The effects of low concentrations of aluminium on the growth and uptake of nitrate-N by white clover

Summary The effects of aluminium (Al³⁺) at concentrations of 0, 25, 50 and 100 μM on the growth of white clover, dependent upon N supplied as NO ₃ ⁻ , were examined in flowing solution culture. Plants were established with a normal nutrient supply for 7 weeks and then grown with carefully controlled pH (at 4.5) and P concentrations, and with 0, 25, 50 or 100 μM Al³⁺ for a further three weeks. There were rapid visual effects (i.e. symptoms of P deficiency and reduction in root extension) and the dry weights of shoots and roots were reduced at 50 and 100 μM. Less than 10% of Al absorbed from solution was transported to the shoots. The uptake of P, and its transport between roots and shoots, were reduced in plants grown with Al. The uptake of NO ₃ ⁻ stopped immediately after the introduction of 50 or 100 μM Al, and was significantly reduced at 25 μM after three weeks. During a second phase of the experiment, plants previously grown at 0, 25, 50 and 100 μM Al, were grown for a further 2 weeks either with NO ₃ ⁻ (with and without 50 μM Al³⁺) or without NO ₃ ⁻ but with inoculation by Rhizobia (and with or without 50 μM Al³⁺). The effects of the previous treatments with Al on N uptake were small during the second phase, but uptake by all plants was restricted when Al was present. Inoculation did not result in nodulation in the second phase when Al³⁺ was present in the solution, but Al already in the plant from the first phase did not prevent nodulation in the absence of Al during the second phase.     

Highly efficient embryogenesis and plant regeneration of tall fescue (Festuca arundinacea Schreb.) from mature seed-derived calli

Summary We report a protocol for efficient plant regeneration of four tall fescue (Festuca arundinacea Schreb.) cultivars (‘Surpro’, ‘Coronado’, ‘Summer Lawn’, and ‘Fawn’) via somatic embryogenesis. Calli were initiated from mature seeds grown on modified Murashige and Skoog (MMS) medium supplemented with 7.0mgl⁻¹ (31.7μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 mgl⁻¹ (0.23 μM) kinetin (Kin). Calli were maintained and proliferated by subculture at monthly intervals on MMS medium containing 4.5 mgl⁻¹ (20.4 μM) 2,4-D and 0.2mgl⁻¹ (0.9 μM) Kin. Somatic embryos (SE) were induced from seed-derived calli on SE-induction medium (MMS supplemented with 2.0 mgl⁻¹ 2,4-D and 0.2mgl⁻¹ Kin). Plantlets were regenerated from somatic embryogenic calli grown on modified SH medium supplemented with 2 mgl⁻¹ Kin. Using this optimized protocol, 78.6–82.3% of mature seeds of all four cultivars produced SE clusters, of which 93.5–95.3% regenerated into plants within 10 wk. The regenerants showed no phenotypic abnormalities.     

Effect of NH4 +:NO3 - ratios on growth and nitrogen uptake by onions

The modelling of ion uptake by plants requires the measurement of kinetic and growth parameters under specific conditions. The objective of this study was to evaluate the effect of nine NH _inf4 ^sup+ :NO _inf3 ^sup− ratios on onions (Allium cepa L.). Twenty-eight to 84 day-old onion plants were treated with NH _inf4 ^sup+ :NO_f3/sup− ratios ranging from 0 to 100% of each ionic species in one mM solutions in a growth chamber. Maximum N influx (I_max) was assessed using the N depletion method. Except at an early stage, ionic species did not influence significantly I_max, the Michaelis constant (K_m) and the minimum concentration for net uptake (C_min). I_max for ammonium decreased from 101 to 59 pmole cm^-2 s^-1 while I_max for nitrate increased from 26 to 54 pmole cm^-2 s^-1 as the plant matured. On average, K_m and C_min values were 14.29 μM, and 5.06 μM for ammonium, and 11.90 μM and 4.54 μM for nitrate, respectively. In general, the effect of NH₄ ⁺:NO₃ ^- ratios on root weight, shoot weight and total weight depended on plant age. At an early stage, maximum plant growth and N uptake were obtained with ammonium as the sole source of N. At later stages, maximum plant growth and N uptake were obtained as the proportion of nitrate increased in the nutrient solution. The was no apparent nutrient deficiency whatever NH₄ ⁺:NO₃ ^- ratio was applied, although ammonium reduced the uptake of cations and increased the uptake of phosphorus. The research was supported by the Natural Sciences and Engineering Research Council of Canada.     

Effect of phytohormones and tungsten on light-induced and nitrate-induced nitrate reductase activity of mustard cotyledons

Mustard (Brassica juncea Coss cv. T-59 ‘Varuna’) seedlings pretreated with gibberellic acid (GA) and kinetin (KiN) were grown in light. In vivo nitrate reductase (NR) activity was estimated and effect of tungsten on light-induced and NO ₃ ^su− -induced NR activity was investigated. Different concentrations of GA did not show any effect on induction of light-induced NR; addition of nitrate promoted in vivo NR activity but no concentration effect of GA was evident. Light-induced NR was promoted by KiN and like in GA treatment, addition of nitrate increased NR activity. Addition of Na-tungstate inhibited NO ₃ ⁻ induced NR while light-induced NR was not much affected in both GA and KiN treated seedlings. The two forms of NRs were further characterized by studying the decay kinetics using Na-tungstate. In light-induced NR, tungstate did not affect NR activity up to 11 h, while at later periods, a slight decay was observed. On the other hand, NO ₃ ⁻ -induced NR activity increased up to 4 h and subsequently a rapid fall was observed. It was therefore apparent that light-induced NR had a very low turnover rate as compared to NO ₃ ⁻ -induced NR. These results further support the earlier conclusion that in mustard seedlings two distinct types of NR enzyme exist and that nitrate requirement for NR induction is not absolute.     

Nitrate and ammonium absorption by plants growing at a sufficient or insufficient level of phosphorus in nutrient solutions

Summary Absorption of nitrate and ammonium was studied in water culture experiments with 4 to 6 weeks old plants of barley (Hordeum vulgare L.), buckwheat (Fagopyrum esculentum L. Moench) and rape (Brassica napus L.). The plants were grown in a complete nutrient solution with nitrate (5.7±0.2 mM) or nitrate (5.6±0.2 mM) + ammonium (0.04±0.02 mM). The pH of the nutrient solution was kept at 5.0 using a pH-stat. It was found that phosphorus deficiency reduced the rate of nitrate uptake by 58±3% when nitrate was the sole N source and by 83±1% when both nitrate and ammonium were present. The reduction occurred even before growth was significantly impeded by P deficiency. The inhibition of the uptake of ammonium was less,i.e. ammonium constituted 10±1% of the total N uptake in the P sufficient plants and 30±5% in the P deficient plants. The reduction of nitrate absorption greatly decreased the difference between the uptake of anions and cations. It is suggested that P deficiency reduced the assimilation of NO ₃ ^– into the proteins, which might cause a negative feedback on NO ₃ ^– influx and/or stimulate NO ₃ ^– efflux.     

Rhizodeposition and C-partitioning of Lolium perenne in axenic culture affected by nitrogen supply and defoliation

This study investigated the effects of N-supply and partial defoliation on C-partitioning, root morphology and soluble rhizodeposition, for Lolium perenne grown in axenic sand culture systems percolated with nutrient solution. Plants were grown for 36 d in nutrient solutions with differing N concentrations (4 mM or 0.02 mM NH₄ ⁺NO₃ ^-), and effects of repeated defoliation to 4 cm were determined. The ‘low N’ supply reduced (P < 0.05) dry matter accumulation, with proportionately increased partitioning to the root systems. Root morphology was also altered at ‘low N’, with development of a finer root system, manifest as increased (P < 0.05) specific root length. Concurrent with these effects on growth of L. perenne, ‘low N’ increased (P < 0.05) exudation of C-compounds from roots on a per g root basis. Defoliation was found to increase exudation (P < 0.05) of soluble compounds for periods of 3-5 d following each cut, at both N-supply rates. The effects of N-supply and defoliation are of importance in understanding the coupling of plant productivity to nutrient cycling in soils with differing N availabilities and for grassland systems which are subject to grazing. This revised version was published online in June 2006 with corrections to the Cover Date.     

The toxicity of cadmium to barley plants as affected by complex formation with humic acid

An ‘alternating solution’ culture method was used to study the effects of chloride ions and humic acid (HA) on the uptake of cadmium by barley plants. The plants were transferred periodically between a nutrient solution and a test solution containing one of four levels of HA (0, 190, 569 or 1710 μg cm⁻³) and one of five levels of Cd (0, 0.5, 1.0, 2.5 or 5.0 μg cm⁻³) in either a 0.006M NaNO₃ or 0.006M NaCl medium. Harvest and analysis of shoots and roots was after nineteen days. The distribution of Cd in the test solutions between Cd²⁺, CdCl⁺ and HA-Cd was determined in a separate experiment by dialysis equilibrium. In the nitrate test solutions Cd uptake was clearly controlled by Cd²⁺ concentration and was therefore reduced by HA complex formation. In the absence of HA, chloride suppressed Cd uptake indicating that Cd²⁺ was the preferred species. However complex formation with Cl⁻ enhanced uptake when HA was present because of an increase in the concentration of inorganic Cd species relative to the nitrate system. The ratio root-Cd/shoot-Cd remained at about 10 across a wide range of shoot-Cd concentrations, from about 3 μg g⁻¹ (sub-toxic) up to 85 μg g⁻¹ (80% yield reduction). The ability of the barley plants to accumulate ‘non-toxic’ Cd in their roots was thus very limited. Humic acid also had no effect on Cd translocation within the plant and the root/shoot weight ratio did not vary with any treatment. At shoot-Cd concentrations in excess of 50 μg g⁻¹, K, Ca, Cu and Zn uptake was reduced, probably the result of root damage rather than a specific ion antagonism. The highest concentration of HA also lowered Fe and Zn uptake and there was a toxic effect with increasing HA concentration at Cd=0. However the lowest HA level, comparable with concentrations found in mineral soil solutions, only reduced yield (in the absence of Cd) by <5% while lowering Cd uptake across the range of Cd concentrations by 66%–25%.     

Macronutrient content in leaves of bean plants (Phaseolus vulgaris,L.) grown with differents rates of N/S as fertilizers

Summary The effect of increasing rates of nitrogen (N) and sulphur (S) as fertilizers on the yield, leaf area and N, P, S, Ca, Mg, NO₃ ⁻ and SO₄ ⁼ content in leaves of bean (Phaseolus vulgaris, L.) were studied in a hydroponic culture experiment under greenhouse conditions. Bean plants responded significantly to all treatments with differents N/S ratios. When plants grew with high N/S ratios, the leaf content of N, Ca and NO₃ ⁻ increased while the content of K, P and SO₄ ⁼ decreased. However, optimal yield and leaf area were not obtained. Optimal leaf and fruit dry matter was obtained at N/S ratio value of 1.41. When lower N/S rates were used, optimal leaf and fruit dry matter was only observed when the leaf N/S ratio was between 15 and 16. At high sulphate levels in the nutrient solution there is no interaction with nitrate which is easily observed, resulting in an increase in yield. An interaction between nitrate and sulphate in the nutrient solution was found at a N/S ratio of 0.81 which produced in leaves a synergic effect between P-K, an antagonistic effect between N-P and N-K and a lower yield. This research was supported by Fundacion ‘Ramon Areces’.     

Effect of temperature and irradiance on the uptake of ammonium and nitrate by <Emphasis Type="Italic">Laurencia brongniartii</Emphasis> (Rhodophyta,Ceramiales)

The effects of temperature (20, 24 and 28 °C) and irradiance (15 and 40 μmol photon m⁻² s⁻¹) on the nitrate and ammonium uptake rates of the subtropical red alga, Laurencia brongniartii, were investigated to prepare for tank cultivation. Nitrate uptake followed saturation kinetics and was faster at higher irradiances and temperatures. In contrast, ammonium uptake was linear over the experimental range and was not affected by an increase in temperature. A parameter, β, was calculated to compare substrate uptake rates of nitrate along the linear portion of the uptake curve with that of ammonium. For nitrate, β was lower at low irradiance and higher at high irradiance (β = 0.007 ± 0.003 and 0.030 ± 0.002 [μmol N L⁻¹ (μmol N g_ww⁻¹ d⁻)⁻¹], respectively). However, β was 0.023 ± 0.002 and 0.034 ± 0.002 [μmol N L⁻¹ (μmol N g_ww⁻¹ d⁻¹)⁻¹] for ammonium, suggesting a preference for ammonium over nitrate.     

Improved method of regeneration of black gram (Vigna mungo L.) through liquid culture

Summary A very rapid and efficient regeneration method of Vigna mungo L. has been established using liquid culture. A highly regenerable explant, viz., young multiple shoots obtained by germinating the seeds in 2 mgl⁻¹ (8.9μM) N⁶-benzyladenine-supplemented Murashige and Skoog (MS) medium, was used as a source of tissue to initiate the liquid culture. The liquid medium consisted of half-strength B5 or MS salts supplemented with MS organics, α-naphthaleneacetic acid (0.1 mgl⁻¹, 0.54μM) and N⁶-benzyladenine (0.5mgl⁻¹, 2.2μM). Transferring the growing tissues to fresh medium every third day resulted in ca. 142% increase in the number of shoot buds produced after 24d. Shoot buds elongated on one-third-strength MS (MS_1/3) semisolid medium and plantlets were obtained by transferring the shoots onto MS_1/3 semisolid medium supplemented with indolebutyric acid (1 mgl⁻¹, 4.9 μM).     

Evidence for a periodic excretion of nitrogen by roots of grass-legume associations

Diurnal variation in ion content of the solution bathing roots of two plants growing together in sand culture was analysed for three pairs of grass-legume species (Lolium multiflorum andTrifolium pratense; Zea mays andGlycine hispida; Avena sativa andVicia sativa) and their monospecific controls. Biomass and nitrogen content of plants were determined. Ion concentration (NO ₃ ⁻ , NO ₂ ⁻ , NH ₄ ⁺ , and K⁺) and pH of root solutions were measured for Lolium-Trifolium plant pairs and controls at 6 hours intervals over 36 h, starting at 8 am within a circadian cycle. Root solutions were regularly depleted in NO ₃ ⁻ by the grasses (Lolium-Lolium control) throughout the cycle. For associations involving the legume (Lolium-Trifolium and Trifolium-Trifolium), NO ₃ ⁻ depletion was followed by NO ₃ ⁻ enrichment at night, from late afternoon to early morning; the enrichment was more marked for the Lolium-Trifolium association. Solutions which did not contain NO ₂ ⁻ ions, were enriched by trace amounts of NH ₄ ⁺ ions, largely depleted in K⁺ and alkalanized for all associations throughout the cycle. Repeating the experiment with the three pairs of species at the vegetative phase of development confirmed the previous results: NO ₃ ⁻ enrichment during the night for associations with legumes. When the experiment was repeated with older plants which had almost completed their flowering stage, depletion only was observed and no NO ₃ ⁻ enrichment. These data suggest that NO ₃ ⁻ enrichment results from N excretion from active nodulated roots of the legume, accounting for the increase in both biomass and nitrogen content of the companion grass in grass-legume association. The quantitative importance and periodicity of nitrogen excretion as well as the origin of nitrate enrichment are discussed.     

Efficient production of protocorm-like bodies and plant regeneration from flower stalk explants of the sympodial orchid <Emphasis Type="Italic">Epidendrum radicans</Emphasis>

Summary A simple and efficient micropropagation method was established for direct protocorm-like body (PLB) formation and plant regeneration from flower stalk internodes of a sympodial orchid, Epidendrum radicans. Small transparent tissues formed on surfaces and cut ends of flower stalk internodes on a modified half-strength Murashige and Skoog basal medium with or without thidiazuron (TDZ) after 1–2 wk of culture. In the light, the transparent tissues enlarged and turned into organized calluses on most of the explants. However, PLBs formed only on a medium supplemened with 0.45 μM TDZ within 2 mo. of culture. Sucrose, NH₄NO₃, and KNO₃ were used in media to test their effects on PLB proliferation and shooting. The best response on number of PLBs per tube was 23.6 at 40 gl⁻¹ sucrose, 825 mgl⁻¹ NH₄NO₃, and 950 mgl⁻¹ KNO₃, and the highest number of PLBs with shoots was found at 10 gl⁻¹ sucrose, 825 mgl⁻¹ NH₄NO₃, and 950 mgl⁻¹ KNO₃. Homogenized PLB tissues produced by blending were used to test the effects of four cytokinins [TDZ, N⁶-benzyladenine (BA), zeatin-riboside, and kinetin] on PLB proliferation and shoot formation. The best responses on number of PLBs per tube, proliferation rate, and number of PLBs with shoots per tube were obtained at 4.44 μM BA, 0.28 μM zeatin-riboside, and 1.39 μM kinetin, respectively. Normal plantlets converted from PLBs on the same TDZ-containing medium after 1 mo. of culture. The optimized procedure required about 12–13 wk from the initiation of PLBs to plantlet formation. The regenerated plants grew well with an almost 100% survival rate when acclimatized in a greenhouse.     

Effect of salinity on phosphate accumulation and injury in soybean

Many soybean [Glycine max (L.) Merr.] genotypes that are grown in solution cultures are highly sensitive to the combination of both salinity and inorganic phosphate (Pi) in the substrate. This effect has been observed on numerous occasions on plants grown in a saline medium that contained a substantial amount of Ca (i.e., CaCl₂/NaCl=0.5 on a molar basis). Because Ca is important in regulating ion transport and membrane permeability, solution culture experiments were designed to examine the effects of various concentrations of Pi and ratios of CaCl₂/NaCl (0 to 0.5 on a molar basis) at a constant osmotic potential (−0.34 MPa) on this adverse interaction. Four soybean cultivars (‘Lee’, ‘Lee 74’ ‘Clark’ and ‘Clark 63’) were tested. No adverse salinity x Pi interaction was found on Lee at any ratio and leaf P and Cl were maintained below 300 and 200 mmol kg⁻¹ dry wt, respectively. Clark, Clark 63 and Lee 74 soybean plants, on the other hand, were severely injured by solution salinity (−0.34 MPa osmotic potential) when substrate Pi was ≥0.12 mM. Reduced substrate Ca did not intensify the salinity x Pi interaction. On the contrary, the onset of injury was hastened and more severe with increased CaCl₂/NaCl ratios in isotonic solutions. Shoot and root growth rates decreased as injury increased. Leaf P concentrations from these cultivars grown in saline solutions with 0.12 mM Pi were excessive (>600 mmol kg⁻¹ dry wt) compared with concentrations commonly found in soybean leaf tissue yet they were independent of the severity of injury. Since leaf Cl increased wiht increased CaCl₂/NaCl ratio, we suspect that the severity of foliar injury was related to the combined effects of excessive P and Cl within the tissue. Lee 74, the only injured cultivar examined that excluded Cl from its leaves, was less sensitive than either Clark cultivar and its injury was characteristically different. Other ion interactions were reported that may have played a role in injury susceptibility.     

Soil N chemistry in oak forests along a nitrogen deposition gradient

Anthropogenic N deposition may change soil conditions in forest ecosystems as demonstrated in many studies of coniferous forests, whereas results from deciduous forests are relatively scarce. Therefore the influence of N deposition on several variables was studied in situ in 45 oak-dominated deciduous forests along a N deposition gradient in southern Sweden, where the deposition ranged from 10 to 20 kg N ha⁻¹ year⁻¹. Locally estimated NO ₋ ³ deposition, as measured with ion-exchange resins (IER) on the soil surface, and grass N concentration (%) were positively correlated with earlier modelled regional N deposition. Furthermore, the δ¹⁵N values of grass and uppermost soil layers were negatively correlated with earlier modelled N deposition. The data on soil NO ₋ ³ , measured with IER in the soil, and grass N concentration suggest increased soil N availability as a result of N deposition. The δ¹⁵N values of grass and uppermost soil layers indicate increased nitrification rates in high N deposition sites, but no large downward movements of NO ₋ ³ in these soils. Only a few sites had NO ₋ ³ concentrations exceeding 1 mg N l⁻¹ in soil solution at 50 cm depth, which showed that N deposition to these acid oak-dominated forests has not yet resulted in extensive leaching of N. The δ¹⁵N enrichment factor was the variable best correlated with NO ₋ ³ concentrations at 50 cm and is thus a variable that potentially may be used to predict leaching of NO ₋ ³ from forest soils.     

Lack of a direct nitrate-trifoliin A interaction in the Rhizobium-clover symbiosis

Summary Nitrate added at critical concentrations to plant growth medium inhibits the infection of legume roots by Rhizobium. The direct interaction, of nitrate and trifoliin A, a Rhizobium-recognition lection from white clover (Trifolium repens L.), was examined as a possible basis for this regulation. Selective molecular ultrafiltration studies to detect ligand-protein interactions showed that radioactive¹³NO₃ ⁻ did not bind directly to trifoliin A when incubated at two molar ratios. Immunoprecipitation of trifoliin A by its homologous antibody was unaffected by 15 mM NO₃ ⁻. In addition, there was no apparent reduction in attachment ofR. trifolii 0403 to root hairs of clover seedings during 1 h of incubation in the presence of 15 mM NO₃ ⁻. These results show that nitrate inhibition of these early steps of the infection process is not due to a direct interaction of nitrate with trifoliin A or its glycosylated receptors.     

Effects of Al on nitrogen (NH 4 + and NO 3 − ) uptake,nitrate reductase activity and proton release in two sorghum cultivars differing in Al tolerance

After growth for 17 to 36 days on nutrient solutions with NH₄NO₃ as nitrogen source (pH 4.2) dry matter of sorghum genotype SC0283 was much less affected by Al (1.5 and 3.0 ppm) than that of genotype NB9040. In the absence of Al both cultivars released protons into the nutrient solution as a result of an excess of cationic nutrients taken up. When Al was present, this proton efflux per unit dry weight increased drastically, especially with the sensitive genotype NB9040. Chemical analysis of plant material and continuous analyses of NO ₃ ⁻ and NH ₄ ⁺ in the nutrient solution indicated, that the Al-induced shift in H⁺-balance of both genotypes could almost completely be attributed to a decreased NO ₃ ⁻ /NH ₄ ⁺ uptake ratio. In vivo nitrate reductase activity (NRA) was reduced in the shoot of NB9040 and to a lesser degree in SC0283. Al-induced decrease in NRA was accompanied by similar percentual decreases in NO ₃ ⁻ tissue concentrations. Therefore this decrease is interpreted as being indirect,i.e., the consequence of the reduced NO ₃ ⁻ uptake of the plants. A direct repression of NRA by Al seems also unlikely because nitrate reductase activity of the roots (where cellular Al-concentrations should be higher than in shoots) was not affected in Al-treated plants of either genotype.     

The Impact of Nitrogen Placement and Tillage on NO, N2O, CH4 and CO2 Fluxes from a Clay Loam Soil

To evaluate the impact of N placement depth and no-till (NT) practice on the emissions of NO, N₂O, CH₄ and CO₂ from soils, we conducted two N placement experiments in a long-term tillage experiment site in northeastern Colorado in 2004. Trace gas flux measurements were made 2–3 times per week, in zero-N fertilizer plots that were cropped continuously to corn (Zea mays L.) under conventional-till (CT) and NT. Three N placement depths, replicated four times (5, 10 and 15 cm in Exp. 1 and 0, 5 and 10 cm in Exp. 2, respectively) were used. Liquid urea–ammonium nitrate (UAN, 224 kg N ha⁻¹) was injected to the desired depth in the CT- or NT-soils in each experiment. Mean flux rates of NO, N₂O, CH₄ and CO₂ ranged from 3.9 to 5.2 μg N m⁻² h⁻¹, 60.5 to 92.4 μg N m⁻² h⁻¹, −0.8 to 0.5 μg C m⁻² h⁻¹, and 42.1 to 81.7 mg C m⁻² h⁻¹ in both experiments, respectively. Deep N placement (10 and 15 cm) resulted in lower NO and N₂O emissions compared with shallow N placement (0 and 5 cm) while CH₄ and CO₂ emissions were not affected by N placement in either experiment. Compared with N placement at 5 cm, for instance, averaged N₂O emissions from N placement at 10 cm were reduced by more than 50% in both experiments. Generally, NT decreased NO emission and CH₄ oxidation but increased N₂O emissions compared with CT irrespective of N placement depths. Total net global warming potential (GWP) for N₂O, CH₄ and CO₂ was reduced by deep N placement only in Exp. 1 but was increased by NT in both experiments. The study results suggest that deep N placement (e.g., 10 cm) will be an effective option for reducing N oxide emissions and GWP from both fertilized CT- and NT-soils.     

Efficient and simple plant regeneration via organogenesis from leaf segment cultures of persimmon (Diospyros kaki thunb.)

Summary An efficient and simple plant regeneration system via organogenesis from leaf segments of persimmon (Diospyros kaki Thunb.) cultivars ‘Fuyu’ and ‘Nishimurawase’ has been developed. The regeneration capacity was influenced by the culture vessels, gelling agents, plant growth regulators, and light conditions. Leaf explants taken from in vitro shoots were cultured on a modified Murashige and Skoog medium (MS1/2N), for 16 wk without transfer to fresh medium. Adventious shoots appeared after 4 and 8 wk in culture of ‘Nishimurawase’ and ‘Fuyu’ tissues, respectively. The culture of leaf explants in Erlenmeyer flasks with medium containing 4 g l⁻¹ agar enhanced shoot formation in comparison to media with increased agar concentrations. Optimal shoot regeneration was obtained with 5 mg l⁻¹ (22.8 μM) zeatin and 0.1 mg l⁻¹ (0.05 μM) indole-3-butyric acid (IBA) for ‘Nishimurawase’, and 10 mg l⁻¹ (45.6 μM) zeatin and 0.1 mg l⁻¹ (0.05 μM) IBA for ‘Fuyn’. Shoot regeneration frequencies in both cultivars were 100%, and shoot numbers per explant reached up to 9.2 for ‘Nishimurawase’ and 2.2 for ‘Fuyu’. Dark incubation during the first 4–5 wk was the most effective condition to successfully influence shoot regeneration in both cultivars. While dark incubation was essential for adventitious shoot formation by ‘Fuyu’, it was only slightly beneficial to ‘Nishimurawase’. More than 80% of the regenerated shoots rooted within 4 wk on hormone-free MS1/2N demium after having been dipped for 30 s in 250 mg l⁻¹ (1.1. mM) IBA solution.     

An adapid primate,sinoadapis, from lufeng,latest miocene,Yunnan, China

The paper deals with a new form ofSinoadapis from the Lufeng hominoid Locality, assigned toSinoadapis shihuibaensis sp. nov. Holotype PA 882 A fragment of right mandible with C,−M₃. Other materials. PA 903 A fragment of left maxilla with P³−M³. PA 959 Left lower tooth row with I₁−P², P₄. PA 902 Right upper tooth row with C′−P³. PA 964 An isolated left I¹. PA 907 An isolated right I³. PA 972 An isolated right M³.     

Uptake of inorganic ions and compatible solutes in cultured mangrove cells during salt stress

Summary Callus of the mangrove plant, Sonneratia alba J. Smith, established from pistils of flower buds were cultured on solid Murashige and Skoog medium supplemented with 0 to 500 mM NaCl. Maximum growth was observed with 50 mM NaCl, and net growth of callus occurred for concentrations up to 200 mM NaCl. At 500 mM NaCl, growth of callus was completely inhibited, although a part of the tissue was still alive after 30 d. Cellular levels of Na⁺ and Cl⁻ were greatly increased by the treatment with NaCl. Uptake of K⁺ was also enhanced and was accompanied by increasing levels of Na⁺ and Cl⁻ so that the Na⁺/K⁺ ratio was almost constant (4.1–4.2) in callus grown with 50–200 mM NaCl. Levels of Mg²⁺ and Ca²⁺ were not changed significantly with 50–200 mM NaCl, whereas levels of free NH ₄ ⁺ , NO ₃ ⁻ and SO ₄ ²⁻ ions, which are convertible to organic compounds, were lowest in callus grown with 50 mM NaCl. The rate of conversion of ¹⁵NH ₄ ⁺ into macromolecules during 30 d culture with 0–100 mM NaCl did not vary greatly, but 200 mM NaCl reduced the biosynthesis of macromolecules from this ion. The highest rate of conversion of ¹⁵NO ₃ ⁻ into macromolecules was observed at 50 mM NaCl. Identification of compatible solutes with NMR-spectroscopy indicated that mannitol is the compatible solute for intact plants of Sonneratia alba, but no accumulation of mannitol was found in calluses, not even in those grown at high concentrations of NaCl.