Ecophysiological Performance of Proteaceae Species From Southern South America Growing on Substrates Derived From Young Volcanic Materials.

Southern South American Proteaceae thrive on young volcanic substrates, which are extremely low in plant-available phosphorus (P). Most Proteaceae exhibit a nutrient-acquisition strategy based on the release of carboxylates from specialized roots, named cluster roots (CR). Some Proteaceae colonize young volcanic substrates which has been related to CR functioning. However, physiological functioning of other Proteaceae on recent volcanic substrates is unknown. We conducted an experiment with seedlings of five Proteaceae (Gevuina avellana, Embothrium coccineum, Lomatia hirsuta, L. ferruginea, and L. dentata) grown in three volcanic materials. Two of them are substrates with very low nutrient concentrations, collected from the most recent deposits of the volcanoes Choshuenco and Calbuco (Chile). The other volcanic material corresponds to a developed soil that exhibits a high nutrient availability. We assessed morphological responses (i.e., height, biomass, and CR formation), seed and leaf macronutrient and micronutrient concentrations and carboxylates exuded by roots. The results show that G. avellana was less affected by nutrient availability of the volcanic substrate, probably because it had a greater nutrient content in its seeds and produced large CR exuding carboxylates that supported their initial growth. Embothrium coccineum exhibited greater total plant height and leaf P concentration than Lomatia species. In general, in all species leaf macronutrient concentrations were reduced on nutrient-poor volcanic substrates, while leaf micronutrient concentrations were highly variable depending on species and volcanic material. We conclude that Proteaceae from temperate rainforests differ in their capacity to grow and acquire nutrients from young and nutrient-poor volcanic substrates. The greater seed nutrient content, low nutrient requirements (only for G. avellana) and ability to mobilize nutrients help explain why G. avellana and E. coccineum are better colonizers of recent volcanic substrates than Lomatia species.

Plants sustain the terrestrial silicon cycle during ecosystem retrogression.

The biogeochemical silicon cycle influences global primary productivity and carbon cycling, yet changes in silicon sources and cycling during long-term development of terrestrial ecosystems remain poorly understood. Here, we show that terrestrial silicon cycling shifts from pedological to biological control during long-term ecosystem development along 2-million-year soil chronosequences in Western Australia. Silicon availability is determined by pedogenic silicon in young soils and recycling of plant-derived silicon in old soils as pedogenic pools become depleted. Unlike concentrations of major nutrients, which decline markedly in strongly weathered soils, foliar silicon concentrations increase continuously as soils age. Our findings show that the retention of silicon by plants during ecosystem retrogression sustains its terrestrial cycling, suggesting important plant benefits associated with this element in nutrient-poor environments.

Phosphorus-acquisition strategies of canola, wheat and barley in soil amended with sewage sludges.

Crops have different strategies to acquire poorly-available soil phosphorus (P) which are dependent on their architectural, morphological, and physiological root traits, but their capacity to enhance P acquisition varies with the type of fertilizer applied. The objective of this study was to examine how P-acquisition strategies of three main crops are affected by the application of sewage sludges, compared with a mineral P fertilizer. We carried out a 3-months greenhouse pot experiment and compared the response of P-acquisition traits among wheat, barley and canola in a soil amended with three sludges or a mineral P fertilizer. Results showed that the P-acquisition strategy differed among crops. Compared with canola, wheat and barley had a higher specific root length and a greater root carboxylate release and they acquired as much P from sludge as from mineral P. By contrast, canola shoot P content was greater with sludge than with mineral P. This was attributed to a higher root-released acid phosphatase activity which promoted the mineralization of sludge-derived P-organic. This study showed that contrasted P-acquisition strategies of crops allows increased use of renewable P resources by optimizing combinations of crop and the type of P fertilizer applied within the cropping system.

Phosphorus nutrition in Proteaceae and beyond.

Proteaceae in southwestern Australia have evolved on some of the most phosphorus-impoverished soils in the world. They exhibit a range of traits that allow them to both acquire and utilize phosphorus highly efficiently. This is in stark contrast with many model plants such as Arabidopsis thaliana and crop species, which evolved on soils where nitrogen is the major limiting nutrient. When exposed to low phosphorus availability, these plants typically exhibit phosphorus-starvation responses, whereas Proteaceae do not. This Review explores the traits that account for the very high efficiency of acquisition and use of phosphorus in Proteaceae, and explores which of these traits are promising for improving the phosphorus efficiency of crop plants.

Convergence of a specialized root trait in plants from nutrient-impoverished soils: phosphorus-acquisition strategy in a nonmycorrhizal cactus.

In old, phosphorus (P)-impoverished habitats, root specializations such as cluster roots efficiently mobilize and acquire P by releasing large amounts of carboxylates in the rhizosphere. These specialized roots are rarely mycorrhizal. We investigated whether Discocactus placentiformis (Cactaceae), a common species in nutrient-poor campos rupestres over white sands, operates in the same way as other root specializations. Discocactus placentiformis showed no mycorrhizal colonization, but exhibited a sand-binding root specialization with rhizosheath formation. We first provide circumstantial evidence for carboxylate exudation in field material, based on its very high shoot manganese (Mn) concentrations, and then firm evidence, based on exudate analysis. We identified predominantly oxalic acid, but also malic, citric, lactic, succinic, fumaric, and malonic acids. When grown in nutrient solution with P concentrations ranging from 0 to 100 µM, we observed an increase in total carboxylate exudation with decreasing P supply, showing that P deficiency stimulated carboxylate release. Additionally, we tested P solubilization by citric, malic and oxalic acids, and found that they solubilized P from the strongly P-sorbing soil in its native habitat, when the acids were added in combination and in relatively low concentrations. We conclude that the sand-binding root specialization in this nonmycorrhizal cactus functions similar to that of cluster roots, which efficiently enhance P acquisition in other habitats with very low P availability.

Carbon trading for phosphorus gain: the balance between rhizosphere carboxylates and arbuscular mycorrhizal symbiosis in plant phosphorus acquisition.

Two key plant adaptations for phosphorus (P) acquisition are carboxylate exudation into the rhizosphere and mycorrhizal symbioses. These target different soil P resources, presumably with different plant carbon costs. We examined the effect of inoculation with arbuscular mycorrhizal fungi (AMF) on amount of rhizosphere carboxylates and plant P uptake for 10 species of low-P adapted Kennedia grown for 23 weeks in low-P sand. Inoculation decreased carboxylates in some species (up to 50%), decreased plant dry weight (21%) and increased plant P content (23%). There was a positive logarithmic relationship between plant P content and the amount of rhizosphere citric acid for inoculated and uninoculated plants. Causality was indicated by experiments using sand where little citric acid was lost from the soil solution over 2 h and citric acid at low concentrations desorbed P into the soil solution. Senesced leaf P concentration was often low and P-resorption efficiencies reached >90%. In conclusion, we propose that mycorrhizally mediated resource partitioning occurred because inoculation reduced rhizosphere carboxylates, but increased plant P uptake. Hence, presumably, the proportion of plant P acquired from strongly sorbed sources decreased with inoculation, while the proportion from labile inorganic P increased. Implications for plant fitness under field conditions now require investigation.

Increased ecological amplitude through heterosis following wide outcrossing in Banksia ilicifolia R.Br. (Proteaceae).

To assess whether wide outcrossing (over 30 km) in the naturally fragmented Banksia ilicifolia R.Br. increases the ecological amplitude of offspring, we performed a comparative greenhouse growth study involving seedlings of three hand-pollinated progeny classes (self, local outcross, wide outcross) and a range of substrates and stress conditions. Outcrossed seedlings outperformed selfed seedlings, with the magnitude of inbreeding depression as high as 62% for seed germination and 37% for leaf area. Wide outcrossed seedlings outperformed local outcrossed seedlings, especially in non-native soils, facilitated in part by an improved capacity to overcome soil constraints through greater root carboxylate exudation. Soil type significantly affected seedling growth, and waterlogging and water deficit decreased growth, production of cluster roots, root exudation and total plant P uptake. Our results suggest that the interaction of narrow ecological amplitude and the genetic consequences of small fragmented populations may in part explain the narrow range of local endemics, but that wide outcrossing may provide opportunities for increased genetic variation, increased ecological amplitude and range expansion.

Rhizosphere carboxylate concentrations of chickpea are affected by soil bulk density.

We investigated whether carboxylate exudation by chickpea (Cicer arietinum L.) was affected by soil bulk density and if this effect was local or systemic. We hypothesised that concentrations of carboxylates would increase with distance from the root apex due to continuous and constitutive accumulation of carboxylates, and that exudate accumulation would be greater in a compacted soil than in a loose soil. Plants were grown in split-root or single cylinders containing loose (1400 kg m (-3)) or compacted (1800 kg m (-3)) soil. Rhizosphere carboxylate concentrations were measured of whole root systems as well as of sections along the root. The root diameter was greatest of plants grown in the compacted soil; however, root diameters were the same for both root halves in the split-root design, whether they grew in loose soil or in compacted soil. Similarly, carboxylate concentrations tended to be lower for the whole root system in the compacted soil, but were the same for both root halves in the split-root design, irrespective of whether the roots were in loose soil or in compacted soil. These results indicate that both root diameter and carboxylate exudation by roots in chickpea is regulated systemically via a signal from the shoot rather than by local signals in the roots. There was no accumulation of carboxylates with increasing distance from the apex, probably because microbial degradation occurred at similar rates as carboxylate exudation. Malonate, previously suggested as deterrent to microorganisms, is likely only a selective deterrent.

Natural skin surface pH is on average below 5, which is beneficial for its resident flora.

Variable skin pH values are being reported in literature, all in the acidic range but with a broad range from pH 4.0 to 7.0. In a multicentre study (N = 330), we have assessed the skin surface pH of the volar forearm before and after refraining from showering and cosmetic product application for 24 h. The average pH dropped from 5.12 +/- 0.56 to 4.93 +/- 0.45. On the basis of this pH drop, it is estimated that the 'natural' skin surface pH is on average 4.7, i.e. below 5. This is in line with existing literature, where a relatively large number of reports (c. 50%) actually describes pH values below 5.0; this is in contrast to the general assumption, that skin surface pH is on average between 5.0 and 6.0. Not only prior use of cosmetic products, especially soaps, have profound influence on skin surface pH, but the use of plain tap water, in Europe with a pH value generally around 8.0, will increase skin pH up to 6 h after application before returning to its 'natural' value of on average below 5.0. It is demonstrated that skin with pH values below 5.0 is in a better condition than skin with pH values above 5.0, as shown by measuring the biophysical parameters of barrier function, moisturization and scaling. The effect of pH on adhesion of resident skin microflora was also assessed; an acid skin pH (4-4.5) keeps the resident bacterial flora attached to the skin, whereas an alkaline pH (8-9) promotes the dispersal from the skin.

The contribution of roots and shoots to whole plant nitrate reduction in fast- and slow-growing grass species.

The hypothesis was tested that slow-growing grass species perform a greater proportion of total plant NO3- reduction in their roots than do fast-growing grasses. Eight grass species were selected that differed in maximum relative growth rate (RGR) and net NO3- uptake rate (NNUR). Plants were grown with free access to nutrients in hydroponics under controlled-environment conditions. The site of in vivo NO3- reduction was assessed by combining in vivo NO3- reductase activity (NRA) assays with biomass allocation data, and by analysing the NO3- to amino acid ratio of xylem sap. In vivo NRA of roots and shoots increased significantly with increasing NNUR and RGR. The proportion of total plant NO3- reduction that occurs in roots was found to be independent of RGR and NNUR, with the shoot being the predominant site of NO3- reduction in all species. The theoretical maximum proportion of whole plant nitrogen assimilation that could take place in the roots was calculated using information on root respiration rates, RGR, NNUR, and specific respiratory costs associated with growth, maintenance and ion uptake. The calculated maximum proportion that the roots can contribute to total plant NO3- reduction was 0.37 and 0.23 for the fast-growing Dactylis glomerata L. and the slow-growing Festuca ovina L., respectively. These results indicate that slow-growing grass species perform a similar proportion of total plant NO3- reduction in their roots to that exhibited by fast-growing grasses. Shoots appear to be the predominant site of whole plant NO3- reduction in both fast- and slow-growing grasses when plants are grown with free access to nutrients.

Regulation of alternative oxidase activity in six wild monocotyledonous species. An in vivo study at the whole root level.

The activity of the alternative pathway is affected by a number of factors, including the level and reduction state of the alternative oxidase (AOX) protein, and the reduction state of the ubiquinone pool. To investigate the significance of these factors for the rate of alternative respiration in vivo, we studied root respiration of six wild monocotyledonous grass species that were grown under identical controlled conditions. The activity of the alternative pathway was determined using the oxygen isotope fractionation technique. In all species, the AOX protein was invariably in its reduced (high activity) state. There was no correlation between AOX activity and AOX protein concentration, ubiquinone (total, reduced, or oxidized) concentration, or the reduction state of the ubiquinone pool. However, when some of these factors are combined in a linear regression model, a good fit to AOX activity is obtained. The function of the AOX is still not fully understood. It is interesting that we found a positive correlation between the activity of the alternative pathway and relative growth rate; a possible explanation for this correlation is discussed. Inhibition of the AOX (with salicylhydroxamic acid) decreases respiration rates less than the activity present before inhibition (i.e. measured with the 18O-fractionation technique).

Can meristematic activity determine variation in leaf size and elongation rate among four Poa species? A kinematic study.

We studied inherent variation in final leaf size among four Poa spp. that live at different elevations. The average final length of leaf 7 of the main stem of the smallest species (Poa alpina) was only one-half that of the largest species (Poa trivialis); it was correlated with leaf elongation rate, but not with the duration of leaf elongation. A faster rate of leaf elongation rate was associated with (a) larger size of the zone of cell expansion, and (b) faster rates of cell production (per cell file) in the meristem, which in turn were due to greater numbers of dividing cells, whereas average cell division rates were very similar for all species (except Poa annua). Also we found that the proliferative fraction equaled 1 throughout the meristem in all species. It was remarkable that rates of cell expansion tended to be somewhat higher in the species with slower growing leaves. We discuss the results by comparing the spatial and material viewpoints, which lead to different interpretations of the role of cell division. Although the presented data do not strictly prove it, they strongly suggest a regulatory role for cell division in determining differences in growth rate among the present four Poa spp.

The alternative oxidase in roots of poa annua after transfer from high-light to low-light conditions.

The activity of the alternative pathway can be affected by a number of factors, including the amount and reduction state of the alternative oxidase protein, and the reduction state of the ubiquinone pool. To investigate the importance of these factors in vivo, we manipulated the rate of root respiration by transferring the annual grass Poa annua L. from high-light to low-light conditions, and at the same time from long-day to short-day conditions for four days. As a result of the low-light treatment, the total respiration rate of the roots decreased by 45%, in vitro cytochrome c oxidase capacity decreased by 49%, sugar concentration decreased by 90% and the ubiquinone concentration increased by 31%, relative to control values. The absolute rate of oxygen uptake via the alternative pathway, as determined using the 18O-isotope fractionation technique, did not change. Conversely, the cytochrome pathway activity decreased during the low-light treatment; its activity increased upon addition of exogenous sugars to the roots. Interestingly, no change was observed in the concentration of the alternative oxidase protein or in the reduction state of the protein. Also, there was no change in the reduction state of the ubiquinone pool. In conclusion, the concentration and activity of the alternative oxidase were not changed, even under severe light deprivation.

Respiratory costs and rate of protein turnover in the roots of a fast-growing (Dactylis glomerata L.) and a slow-growing (Festuca ovina L.) grass species.

Protein turnover is generally regarded as one of the most important maintenance processes in plants in terms of energy requirements. In this study, the contribution of protein turnover to the respiratory costs for maintenance in the roots of two grass species, the fast-growing D. actylis glomerata L. and the slow-growing F. estuca ovina L., is evaluated. Plants were grown under controlled-environment conditions in a nutrient solution to which NO(3)- was added at a relative addition rate of 0.2 and 0.1 mol N mol(-1) N already present in the plant d(-1) for D. glomerata and F. ovina, respectively, so as to obtain a steady exponential growth rate close to the plants' maximum relative growth rate. Pulse-chase labelling with (14)C-leucine was used to determine the rate of protein turnover in the grass roots. The rate of turnover of the total protein pool did not differ significantly between the two species. The protein degradation constant in D. glomerata and F. ovina was 0.156 and 0.116 g protein g(-1) protein d(-1), respectively, which corresponds with a total protein half-life of 4 d and 6 d. Assuming specific respiratory costs for protein turnover of 148 mmol ATP g(-1) protein, the estimated respiratory costs for protein turnover in the roots were 2.8 and 2.4 mmol ATP g(-1) root DM d(-1) in D. glomerata and F. ovina, respectively. Both the fast- and the slow-growing grass spent between 22-30% of their daily ATP production for maintenance on protein turnover, which corresponds to 11-15% of the total root ATP production per day. Note that the data presented in this abstract are based on the assumption that 50% recycling of the (14)C-labelled leucine took place in the roots of both grass species.

Leaf respiration of snow gum in the light and dark. Interactions between temperature and irradiance.

We investigated the effect of temperature and irradiance on leaf respiration (R, non-photorespiratory mitochondrial CO(2) release) of snow gum (Eucalyptus pauciflora Sieb. ex Spreng). Seedlings were hydroponically grown under constant 20 degrees C, controlled-environment conditions. Measurements of R (using the Laisk method) and photosynthesis (at 37 Pa CO(2)) were made at several irradiances (0-2,000 micromol photons m(-2) s(-1)) and temperatures (6 degrees C-30 degrees C). At 15 degrees C to 30 degrees C, substantial inhibition of R occurred at 12 micromol photons m(-2) s(-1), with maximum inhibition occurring at 100 to 200 micromol photons m(-2) s(-1). Higher irradiance had little additional effect on R at these moderate temperatures. The irradiance necessary to maximally inhibit R at 6 degrees C to 10 degrees C was lower than that at 15 degrees C to 30 degrees C. Moreover, although R was inhibited by low irradiance at 6 degrees C to 10 degrees C, it recovered with progressive increases in irradiance. The temperature sensitivity of R was greater in darkness than under bright light. At 30 degrees C and high irradiance, light-inhibited rates of R represented 2% of gross CO(2) uptake (v(c)), whereas photorespiratory CO(2) release was approximately 20% of v(c). If light had not inhibited leaf respiration at 30 degrees C and high irradiance, R would have represented 11% of v(c). Variations in light inhibition of R can therefore have a substantial impact on the proportion of photosynthesis that is respired. We conclude that the rate of R in the light is highly variable, being dependent on irradiance and temperature.

Presymptomatic visualization of plant-virus interactions by thermography.

Salicylic acid (SA), produced by plants as a signal in defense against pathogens, induces metabolic heating mediated by alternative respiration in flowers of thermogenic plants, and, when exogenously applied, increases leaf temperature in nonthermogenic plants. We have postulated that the latter phenomenon would be detectable when SA is synthesized locally in plant leaves. Here, resistance to tobacco mosaic virus (TMV) was monitored thermographically before any disease symptoms became visible on tobacco leaves. Spots of elevated temperature that were confined to the place of infection increased in intensity from 8 h before the onset of visible cell death, and remained detectable as a halo around the ongoing necrosis. Salicylic acid accumulates during the prenecrotic phase in TMV-infected tobacco and is known to induce stomatal closure in certain species. We show that the time course of SA accumulation correlates with the evolution of both localized thermal effect and stomatal closure. Since the contribution of leaf respiration is marginal, we concluded that the thermal effect results predominantly from localized, SA-induced stomatal closure. The presymptomatic temperature increase could be of general significance in incompatible plant-pathogen interactions.

Enhanced expression and activation of the alternative oxidase during infection of Arabidopsis with Pseudomonas syringae pv tomato.

Cyanide-resistant ("alternative") respiration was studied in Arabidopsis during incompatible and compatible infection with Pseudomonas syringae pv tomato DC3000. Total leaf respiration increased as the leaves became necrotic, as did the cyanide-resistant component that was sensitive to salicylhydroxamic acid. Infiltration of leaves with an avirulent strain rapidly induced alternative oxidase (AOX) mRNA, whereas the increase was delayed in the compatible combination. The increase in mRNA correlated with the increase in AOX protein. Increased expression was confined to the infected leaves, in contrast to the pathogenesis-related protein-1, which was induced systemically. Virtually all of the AOX protein was in the reduced (high-activity) form. Using transgenic NahG and mutant npr1-1 and etr1-1 plants, we established that the rapid induction of the AOX was associated with necrosis and that ethylene, but not salicylic acid, was required for its induction. Increased pyruvate levels in the infected leaves suggested that increased substrate levels were respired through the alternative pathway; however, in the control leaves and the infected leaves, respiration was not inhibited by salicylhydroxamic acid alone. Increased respiration appeared to be associated primarily with symptom expression rather than resistance reactions.

The role of the alternative oxidase in stabilizing the in vivo reduction state of the ubiquinone pool and the activation state of the alternative oxidase

A possible function for the alternative (nonphosphorylating) pathway is to stabilize the reduction state of the ubiquinone pool (Qr/Qt), thereby avoiding an increase in free radical production. If the Qr/Qt were stabilized by the alternative pathway, then Qr/Qt should be less stable when the alternative pathway is blocked. Qr/Qt increased when we exposed roots of Poa annua (L.) to increasing concentrations of KCN (an inhibitor of the cytochrome pathway). However, when salicylhydroxamic acid, an inhibitor of the alternative pathway, was added at the same time, Qr/Qt increased significantly more. Therefore, we conclude that the alternative pathway stabilizes Qr/Qt. Salicylhydroxamic acid increasingly inhibited respiration with increasing concentrations of KCN. In the experiments described here the alternative oxidase protein was invariably in its reduced (high-activity) state. Therefore, changes in the reduction state of the alternative oxidase cannot account for an increase in activity of the alternative pathway upon titration with KCN. The pyruvate concentration in intact roots increased only after the alternative pathway was blocked or the cytochrome pathway was severely inhibited. The significance of the pyruvate concentration and Qr/Qt on the activity of the alternative pathway in intact roots is discussed.

Leaf Respiration in Light and Darkness (A Comparison of Slow- and Fast-Growing Poa Species).

We investigated whether leaf dark respiration (nonphotorespiratory mitochondrial CO2 release) is inhibited by light in several Poa species, and whether differences in light inhibition between the species are related to differences in the rate of leaf net photosynthesis. Four lowland (Poa annua L., Poa compressa L., Poa pratensis L., and Poa trivialis L.), one subalpine (Poa alpina L.), and two alpine (Poa costiniana Vick. and Poa fawcettiae Vick.) Poa species differing in whole plant relative growth rates were grown under identical controlled conditions. Nonphotorespiratory mitochondrial CO2 release in the light (Rd) was estimated according to the Laisk method. Photosynthesis was measured at ambient CO2 partial pressure (35 Pa) and 500 [mu]mol photons m-2 s-1. The rate of photosynthesis per unit leaf mass was positively correlated with the relative growth rate, with the slow-growing alpine Poa species exhibiting the lowest photosynthetic rates. Rates of both Rd and respiration in darkness were also substantially lower in the alpine species. Nonphotorespiratory CO2 release in darkness was higher than Rd in all species. However, despite some variation between the species in the level of light inhibition of respiration, no relationship was observed between the level of inhibition and the rate of photosynthesis. Similarly, the level of inhibition was not correlated with the relative growth rate. Our results support the suggestion that rates of leaf respiration in the light are closely associated with rates in darkness.