period, suggestingthat the treatment resulted in impaired stomatal action. Allthe effects mentioned were more pronounced after treatment withBA, compared to kinetin. In spite of their magnitude, the observedchanges in leaf structure and function did not seem to havean important effect on total leaf diffusion resistance to carbondioxide during the course of the light period. Key words: Cytokinins, Leaf architecture     

Blue Light Pulse-Induced Transient Changes of Electric Potential and Turgor Pressure in the Motor Cells of Phaseolus vulgaris L.

Blue light induces both depolarization of membrane potentialin the motor cell and turgor movement in the laminar pulvinusof bean plant. This paper describes the changes of electricpotential and turgor pressure induced in Phaseolus vulgarisL. by blue light pulses. A transient depolarization of membranepotential as large as 40 mV was induced by a short pulse of15 s blue light in motor cells of the laminar pulvinus. Thischange was not an action potential because of the absence ofa refractory period and threshold. The magnitudes of the responsewere dependent on the fluence of light. The response was long-lived,indicating that continuous input of light energy is not requiredfor a sustained response. The potential change was always followedby a transient turgor movement of the pulvinus. A molecular mechanism similar to a model postulated for theblue light response of stomata may operate in the motor cell.However, the direction of the electrical response to blue light(depolarization) in the motor cell was the opposite of thatin the guard cell (hyperpolarization). Turgor change of themotor cell by blue light was also opposite in direction (decrease). (Received February 19, 1988; Accepted June 28, 1988)     

The Effects of Light Breaks in the Dark Period on Flower-bud Development in Varieties of Phaseolus vulgaris L.

A light break imposed in the middle of, or more effectively,two-thirds of the way through a 13 h dark period inhibited thedevelopment of the first initiated flower buds and reduced theproduction of open flowers in two daylength-sensitive varietiesof Phaseolus vulgaris. The effects were similar to those ofa long photoperiod applied continuously and therefore provideclear evidence of the importance of the dark period in mediatingthe effects of daylength on flower-bud development in the twovarieties. Phaseolus vulgaris, bean flower-bud development, photopcriodism     

Light and Hormone Interaction in Apical Dominance in Phaseolus vulgaris L.

Gibberellic acid (GA₂), kinetin, and indole-3yl-acetic acid(IAA) each at four concentrations (0, 0.5, 5, and 50 µM)were applied alone and in all possible combinations to rootsof Phaseolus vulgaris L. grown under four different light regimes(7000, 14 000, 21 000, and 28 000 lx). GA₃ increased growthof main stem and laterals but reduced apical dominance, especiallyin the absence of, or at low kinetin concentrations. A highlevel of kinetin lowered GA₃ induced growth of main stems and,to a lesser extent, laterals. Kinetin greatly reduced apicaldominance, especially in the absence of, or at low GA₃ concentrations.IAA slightly reduced growth of main stems and laterals and slightlyincreased apical dominance. Generally the magnitude of the IAAeffects were less than those of GA₃ or kinetin and there wereless interaction between IAA and other factors than betweenGA₃ or kinetin and other factors. Light affected growth of bothmain stem and laterals but the effect was dependent on GA₃ andkinetin levels and the interactions were complex. Generallya hormone balance seems to be operative with gibberellin-promotinggrowth of main stem and laterals and cytokinins and possiblyauxins preventing excessive elongation. Differential responsesbetween main stem and lateral may be due to different localhormone concentrations and over-all responses may be temperedby light intensity.     

Leaf Growth in Phaseolus vulgaris: 2. Temperature Effects and the Light Factor

Results are reported for two experiments in which bean plantswere grown in a range of daylengths from i hour to continuousillumination at temperatures of 15^° C and 25^° C. Markeddifferences in total leaf area and in areas of individual leaveswere found, and these are related in a quadratic fashion withtotal radiation received per day. With high levels of radiationa decrease in leaf area was noted, and this is not explicablein terms of a simple nutritional hypothesis. There are pronouncedeffects of total radiation on specific leaf area but the mechanismby which these effects are brought about is not clear. The mostimportant influence of temperature on the expansion of the leafsurface appear to be: firstly, by controlling the rate at whichleaves unfold from the stem apex; and secondly, by controllingthe partition of dry matter between the leaves and the restof the plant. No effects of daylength on vegetative developmentwere found.     

Temperature Effects on Mitochondrial Respiration in Phaseolus acutifolius A. Gray and Phaseolus vulgaris L

Electron transport, using succinate as a substrate, was measured polarographically in mitochondria isolated from Phaseolus vulgaris and P. acutifolius plants at 25°C and 32°C. Mitochondria isolated from P. vulgaris plants grown at 32°C had reduced electron transport and were substantially uncoupled. Growth at 32°C had no effect on electron transport or oxidative phosphorylation in P. acutifolius compared to 25°C grown plants. Mitochondria isolated from 25°C grown P. vulgaris plants measured at 42°C were completely uncoupled. Similarly treated P. acutifolius mitochondria remained coupled. The uncoupling of P. vulgaris was due to increased proton permeability of inner mitochondrial membrane. The alternative pathway was more sensitive to heat than the regular cytochrome pathway. At 42°C, no alternative pathway activity was detected. The substantially greater heat tolerance of P. acutifollus compared to P. vulgaris mitochondrial electron transport suggests that mitochondrial sensitivity to elevated temperatures is a major limitation to growth of P. vulgaris at high temperatures and is an important characteristic conveying tolerance in P. acutifolius.     

Growth, Pigment Synthesis, and Ultrastructural Responses of Phaseolus vulgaris L. cv. Blue Lake to Intermittent and Flashing Light

Growing bean plants (Phaseolus vulgaris L. cv. Blue Lake) on cycles of 1 minute light-1 minute dark or 5 minutes light-5 minutes dark, providing an integrated 12 hours light-12 hours dark per day for each set of plants, led to production after 21 days of new leaves low or lacking in chloroplast pigments. Subsequently, dry weight increase was sharply cut. Leaf area was affected by the light regimes after the second week of growth. By the fourth week, plants on the 1 minute light-1 minute dark cycle showed about one-half the leaf area of the controls. Shoot growth was favored over root growth to the greatest degree on the 1 minute light-1 minute dark regimes. Chlorophyll a/b ratios were close to 3.0 in all of the intermittent light regimes, but the total amounts of chlorophyll in milligrams per primary leaf were higher from day 9 to day 23 for the 12 hour light-12 hours dark controls than for other plants.

Although they produced chlorophyll, the plants receiving 1 or 2 milliseconds per second of light continued to lose weight at the same rate as the dark controls; thus, it is assumed there was no net photosynthesis. Plants receiving flashing light allocated significantly more food reserves from the seed to roots than did dark controls. Total chlorophyll formation was significantly accelerated by 2 milliseconds per second light. With 1 millisecond per second light, it took 5 days longer to achieve the same level of chlorophyll. After the 18th day, there was a steady decline in chlorophyll, b degrading more rapidly than a.

It is thought that several light-driven reactions are involved in the observed pigment synthesis, photosynthesis, food allocation, and growth of bean. Some of these reactions may be cyclic and others linear. Collectively, they must reach a harmonic point for normal metabolism and development to occur. Because time courses for each of these reactions are different, the intermittent and flashing light technique offers the possibility of individually studying some of the key light-driven reactions.

     

Effects of simulated acid rain on the photosynthetic characteristics of Phaseolus vulgaris L.

A single treatment with a low pH solution of bean plants led to serious changes in the net photosynthetic rate (P _N) as well as in various parameters of photosystem 2 (PS2) activity. A considerable suppression of P _N was established already in the first hours (3 and 5) following the acid treatment (pH 2.4-1.8). The period of strong inhibition of CO₂ uptake and photochemical activity was followed by the period of recovery (24-72 h). At a single spraying, pH values exceeding 2.0 did not lead to irreversible damages of the photosynthetic apparatus. The damages resulting from treatments with pH 2.0 and 1.8 were on the threshold of irreversible ones and were the cause of faster ageing. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effects of simulated acid rain on the photosynthetic characteristics of Phaseolus vulgaris L.

A single treatment with a low pH solution of bean plants led to serious changes in the net photosynthetic rate (P _N) as well as in various parameters of photosystem 2 (PS2) activity. A considerable suppression of P _N was established already in the first hours (3 and 5) following the acid treatment (pH 2.4-1.8). The period of strong inhibition of CO₂ uptake and photochemical activity was followed by the period of recovery (24-72 h). At a single spraying, pH values exceeding 2.0 did not lead to irreversible damages of the photosynthetic apparatus. The damages resulting from treatments with pH 2.0 and 1.8 were on the threshold of irreversible ones and were the cause of faster ageing.     

Physiological and Anatomical Effects of Growth Temperature on Phaseolus vulgaris L. Cultivars

Two Phaseolus vulgaris L. cultivars were grown at 20/15, 25/20,and 30/25 °C day/night temperatures in growth chambers witha 16 h thermoperiod corresponding to the photoperiod. When thefirst trifoliolate leaf was fully expanded rates of CO₂ exchange(CER) were measured at 27 °C and saturating light usinginfrared gas analysis. Stomatal (r_s) and mesophyll resistances,CO₂ compensation points, activities of the enzymes ribulosebisphosphate carboxylase (RuBPCase), glycolate oxidase (GAO),malate dehydrogenase (MDH), and fructose-1, 6 diphosphate (FDP),chlorophyll content, Hill activities, and leaf anatomy at boththe light and electron microscope level were also investigatedin these leaves. Rates of CO₂ exchange in the light, transpiration rate, andchlorophyll content increased with increasing growth temperaturewhile leaf thickness, specific leaf weight, RuBPCase activity,compensation point, and stomatal resistance decreased. Mesophyllresistance also decreased when calculated assuming zero chloroplastCO₂ concentration (rm, o), but not when calculated assuminga chloroplast CO₂ concentration equal to the CO₂ compensationconcentration (rm, g). Average leaf size was maximal in 25/20°C plants while dark respiration, MDH activity, stomataldensity, and starch were minimal. The activities of GAO andFDP and Hill activity were not affected by temperature pretreatment.     

Characterization of a membrane-associated apoplastic lipoxygenase in Phaseolus vulgaris L.

An extracytoplasmic 86.7 kDa protein was isolated from intercellular washing fluids (IWF) of Phaseolus vulgaris etiolated hypocotyls. Micro sequencing of tryptic peptides of the 86.7 kDa protein revealed 100% identity with a bean lipoxygenase (LOX) protein fragment. Purified P87-LOX exhibited LOX activity characterized by an optimal pH of 6.0 and linolenic acid as an optimal substrate, and was classified as a 13-LOX with respect to its positional specificity of linoleic acid oxygenation. A protein identical to P87-LOX, as determined by MALDI-TOF analysis and biochemical characterization, was purified from hypocotyl microsomes. Immunoblot analysis showed that P87-LOX is present in plasma membrane-enriched fractions, from which it was solubilized using high ionic strength buffers. These observations suggest that P87-LOX is a peripheral protein associated to the apoplastic face of the plasma membrane.     

The influence of aluminium availability on phosphate uptake in Phaseolus vulgaris L. and Phaseolus lunatus L.

Aluminium toxicity is one of the major limiting factors of crop productivity on acid soils. High levels of available aluminium in soil may induce phosphorus deficiency in plants. This study investigates the influence of Aluminium (Al) on the phosphate (P_i) uptake of two Phaseolus species, Phaseolus vulgaris L. var. Red Kidney and Phaseolus lunatus L. The two bean species were treated first with solutions of Al at different concentrations (0, 25, 50 and 100 μM, pH 4.50) and second with solutions of P_i (150 μM) at pH 4.50. The higher the Al concentration the higher the Al concentration sorbed but P. vulgaris L var. Red Kidney adsorbed significantly more Al than P. lunatus L. Both species released organic acids: P. vulgaris L var. Red Kidney released fumaric acid and P. lunatus L. fumaric and oxalic acids which could have hindered further Al uptake.The two bean species showed a sigmoid P_i uptake trend but with two different mechanisms. P. vulgaris L var. Red Kidney showed a starting point of 3 h whereas P. lunatus L. adsorbed P_i immediately within the first minutes. In addition, P. vulgaris L var. Red Kidney presented significantly higher P_i uptake (higher uptake rate ‘k’ and higher maximum adsorption ‘a’ of the kinetic uptake model). The Al treatments did not significantly influence P_i uptake. Results suggest that P. lunatus L. might adopt an external Al detoxification mechanism by the release of oxalic acid. P. vulgaris L var. Red Kidney on the other hand seemed to adopt an internal detoxification mechanism even if the Al sorbed is poorly translocated into the shoots. More detailed studies will be necessary to better define Al tolerance and/or resistance of Phaseolus spp.     

Photosynthesis, Carbohydrate Metabolism, and Export in Beta vulgaris L. and Phaseolus vulgaris L. during Square and Sinusoidal Light Regimes

Rates of photosynthesis, sucrose synthesis, starch accumulation and degradation were measured in sugar beet (Beta vulgaris L.) and bean (Phaseolus vulgaris L.) plants under a square-wave light regime and under a sinusoidal regime that simulated the natural daylight period. Photosynthesis rate increased in a measured manner in direct proportion to the increasing light level. In contrast to this close correspondence between photosynthesis and light, a lag in photosynthesis rate was seen during the initial hour under square-wave illumination. The leaf appeared to be responding to limits set by carbon metabolism rather than by gas exchange or light reactions. Under the sinusoidal regime starch degradation occurred during the first and last 2 hours of the photoperiod, likely in response to photosynthesis rate rather than directly to light level. Starch broke down when photosynthesis was below a threshold rate and accumulated above this rate. Under square-wave illumination, accumulation of starch did not begin until irradiance was at full level for an hour or more and photosynthesis was at or near its maximum. Under a sinusoidal light regime, sucrose synthesis rate comprised carbon that was newly fixed throughout the day plus that from starch degradation at the beginning and end of the day. Synthesis of sucrose from recently fixed carbon increased with increasing net carbon fixation rate while its formation from degradation of starch decreased correspondingly. The complementary sources of carbon maintained a relatively steady rate of sucrose synthesis under the changing daytime irradiance.     

Effects of radiation quality on the opening of stomata in intact Phaseolus vulgaris L. leaves

In intact French bean (Phaseolus vulgaris L.) leaves blue radiation enhanced opening of stomata both when it was used individually and when it was used as preirradiation before "white light" irradiation. Effects of red radiation were just the contrary.