Effects of Sodium Azide on Sodium Fluxes in Frog Striated Muscle

Unidirectional Na fluxes from frog''s striated muscle were measured in the presence of 0 to 5 mM sodium azide. With azide concentrations of 2 and 5 mM the Na efflux was markedly stimulated; the Na efflux with 5 mM azide was about 300 per cent greater than normal. A similar increase was present when all but the 5.0 mM sodium added with azide was replaced by choline. 10^-5 M strophanthidin abolished the azide effect on Na²⁴ efflux. Concentrations of azide of 1.0 mM or less had no effect on Na efflux. The Na influx, on the other hand, was only increased by 41 per cent in the presence of 5 mM NaN₃. From these findings it is concluded that the active transport of Na is stimulated by the higher concentrations of azide. The hypothesis is advanced that the active transport of Na is controlled by the transmembrane potential and that the stimulation of Na efflux is produced as a consequence of the membrane depolarization caused by the azide.     

Effects of Salinity, Diurnal Cycle and Age on Nucleotide Pools of Bean Leaves

In the present study, we investigated the relationship betweensalt stress and nucleotide levels in the shoot of Phaseolusvulgaris L. cv. Stringless Green Pod to determine if reducedgrowth was correlated with reduced nucleotide levels. Overallfresh weight of 25-d-old plants after having been on full salttreatment for 7 d was 33% lower compared with untreated plants.Shoot fresh weight decreased by 40% compared with 22% for theroots thus increasing the root to shoot ratio from 0·7to 0·9. We examined young and juvenile leaves as wellas mature leaves in order to compare growing tissue to fullyenlarged tissue. To ascertain whether the effects of salt stresson nucleotide pools were more severe during the day than atnight, we studied the combined effects of diurnal cycle andsalt stress on these nucleotide pools. Salt treatment selectivelyaffected certain nucleotide pools with the adenine nucleotides(AdN) being the most affected. We found large diurnal fluctuationsof AdN pools in all leaves. During the day, AMP and ADP increasedwhile ATP decreased. The sum, ATP + ADP, tended to remain constantand in mature leaves total AdN increased with AMP, an indicationof net synthesis. At night, ATP increased in all leaves. However,salt stress prevented this night-time increase in mature leaveswhile enhancing it in juvenile and young leaves. In the daytime,salt stress caused a nearly 2-fold increase in AMP of youngleaves and a large increase in the adenylate kinase mass actionratio (K). At night, the excess AMP disappeared with no changein total AdN. It is clear from these results that salt stressdid not reduce shoot growth by depleting ATP in growing leaves.It did, however, reduce the ATP level of mature leaves and perhapstheir ability to supply essential metabolites for growing regions. Key words: Phaseolus, nucleotides, salt stress, salinity, growth     

Effects of 6N-Benzyladenine on Primary Leaves of Intact Bean Plants and on their Sodium Absorption Capacity

The application of ⁶N-benzyladenine (BA) to primary bean-leaves attached to the intact plant resulted in increased leaf area. This was due to an extension of the duration of the period of leaf expansion. The dry-weight percentage of untreated leaves decreased continuously from emergence to abscission. In BA treated leaves this decrease stopped with the cessation of leaf expansion. Net chlorophyll synthesis occurs in untreated leaves as long as they continue to expand, shortly afterwards net chlorophyll decomposition starts. Benzyladenine treatments diminished the rate of chlorophyll synthesis immediately after its first application, but prevented the loss of chlorophyll afterwards. The abscission of primary leaves was also delayed by BA. During the period of rapid leaf expansion in slices cut from BA treated leaves, sodium absorption rates were lower than in those from untreated leaves. From the age of 14 days after sowing, till abscission time of untreated primary leaves, the BA treatment did not significantly affect sodium absorption rates by leaf slices.     

The Effect of Age on the Phytochrome-Mediated Chlorophyll Formation in Dark-grown Bean Leaves

The effect of red and far-red treatment on chlorophyll synthesis in dark-grown bean leaves was studied at various ages. Although the effect was pronounced in the old leaves, no effect was observed in the young ones (4 days old). In the 5-day old leaves a measurable effect of red light pretreatment can be observed, whereas the far-red reversal effect was not observed. — The length of the dark period between the red pretreatment and the continuous illumination is also age dependent. Leaves older than 6 days show a maximum at about six hours, while in the young leaves the red light effect increases with the time of dark incubation up to the 24 hours tested. — The reversal effect of far-red light on protochlorophyllide regeneration was also examined. The far-red light has no reversal effect on leaves younger than 6 days old, while on the old leaves it has such an effect.     

Effects of Heavy Metals on Stomatal Movements in Broad Bean Leaves

How do heavy metals affect stomatal movements and whether water channels are involved in stomatal movements was investigated in broad bean (Vicia faba L.) leaves. Three-week old fully expanded leaves were harvested. Leaf epidermis was peeled off and soaked in the Mes–KOH buffer containing the salts of heavy metals. Stomatal aperture was measured under the microscope. The tested heavy metal ions, such as Hg²⁺, Pb²⁺, Zn²⁺, and La³⁺, partly inhibited stomatal opening in light or closing in darkness at submillimolar concentrations, while K⁺, Na⁺ and Mg²⁺ had no visible effects on stomatal movements. As compared to La³⁺, Hg²⁺ affected stomatal movements more significantly. Stomatal movements were almost completely inhibited under a combined Hg²⁺ and La³⁺ treatment. Apparently, La³⁺, a Ca²⁺ channel blocker, inhibits the changes in the cytosolic Ca²⁺ concentration in guard cells, thus affecting stomatal movements. The inhibitory effect of Hg²⁺ on stomatal movements may be explained by the inhibition of water channels. Like Hg²⁺, Zn²⁺ and Pb²⁺ interfered with stomatal movements. It is concluded that heavy metals at submillimolar concentrations inhibit stomatal movements. They may affect water fluxes through guard cell membranes in different ways, i.e., Hg²⁺, Pb²⁺, and Zn²⁺ inhibit water channels, whereas La³⁺ block ion channels. Water channels may be involved in stomatal movements by regulating water fluxes and play a dominant and primary role in stomatal movements.     

Effects of External Potassium and Strophanthidin on Sodium Fluxes in Frog Striated Muscle

Unidirectional Na fluxes in isolated fibers from the frog''s semitendinosus muscle were measured in the presence of strophanthidin and increased external potassium ion concentrations. Strophanthidin at a concentration of 10^-5 M inhibited about 80 per cent of the resting Na efflux without having any detectable effect on the resting Na influx. From this it is concluded that the major portion of the resting Na efflux is caused by active transport processes. External potassium concentrations from 2.5 to 7.5 mM had little effect on resting Na efflux. Above 7.5 mM and up to 15 mM external K, the Na efflux was markedly stimulated; with 15 mM K the Na influx was 250 to 300 per cent greater than normal. On the other hand, Na influx was unchanged with 15 mM K. The stimulated Na efflux with the higher concentrations was not appreciably reduced when choline or Li was substituted for external Na, but was completely inhibited by 10^-5 M strophanthidin. From these findings it is concluded that the active transport of Na is stimulated by the higher concentrations of K. It is postulated that this effect on the Na "pump" is produced as a result of the depolarization of the muscle membranes and is related to the increased metabolism and heat production found under conditions of high external K.     

Effects of Bean Leaf Age on Pathogenicity by Botrytis fabae

Leaves from field bean plants grown out of doors were inoculated with conidia of B. fabae immediately after detaching from stems. The oldest leaves developed more lesions than youngest ones, although they were not chlorotic. On intact plants at high humidities, established lesions on young leaves increased in size at only half the rate of those on old. but still green leaves. Seven days after inoculation a higher proportion of lesions on old leaves bore conidia than those on young leaves, but leaf age had no significant effect on numbers of conidia per mm² of lesion area. Young leaflets from bean plants grown in a controlled environment or in the field challenged with β. cinerea accumulated more phytoalexins than did old ones.     

Compartmentation of Assimilate Fluxes in Leaves

Abstract: Sugar levels in the apoplast of assimilate exporting leaves were studied in two groups of plant species with contrasting structures of companion cells in minor veins. These species are termed either "symplastic" (with intermediary cells) or "apoplastic" (with transfer or ordinary cells). Sugars were measured in intercellular washing fluid after extracting the apoplast by an infiltration-centrifugation technique. During the course of a day, sugar contents in the apoplast were, in general, lower in species with intermediary cells than in species with transfer or ordinary cells. In "symplastic" species, apoplastic sucrose concentrations were between 0.3 and 1 mM. In "apoplastic" species with transfer cells, they ranged between 2 and 6 mM. Apoplastic hexose contents were between 0.3 and 1 mM irrespective of presumed transport mode. "Symplastic" and "apoplastic" plants differed markedly in their response to a'translocation block. In "symplastic" plants, inhibition of assimilate export left apoplastic concentrations of sucrose and hexoses unchanged, whereas in "apoplastic" plants sugar levels increased, the maximal increase being observed with sucrose. In these plants, concentrations of sucrose were two to six times higher in the apoplast under export inhibition than in control leaves. The data suggest a different role of the leaf apoplast in the compartmentation and export of assimilates in the two plant groups under study.     

The Effects of Ouabain on Sodium and Potassium Fluxes in Excised Root Tissue 2

Washed carrot slices take up more K than Na during accumulationof C1 salts from a mixed salt solution. Ouabain does not affectthe overall influx of labelled K or Na to the vacuole, but inhibitsthe efflux of labelled Na from the vacuole. The simplest hypothesisis the ouabain inhibits a Na efflux pump at the plasmalemma.Some suggestions to explain the transient changes in Na effluxafter the addition of ouabain are put forward.     

Effects of Enhanced UV- B Radiation on Protein Metabolism of Bean Leaves

Bean ( Phaseolus vulgaris L. ) seedlings were grown in a greenhouse condition under UV-B (280 ~ 320 nm) radiation and an UV-B-free control condition. At early stage of bean growth, UV-B radiation(UV- BBE: 16 kJ·m-2·d-1 ) increased the synthetic rate of protein in bean leaves. UV-B radiation decreased the synthetic rate of protein and the content of soluble protein, increased the activities of caseolytic enzymes and the content of total amino acids in bean leaves at later growth stage. The results of SDS-PAGE indicated that UV-B radiation increased the contents of polypeptide in a range of 99 kD, 88 kD, 76 kD, 42 kD, 35 kD and 33 kD at the early stage, and increased that of 10 ~ 14 kD, 29 kD, 33 kD and 35 kD at later growth stage. It is suggested that the increase of these polypeptides is an adaptation response of plants to the enhanced UV-B radiation.     

Endogenous Regulation of Photosynthetic Rate in Primary Leaves of Bean (Phaseolus vulgaris L.)

The rate of photosynthesis and/or dry matter production wasstudied in fully-expanded primary leaves of bean (Phaseolusvulgaris cv. Bulgarian) plants which had been subjected to varioussurgical and hormonal treatments. Between 30 and 40 per centof the assimilates produced by the primary leaves, over a 4-dayperiod starting with expansion of the first trifoliate leaf,were diverted to the growing shoot above the insertion of theprimary leaves. In detopped plants (i.e. lacking all leaves,stem and buds above insertion of primary leaves), both the rateof net photosynthesis (NP) of the primary leaves 4 days afterdetopping, and the mean net assimilation rate (NAR) over thisinterval, did not differ significantly from those of intactplants. The assimilate normally diverted to the top in intactplants was distributed between the remaining organs of the detoppedplant. When translocation of assimilates from the primary leaveswas stopped by girdling their petioles, both NAR and NP wereas in untreated control plants after a 2-day period. The assimilatesproduced during that period accumulated in the mesophyll chlorenchymain the form of starch granules. Intact plants supplied withGA₃, or IAA, through the primary leaves as well as detoppedplants supplied with IAA through the stump, differed from untreatedcontrol plants in the pattern of distribution of the assimilatesproduced: IAA favoured dry-matter accumulation in the roots,while GA₃ favoured the tops. Nevertheless, neither NP, nor NARdiffered significantly from the corresponding controls.     

Fluxes of Sodium and Potassium in Acetabularia mediterranea

Sodium efflux in Acetabularia mediterranea occurs against agradient of electrochemical potential and is a light-stimulated,temperature-sensitive process; it is not sensitive to the uncouplerCCCP. Sodium influx is stimulated in CCCP and at low temperature.Potassium influx is temperature- and uncoupler-sensitive, butis not light-stimulated. Tracer K efflux shows complex kinetics,which cannot be explained by any arrangement of intracellularcompartments; it appears to be stimulated at low temperatureand is insensitive to light and uncouplers. There is no evidencefor any chemical linkage between fluxes of Na⁺, K⁺, or Cl^–.It is concluded that Na^– efflux at the plasmalemma isan active process, but no consistent explanation can be advancedto account for the results of K⁺ flux measurements.     

Development of the Primary Photochemical Apparatus of Photosynthesis during Greening of Etiolated Bean Leaves

Seven-day-old dark-grown bean leaves were greened under continuous light. The amount of chlorophyll, the ratio of chlorophyll a to chlorophyll b, the O₂ evolving capacity and the primary photochemical activities of Photosystem I and Photosystem II were measured on the leaves after various times of greening. The primary photochemical activities were measured as the photo-oxidation of P₇₀₀, the photoreduction of C-550, and the photo-oxidation of cytochrome b₅₅₉ in intact leaves frozen to −196 C. The results indicate that the reaction centers of Photosystem I and Photosystem II begin to appear within the first few minutes and that Photosystem II reaction centers accumulate more rapidly than Photosystem I reaction centers during the first few hours of greening. The very early appearances of the primary photochemical activity of Photosystem II was also confirmed by light-induced fluorescence yield measurements at −196 C.     

Starch Accumulation and Photosynthetic Activity in Primary Leaves of Bean (Phaseolus vulgaris L.)

The effects of starch accumulation on photosynthesis and chloroplastultrastructure were studied in primary leaves of bean (Phaseolusvulgaris L. cv. Bulgarian). De-topping the shoot above the primaryleaf node, caused over an 8-day period, a considerable increasein the photosynthetic activity of the primary leaves, despitethe fact that a large quantity of starch had accumulated intheir chloroplasts. The accumulation of starch was greater inthe chloroplasts of spongy cells in comparison with that ofthe palisade cells. Initiation of starch grains was observedmainly in the peripheral part of the chloroplast, distant fromthe cell wall. As a result, most of the starch was accumulatedclose to the inner part of the cell, leaving a considerablemass of the chloroplast near the cell wall free of starch. Theaccumulation of starch was accompanied by the destruction, deformationand disorientation of grana and thylakoids. It is concludedthat the accumulation of starch is not inevitably a limitingfactor in photosynthesis and the results cast doubt on the hypothesisthat starch accumulation or dissipation is the main factor involvedin the regulation of photosynthesis. Phaseolus vulgaris L, bean, photosynthesis, starch accumulation, chloroplast ultrastructure     

Evidence for a Tumor-associated Factor Active in the Promotion of Crown-gall Tumor Growth on Primary Pinto Bean Leaves

The growth of crown-gall tumors on primary pinto bean leaves (Phaseolus vulgaris L. cv. “Pinto”) between day 3 and day 6 after inoculation was found to be proportional to the number of tumors on the leaves. Similar differences observed in the growth of tumors induced by adenine, methionine and asparagine requiring mutants of Agrobacterium tumefaciens (Smith and Town.) Conn appear to be due to the same phenomenon. Tumors induced by these auxotrophs thus show no obvious growth differences from those induced by the prototrophic strain despite the lower specific infectivity and the existence of a mutational lesion in these bacteria. A diffusible growth factor(s) produced by the tumor tissue is proposed to account for the relation between tumor number and early tumor growth.