Michael D. Cramer

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.

The influence of NO3- and NH4+ nutrition on the carbon and nitrogen partitioning characteristics of wheat (Triticum aestivum L.) and maize (Zea mays L.) plants

The carbon and nitrogen partitioning characteristics of wheat (Triticum aestivum L.) and maize (Zea mays L.) grown hydroponically at a constant pH on either 4 mM or 12 mM NO3- or NH4+ nutrition were investigated using either 14C or 15N techniques. Greater allocation of 14C to amino-N fractions occurred at the expense of allocation of 14C to carbohydrate fractions in NH4+-compared to NO3--fed plants. The [14C]carbohydrate:[14C]amino-N ratios were 1.5-fold and 2.0-fold greater in shoots and roots respectively of 12 mM NO3--compared to 12 mM NH4+-fed wheat. In both 4 mM and 12 mM N-fed maize the [14C]carbohydrate:[14C]amino-N ratios were approximately 1.7-fold and 2.0-fold greater in shoots and roots respectively of NO3--compared to NH4+-fed plants. Similar results were observed in roots of wheat and maize grown in split-root culture with one root-half in NO3--and the other in NH4+-containing nutrient media. Thus the allocation of carbon to the amino-N fractions occurred at the expense of carbohydrate fractions, particularly within the root. Allocation of 14N and 15N within separate sets of plants confirmed that NH4--fed plants accumulated more amino-N compounds than NO3--fed plants. Wheat roots supplied with 15NH4+ for 8 h were found to accumulate 15NH4+ (8.5 μg 15N g-1 h-1) whereas in maize roots very little 15NH4+ accumulated (1.5 μg 15N g-1 h-1)It is proposed that the observed accumulation of 15NH4+ in wheat roots in these experiments is the result of limited availability of carbon within the roots of the wheat plants for the detoxification of NH4+, in contrast to the situation in maize. Higher photosynthetic capacity and lower shoot: root ratios of the C4 maize plants ensure greater carbon availability to the root than in the C3 wheat plants. These differences in carbon and nitrogen partitioning between NO3--and NH4+-fed wheat and maize could be responsible for different responses of wheat and maize root growth to NO3- and NH4+ nutrition.

photosynthesis and sink activity of wasp-induced galls in acacia pycnantha

Although insect galls are widely known to influence source-sink relationships in plants, the relationship between photosynthesis and gall activity has not been extensively studied. In this study we used 14CO2, photosynthesis, and respiration measurements to examine the capacity of bud galls induced by the wasp Trichilogaster signiventris (Pteromalidae) as carbon sinks in Acacia pycnantha. Galls of this species develop either in vegetative or reproductive buds, depending on the availability of tissues at different times of the year, and effectively eliminate seed production by the plant. Photosynthetic rates in phyllodes subtending clusters of galls were greater than rates in control phyllodes, a result we attributed to photosynthesis compensating for increased carbon demand by the galls. Contrary to previous studies, we found that photosynthesis within galls contributed substantially to the carbon budgets of the galls, particularly in large, mature galls, which exhibited lower specific respiration rates allowing for a net carbon gain in the light. To determine the sink capacity and competitive potential of galls, we measured the proportion of specific radioactivity in galls originating from either vegetative or reproductive buds and found no difference between them. The proportion of the total amount of phyllode-derived 14C accumulated in both clustered and solitary galls was less than that in fruits. Galls and fruits were predominantly reliant on subtending rather than on distant phyllodes for photosynthate. Solitary galls that developed in vegetative buds constituted considerably stronger sinks than galls in clusters on inflorescences where there was competition between galls or fruits for resources from the subtending phyllode. Wasps developing in solitary vegetative galls were correspondingly significantly larger than those from clustered galls. We conclude that, in the absence of inflorescence buds during summer and fall, the ability of the wasps to cause gall formation in vegetative tissues tempers intraspecific competition and substantially increases the availability of plant resources for the development of wasps in such galls.

Nitrogen regulation of transpiration controls mass-flow acquisition of nutrients

To test whether N regulates transpiration, Phaseolus vulgaris was grown with N placed at one of six distances behind a root-impenetrable mesh whilst control plants intercepted the N-source. N-availability regulated transpiration-driven mass-flow of nutrients from soil zones that were inaccessible

Convergence of soil nitrogen isotopes across global climate gradients

Quantifying global patterns of terrestrial nitrogen (N) cycling is central to predicting future patterns of primary productivity, carbon sequestration, nutrient fluxes to aquatic systems, and climate forcing. With limited direct measures of soil N cycling at the global scale, syntheses of the 15N:14N ratio of soil organic matter across climate gradients provide key insights into understanding global patterns of N cycling. In synthesizing data from over 6000 soil samples, we show strong global relationships among soil N isotopes, mean annual temperature (MAT), mean annual precipitation (MAP), and the concentrations of organic carbon and clay in soil. In both hot ecosystems and dry ecosystems, soil organic matter was more enriched in 15N than in corresponding cold ecosystems or wet ecosystems. Below a MAT of 9.8°C, soil δ15N was invariant with MAT. At the global scale, soil organic C concentrations also declined with increasing MAT and decreasing MAP. After standardizing for variation among mineral soils in soil C and clay concentrations, soil δ15N showed no consistent trends across global climate and latitudinal gradients. Our analyses could place new constraints on interpretations of patterns of ecosystem N cycling and global budgets of gaseous N loss.

Phosphate as a limiting resource: introduction

Phosphorus (P) is commonly a limiting nutrient for both terrestrial and aquatic productivity, with the consequence that it is considered important in determining the biodiversity and biomass of natural ecosystems. P-limitation of terrestrial plants is not a recent development. Karandashov and Bucher (2005) argued that the evolutionary transition of plants from the aquatic to terrestrial habitats was contingent on the presence of arbuscular mycorrhiza, which facilitated P acquisition. As a consequence of persistent P-limitations, terrestrial plants have evolved a wide range of P acquisition strategies, including mechanisms that increase the range of chemical forms that can be accessed, the range of concentrations that can be taken up and the effective absorbing area of the roots (Lambers et al. 2008). Likewise, plants have a range of strategies for conserving P that enable them to persist in P-deficient habitats, including sclerophyllous leaves and serotinous cones. In order to ensure successful recruitment in P-limited environments they also produce P-rich seeds (e.g. Groom and Lamont 2010, this volume). P is also often a limiting nutrient in agriculture (e.g. Sanchez 2010), deficiency being redressed through applications of P-fertilisers (globally ca. 20×10 kg P annum; Smit et al. 2009). Alarmingly, the finite global stocks of P (2,400×10 kg P) are likely to be depleted within 125 years (Smit et al. 2009; Vaccari 2009). Anthropogenic modification of the global P cycle by fertiliser use as well as waste streams and detergent use have effectively doubled global P cycling since the mid-19th century (Filippelli 2002). A consequence is that many natural ecosystems are threatened by super-abundance of a formerly limiting resource, with resulting biodiversity losses (e.g. Tilman et al. 2001). This is likely to be especially true in systems where P was formerly most-limiting, such as in Mediterranean terrestrial ecosystems (Sala et al. 2000), oligotrophic lakes (Schindler et al. 2008) and nutrient-impoverished oceans (Rabalais et al. 2008). Apart from the direct biodiversity consequences of P-eutrophication, release from P-limitation combined with increased atmospheric CO2 concentration and N deposition may exacerbate the loss of biodiversity.

Biological Nitrogen Fixation is not a Major Contributor to the Nitrogen Demand of a Commercially Grown South African Sugarcane Cultivar

It has previously been reported that endophytic diazotrophic bacteria contribute significantly to the nitrogen budgets of some graminaceous species. In this study the contribution of biological nitrogen fixation to the N-budget of a South African sugarcane cultivar was evaluated using 15N natural abundance, acetylene reduction and 15N incorporation. Plants were also screened for the presence of endophytic diazotrophic bacteria using acetylene reduction and nifH-gene targeted PCR with the pure bacterial strains. 15N natural abundance studies on field-grown sugarcane indicated that the plants did not rely extensively on biological nitrogen fixation. Furthermore, no evidence was found for significant N2-fixation or nitrogenase activity in field-grown or glasshouse-grown plants using 15N incorporation measurements and acetylene reduction assays. Seven endophytic bacterial strains were isolated from glasshouse-grown and field-grown plants and cultured on N-free medium. The diazotrophic character of these seven strains could not be confirmed using acetylene reduction and PCR screening for nifH. Thus, although biological nitrogen fixation may occur in South African sugarcane varieties, the contribution of this N-source in the tested cultivar was not significant.

Are Namibian “Fairy Circles” the Consequence of Self-Organizing Spatial Vegetation Patterning?

Causes of over-dispersed barren “fairy circles” that are often surrounded by ca. 0.5 m tall peripheral grasses in a matrix of shorter (ca. 0.2 m tall) grasses in Namibian grasslands remain mysterious. It was hypothesized that the fairy circles are the consequence of self-organizing spatial vegetation patterning arising from resource competition and facilitation. We examined the edaphic properties of fairy circles and variation in fairy circle size, density and landscape occupancy (% land surface) with edaphic properties and water availability at a local scale (<50 km) and with climate and vegetation characteristics at a regional scale. Soil moisture in the barren fairy circles declines from the center towards the periphery and is inversely correlated with soil organic carbon, possibly indicating that the peripheral grass roots access soil moisture that persists into the dry season within fairy circles. Fairy circle landscape occupancy is negatively correlated with precipitation and soil [N], consistent with fairy circles being the product of resource-competition. Regional fairy circle presence/absence is highly predictable using an empirical model that includes narrow ranges of vegetation biomass, precipitation and temperature seasonality as predictor variables, indicating that fairy circles are likely a climate-dependent emergent phenomenon. This dependence of fairy circle occurrence on climate explains why fairy circles in some locations may appear and disappear over time. Fairy circles are only over-dispersed at high landscape occupancies, indicating that inter-circle competition may determine their spacing. We conclude that fairy circles are likely to be an emergent arid-grassland phenomenon that forms as a consequence of peripheral grass resource-competition and that the consequent barren circle may provide a resource-reservoir essential for the survival of the larger peripheral grasses and provides a habitat for fossicking fauna.

Defoliation depletes the carbohydrate reserves of resprouting Acacia saplings in an African savanna

Over the past century there has been a global trend towards tree expansion and densification in rangelands and savannas. This phenomenon is commonly referred to as bush encroachment. In South Africa Acacia karroo is one of the key species responsible for bush encroachment. It has been suggested that the combination of fire and browsing might limit bush encroachment by A. karroo more effectively than either browsing or fire alone. We hypothesized that these repeated disturbances progressively deplete root carbohydrates and compromise resprouting ability. This was tested by burning and then manually defoliating A. karroo once a month for 1 year. Manual defoliation did not inhibit the rapid shoot elongation after topkill of A. karroo saplings. During this initial phase, the growth of the new shoots of A. karroo was dependent more on mobilised root reserves than on photoassimilates from the new shoots. Frequent manual defoliation of resprouting A. karroo saplings prevented the replenishment of starch reserves. We suggest a mechanism for how the interaction of browsing and fire can suppress and perhaps reverse bush encroachment in African savannas. Saplings that have reduced starch reserves at the end of dry season due to browsing will struggle to resprout if they are burnt. Even if they do not die, they will be less able to escape fire damage in the next fire than if they had been able to resprout unimpeded.

How succulent leaves of Aizoaceae avoid mesophyll conductance limitations of photosynthesis and survive drought

In several taxa, increasing leaf succulence has been associated with decreasing mesophyll conductance (g M) and an increasing dependence on Crassulacean acid metabolism (CAM). However, in succulent Aizoaceae, the photosynthetic tissues are adjacent to the leaf surfaces with an internal achlorophyllous hydrenchyma. It was hypothesized that this arrangement increases g M, obviating a strong dependence on CAM, while the hydrenchyma stores water and nutrients, both of which would only be sporadically available in highly episodic environments. These predictions were tested with species from the Aizoaceae with a 5-fold variation in leaf succulence. It was shown that g M values, derived from the response of photosynthesis to intercellular CO2 concentration (A:C i), were independent of succulence, and that foliar photosynthate δ13C values were typical of C3, but not CAM photosynthesis. Under water stress, the degree of leaf succulence was positively correlated with an increasing ability to buffer photosynthetic capacity over several hours and to maintain light reaction integrity over several days. This was associated with decreased rates of water loss, rather than tolerance of lower leaf water contents. Additionally, the hydrenchyma contained ~26% of the leaf nitrogen content, possibly providing a nutrient reservoir. Thus the intermittent use of C3 photosynthesis interspersed with periods of no positive carbon assimilation is an alternative strategy to CAM for succulent taxa (Crassulaceae and Aizoaceae) which occur sympatrically in the Cape Floristic Region of South Africa.