Evidence for phytochrome involvement in light-mediated stomatal movement in Phaseolus vulgaris L.

Observations made with primary leaves of Phaseolus vulgaris L. demonstrated that phytochrome modulates light-induced stomatal movement. Removal of the far-red-absorbing form of the pigment (Pfr) with far-red (FR) radiation decreased the time required by the stomata to reach maximal opening following a dark-to-light transition; this effect of FR was fully reversible with red. Removal of Pfr with FR also decreased the time required to reach maximal closure following a light-to-dark transition, and the rate of closure was dependent on the final irradiation treatment before darkness. No evidence was found for phytochrome involvement in determining stomatal aperture under constant conditions of either darkness of light.Abbreviations and symbols Chl chlorophyll - D darkness - FR far-red - phytochrome photostationary state - Pfr, Pr FR- and R-absorbing forms of phytochrome, respectively - R red     

Chilling and age related changes in the free sterol composition of Phaseolus vulgaris L. primary leaves

Chill-stressed or aged bean (Phaseolus vulgaris cv. 194) plants showed relative increase in the campesterol content of the primary leaves. Sitosterol/stigmasterol ratio declined significantly 24 h following transfer of chilled plants to warmer conditions. Sitosterol/stigmasterol varied markedly according to plant age, showing a decline during the leaf greening phase but an increase by the senescent (leaf-yellowing) phase.     

Spatial and temporal variation in gas exchange over the lower surface of Phaseolus vulgaris L. primary leaves

This paper describes spatio-temporal variation in gas exchange over thelower surface of primary leaves of glasshouse-grown Phaseolusvulgaris L. plants. Simultaneous measurements of assimilationand water vapour conductance were made with a small area cuvette attachedto an infra-red gas analyser. The plants were kept in glass chambers sothat the external gaseous environment could be controlled. Observations arereported from four half hour periods during a day in which the ambientPPFD, while variable, was close to saturating for photosynthesis.'Snapshot' measurements of gas exchange were made at 20 positions on theleaf surface using a stratified random out-to-in strategy, which avoideddisturbance of yet-to-be-measured sites. Data were mapped using the'Unimap' cartographic program. For any given measurement period, gasexchange varied greatly over the leaf surface: typically, net assimilation(A) varied by over 4-fold and leaf conductance(gi) by over 3-fold. Estimated intracellular pressuresof CO2 and leaf temperatures showed less relative variation both in spaceand time. Comparing measurement periods, the spatial patterns of variationin A and gi were dissimilar.Moreover, at different sites on the leaf, the trends in a given variablecould be in opposite directions, while external conditions were relativelyconstant. Although the correlation between A andgi was significant overall, there was a large degreeof scatter in the data and zones of high gi oftencorresponded to areas of low A. Depending on the basisof calculation, A was as much as 63% lower than avalue predicted on the basis of steady-state measurements for theappropriate mean irradiance. It was not possible to deduce from therelationships between pairs of variables which factors were most importantin determining A and gi at anygiven time or space, but gi did not appear to be theonly factor limiting A. It is hypothesized that theobserved variation in gas exchange, the lack of close correspondencebetween gi and A and thereduction in photosynthesis compared with the apparent potential value areall phenomena that arise from differences in the induction times for thesevariables following changes in conditions, interacting with other factorsassociated with position on the leaf.     

The effect of environmental heterogeneity on clonal behaviour of Prunella vulgaris L.

Melinis minutiflora Beauv. (Poaceae) is an African grass that is invading mid-elevation Trachypogon savannas in Venezuela. The objective of this study was to investigate the influence of soil fertility, competition and soil disturbance in facilitating Melinis' invasion and growth in these savanna sites. We manipulated soil fertility by adding nitrogen (+N), phosphorus and potassium (+PK), or nitrogen, phosphorus, and potassium (+NPK). We simultaneously manipulated the competitive environment by clipping background vegetation. In a separate experiment, we mechanically disrupted the soil to simulate disturbance. We hypothesized that germination and growth were bottlenecks to early establishment in undisturbed savanna, but that disturbance would alleviate those bottlenecks. We measured Melinis seed germination and subsequent establishment by adding seeds to all plots. We examined Melinis growth by measuring biomass of Melinis seedling transplants, 11 months after they were placed into treatment plots. Germination and establishment of Melinis from seed was extremely low. Of the 80,000 seeds applied in the experiment, only 28 plants survived the first growing season. Mortality of Melinis seedling transplants was lowest in PK fertilized plots, but in the absence of PK mortality increased with N additions and clipping. By contrast, fertilization of the savanna with NPK greatly increased Melinis seedling biomass and this effect was greatly enhanced when competition was reduced (e.g. clipping). Melinis transplant growth responded strongly to soil disturbance- a response not fully explained by removal of competitors (clipping) or changes in soil nutrients and moisture. We suspect that disruption of the soil structure allowed for greater root proliferation and subsequent plant growth. We believe that native savanna is relatively resistant to Melinis invasion, since Melinis seedlings persisted in intact savanna but exhibited little or no growth during the first year. The significant enhancement of Melinis seedling growth with clipping and nutrient additions suggests that low soil nutrients and the presence of native savanna species are important factors in the ability of native savanna to resist Melinis establishment. However, the potential for Melinis growth increases enormously with soil disturbance.     

Dihydrozeatin riboside,a minor cytokinin from the leaves of Phaseolus vulgaris L.

Dihydrozeatin riboside has been identified in the leaves of decapitated bean plants by Sephadex LH20 chromatography and combined gas chromatography-mass spectrometry. The relationship between the cytokinins isolated and identified from this system and those previously reported in Phaseolus is discussed.Abbreviations DHZ dihydrozeatin - DHZOG dihydrozeatin-O--D-glucoside - DHZR dihydrozeatin riboside - GCMS combined gas chromatography-mass spectrometry - GLC gas-liquid chromatography - TIC total ion current - TMS trimethylsilyl - Z zeatin - ZR zeatin riboside     

Light-stimulated cell expansion in bean (Phaseolus vulgaris L.) leaves

The quantity and quality of light required for light-stimulated cell expansion in leaves of Phaseolus vulgaris L. have been determined. Seedlings were grown in dim red light (RL; 4 micromoles photons m-2 s-1) until cell division in the primary leaves was completed, then excised discs were incubated in 10 mM sucrose plus 10 mM KCl in a variety of light treatments. The growth response of discs exposed to continuous white light (WL) for 16 h was saturated at 100 micromoles m-2 s-1, and did not show reciprocity. Extensive, but not continuous, illumination was needed for maximal growth. The wavelength dependence of disc expansion was determined from fluence-response curves obtained from 380 to 730 nm provided by the Okazaki Large Spectrograph. Blue (BL; 460 nm) and red light (RL; 660 nm) were most effective in promoting leaf cell growth, both in photosynthetically active and inhibited leaf discs. Far-red light (FR; 730 nm) reduced the effectiveness of RL, but not BL, indicating that phytochrome and a separate blue-light receptor mediate expansion of leaf cells.     

Role of acid efflux during growth promotion of primary leaves of Phaseolus vulgaris L. by hormones and light

The white-light-(WL) induced enlargement of dicotyledonous leaf cells is known to occur via an acid-growth mechanism; i.e., WL causes leaf cells to excrete protons which lead to an increase in wall extensibility and thus cell enlargement. Gibberellic acid (GA₃) and N⁶-benzyladenine (BA) also induce leaf cell enlargement. To see if they also act via acid-induced cell wall loosening, a comparison has been made of WL-, GA₃-and BA-induced growth of strips, taken from primary leaves of bean (Phaseolus vulgaris L.) plants raised in continuous red light for 10 d. White light, GA₃ and BA all increased wall extensibility as measured by the Instron technique, and this change preceded the increase in growth rate. However, whereas WL induced significant proton excretion, neither GA₃ nor BA caused any acidification of the apoplast. Furthermore, neutral buffers, which effectively inhibited the growth induced by WL, were without effect on growth promoted by either GA₃ or BA. These results indicate that while WL, GA₃ and BA all initiate growth in bean leaves by altering cell-wall properties, GA₃ and BA do so through some wall loosening mechanism other than wall acidification. Neither gibberellin nor cytokinin is likely to play a major role in light-induced cell enlargement of dicotyledonous leaves.Abbreviations BA N^o-benzyladenine - FC fusicoccin - GA₃ gibberellic acid - RL red light - SK medium 10 mM sucrose+10mM KCl - WL white light     

Apical Dominance in Phaseolus vulgaris L.

Although determinations of the ABA content of lateral buds ofPhaseolus vulgaris revealed no difference between decapitatedand intact control plants in the first 12 h following decapitation,a relative decrease in the ABA content of lateral buds of decapitatedplants was detectable 24 h following decapitation. Shoot decapitationwas also observed to result in a decrease in the ABA contentof stem tissue. The application of IAA to the stem of decapitatedplants prevented these changes and increased the ABA contentof stem tissue relative to that of intact plants. The levelsof IAA and ABA were also determined in the stem tissue fromthe nodes of intact bean plants. The possible interdependenceof these two plant hormones was further investigated by a studyof [2_–14ClABA metabolism. The results are discussed inrelation to the possible role of these hormones in apical dominance. Key words: Apical dominance, Abscisic acid, Indole-3-acetic acid     

Cytokinin metabolism in Phaseolus vulgaris L.

The major cytokinins in stems of decapitated, disbudded bean plants have been identified by enzymic degradation, Sephadex LH20 and reversed phase high performance liquid chromatography, and by combined gas chromatography-mass spectrometry as 6-(4-hydroxy-3-methylbut-trans-2-enylamino)-9--D-ribofuranosylpurine (zeatin riboside), 6-(4-hydroxy-3-methylbutylamino)-9--D-ribofuranosylpurine (dihydrozeatin riboside), and the 5-phosphates of these compounds (zeatin ribotide and dihydrozeatin ribotide). Minor cytokinins in this tissue were tentatively identified as dihydrozeatin-O--D-glucoside and zeatin ribotide-O--D-glucoside. [8-¹⁴C-]Dihydrozeatin appeared to be rapidly metabolized to dihydrozeatin ribotide when supplied to segments of stems from decapitated plants. These results are discussed in relation to the metabolism and distribution of cytokinins in the whole plant.Abbreviations TEAB triethyl ammonium bicarbonate - UV ultra-violet - GCMS gas chromatography-mass spectrometry - HPLC high performance liquid chromatography - TMS trimethyl silyl     

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. This revised version was published online in June 2006 with corrections to the Cover Date.     

Plastid development in primary leaves of Phaseolus vulgaris: The light-induced development of the chloroplast cytochromes

Summary Etioplasts obtained from the primary leaves of dark-grown bean plants contained cytochromes f, b-559_LP and b-563 in a molar ratio of approximately 1.0:2.0:1.5. On illumination of the plants there was a lag of between 10 and 15 h before these cytochromes increased in amount, but after 48 h they had increased from 6- to 10-fold on a per plastid basis. The presence of cytochrome b-559_HP in the plastids was first detected after 15 h of illumination, which coincided with the commencement of grana formation and the onset of a number of photosynthetic reactions in the greening leaves. After 48 h of illumination the molar ratio for cytochromes f, b-559_HP, b-559_LP and b-563 was 1.0:1.2:2.8:2.6.Agranal chloroplasts formed by the exposure of dark-grown plants to intense light flashes contained high amounts of cytochromes f, b-559_LP and b-563 but cytochrome b-559_HP could not be detected.As the light-induced formation of cytochromes f, b-559_LP and b-563 was substantially inhibited by D-threo chloramphenicol, but not by the L-threo isomer, it seems likely that their formation was dependent on 70S ribosomes. Both chloramphenicol isomers gave plastids which lacked cytochrome b-559_HP.     

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.     

Tissue Cultures of Phaseolus vulgaris L.

Abstract

Callus production and plant regeneration from different explants of Phaseolus vulgaris L. cv. Giza are reported. Calli cultures were induced from leaf, hypocotyl, embryo and root explants. Rapid growth of callus was achieved by leaf explants cultured on MS salts, B5 vitamins and supplemented with 2,4— dichlorophenoxyacetic acid (2, 4—D)+0.5 mg/l kinetin (kin). Addition of casein hydrolysate at 2 g/l to maintenance medium enhanced callus growth and hindered the early appearance of necrotic parts. This report also provides a detailed method for production of multiple shoots directly from the wounded edges of immature cotyledon explant via organogenesis on 1 mg/l benzyladenine (BA) or indirectly on 0.5 mg/l naphthaleneacetic acid (NAA)+2 mg/l BA. The regeneration of bean plants through the two ways described here (direct or indirect) may be of use in genetic improvement of bean.     

Effect of nitrogen nutrition on the carbohydrate repression of photosynthesis in leaves of Phaseolus vulgaris L.

When carbohydrates accumulate in leaves, photosynthesis is repressed. Limited nitrogen nutrition is thought to enhance this repressing effect. However, the interaction between carbohydrate and nitrogen limitation in leaf photosynthesis has not been examined intensively. In this study, we grew Phaseolus vulgaris L. plants at three different nitrogen levels, and examined the effects of sucrose feeding to the roots on the nitrogen content, carbohydrate content and photosynthetic properties of the primary leaves. Nitrogen content and photosynthetic rate were lower and the carbohydrate content was greater in plants grown with limited nitrogen than in well-fertilized plants. Sucrose feeding to the plants increased carbohydrate content and decreased photosynthetic rate and nitrogen content. The increase in carbohydrate content and the decreases in nitrogen content and photosynthetic rate occurred at the same time, and the negative relationship between the carbohydrate content and photosynthetic rate did not differ among nitrogen nutrition levels. These results show that carbohydrate accumulation in the leaves leads to a decrease in photosynthetic rate. At low nitrogen nutrition levels, carbohydrates accumulated markedly, which accelerated this effect. It appears that the nitrogen nutrition level influences leaf photosynthesis through changing the carbohydrate level rather than through modifying sensitivity of the leaf to the carbohydrate level.     

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.     

Effects of salt stress on the growth,ion content,stomatal behaviour and photosynthetic capacity of a salt-sensitive species,Phaseolus vulgaris L.

Phaseolus vulgaris (cv. Hawkesbury Wonder) was grown over a range of NaCl concentrations (0–150 mM), and the effects on growth, ion relations and photosynthetic performance were examined. Dry and fresh weight decreased with increasing external NaCl concentration while the root/shoot ratio increased. The Cl^- concentration of leaf tissue increased linearly with increasing external NaCl concentration, as did K⁺ concentration, although to a lesser degree. Increases in leaf Na⁺ concentration occurred only at the higher external NaCl concentrations (100 mM). Increases in leaf Cl^- were primarily balanced by increases in K⁺ and Na⁺. X-ray microanalysis of leaf cells from salinized plants showed that Cl^- concentration was high in both the cell vacuole and chloroplast-cytoplasm (250–300 mM in both compartments for the most stressed plants), indicating a lack of effective intracellular ion compartmentation in this species. Salinity had little effect on the total nitrogen and ribulose-1,5-bisphosphate (RuBP) carboxylase (EC 4.1.1.39) content per unit leaf area. Chlorophyll per unit leaf area was reduced considerably by salt stress, however. Stomatal conductance declined substantially with salt stress such that the intercellular CO₂ concentration (C _i) was reduced by up to 30%. Salinization of plants was found to alter the ¹³C value of leaves of Phaseolus by up to 5 and this change agreed quantitatively with that predicted by the theory relating carbon-isotope fractionation to the corresponding measured intercellular CO₂ concentration. Salt stress also brought about a reduction in photosynthetic CO₂ fixation independent of altered diffusional limitations. The initial slope of the photosynthesis versus C _i response declined with salinity stress, indicating that the apparent in-vivo activity of RuBP carboxylase was decreased by up to 40% at high leaf Cl^- concentrations. The quantum yield for net CO₂ uptake was also reduced by salt stress.Abbreviations and symbols A net CO₂ assimilation rate - C _a ambient CO₂ concentration - C _i intercellular CO₂ concentration - RuBP ribulose-1,5-bisphosphate - ¹³C ratio of ¹³C to ¹²C relative to standard limestone     

The effect of 6-benzyladenine derivatives on the rooting of Phaseolus vulgaris L. primary leaf cuttings

The effects of 6-benzyladenine (BA), its 3-glucoside (BA-3-G) and 9-glucoside (BA-9-G), and its riboside (BA-R) on the rooting of primary leaf explants of Phaseolus vulgaris L. were compared, alone or in combination with indole-3-butyric acid (IBA). At all except the lowest concentration used (4.4 × 10^–7 M), BA and BA-3-G reduced the number of roots and their averge length relative to a distilled water control. Addition of IBA (4.4 × 10^–5 M) increased rooting beyond the control, but not to the level of IBA alone. This contrasted with BA-9-G where a response very similar to that of the control was recorded. The riboside of BA was more inhibitory than the free base, its effect extending to the lowest concentration used (4.4 × 10^–7 M).     

The hormonal regulation of bud outgrowth in Phaseolus vulgaris L.

Summary Aqueous solutions of indole acetic acid, kinetin, gibberellic acid and abscisic acid were applied singly and in combination to the decapitated stem stump of Phaseolus seedlings. Application of indole acetic acid will not completely replace the intact stem apex with regard to the inhibition of lateral bud extension. The greatest inhibition of bud growth is obtained when indole acetic acid is applied in combination with both kinetin and abscisic acid. Treatment with gibberellic acid causes massive bud growth even in the presence of indole acetic acid, kinetin and abscisic acid. Although both abscisic acid and kinetin have only a slight promoting effect on bud outgrowth when applied singly, these hormones will modify the effects of indole acetic acid and gibberellic acid.