Hakea Prostrata Research Articles

A root trait accounting for the extreme phosphorus sensitivity of Hakea prostrata (Proteaceae)

ABSTRACTProteaceae are adapted to acquire P from nutrient‐impoverished soils; many function at very low leaf P levels, but are killed by P fertilization. Phosphorus toxicity develops at a remarkably low external P concentration. Previous studies have described P toxicity in Proteaceae, but the physiological basis for it remained unclear. The aim of the present study was to elucidate the physiological basis of P toxicity in Hakea prostrata R. Br. (Proteaceae). Triticum aestivum L. (Gramineae), Medicago truncatula Gaertn., Lupinus albus L. (both Fabaceae) and Hakea prostrata R.Br. were grown in solution at a range of P concentrations (0–1000 mmol P m−3), and determined net P‐uptake rates at 5 (all species) and 50 mmol P m−3 (H. prostrata only). With the exception of H. prostrata, net P‐uptake rates were fastest for plants grown without added P. Down‐regulation occurred for T. aestivum, M. truncatula and L. albus when the P concentration during growth was increased from 0 to 0.8 mmol P m−3, whereas in H. prostrata rates decreased only for plants grown at 10 mmol P m−3 or more. The leaf [P] at which P toxicity occurred in H. prostrata exceeded 10 mg g−1 dry matter, similar to that for crop species. The low capacity to reduce P uptake in response to increased supply offers a physiological explanation for the extreme sensitivity to P supply in H. prostrata, and possibly other Proteaceae.

Developmental physiology of cluster-root carboxylate synthesis and exudation in harsh hakea. Expression of phosphoenolpyruvate carboxylase and the alternative oxidase.

Harsh hakea (Hakea prostrata R.Br.) is a member of the Proteaceae family, which is highly represented on the extremely nutrient-impoverished soils in southwest Australia. When phosphorus is limiting, harsh hakea develops proteoid or cluster roots that release carboxylates that mobilize sparingly soluble phosphate in the rhizosphere. To investigate the physiology underlying the synthesis and exudation of carboxylates from cluster roots in Proteaceae, we measured O2 consumption, CO2 release, internal carboxylate concentrations and carboxylate exudation, and the abundance of the enzymes phosphoenolpyruvate carboxylase and alternative oxidase (AOX) over a 3-week time course of cluster-root development. Peak rates of citrate and malate exudation were observed from 12- to 13-d-old cluster roots, preceded by a reduction in cluster-root total protein levels and a reduced rate of O2 consumption. In harsh hakea, phosphoenolpyruvate carboxylase expression was relatively constant in cluster roots, regardless of developmental stage. During cluster-root maturation, however, the expression of AOX protein increased prior to the time when citrate and malate exudation peaked. This increase in AOX protein levels is presumably needed to allow a greater flow of electrons through the mitochondrial electron transport chain in the absence of rapid ATP turnover. Citrate and isocitrate synthesis and accumulation contributed in a major way to the subsequent burst of citrate and malate exudation. Phosphorus accumulated by harsh hakea cluster roots was remobilized during senescence as part of their efficient P cycling strategy for growth on nutrient impoverished soils.

Effects of external phosphorus supply on internal phosphorus concentration and the initiation, growth and exudation of cluster roots in Hakea prostrata R.Br.

The response of internal phosphorus concentration, cluster-root initiation, and growth and carboxylate exudation to different external P supplies was investigated in Hakea prostrata R.Br. using a split-root design. After removal of most of the taproot, equal amounts of laterals were allowed to grow in two separate pots fastened together at the top, so that the separate root halves could be exposed to different conditions. Plants were grown for 10 weeks in this system; one root half was supplied with 1 μM P while the other halves were supplied with 0, 1, 25 or 75 μM P. Higher concentrations of P supplied to one root half significantly increased the P concentration of those roots and in the shoots. The P concentrations in root halves supplied with 1 μM P were invariably low, regardless of the P concentration supplied to the other root half. Cluster root initiation was completely suppressed on root halves supplied with 25 or 75 μM P, whereas it continued on the other halves supplied with 1 μM P indicating that cluster-root initiation was regulated by local root P concentration. Cluster-root growth (dry mass increment) on root halves supplied with 1 μM P was significantly reduced when the other half was either deprived of P or supplied with 25 or 75 μM P. Cluster-root growth was favoured by a low shoot P status at a root P supply that was adequate for increased growth of roots and shoots without increased tissue P concentrations. The differences in cluster-root growth on root halves with the same P supply suggest that decreased cluster-root growth was systemically regulated. Carboxylate-exudation rates from cluster roots on root halves supplied with 1 μM P were the same, whether the other root half was supplied with 1, 25 or 75 μM P, but were approximately 30 times faster when the other half was deprived of P. Estimates of root P-uptake rates suggest a rather limited capacity for down-regulating P uptake when phosphate was readily available.

Exudation of carboxylates in Australian Proteaceae: chemical composition

AbstractRoots of a wide range of plant species exude carboxylates, such as citrate, into the rhizosphere. In the present study, seedlings of a range of Australian Banksia, Hakea and Dryandra species (Proteaceae) were assayed for their exudation of carboxylates. All of these species (Hakea prostrata, Hakea undulata, Hakea petiolaris, Hakea baxteri, Banksia grandis, Banksia prionotes, Banksia occidentalis and Dryandra sessilis) form cluster roots when grown in nutrient solution with a low phosphate concentration. Exudation of carboxylates was studied for cluster roots and non‐cluster roots separately, and for the entire root system. Cluster roots of these Proteaceae exuded malate, malonate, lactate, acetate, maleate, citrate, fumarate, cis‐ and trans‐aconitate. The relative contributions of each of these carboxylates differed between species. Malate, malonate, lactate, citrate and trans‐aconitate, however, were invariably present in large proportions of total carboxylate exudation. Non‐cluster roots of H. prostrata exuded a spectrum of carboxylates (mainly malonate, lactate and citrate), which differed somewhat from the exudation pattern of cluster roots (mainly malate, malonate, lactate and citrate). The rate of exudation for cluster roots of the seven species was approximately 1·6 nmol g−1 FM s−1, which is considerably higher than that reported for a variety of crop and native species that do or do not form cluster roots. Contrary to what occurs in the cluster roots of Lupinus albus, which release carboxylates accompanied by protons so that the rhizosphere is acidified, the present Proteaceae exude the carboxylates as anions without concomitant proton release. The role of carboxylates in the mobilization of phosphate and other nutrients from soil is discussed.

Capillary electrophoresis for the determination of major amino acids and sugars in foliage: application to the nitrogen nutrition of Sclerophyllous species.

Amino acids and sugars are probably the most commonly measured solutes in plant fluids and tissue extracts. Chromatographic techniques used for the measurement of such solutes require complex derivatization procedures, analysis times are long and separate analyses are required for sugars and amino acids. Two methods were developed for the analysis of underivatized sugars and amino acids by capillary electrophoresis (CE). Separation of a range of sugars and amino acids was achieved in under 30 min, with good reproducibility and linearity. In general, there was close agreement between amino acid analyses by CE and HPLC with post-column derivatization. An alternative, more rapid method was optimized for the common neutral sugars. Separation of a mixture of fructose, glucose, sucrose, and fucose (internal standard) was achieved in less than 5 min. How the source of N applied (nitrate or ammonium) and its concentration (8.0 or 0.5 mM) affects the amino acid and sugar composition of leaves from Banksia grandis Willd. and Hakea prostrata R. Br. was investigated. The amino acid pool of Banksia and Hakea were dominated by seven amino acids (aspartic acid, glutamic acid, asparagine, glutamine, serine, proline, and arginine). Of these, asparagaine and glutamine dominated at low N-supply, whereas at high N-supply the concentration of arginine increased and dominated amino-N. Plants grown with nitrate had a greater concentration of proline relative to plants with ammonium. In Banksia the concentration of amides was greatest and arginine least with a nitrate N-source, whereas in Hakea amides were least and arginine greatest with nitrate N-source. The concentration of sugars was greater in Banksia than Hakea and in both species at greater N-supply.

Is photosynthesis related to concentrations of nitrogen and Rubisco in leaves of Australian native plants?

The relationships among light-saturated photosynthesis and concentrations of nitrogen and ribulose-1,5- bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) in Australian native plants are poorly known, primarily due to the difficulty of extracting and analysing Rubisco from such species. Rubisco may be rapidly quantified in crude extracts of plant tissue by capillary electrophoresis (CE); however, the presence of phenolic compounds in many Australian native plants limits the use of these methods. The addition of insoluble polyvinylpolypyrrolidone (PVPP) during leaf extractions effectively removed phenols permitting quantitation of Rubisco. Relationships among maximum rates of photosynthesis and concentrations of nitrogen and Rubisco were then investigated in ten species native to Australia. Total nitrogen and the major pools of N in foliage varied greatly between species. Equally, within species N-partitioning was highly plastic, as affected by different concentrations and forms of N applied in sand culture (0.5 or 8 mM, nitrate or ammonium). In Hakea prostrata, for example, the proportion of total N present as soluble proteins varied between 43 and 71%, while the proportion of total N present as Rubisco N ranged between 9.4 and 30.0%, and the contribution of Rubisco to soluble proteins varied between 21 and 42%. The measured concentration of Rubisco varied between 40% and 600% of that estimated from enzyme kinetics and measured rates of photosynthesis. Generally there was a large ‘excess’ of Rubisco, and in only two cases was the measured concentration of Rubisco significantly less than predicted. Total N, soluble protein and Rubisco concentrations were poorly related to maximum rates of photosynthesis, while the relationship between photosynthesis and Rubisco was worse than that with N, primarily due to the plants’ variable over-investment in Rubisco.

Factors affecting the distribution of proteoid roots within the root systems of two Hakea species

The proteoid roots of Hakea prostrata and H. laurina are concentrated in the surface soil horizons, even though the root systems penetrate to much greater depths. The relationships of a number of soil and plant factors to proteoid root occurrence in a given portion of the root system were examined. Pockets of humus-rich soil in any part of the root system greatly increased the proteoid root concentration in that region. The following factors, listed in their apparent order of importance, were analysed: local concentration of parent roots, local level of soil organic matter, local nitrogen availability, shoot growth, nitrogen concentration of the shoots, vertical distance of the region from the soil surface, local availability of calcium, magnesium, and potassium, local bulk density and certain other physical factors, nutrient status of the rest of the root system, horizontal distance of the region from the centre of the plant, relative maturity of parent roots in the region, and local soil pH and certain other chemical factors. The nitrogen component of soil regions high in organic matter largely accounted for their higher non-proteoid root concentration, smaller proteoid root size, greater number of laterals, and longer roots per unit weight, but not their much greater number of proteoid roots per unit total root weight. This suggests that other factors are also involved in proteoid root formation.