Quantifying the influence of water deficit on root and shoot growth in wheat using X-ray Computed Tomography.

The potential increased frequency and severity of drought associated with environmental change represents a significant obstacle to efforts aimed at enhancing food security due to its impact on crop development, and ultimately, yield. Our understanding of the impact of drought on crop growth in terms of plant aerial tissues is much more advanced than knowledge of the below-ground impacts. We undertook an experiment using X-ray Computed Tomography that aimed to support measurements of infrared gas exchange from plant shoots with quantification of 3D root architecture traits and the associated soil structural characteristics. Winter wheat (cv. Zebedee) was assessed at two early growth stages (14 and 21 days) under four water treatments (100, 75, 50 and 25 % of a notional field capacity (FC) and across two soil types (sandy loam and clay loam)). Plants generally grew better (to a larger size) in sandy loam soil as opposed to clay loam soil, most likely due to the soil structure and the associated pore network. All plants grew poorly under extreme water stress and displayed optimal growth at 75 % of FC, as opposed to 100 %, as the latter was most likely too wet. The optimal matric potential for root and shoot growth, inferred from the water release curve for each soil type, was higher than that for photosynthesis, stomatal conductance and transpiration suggesting root and shoot growth was more affected by soil water content than photosynthesis-related characteristics under water deficit conditions. With incidences of drought likely to increase, identification of wheat cultivars that are more tolerant of these conditions is important. Studies that consider the impact of water stress on both plant shoots and roots, and the role of the soil pore system such as this offer considerable potential in supporting these efforts.

A comparison between water uptake and root length density in winter wheat: effects of root density and rhizosphere properties.

BACKGROUND AND AIMS: We aim to quantify the variation in root distribution in a set of 35 experimental wheat lines. We also compared the effect of variation in hydraulic properties of the rhizosphere on water uptake by roots. METHODS: We measured the root length density and soil drying in 35 wheat lines in a field experiment. A 3D numerical model was used to predict soil drying profiles with the different root length distributions and compared with measured soil drying. The model was used to test different scenarios of the hydraulic properties of the rhizosphere. RESULTS: We showed that wheat lines with no detectable differences in root length density can induce soil drying profiles with statistically significant differences. Our data confirmed that a root length density of at least 1 cm/cm3 is needed to drain all the available water in soil. In surface layers where the root length density was far greater than 1 cm/cm3 water uptake was independent of rooting density due to competition for water. However, in deeper layers where root length density was less than 1 cm/cm3, water uptake by roots was proportional to root density. CONCLUSION: In a set of wheat lines with no detectable differences in the root length density we found significant differences in water uptake. This may be because small differences in root density at depth can result in larger differences in water uptake or that the hydraulic properties of the rhizosphere can greatly affect water uptake.

Brachiaria species influence nitrate transport in soil by modifying soil structure with their root system.

Leaching of nitrate from fertilisers diminishes nitrogen use efficiency (the portion of nitrogen used by a plant) and is a major source of agricultural pollution. To improve nitrogen capture, grasses such as brachiaria are increasingly used, especially in South America and Africa, as a cover crop, either via intercropping or in rotation. However, the complex interactions between soil structure, nitrogen and the root systems of maize and different species of forage grasses remain poorly understood. This study explored how soil structure modification by the roots of maize (Zea maize), palisade grass (Brachiaria brizantha cv. Marandu) and ruzigrass (Brachiaria ruziziensis) affected nitrate leaching and retention, measured via chemical breakthrough curves. All plants were found to increase the rate of nitrate transport suggesting root systems increase the tendency for preferential flow. The greater density of fine roots produced by palisade grass, subtly decreased nitrate leaching potential through increased complexity of the soil pore network assessed with X-ray Computed Tomography. A dominance of larger roots in ruzigrass and maize increased nitrate loss through enhanced solute flow bypassing the soil matrix. These results suggest palisade grass could be a more efficient nitrate catch crop than ruzigrass (the most extensively used currently in countries such as Brazil) due to retardation in solute flow associated with the fine root system and the complex pore network.

Multiple abiotic stress, nitrate availability and the growth of wheat.

In the field, wheat experiences a combination of physical and nutrient stresses. There has been a tendency to study root impedance and water stress in separation and less is known about how they might interact. In this study, we investigated the effect of root impedance on the growth of three wheat varieties (Cadenza, Xi19 and Battalion) at different levels of nitrate availability, from 0-20 mM nitrate, in sand culture. This model system allows soil strength to be increased while maintaining adequate water availability. In a separate pot experiment, we grew the same wheat varieties in a loamy sand where soil was allowed to dry sufficiently to both reduce water potential and increase root impedance. This pot experiment also had a range of nitrate availabilities 0-20 mM nitrate. Once the seedlings were established we limited water supply to apply a matric potential of approximately -200 kPa to the roots. Soil drying increased the penetrometer resistance from approximately 300 kPa to more than 1 MPa. There were differences between the two experimental systems; growth was smaller in the soil-based experiment compared to the sand culture. However, the effects of the experimental treatment, root impedance or water withholding, relative to the control were comparable. Our data confirmed that leaf elongation in Cadenza (carrying the tall Rht allele) was the most sensitive to root impedance. Leaf stunting occurred irrespective of nitrate availability. Leaf elongation in the Xi19 and Battalion (carrying the semi-dwarf Rht allele) was less sensitive to root impedance and drought than Candenza. We suggest that the critical stress in a pot experiment where the soil was allowed to dry to approximately -200 kPa was root impedance and not water availability.

Assessing the long-term effects of zero-tillage on the macroporosity of Brazilian soils using X-ray Computed Tomography.

Zero-tillage (ZT) is being increasingly adopted globally as a conservationist management system due to the environmental and agronomic benefits it provides. However, there remains little information on the tillage effect on soil pore characteristics such as shape, size and distribution, which in turn affect soil physical, chemical and biological processes. X-ray micro Computed Tomography (µCT) facilitates a non-destructive method to assess soil structural properties in three-dimensions. We used X-ray µCT at a resolution of 70 µm to assess and calculate the shape, size and connectivity of the pore network in undisturbed soil samples collected from a long-term experiment (~30 years) under zero tillage (ZT) and conventional tillage (CT) systems in Botucatu, Southeastern Brazil. In both systems, a single, large pore (>1000 mm3) typically contributed to a large proportion of macroporosity, 91% in CT and 97% in ZT. Macroporosity was higher in ZT (19.7%) compared to CT (14.3%). However the average number of pores was almost twice in CT than ZT. The largest contribution in both treatments was from very complex shaped pores, followed by triaxial and acircular shaped. Pore connectivity analysis indicated that the soil under ZT was more connected that the soil under CT. Soil under CT had larger values of tortuosity than ZT in line with the connectivity results. The results from this study indicate that long-term adoption of ZT leads to higher macroporosity and connectivity of pores which is likely to have positive implications for nutrient cycling, root growth, soil gas fluxes and water dynamics.

The emergent rhizosphere: imaging the development of the porous architecture at the root-soil interface.

The rhizosphere is the zone of soil influenced by a plant root and is critical for plant health and nutrient acquisition. All below ground resources must pass through this dynamic zone prior to their capture by plant roots. However, researching the undisturbed rhizosphere has proved very challenging. Here we compare the temporal changes to the intact rhizosphere pore structure during the emergence of a developing root system in different soils. High resolution X-ray Computed Tomography (CT) was used to quantify the impact of root development on soil structural change, at scales relevant to individual micro-pores and aggregates (µm). A comparison of micro-scale structural evolution in homogenously packed soils highlighted the impacts of a penetrating root system in changing the surrounding porous architecture and morphology. Results indicate the structural zone of influence of a root can be more localised than previously reported (µm scale rather than mm scale). With time, growing roots significantly alter the soil physical environment in their immediate vicinity through reducing root-soil contact and crucially increasing porosity at the root-soil interface and not the converse as has often been postulated. This 'rhizosphere pore structure' and its impact on associated dynamics are discussed.

Region-specific associations between sex, social status, and oxytocin receptor density in the brains of eusocial rodents.

Naturally occurring variations in neuropeptide receptor distributions in the brain contribute to numerous mammalian social behaviors. In naked mole-rats, which live in large social groups and exhibit remarkable reproductive skew, colony-related social behaviors vary with reproductive status. Here we examined whether variation in social status is associated with variations in the location and/or density of oxytocin binding in this species. Autoradiography was performed to assess forebrain oxytocin receptor (OTR) densities in breeding and non-breeding naked mole-rats of both sexes. Overall, males exhibited higher OTR binding in the medial amygdala in comparison to females. While there were no main effects of reproductive status in any region, a sex difference in OTR binding in the nucleus accumbens was mediated by status. Specifically, breeding males tended to have more OTR binding than breeding females in the nucleus accumbens, while no sex difference was observed in subordinates. These effects suggest that oxytocin may act in a sex- and region-specific way that corresponds to reproductive status and associated social behaviors.

Quantification of eggshell microstructure using X-ray micro computed tomography.

1. X-ray microcomputed tomography can be used to produce rapid, fully analysable, three-dimensional images of biological and other materials without the need for complex or tedious sample preparation and sectioning. We describe the use of this technique to visualise and analyse the microstructure of fragments of shell taken from three regions of chicken eggs (sharp pole, blunt pole and equatorial region). 2. Two- and three-dimensional images and data were obtained at a resolution of 1.5 microns. The images were analysed to provide measurements of shell thickness, the spacial density of mammillary bodies, the frequency, shape, volume and effective diameter of individual pore spaces, and the intrinsic sponginess (proportion of non-X-ray dense material formed by vesicles) of the shell matrix. Measurements of these parameters were comparable with those derived by traditional methods and reported in the literature. 3. The advantages of using this technology for the quantification of eggshell microstructural parameters and its potential application for commercial, research and other purposes are discussed.

Social regulation of adult neurogenesis in a eusocial mammal.

The present study examined the effects of social status on adult neurogenesis in an extreme cooperative breeder: the naked mole rat. These animals live in large colonies of up to 300 individuals, with a strict reproductive dominance hierarchy; one female and one to three males breed, and all other members are socially subordinate and reproductively suppressed. We examined the effects of social and gonadal cues on doublecortin (DCX; a marker for immature neurons) immunoreactivity in the dentate gyrus (DG), piriform cortex (PCx) and basolateral amygdala (BLA) by comparing dominant breeding animals to non-breeding subordinates from intact colonies. We also examined DCX expression in subordinate animals that had been removed from their colony and paired with an opposite- or same-sex conspecific for 6months. Compared to subordinates, dominant breeders had significantly reduced DCX immunoreactivity in all brain areas, with BLA effects confined to females. By contrast, the effects of same- versus opposite-sex housing were region-specific. In the DG and PCx, more DCX immunoreactivity was observed for opposite- than same-sex-paired subordinates. Conversely, same-sex-paired females had more DCX immunoreactivity than opposite-sex-paired females in the BLA. Gonadectomy did not affect DCX expression in opposite-sex-paired animals, and no significant relationships between gonadal steroids and DCX immunoreactivity were detected, suggesting that group differences in neurogenesis are independent of gonadal hormones. The apparent lower neurogenic capacity displayed by breeders contrasts previous reports on neurogenesis and social rank, challenging the conventional view that subordination is stressful and impairs neurogenesis. Future work will clarify whether the present findings can be attributed to status-dependent differences in stress, behavioral plasticity, or life stage.

Social condition and oxytocin neuron number in the hypothalamus of naked mole-rats (Heterocephalus glaber).

The naked mole-rat is a subterranean colonial rodent. In each colony, which can grow to as many as 300 individuals, there is only one female and 1-3 males that are reproductive and socially dominant. The remaining animals are reproductively suppressed subordinates that contribute to colony survival through their cooperative behaviors. Oxytocin is a peptide hormone that has shown relatively widespread effects on prosocial behaviors in other species. We examined whether social status affects the number of oxytocin-immunoreactive neurons in the paraventricular nucleus and the supraoptic nucleus by comparing dominant breeding animals to subordinate non-breeding workers from intact colonies. We also examined these regions in subordinate animals that had been removed from their colony and paired with an opposite- or same-sex conspecific for 6 months. Stereological analyses indicated that subordinates had significantly more oxytocin neurons in the paraventricular nucleus than breeders. Animals in both opposite- and same-sex pairs showed a decreased oxytocin neuron number compared to subordinates suggesting that status differences may be due to social condition rather than the reproductive activity of the animal per se. The effects of social status appear to be region specific as no group differences were found for oxytocin neuron number in the supraoptic nucleus. Given that subordinate naked mole-rats are kept reproductively suppressed through antagonism by the queen, we speculate that status differences are due either to oxytocin's anxiolytic properties to combat the stress of this antagonism or to its ability to promote the prosocial behaviors of subordinates.

Assessing the effectiveness of actions to mitigate nutrient loss from agriculture: a review of methods.

Diffuse nutrient loss from agriculture is degrading surface and groundwater quality throughout Europe, leaving water bodies at risk of not reaching targets set by the Water Framework Directive (WFD). Mitigation methods to reduce diffuse agricultural nutrient loss need to be implemented where water bodies have been identified as at risk of not reaching good status by 2015. Though the effectiveness of individual mitigation methods has usually been assessed in controlled experiments, it is necessary to quantify impact under a wider range of environmental and agricultural conditions and at the catchment scale to ensure that action taken now will be sufficient to meet WFD targets. Due to catchment buffering and long transit times (>50 years), it is unlikely that responses to intervention will be observed by 2015 in many water bodies. In this review, we compare the attributes and usefulness of different approaches (direct measurement, nutrient budgeting, risk assessment and modelling) to assess the efficacy of actions to mitigate sources and transport of nitrogen (N) and phosphorus (P) from agricultural land to water. Modelling and 'measured changes in farm activity' through budgeting avoid the time lags associated with direct measurement and enable rapid evaluations of different options before implementation. Budgeting approaches using on-farm data also provide a simpler, more communicable means of assessment but currently fail to consider the timing and transport aspects of mitigation and assume a direct causal relationship between potential and actual nutrient loss. Risk assessment and modelling applications are potentially more comprehensive and able to better reflect choice of mitigation at a range of scales, but assessments demand increased availability of data, and there is a large degree of uncertainty associated with their spatial and temporal dimensions which is difficult to validate adequately. The failings of individual approaches suggest that these assessment methods should be integrated to maximise their potential usefulness and positive attributes. This will enable nutrient inputs to be utilised most efficiently at broad scales and site specific actions to reduce nutrient transport and delivery can be targeted most cost-effectively at smaller scales. Such an integrated approach will also more effectively engage and involve the farmer in what must be an iterative process.

Micromorphological investigations into root penetration in a landfill mineral cap, Hertfordshire, UK.

A micromorphological study of the soil fabric of a landfill mineral cap in Hertfordshire, UK is described. The study was undertaken to examine the nature of the cap and roots within it, and whether micromorphology could help in the explanation for root penetration. The results from thin section description supported by macro and micro image analysis clearly demonstrated that rooting was associated with zones of weakness in the cap, due to the heterogeneous particle/pore size distribution, low bulk density and presence of organic matter. The study of the material fabric also suggested evidence of pedological activity within the cap, further indicating that the mineral cap had not been engineered sufficiently to prevent the risk of root penetration. A greater uniformity of particle size within the cap material with a reduced clay and stone fraction were identified as potential solutions to avoid the extent of root penetration. The study demonstrated the value of observing and quantifying the undisturbed fabric of the micro structure in the examination of mineral cap fabric, and diagenetic and pedological processes acting on it.

Quantification of pore clogging characteristics in potential permeable reactive barrier (PRB) substrates using image analysis.

Permeable reactive barriers (PRBs) are now an established approach for groundwater remediation. However, one concern is the deterioration of barrier material performance due to pore clogging. This study sought to quantify the effect of pore clogging on the alteration of the physical porous architecture of two novel potential PRB materials (clinoptilolite and calcified seaweed) using image analysis of SEM-derived images. Results after a water treatment contaminated with heavy metals over periods of up to 10 months identified a decrease in porosity from c. 22% to c. 15% for calcified seaweed and from c. 22% to c. 18% for clinoptilolite. Porosity was reduced by as much as 37% in a calcified seaweed column that clogged. The mean pore size (2D) of both materials slightly decreased after water treatment with c. 11% reduction in calcified seaweed and c. 7% reduction in clinoptilolite. An increase in the proportion of crack-shaped pores was observed in both materials after the contaminated water treatment, most noticeably in the bottom of columns where contaminated water first reacted with the material. The distribution of pores (within a given image) derived from the distance transform indicated the largest morphological differences in materials was recorded in calcified seaweed columns, which is likely to impact significantly on their performance as barrier materials. The magnitude of porosity reduction over a short time period in relation to predicted barrier longevity suggest these and similar materials may be unsuited for barrier installation in their present form.