Zinc tolerance in populations ofDeschampsia cespitosa (Gramineae) beneath electricity pylons

Patterns of zinc tolerance were examined in eightDeschampsia cespitosa (L.)Beauv. populations from normal and zinc-contaminated soils, using solution culture methods. Zinc-tolerant populations have evolved beneath pylons, and their tolerance patterns (degree, variance, heritability) vary. Tolerance is genetically based in all of them. In contrast to the cases of other species previously reported, some within-population differences are considerable, and may be due to both heterogeneity of the soils zinc contents, various gene combinations and gene recombinations between genotypes. One control population contains many tolerant plants, an original result which is discussed.     

Polymorphism and physiology of arsenate tolerance in Holcus lanatus L. from an uncontaminated site

The polymorphism of arsenate tolerance in a Holcus lanatus L. population from an uncontaminated soil was investigated and a high percentage of tolerant individuals (65%) was found in the population studied. Influx of arsenate was highly correlated to arsenate tolerance within the population, with the most tolerant individuals having the lowest rates of arsenate influx. Isotherms for the high affinity arsenate uptake systems were determined in six tolerant and six non-tolerant genotypes. Tolerant plants had the lowest rates of arsenate influx. This was achieved by adaptation of the V_max of arsenate influx with the V_max of the high affinity uptake system saturating at lower substrate concentrations in the tolerant plants. The polymorphism is discussed with relation to adaptation to the extreme environments to which the plants are subjected on mine-spoil soils.     

Stem tissue phosphorus as an index of the phosphorus status of Banksia ericifolia L. f.

The effects of P fertilizer rate on shoot growth and the total P concentration of the whole shoot, new and mature leaves, symptom leaves and stems of Banksia ericifolia L. f., a P-sensitive species, were investigated in a six month greenhouse pot experiment. Shoot dry weight of plants growing in an Australian sedge peat, coarse sand and perlite potting mix (1:1:1) increased with up to 100 mg P L⁻¹ supplied as a six month controlled release P (0:18:0) fertilizer, but was reduced by toxicity at the highest application rate (200 mg P L⁻¹). Plants receiving this treatment developed chlorotic new and mature leaves. Leaf symptoms observed at rates of 60–100 mg P L⁻¹ were confined to old leaves and were related to the P concentration of the shoot. Growth was not affected at these rates. The P concentration of stems was strongly influenced by P supply. This tissue acted as a sink for excess P, helping to regulate the P concentration of leaves. The approximate range of P concentrations in stem tissue, associated with greater than 90% of maximum shoot dry weight, was 0.5–1.5 g P kg⁻¹ tissue dry weight. This was greater than that calculated for mature leaves (0.5–0.8 g kg⁻¹) or for whole shoots (0.5–1.2 g kg⁻¹). This wider range, and the capacity to store P in excess to requirement, makes the stem a better index tissue for plant P status than either leaves or whole shoots. This revised version was published online in June 2006 with corrections to the Cover Date.     

Plantago lanceolata L. and Rumex acetosella L. differ in their utilisation of soil phosphorus fractions

To establish relationships between soil phosphorus (P) fractions and leaf P, a mycorrhizal species (Plantago lanceolata L.) was compared with a typically non-mycorrhizal species (Rumex acetosella L.) in a glasshouse experiment. The plants were grown in 40 soils from non-fertilised, abandoned pastures or abandoned arable fields and leaf P concentration were found to be related to various soil P fractions after six weeks of growth. The differences in the P fractions in soil can account for a large share of the variation in leaf P concentration in both species, but the two species differed in their utilisation of P fractions. Leaf P concentration of R. acetosella was more related to extractable soil P than that of P. lanceolata. Rumex acetosella showed a higher maximum P concentration. The P fractions accounting for the largest share of the variation in leaf P concentration was the Bray 1 extractable and the weak oxalate (1 mM) extractable P, and for P. lanceolata also the Na₂SO₄+NaF extractable P fraction. P extracted with these methods accounted for up to 80% of the variation in P concentration in leaves of R. acetosella and 65% of the variation in leaves of P. lanceolata. More P extractable with weak oxalate, Na₂SO₄+NaF and strong oxalate (50 mM) was released from the soil than was taken up by the plants during the experimental period. The Bray 1 extractable P fraction, however, decreased in both unplanted and planted soils. Phosphatase release was not induced in any of the plants during the experimental period, indicating that they were not mobilising soil organic P. However, some of the methods extracted a large share of the organic P and still explained much of the variation in leaf P concentration. Mycorrhizal colonisation of P. lanceolata was inversely related to the extractable soil P. The consistently fast P uptake of R. acetosella indicates that this species have a high demand for P. The differences in P utilisation between R. acetosella and P. lanceolata could be caused by their different mycorrhizal status.     

Effects of arsenate and phosphate on their accumulation by an arsenic-hyperaccumulator Pteris vittata L.

Arsenate and phosphate interactions are important for better understanding their uptake and accumulation by plant due to their similarities in chemical behaviors. The present study examined the effects of arsenate and phosphate on plant biomass and uptake of arsenate and phosphate by Chinese brake (Pteris vittata L.), a newly-discovered arsenic hyperaccumulator. The plants were grown for 20 weeks in a soil, which received the combinations of 670, 2670, or 5340 mol kg^–1 arsenate and 800, 1600, or 3200 mol kg^–1 phosphate, respectively. Interactions between arsenate and phosphate influenced their availability in the soil, and thus plant growth and uptake of arsenate and phosphate. At low and medium arsenate levels (670 and 2670 mol kg^–1), phosphate had slight effects on arsenate uptake by and growth of Chinese brake. However, phosphate substantially increased plant biomass and arsenate accumulation by alleviating arsenate phytotoxicity at high arsenate levels (5340 mol kg^–1). Moderate doses of arsenate increased plant phosphate uptake, but decreased phosphate concentrations at high doses because of its phytotoxicity. Based on our results, the minimum P/As molar ratios should be at least 1.2 in soil solution or 1.0 in fern fronds for the growth of Chinese brake. Our findings suggest that phosphate application may be an important strategy for efficient use of Chinese brake to phytoremediate arsenic contaminated soils. Further study is needed on the mechanisms of interactive effects of arsenate and phosphate on Chinese brake in hydroponic systems.     

Effect of silicate on the growth and arsenate uptake by rice (Oryza sativa L.) seedlings in solution culture

A solution culture experiment was conducted to investigate the effect of silicate on the yield and arsenate uptake by rice. Rice seedlings (Oryza sativaL. cv. Weiyou 77) were cultured in modified Hoagland nutrient solution containing three arsenate levels (0, 0.5 and 1.0 mg L ^–1 As) and four silicate levels (0, 14, 28 and 56 mg L ^–1 Si). Addition of Si significantly increased shoot dry weight (P=0.001) but had little effect on root dry weight (P=0.43). Addition of As had no significant effect on shoot dry weight (P=0.43) but significantly increased root dry weight (P=0.01). Silicon concentrations in shoots and roots increased proportionally to increasing amounts of externally supplied Si (P < 0.001). The presence of As in the nutrient solution had little effect on shoot Si concentration (P=0.16) but significantly decreased root Si concentration (P=0.005). Increasing external Si concentration significantly decreased shoot and root As concentrations and total As uptake by rice seedlings (P <0.001). In addition, Si significantly decreased shoot P concentration and shoot P uptake (P <0.001). The data clearly demonstrate a beneficial effect of Si on the growth of rice seedlings. Addition of Si to the growth medium also inhibited the uptake of arsenate and phosphate by the rice seedlings.     

The use of indigenous plant species and calcium phosphate for the stabilization of highly metal-polluted sites in southern Poland

Highly metal-polluted (Pb, Cd, Zn) soil from a non-ferrous mine and smelter site in southern Poland, further referred to as Waryski soil, was used to test indigenous plant species for stabilization effectiveness of heavy metals in soils. Results of pilot investigations with commercially available cultivars of plant species showed that these cultivars could not grow on this highly polluted soil even with the application of soil amendments to stabilize the heavy metals. Based on these results, mesocosm and field experiments with an indigenous, metal-tolerant ecotype of Deschampsia cespitosa from the Waryñski site were carried out. The mesocosm experiment showed that applications of calcium phosphate (3.8% w/w) as a heavy metal-stabilizing amendment decreased Cd and Zn concentrations 2 and 3-fold respectively in leachates, whereas lead content was not significantly changed. This decrease in the concentration of heavy metals in leachates was correlated with a lower accumulation of Pb, Cd and Zn in the roots and shoots of D. cespitosa, ecotype Waryñski. In the field experiment, lower accumulations of Cd in roots and shoots and Zn in shoots in the amendment added plot were observed during the second year of investigations. In the first growing season, D. cespitosa plant cover in the amendment enriched mesocosms ranged from 95 to 100%, compared to 10% in mesocosms without calcium phosphate. In the second year of the experiment, in non-amendment enriched mesocosms D. cespitosa was substituted with Cardaminopsis arenosa(95% cover). C. arenosa is an undesirable species for phytostabilization, as it accumulates high amounts of zinc and cadmium in its shoots, even thought it provided better growth cover in not amended soils. However, in amended mesocosms, soil surface cover by D. cespitosa was still very high (90%). Similar results were obtained in field experiments. Addition of calcium phosphate to the soil also resulted in excellent D. cespitosa root system development when compared to soils without amendment. In amended mesocosms, high plant cover and root system development significantly decreased the volume of leachates and improved water retention. These results indicate that the use of D. cespitosa, ecotype Waryski in combination with calcium phosphate as a heavy metals immobilizing agent is sufficient to restore a dense vegetative cover to highly heavy metal-polluted soil.     

Nitrate nutrition ofDeschampsia flexuosa (L.) Trin. in relation to nitrogen deposition in Sweden

Summary Current and maximally induced nitrate reductase activity (NRA), total-N, nitrate, K, P, Ca, Mg, Mo and sucrose in leaves ofDeschampsia flexuosa was measured three times during the vegetation period in forests along a deposition gradient (150 km) in south Sweden, in north Sweden where the nitrogen deposition is considerably lower, and at heavily N-fertilized plots. In addition, the interaction between nitrogen nutrition and light was studied along transects from clearings into forest in both south and north Sweden. Plants from sites with high nitrogen deposition had elevated current NRA compared to plants from less polluted sites, indicating high levels of available soil nitrate at the former. Current NRA and total N concentration in grass from sites with high deposition resembled those found at heavily N-fertilized plots. Under such circumstances, the ratio current NRA: maximally induced NRA as well as the concentration of nitrate was high, while the concentration of sucrose was low. This suggests that the grass at these sites was already utilizing a large portion of its capacity to assimilate nitrate. Light was found to play an important role in the assimilation of nitrate; leaf concentration of sucrose was found to be negatively correlated with both nitrate and total N. Consequently, grass growing under dense canopies in south Sweden is not able to dilute N by increasing growth. The diminished capacity of the grass to assimilate nitrate will increase leaching losses of N from forests approaching N saturation.     

Early responses to cadmium of two poplar clones that differ in stress tolerance

Soil cadmium (Cd) contamination is becoming a matter of great global concern. The identification of plants differentially sensitive to Cd excess is of interest for the selection of genotype adaptive to grow and develop in polluted areas and capable of ameliorating or reducing the negative environmental effects of this toxic metal. The two poplar clones I-214 (Populus × canadensis) and Eridano (Populus deltoides × maximowiczii) are, respectively, tolerant and sensitive to ozone (O₃) exposure. Because stress tolerance is mediated by an array of overlapping defence mechanisms, we tested the hypothesis that these two clones differently sensitive to O₃ stress factor also exhibit different tolerance to Cd. With this purpose, an outdoor pot experiment was designed to study the responses of I-214 and Eridano to the distribution of different Cd solutions enriched with CdCl₂ (0, 50 and 150 μM) for 35 days. Changes in leaf area, biomass allocation and Cd uptake, photosynthesis, chlorophyll fluorescence, leaf concentration of nutrients and pigments, hydrogen peroxide (H₂O₂) and nitric oxide (NO) production and thiol compounds were investigated. The two poplar clones showed similar sensitivity to excess Cd in terms of biomass production, photosynthesis activity and Cd accumulation, though physiological and biochemical traits revealed different defence strategies. In particular, Eridano maintained in any Cd treatment the number of its constitutively wider blade leaves, while the number of I-214 leaves (with lower size) was reduced. H₂O₂ increased 4.5- and 13-fold in I-214 leaves after the lowest (L) and highest (H) Cd treatments, respectively, revealing the induction of oxidative burst. NO, constitutively higher in I-214 than Eridano, progressively increased in both clones with the enhancement of Cd concentration in the substrate. I-214 showed a more elevated antioxidative capacity (GSH/GSSG) and higher photochemical efficiency of PSII (F_v/F_m) and de-epoxidation degree of xantophylls-cycle (DEPS). The glutathione pool was not affected by Cd treatment in both clones, while non-protein thiols and phytochelatins were reduced at L Cd treatment in I-214. Overall, these two clones presented high adaptability to Cd stress and are both suitable to develop and growth in environments contaminated with this metal, thus being promising for their potential use in phytoremediation programmes.     

Effects of silicon supply on apoplastic manganese concentrations in leaves and their relation to manganese tolerance in cowpea (Vigna unguiculata (L.) Walp.)

The effects of silicon (Si) supply on manganese (Mn) toxicity symptoms and Mn and Si concentrations in the leaf apoplast in a Mn-sensitive cowpea cultivar (Vigna unguiculata (L.) Walp. cv. TVu 91) were investigated in solution culture experiments. When 1.44 mM Si was supplied concurrently with 50 M Mn, the Mn toxicity symptoms were clearly avoided without decreasing the total Mn concentration. On the other hand, the symptoms were not completely alleviated when the plants were pretreated with 1.44 mM Si and then exposed to 50 M Mn without concurrent Si supply. Plants of both of these treatments exhibited lower Mn concentrations in the apoplastic washing fluids but higher amounts of adsorbed Mn on the cell walls than the plants treated with 50 M Mn without Si supply. However, the difference in Mn concentration between plants with continuous and interrupted Si supply was not significant. Moreover, the Mn concentration in the apoplastic washing fluids of the plants with continuous supply of 1.44 mM Si and 50 M Mn and not showing Mn toxicity symptoms was higher than that of the plants grown at 10 M Mn without Si supply which showed distinct Mn toxicity symptoms. These results show that Si supply alleviates Mn toxicity not only by decreasing the concentration of soluble apoplastic Mn through the enhanced adsorption of Mn on the cell walls. A role of the soluble Si in the apoplast in the detoxicification of apoplastic Mn is indicated.     

Uptake kinetics and assimilation of phosphorus byCatenella nipaeandUlva lactucacan be used to indicate ambient phosphate availability

Uptake, assimilation and compartmentation of phosphate were studied in the opportunist green macroalgaUlva lactucaand the estuarine red algal epiphyteCatenella nipae. The Michaelis–Menten model was used to describe uptake rates of inorganic phosphate (P_i) at different concentrations. Maximum uptake rates (V _max) of P-starved material exceededV _maxof P-enriched material; this difference was greater forC. nipae. Uptake and allocation of phosphorus (P) to internal pools was measured using trichloroacetic acid (TCA) extracts and³²P. Both species demonstrated similar assimilation paths: when P-enriched, most³²P accumulated as free phosphate. When unenriched,³²P was rapidly assimilated into the TCA-insoluble pool.C. nipaeconsistently assimilated more³²P into this pool thanU. lactuca, indicatingC. nipaehas a greater P-storage capacity. In both species,³²P release data showed two internal compartments with very different biological half-lives. The rapidly exchanging compartment had a short half-life of 2 to 12 min, while the slowly exchanging compartment had a much longer half-life of 12 days in P-starvedC. nipaeor 4 days in P-starvedU. lactuca. In both species, the slowly exchanging compartment accounted for more than 90% of total tissue.U. lactucaandC. nipaeresponded differently to high external P_i.U. lactucarapidly took up P_i, transferring this P_iinto tissue phosphate and TCA-soluble P in a few hours (90% of total P).C. nipaetook up P_iat lower rates and stored much of this P in less mobile TCA-insoluble forms. Long-term storage of refractory forms of P makesC. nipaea useful bioindicator of the prevailing conditions of P_iavailability over at least the previous 7 days, whereas the P-status ofU.lactucamay reflect conditions over no more than the previous few hours or days.C. nipaeis a more useful bioindicator for P status of estuarine and marine waters thanU. lactuca.     

The use of immunological tolerance to investigate B lymphocyte replacement kinetics in chickens

A simple mathematical model has been derived, describing the irreversible inactivation of immature B cells by high doses of antigen during induction of tolerance, and the antigen-independent replacement of B cells by differentiation of their precursors. The latter leads to recovery from tolerance, the rate of which can be used to assess the rate of B cell replacement in experiments. The model has been compared with experimental tolerance to human albumin in newly hatched chickens.(1) It has been shown that this tolerance cannot be explained only by elimination of B cells but (2) the computed rate of B cell replacement agreed with the experimental rate assessed by immunization of tolerant chickens with a cross-reacting antigen. (3) In order to further verify the model, additional experiments to test the rate of B cell replacement were suggested by the model.     

Effects of plant species on phosphorus availability in a range of grassland soils

Vegetative conversion from grass to forest may influence soil nutrient dynamics and availability. A short-term (40 weeks) glasshouse experiment was carried out to investigate the impacts of ryegrass (Lolium perenne) and radiata pine (Pinus radiata) on soil phosphorus (P) availability in 15 grassland soils collected across New Zealand using ³³P isotopic exchange kinetics (IEK) and chemical extraction methods. Results from this study showed that radiata pine took up more P (4.5–33.5 mg P pot^–1) than ryegrass (1.1–15.6 mg pot^–1) from the soil except in the Temuka soil in which the level of available P (e.g., E _1min P_i, bicarbonate extractable P_i) was very high. Radiata pine tended to be better able to access different forms of soil P, compared with ryegrass. There were no significant differences in the level of water soluble P (Cp, intensity factor) between soils under ryegrass and radiata pine, but the levels of Cp were generally lower compared with original soils due to plant uptake. The growth of both ryegrass and radiata pine resulted in the redistribution of soil P from the slowly exchangeable P_i pool (E _{> 10m} P_i, reduced by 31.8% on the average) to the rapidly exchangeable P_i (E _1min-1d P_i, E _1d-10m P_i) pools in most soils. The values of R/r ₁ (the capacity factor) were also generally greater in most soils under radiata pine compared with ryegrass. Specific P mineralisation rates were significantly greater for soils under radiata pine (8.4–21.9%) compared with ryegrass (0.5–10.8%), indicating that the growth of radiata pine enhanced mineralisation of soil organic P. This may partly be ascribed to greater root phosphatase activity for radiata pine than for ryegrass. Plant species × soil type interactions for most soil variables measured indicate that the impacts of plant species on soil P dynamics was strongly influenced by soil properties.     

Agrobacterium-mediated genetic transformation of the desiccation tolerant resurrection plant Ramonda myconi (L.) Rchb.

In this paper we describe the first procedure for Agrobacterium tumefaciens-mediated genetic transformation of the desiccation tolerant plant Ramonda myconi (L.) Rchb. Previously, we reported the establishment of a reliable and effective tissue culture system based on the integrated optimisation of antioxidant and growth regulator composition and the stabilisation of the pH of the culture media by means of a potassium phosphate buffer. This efficient plant regeneration via callus phase provided a basis for the optimisation of the genetic transformation in R. myconi. For gene delivery, both a standard (method A) and a modified protocol (method B) have been applied. Since the latter has previously resulted in successful transformation of another resurrection plant, Craterostigma plantagineum, an identical protocol was utilized in transformation of R. myconi, as this method may prove general for dicotyledonous resurrection plants. On this basis, physical and biochemical key variables in transformation were evaluated such as mechanical microwounding of plant explants and in vitro preinduction of vir genes. While the physical enhancement of bacterial penetration was proved to be essential for successful genetic transformation of R. myconi, an additional two-fold increase in the transformation frequency was obtained when the above physical and biochemical treatments were applied in combination. All R ₀ and R ₁ transgenic plants were fertile, and no morphological abnormalities were observed on the whole-plant level. Collaborator via a fellowship under the OECD Co-operative Research Programme: Biological Resource Management for Sustainable Agriculture Systems     

Carbon discrimination,water-use efficiency,nitrogen and phosphorus nutrition of the host / mistletoe pair Eucalyptus behriana F. Muell and Amyema miquelii (Lehm. ex Miq.) Tiegh. at permanently low plant water status in the field

Summary Under conditions where both plants had permanently low water status, the mistletoe, Amyema miquelii (Lehm. ex Miq.) Tiegh., had lower nitrogen contents in leaf tissue than its host, Eucalyptus behriana F. Muell. The parasite transpired less than its host which is consistent with the hypothesis that mistletoe transpiration acts as a nitrogen gathering mechanism. Nitrogen and phosphorus contents were generally low in both plants; they were positively correlated, and mistletoes reduced nutrient contents of infested hosts. The carbon discrimination ratio, ¹³C (a measure of water-use efficiency) of each plant was within the range reported for other mistletoes and their hosts. Although it did not differ significantly between host and parasite it indicated lower water-use efficiency in the mistletoe. For the nitrogen content of host leaves the gradient within the pair, (¹³C), is much lower compared to the correlation given by Ehleringer et al. (1985). It is concluded that at permanently low water status on nitrogen and phosphorus deficient soils a water-saving strategy accompanied with slow growth is more appropriate for both mistletoe and host.