Sodium as nutrient and toxicant

Background and Scope

Because of the crucial role coarse roots (>2 mm diameter) play in plant functions and terrestrial ecosystems, detecting and quantifying the size, architecture, and biomass of coarse roots are important. Traditional excavation methods are labor intensive and destructive, with limited quantification and repeatability of measurements over time. As a nondestructive geophysical tool for delineating buried features in shallow subsurface, ground penetrating radar (GPR) has been applied for coarse root detection since 1999. This article reviews the state-of-knowledge of coarse root detection and quantification using GPR, and discusses its potentials, constraints, possible solutions, and future outlooks. Some useful suggestions are provided that can guide future studies in this field.

Conclusions

The feasibility and accuracy of coarse root investigation by GPR have been tested in various site conditions (mostly in controlled conditions or within plantations) and for different plant species (mostly tree root systems). Thus far, single coarse root identification and coarse root system mapping have been conducted using GPR, including roots under pavements in urban environment. Coarse root diameter and biomass have been estimated from indexes extracted from root GPR radargrams. Coarse root development can be observed by repeated GPR scanning over time. Successful GPR-based coarse root investigation is site specific, and only under suitable conditions can reliable measurements be accomplished. The best quality of root detection by GPR is achieved in well-drained and electrically-resistive soils (such as sands) under dry conditions. Numerous factors such as local soil conditions, root electromagnetic properties, and GPR antenna frequency can impact the reliability and accuracy of GPR detection and quantification of coarse roots. As GPR design, data processing software, field data collection protocols, and root parameters estimation methods are continuously improved, this noninvasive technique could offer greater potential to study coarse roots.     

Impact of root water content on root biomass estimation using ground penetrating radar: evidence from forward simulations and field controlled experiments

Background and aims

The GPR indices used for predicting root biomass are measures of root radar reflectance. However, root radar reflectance is highly correlated with root water content. The objectives of this study are to assess the impact of root water content on GPR-based root biomass estimation and to develop more reliable approaches to quantify root biomass using GPR.

Methods

Four hundred nine roots of five plant species in a sandy area of northern China were examined to determine the general water content range of roots in sandy soils. Two sets of GPR simulation scenarios (including 492 synthesized radargrams in total) were then conducted to compare the changes of root radar signal and the accuracies of root biomass estimation by GPR at different root gravimetric water content levels. In the field, GPR transects were scanned for Ulmus pumila roots buried in sandy soils with three antenna center frequencies (0.5, 0.9, and 2.0 GHz). The performance of two new GPR-based root biomass quantification approaches (one using time interval GPR index and the other using a non-linear regression model) was then tested.

Results

All studied roots exhibited a broad range of gravimetric water content (>125 %), with the water contents of most roots ranging from 90 % to 150 %. Both field experiments and forward simulations indicated that 1) waveforms of root radar reflection, radar-reflectance related GPR indices, and root biomass estimation accuracy were all affected by root water content; and 2) using time interval index and establishing a nonlinear regression model of root biomass on GPR indices improved the accuracy of root biomass estimation, decreasing the prediction error (RMSE) by 4 to 30 % under field conditions.

Conclusions

The magnitude of GPR indices depends on both root biomass and root water content, and root water content affects root biomass estimation using GPR indices. Using a linear regression model of root biomass on radar-reflectance related GPR index for root biomass estimation would only be feasible for roots with a relative narrow range of water content (e.g., when gravimetric water contents of studied roots vary within 20 %). Appropriate GPR index and regression models should be selected based on the water content range of roots. The new protocol of root biomass quantification by GPR presented in this study improves the accuracy of root biomass estimation.     

Forward simulation of root’s ground penetrating radar signal: simulator development and validation

Background and aims

It remains unclear how the limiting factors (e.g., root size, root water content, spacing between roots, and soil water content) affect root investigation using ground penetrating radar (GPR). The objective of this study is to develop a theoretical forward simulation protocol of synthesizing root’s GPR signal and test the feasibility of our proposed simulation protocol in evaluating the impacts of limiting factors on GPR-based root detection and quantification.

Methods

The proposed forward simulation protocol was developed by integrating several existing numerical models, such as the Root Composition Model, the Root Dielectric Constant Model, the Root Electrical Conductivity Model, the Soil Dielectric Constant Model, the Soil Electrical Conductivity Model, and a newly-established model (Root Length-Biomass Model). Resolution and GPR index obtained from both field collected radargrams and corresponding simulations were compared to validate the accuracy of simulation.

Results

Simulated radargrams exhibit similar resolution with that of the in situ collected. The same trends of root radar signals against different levels of root size, root water content, interval between roots, root depth, and antenna frequency were observed on both in situ radargrams and simulated radargrams. Strong correlations (correlation coefficients ranging from 0.87 to 0.96) were found between GPR indices extracted from the simulated data and those from the field collected data.

Conclusions

Our proposed forward simulation is effective for assessing the impacts of limiting factors on root detection and quantification using GPR. This forward simulation protocol can be used to provide guidance for in situ GPR root investigation and can predict the accuracy of GPR-based root quantification under site-specific conditions.     

Leaf litter thickness,but not plant species,can affect root detection by ground penetrating radar

Aim

Ground penetrating radar (GPR), a nondestructive tool that can detect coarse tree roots, has not yet become a mature technology for use in forests. In this study, we asked two questions concerning this technology: (i) Does the leaf litter layer influence root detection and major indices based on the time interval between zero crossings (T) and the amplitude area (A)? (ii) Can GPR images discriminate roots of different plant species?

Methods

Roots buried in a sandy bed, which was covered with different thicknesses of leaf litter, were scanned using a 900 MHz GPR antenna. Roots of four plant species in the bed were also scanned.

Results

Leaf litter decreased root reflections without distorting the shape of the hyperbolas in the radar profile. A values decreased with increasing litter thickness, whereas T was independent of litter thickness. For all species combined, GPR indices were significantly correlated with root diameter.

Conclusions

Leaf litter dramatically decreased root detection, but the influence of the litter could be ignored when the sum of T for all reflection waveforms (ΣT) is adopted to estimate root diameter. To use A values to detect roots, litter should be removed or equalized in thickness. Radar profiles could not reliably differentiate among roots belonging to plants of different species.

     

Effect of species, root branching order and season on the root traits of 13 perennial grass species

Aims

Inter-specific comparisons of plant traits may vary depending on intra-specific variation. Here we examine the impact of root branching order and season on key functional root traits for grass species. We also compare root traits among co-existing grass species as a step towards defining root trait syndromes.

Methods

Monocultures of 13 grass species, grown under field conditions and subjected to intensive management, were used to record root trait values for coarse roots (1st order, >0.3?mm), fine roots (2nd and 3rd orders, <0.2?mm) and mixed root samples over three growing seasons.

Results

Branching order and species had a significant effect on root trait values, whereas season showed a marginal effect. The diameter of coarse roots was more variable than that of fine roots and, as expected, coarse roots had higher tissue density and lower specific root length values than fine roots. Principal component analysis run on eight root traits provided evidence for two trait syndromes related to resource acquisition and conservation strategies across grass species.

Conclusions

Our data show that root branching order is the main determinant of root trait variation among species. This highlights the necessity to include the proportion of fine vs coarse roots when measuring traits of mixed root samples.     

archiDART: an R package for the automated computation of plant root architectural traits

Background and aims

In order to analyse root system architectures (RSAs) from captured images, a variety of manual (e.g. Data Analysis of Root Tracings, DART), semi-automated and fully automated software packages have been developed. These tools offer complementary approaches to study RSAs and the use of the Root System Markup Language (RSML) to store RSA data makes the comparison of measurements obtained with different (semi-) automated root imaging platforms easier. The throughput of the data analysis process using exported RSA data, however, should benefit greatly from batch analysis in a generic data analysis environment (R software).

Methods

We developed an R package (archiDART) with five functions. It computes global RSA traits, root growth rates, root growth directions and trajectories, and lateral root distribution from DART-generated and/or RSML files. It also has specific plotting functions designed to visualise the dynamics of root system growth.

Results

The results demonstrated the ability of the package’s functions to compute relevant traits for three contrasted RSAs (Brachypodium distachyon [L.] P. Beauv., Hevea brasiliensis Müll. Arg. and Solanum lycopersicum L.).

Conclusions

This work extends the DART software package and other image analysis tools supporting the RSML format, enabling users to easily calculate a number of RSA traits in a generic data analysis environment.

     

Root orientation can affect detection accuracy of ground-penetrating radar

Aim

Ground-penetrating radar (GPR) has been applied to detect coarse tree roots. The horizontal angle of a root crossing a scanning line is a factor that affects both root detection and waveform parameter values. The purpose of this study was to quantitatively evaluate the influence of root orientation (x, degree) on two major waveform parameters, amplitude area (A, dB × ns) and time interval between zero crossings (T, ns).

Methods

We scanned four diameter classes of dowels in a sandy bed as simulated roots using a 900 MHz antenna from multiple angles to clarify the relationships between the parameters and x.

Results

Angle x strongly affected reflection images and A values. The variation in A(x) fitted a sinusoidal waveform, whereas T was independent of x. The value of A scanning at 90° was estimated by A values of arbitrary x in two orthogonal transects. The sum of T in all reflected waveforms showed a significant linear correlation with dowel diameter.

Conclusions

We clarified that root orientation dramatically affected root detection and A values. The sum of T of all reflected waveforms was a suitable parameter for estimating root diameter. Applying grid transects can overcome the effects of root orientation.     

Effects of litter types, microsite and root diameters on litter decomposition in Pinus sylvestris plantations of northern China

Background and aims

Litter decomposition is a major process in the carbon (C) flow and nutrient cycling of terrestrial ecosystems, but the effects of litter type, microsite, and root diameter on decomposition are poorly understood.

Methods

Litterbags were used to examine the decomposition rate of leaf litter and roots at three soil depths (5, 10 and 20 cm) over a 470-day period in Pinus sylvestris plantations in northern China.

Results

Leaves and the finest roots decomposed more quickly at 5 cm depth and coarser roots (>1-mm) decomposed more quickly at 10 and 20 cm depth. Roots generally decomposed more quickly than leaf litter, except at 5 cm deep; leaves decomposed more quickly than the coarsest roots (>5-mm). Root decomposition was strongly influenced by root diameter. Leaves experienced net nitrogen (N) immobilization and coarse roots (>2-mm) experienced more N release than fine roots. Significant heterogeneity was seen in N release for fine-roots (<2-mm) with N immobilization occurring in smaller (0.5–2-mm) roots and N release in the finest roots (<0.5-mm).

Conclusions

Soil depth of litter placement significantly influenced the relative contribution of the decomposition of leaves and roots of different diameters to carbon and nutrient cycling.     

Visualization of grapevine root colonization by the Saharan soil isolate Saccharothrix algeriensis NRRL B-24137 using DOPE-FISH microscopy

Background and aim

There is currently a gap of knowledge regarding whether some beneficial bacteria isolated from desert soils can colonize epi- and endophytically plants of temperate regions. In this study, the early steps of the colonization process of one of these bacteria, Saccharothrix algeriensis NRRL B-24137, was studied on grapevine roots to determine if this beneficial strain can colonize a non-natural host plant. An improved method of fluorescence in situ hybridization (FISH), the double labeling of oligonucleotide probes (DOPE)-FISH technique was used to visualize the colonization behavior of such bacteria as well as to determine if the method could be used to track microbes on and inside plants.

Methods

A probe specific to Saccharothrix spp. was firstly designed. Visualization of the colonization behavior of S. algeriensis NRRL B-24137 on and inside roots of grapevine plants was then carried out with DOPE-FISH microscopy.

Results

The results showed that 10 days after inoculation, the strain could colonize the root hair zone, root elongation zone, as well as root emergence sites by establishing different forms of bacterial structures as revealed by the DOPE-FISH technique. Further observations showed that the strain could be also endophytic inside the endorhiza of grapevine plants.

Conclusions

Taking into account the natural niches of this beneficial strain, this study exemplifies that, in spite of its isolation from desert soil, the strain can establish populations as well as subpopulations on and inside grapevine plants and that the DOPE-FISH tool can allow to detect it.     

Application of X-ray computed tomography to quantify fresh root decomposition in situ

Background and aims

Much of our understanding of plant root decomposition and related carbon cycling come from mass loss rates calculated from roots buried in litter bags. However, this may not reflect what actually happens in the soil, where the interactions between root and soil structure presents a more complex physico-chemical environment compared to organic matter isolated in a porous bag buried in disturbed soil. This work investigates the potential of using X-ray micro-computed tomography (CT) to measure root decomposition in situ.

Methods

Roots of Vicia faba L. were excised from freshly germinated seeds, buried in re-packed soil cores and cores incubated for 60 days. Changes in root volume and surface area were measured using repeated scans. Additional samples were destructively harvested and roots weighed to correlate root mass with root volume. The method was further applied to an experiment to investigate the effects of soil bulk density and soil moisture on root decomposition.

Results

Root volume (X-ray CT) and root mass (destructive harvest) decreased by 90 % over the 60 day incubation period, by which stage, root volume and mass had stabilised. There was a strong correlation (R ²?=?0.97) between root volume and root mass.

Conclusions

X-ray CT visualization and analysis provides a unique toolbox to understand root decomposition in situ.     

Root growth in biopores—evaluation with in situ endoscopy

Background and aims

The significance of biopores for nutrient acquisition from the subsoil depends on root-soil contact, which in turn is influenced by root architecture. The aim of this study was to detect differences regarding the architecture and root-soil contact of homorhizous barley roots (Hordeum vulgare L.) and allorhizous oilseed rape roots (Brassica napus L.) growing in biopores.

Methods

In situ endoscopy was used as a technique that allows non-destructive display of pore wall characteristics and root morphology inside large biopores under field conditions.

Results

For both crops, about 85 % of all roots did establish contact to the pore wall. However, according to their different root architecture, the two crops varied in their strategy of resource acquisition: While barley was characterized by thin vertical or ingrowing roots, most of them in direct contact to the pore wall, oilseed rape established contact to the pore wall predominantly via lateral roots.

Conclusions

Root morphological and pore wall assessment with in situ endoscopy in combination with detailed studies of soil biochemical and soil physical parameters of the pore wall is considered an essential prerequisite for more precise future modelling of nutrient acquisition and uptake.     

Spatial and temporal dynamics of hotspots of enzyme activity in soil as affected by living and dead roots—a soil zymography analysis

Aims

Hotspots of enzyme activity in soil strongly depend on carbon inputs such as rhizodeposits and root detritus. In this study, we compare the effect of living and dead Lupinus polyphyllus L. roots on the small-scale distribution of cellulase, chitinase and phosphatase activity in soil.

Methods

Soil zymography, a novel in situ method, was used to analyze extracellular cellulase, chitinase and phosphatase activity in the presence of i. living L. polyphyllus roots prior to shoot cutting and ii. dead/dying roots 10, 20 and 30 days after shoot cutting.

Results

After shoot cutting, cellulase and chitinase activities increased and were highest at the root tips. The areas of high cellulase and phosphatase activity extend up to 55 mm away from the root. Moreover, we observed microhotspots of cellulose, chitinase, and phosphatase activity up to 60 mm away from the next living root. The number and activity of microhotspots of chitinase activity was maximal 10 days after shoot cutting.

Conclusions

The study showed that young root detritus stimulates enzyme activities stronger than living roots. Soil zymography allowed identification of microhotspots of enzyme activity up to several cm away from living and dying roots, which most likely were caused by arbuscular mycorrhizal fungi.     

A comparison study of Agrobacterium-mediated transformation methods for root-specific promoter analysis in soybean

Key message

Both in vitro and in vivo hairy root transformation systems could not replace whole plant transformation for promoter analysis of root-specific and low-P induced genes in soybean.

Abstract

An efficient genetic transformation system is crucial for promoter analysis in plants. Agrobacterium-mediated transformation is the most popular method to produce transgenic hairy roots or plants. In the present study, first, we compared the two different Agrobacterium rhizogenes-mediated hairy root transformation methods using either constitutive CaMV35S or the promoters of root-preferential genes, GmEXPB2 and GmPAP21, in soybean, and found the efficiency of in vitro hairy root transformation was significantly higher than that of in vivo transformation. We compared Agrobacterium rhizogenes-mediated hairy root and Agrobacterium tumefaciens-mediated whole plant transformation systems. The results showed that low-phosphorous (P) inducible GmEXPB2 and GmPAP21 promoters could not induce the increased expression of the GUS reporter gene under low P stress in both in vivo and in vitro transgenic hairy roots. Conversely, GUS activity of GmPAP21 promoter was significantly higher at low P than high P in whole plant transformation. Therefore, both in vitro and in vivo hairy root transformation systems could not replace whole plant transformation for promoter analysis of root-specific and low-P induced genes in soybean.     

Radial force development during root growth measured by photoelasticity

Background and aims

The radial growth of roots largely affects and reorganizes the porous or crack networks of soils and substrates. We studied the consequences of a radial steric constriction on the root growth and the feedback force developed by the root on the solid phase.

Methods

We developed an original method of photoelasticity to measure in situ root forces. By changing the gap width (0.5 to 2.3?mm) between two photoelastic disks we applied variable radial constrictions to root growth and simultaneously measured the corresponding radial forces. Changes in morphology and forces of primary roots of chick pea (Cicer arietinum L.) seedlings were recorded by time-lapse imaging every 24?min up to 5?days.

Results

The probability of root entering the gap depended on the gap size but was also affected by circumnutation. Compared to non-constrained root controls, no significant morphological change (elongation, diameter) was measured outside the gap zone. Inside the gap zone, outer cortex cells were compressed, the central cylinder was unaffected. Radial forces were increasing with time but no force levelling was observed even after 5?days.

Conclusions

Radial constrictions applied to roots did not significantly reduce their growth. The radial force was related to the root strain in the gap.     

Spatio-temporal mapping of local soil pH changes induced by roots of lupin and soft-rush

Aims

The rhizosphere is a dynamic system strongly influenced by root activity. Roots modify the pH of their surrounding soil causing the soil pH to vary as a function of distance from root surface, location along root axes, and root maturity. Non-invasive imaging techniques provide the possibility to capture pH patterns around the roots as they develop.

Methods

We developed a novel fluorescence imaging set up and applied to the root system of two lupin (Lupinus albus L., Lupinus angustifolius L.) and one soft-rush (Juncus effusus L.) species. We grew plants in glass containers filled with soil and equipped with fluorescence sensor foils on the container side walls. We gained highly-resolved data on the spatial distribution of H⁺ around the roots by taking time-lapse images of the samples over the course of several days.

Results

We showed how the soil pH in the vicinity of roots developed over time to different values from that of the original bulk soil. The soil pH in the immediate vicinity of the root surface varied greatly along the root length, with the most acidic point being at 0.56–3.36 mm behind the root tip. Indications were also found for temporal soil pH changes due to root maturity.

Conclusion

In conclusion, this study shows that this novel optical fluorescence imaging set up is a powerful tool for studying pH developments around roots in situ.     

Comment on: “root orientation can affect detection accuracy of ground-penetrating radar”

Introduction

In a recent paper, Tanikawa et al. Plant Soil 373:317–327, (2013) reported a considerable impact of root orientation on the accuracy of root detection and root diameter estimation by ground-penetrating radar (GPR).

Methods

In Tanikawa et al. Plant Soil 373:317–327, (2013), buried root samples in a sand box were scanned from multiple cross angles between root orientation and GPR transecting line under controlled conditions. Changes in radar waveform parameter of roots to different cross angles were investigated.

Results

Tanikawa et al. Plant Soil 373:317–327, (2013) clarified that 1) the variation in amplitude area (a signal strength related waveform parameter) to different cross angles fitted a sinusoidal waveform; and 2) the impact of root orientation on root diameter estimation by GPR could be mathematically corrected by applying a grid transect survey. However, we found that the quantitative relationship established in Tanikawa et al. Plant Soil 373:317–327, (2013) between amplitude area and cross angle was incorrect, and the application of a grid transect survey still underestimated root diameter.

Conclusion

The change in amplitude area to cross angle between transecting line and root orientation fits a sinusoidal waveform but different to that reported in Tanikawa et al. Plant Soil 373:317–327, (2013). The polarization of GPR wave may explain such sinusoidal variation in amplitude area to cross angle. The effect of root orientation on GPR-based root diameter estimation remains to be calibrated.     

Biomechanics of nodal, seminal and lateral roots of barley: effects of diameter, waterlogging and mechanical impedance

Background and aims

Biomechanical properties of cereal root systems largely control both resistance to root lodging and their ability to stabilise soil. Abiotic stresses can greatly modify root system growth and form. In this paper the effect of waterlogging and moderate mechanical impedance on root biomechanics is studied for both lateral roots and the main axes of barley.

Methods

Barley (Hordeum vulgare) plants were subjected to transient water-logging and moderate mechanical impedance in repacked soil columns. Roots were excavated, separated into types (nodal, seminal or lateral) and tested in tension to measure strength and elastic modulus.

Results

Water-logging and mechanical impedance substantially changed root system growth whilst root biomechanical properties were affected by waterlogging. Root strength was generally greater in thin roots and depended on root type. For example, seminal roots 0.4–0.6 mm in diameter were approximately seven times stronger and five times stiffer than lateral roots of the same diameter when mechanically impeded. Root sample populations typically exhibited negative power-law relationships between root strength and diameter for all root types. Mechanical impedance slowed seminal root elongation by approximately 50 % and resulted in a 15 % and 11 % increase in the diameter of in nodal and seminal roots respectively. Power-law relationships between root diameter and root biomechanical properties corresponded to the different root types. Coefficients for between root diameter, strength and elastic modulus improved when separated by root type, with R² values increasing in some roots from 0.05 to 0.71 for root strength and 0.08 to 0.74 for elastic modulus.

Conclusions

Moderate mechanical impedance did not influence the tensile strength of roots, but, waterlogging diminished the relationship between root strength and diameter. Separation of root type improved predictions of root strength and elastic modulus using power-law regressions.     

Longitudinal variation in cadmium influx in intact first order lateral roots of sunflower (Helianthus annuus. L)

Aims

Contamination of sunflower (Helianthus annuus L.) by cadmium (Cd) is a concern for food and feed safety as this species accumulates Cd to a greater extent than other crops. We examined the relationships between root architecture and Cd²⁺ uptake by roots.

Methods

We determined and mathematically modelled the longitudinal variation of Cd²⁺ influx in first order roots of sunflower grown in hydroponics by using short-term exposure to ¹⁰⁹Cd-labelled solutions (0.8 to 500 nM). Thereafter, by taking into account the longitudinal variation of the influx, we simulated the uptake of Cd²⁺ for 24 h by cohorts of roots characterised by various architectural characteristics.

Results

Cd²⁺ influx at the root tip was on average 2.9 times that of the basal region close to the taproot. The simulations indicated that the total Cd²⁺ uptake by root cohorts mainly depends on 1/ the root diameter and the number of roots, 2/ the value of the Cd²⁺ influx at the basal region 3/ the stronger influx at the root tip.

Conclusion

Considering a higher Cd²⁺ influx at the root tip may be important to understand the relationship between root architecture and Cd²⁺ uptake by the root system.     

Potential and actual root growth variations in root systems: modeling them with a two-step stochastic approach

Background and aims

Root elongation is essential in the determination of the root system architecture (RSA). Using experimental data, we show how it varies in the RSA and suggest a new and simple modeling approach to predict these variations.

Methods

We analyzed variation in elongation on data from pot-grown plants belonging to two different species (Helianthus annuus L. and Noccaea caerulescens (J.Presl & C.Presl) F.K.Mey). A stochastic model was designed with two successive steps to quantify and simulate these variations. The first step is the definition of a growth potential, reflected by the apical diameter, and depending on the size of the mother root. The second step, during elongation, describes the dynamic evolution of the meristem and its interaction with soil constraints.

Results

The species exhibited differences in their structured variations and very large residual (pseudo-random) variability in elongation rate and final length. The two-step model allowed us to summarize these species characteristics, and to show the interest of considering the stochastic aspects of root growth to correctly simulate the RSA.

Conclusions

Apart from being a more realistic way of simulating root development, this type of model raises new questions regarding the representation of root soil interactions during elongation.