Effects of species and soil‐nitrogen availability on root system architecture traits – study on a set of weed and crop species

Better managing crop : weed competition in cropping systems while reducing both nitrogen and herbicide inputs is a real challenge that requires a better understanding of crop and weed root architecture in relation to soil‐nitrogen availability. An original approach was used which considered the parameters of a simulation model of root architecture as traits to analyse (a) the interspecific diversity of root system architecture, and (b) its response to soil‐nitrogen availability. Two greenhouse experiments were conducted using three crop and nine weed species grown at two contrasted concentrations of soil‐nitrogen availability. Plant traits were measured to characterise both overall plant growth and root architecture, with a focus on primary root emergence, root elongation and branching. The studied root traits varied among species (from a twofold to a fourfold factor, depending on the trait), validating their use as indicators to analyse the interspecific variability of root architecture. The largest interspecies differences were for two traits: ‘maximal apical root diameter’ and ‘interbranch distance’ (distance between two successive laterals on the same root). Conversely, most of the studied root traits varied little with soil‐nitrogen availability (from no variation to a 1.1‐fold factor, depending on the trait) even though soil‐nitrogen availability varied with a 17‐fold factor and impacted the overall shoot and root biomass. So, the root traits used in this article are stable whatever soil‐nitrogen availability. As they reflect processes underlying root system architecture, this low effect of nitrogen suggests that the rules governing root architecture are little affected by plant nitrogen status and soil‐nitrogen availability. We propose that the determinants of differences in root system architecture between soils with contrasted nitrogen availability mainly originate from differences in the amount of carbon allocated to and within the root system. Characterising each plant species by a combination of root traits gave insights regarding the potential species competitive ability for soil resources in agroecosystems.     

Root elongation rate is accounted for by intercepted PPFD and source-sink relations in field and laboratory-grown sunflower

The existence of relationships between intercepted photo-synthetic photon flux density (PPFD) and growth of individual organs is somewhat controversial. We have tested whether such relationships could account for the natural variability in elongation rates of taproot and secondary roots of sunflower (from 2 to 135 mm d⁻¹), in field and laboratory conditions. Elongation of taproot and secondary roots was recorded daily through windows in the field. A range of PPFD was obtained by following day-to-day natural fluctuation for three contrasting growing periods, and by shading part of the plants under study. A parallel experiment was carried out in a growth chamber with contrasting light intensities and with a ¹⁴CO₂ labelling experiment. After the two-leaf stage, i.e. when the contribution of photosynthetic carbon became appreciable in root growth, daily root elongation rate was closely linked to the PPFD intercepted from 36 to 12 h before the measurement of root elongation. Curvilinear relationships applied to plants grown in the field as well as in a growth chamber, and to shaded plants as well as to plants subjected to day-to-day changes in intercepted PPFD. For a given intercepted PPFD, the taproot elongated faster than secondary roots, and secondary roots originating near the base of the taproot elongated faster than those originating near the apex. The elongation rate of any secondary root apex was accounted for (r= 0.77) by the ratio of intercepted PPFD to the distance between the apex and the base of the taproot. No relationships between intercepted PPFD and elongation rate were observed before the two-leaf stage, when the CO₂ labelling experiment suggests that carbon essentially originates from the seed. Therefore, this study suggests a role for source-sink relations in the distribution of elongation between apices and a role for carbon nutrition in day-to-day variations of root elongation rate. Precise mechanisms explaining this behaviour remain to be investigated.     

Phosphate Availability Alters Lateral Root Anatomy and Root Architecture of Fraxinus mandshurica Rupr. Seedlings

Plants have evolved some mechanisms to maximize the efficiency of phosphorus acquisition.Changes in root architecture are one such mechanism. When Fraxinus mandshurica Rupr. seedlings were grown under conditions of low phosphorus availability, the length of cells in the meristem zone of the lateral roots was longer, but the length of cells in the elongation and mature zones of the lateral roots was shorter,compared with seedlings grown under conditions of high phosphorus availability. The elongation rates of primary roots increased as phosphorus availability increased, but the elongation rates of the branched zones of the primary roots decreased. The number of lateral root primordia and the length of the lateral roots decreased as phosphorus availability increased. The topological index (altitude slope) decreased as phosphorus availability increased, suggesting that root architecture tended to be herringbone-like when seedlings were grown under conditions of low phosphate availability. Herringbone-like root systems exploit nutrients more efficiently, but they have higher construction costs than root systems with a branching pattern.     

Why are laterals less affected than main axes by homogeneous unfavourable physical conditions? A model-based hypothesis

When plants develop in strong soils, growth of the root system is generally depressed. However, branching and elongation of branches are often less affected than growth of the main axes, whenever the whole root system encounters even-impeded conditions. On the basis of a model simulating root growth and architecture as related to assimilate availability, we propose a simple hypothesis to explain such behaviour. In the model, growth of each root depends on its own elongation potential, which is estimated by its apical diameter. The potential elongation rate–apical diameter relationship is the same for all the roots of the system and is described by a monomolecular function. Our hypothesis is that the effect of soil strength can be simulated by introducing an impedance factor in the definition of root maximum potential elongation rate, common to the whole root system. When such impedance factor is applied, it affects more the potential of larger roots (main axes) than that of thinner roots (secondary and tertiary branches). Simulations provided in high impedance conditions led to root systems characterised by short taproots, whereas growth of secondary roots was unaffected and growth of tertiary roots was enhanced. Actual branching density was also higher, although branching rules have been unchanged. Such simulated systems where similar to that observed in strong soils. Friction laws or pore size can be involved in the larger reduction of the potential growth of main axes. Moreover, when growth of main axes is restricted, assimilate availability becomes higher for branches and that could explain that their growth could be increased in a homogeneous strong soil. This revised version was published online in June 2006 with corrections to the Cover Date.     

Phosphate Availability Alters Lateral Root Anatomy and Root Architecture of Fraxinus mandshurica Rupr. Seedlings

Plants have evolved some mechanisms to maximize the efficiency of phosphorus acquisition. Changes in root architecture are one such mechanism. When Fraxinus mandshurica Rupr. seedlings were grown under conditions of low phosphorus availability, the length of cells in the meristem zone of the lateral roots was longer, but the length of cells in the elongation and mature zones of the lateral roots was shorter,compared with seedlings grown under conditions of high phosphorus availability. The elongation rates of primary roots increased as phosphorus availability increased, but the elongation rates of the branched zones of the primary roots decreased. The number of lateral root primordia and the length of the lateral roots decreased as phosphorus availability increased. The topological index (altitude slope) decreased as phosphorus availability increased, suggesting that root architecture tended to be herringbone-like when seedlings were grown under conditions of low phosphate availability. Herringbone-like root systems exploit nutrients more efficiently, but they have higher construction costs than root systems with a branching pattem.     

Patterns of variability in the diameter of lateral roots in the banana root system

The relative importance of root system structure, plant carbon status and soil environment in the determination of lateral root diameter remains unclear, and was investigated in this study. Banana (Musa acuminata) plants were grown at various moderate levels of soil compaction in two distinct experiments, in a field experiment (FE) and in a glasshouse experiment (GE). Radiant flux density was 5 times lower in GE. The distribution of root diameter was measured for several root branching orders. Root diameters ranged between 0.09 and 0.52 mm for secondary roots and between 0.06 and 0.27 mm for tertiary roots. A relationship was found between the diameter of the parent bearing root and the median diameter of its laterals, which appears to be valid for a wide range of species. Mean lateral root diameter increased with distance to the base of the root and decreased with branching density [number of lateral roots per unit length of bearing root (cm(-1))]. Typical symptoms of low light availability were observed in GE. In this case, lateral root diameter variability was reduced. Although primary root growth was affected by soil compaction, no effects on lateral root diameter were observed.     

Statistical modeling of nitrogen-dependent modulation of root system architecture in Arabidopsis thaliana

Plant root development is strongly affected by nutrient availability. Despite the importance of structure and function of roots in nutrient acquisition,statistical modeling approaches to evaluate dynamic and temporal modulations of root system architecture in response to nutrient availability have remained as widely open and exploratory areas in root biology. In this study,we developed a statistical modeling approach to investigate modulations of root system architecture in response to nitrogen availability. Mathematical models were designed for quantitative assessment of root growth and root branching phenotypes and their dynamic relationships based on hierarchical con figuration of primary and lateral roots formulating the fishbone-shaped root system architecture in Arabidopsis thaliana. Time-series datasets reporting dynamic changes in root developmental traits on different nitrate or ammonium concentrations were generated for statistical analyses. Regression analyses unraveled key parameters associated with:(i) inhibition of primary root growth under nitrogen limitation or on ammonium;(ii) rapid progression of lateral root emergence in response to ammonium; and(iii) inhibition of lateral root elongation in the presence of excess nitrate or ammonium. This study provides a statistical framework for interpreting dynamic modulation of root system architecture,supported by metaanalysis of datasets displaying morphological responses of roots to diverse nitrogen supplies.     

Apical diameter and branching density affect lateral root elongation rates in banana

Thick roots elongate faster than thinner ones. However, within one species, the growth achieved by roots of a given diameter can be very variable, and root diameter can only be considered as a determinant of root potential elongation rate. As root elongation is highly correlated to carbon availability, it can be hypothesized that local competition for resources, expressed as the number of lateral roots per unit length (i.e. the branching density), modulates root elongation. Using novel methods in field conditions, we have estimated apical diameters, elongation rates and growth durations of nearly 3500 banana lateral roots, in a field experiment with high radiations and a shaded glasshouse experiment with low radiations. Apical diameters and branching densities were lower in the experiment with low radiation, but elongation rates were higher. In both experiments, mean elongation rates of first-order laterals and thick second-order laterals were negatively correlated with bearing root branching densities. It is hypothesized that, even though apical diameters were lower, low branching densities in the shaded glasshouse allowed enhanced lateral root elongation. In both experiments, second-order laterals elongated more slowly than first-order laterals of similar diameter. A specific effect of root order, independent of branching density and apical diameter, contributed to explain these slow second-order lateral elongation rates. Most lateral roots elongated between 9 and 21 days and growth duration was mainly correlated with root diameter.     

Phosphate availability regulates root system architecture in Arabidopsis

Plant root systems are highly plastic in their development and can adapt their architecture in response to the prevailing environmental conditions. One important parameter is the availability of phosphate, which is highly immobile in soil such that the arrangement of roots within the soil will profoundly affect the ability of the plant to acquire this essential nutrient. Consistent with this, the availability of phosphate was found to have a marked effect on the root system architecture of Arabidopsis. Low phosphate availability favored lateral root growth over primary root growth, through increased lateral root density and length, and reduced primary root growth mediated by reduced cell elongation. The ability of the root system to respond to phosphate availability was found to be independent of sucrose supply and auxin signaling. In contrast, shoot phosphate status was found to influence the root system architecture response to phosphate availability.     

Estimating root elongation rates from morphological measurements of the root tip

To measure the elongation rate of individual roots in soil remains a challenge. A novel method for estimating elongation rates of excavated roots is presented. Morphological markers are identified along the tip of excavated roots, and their distance relative to the apex is measured. These markers correspond to developmental stages which follow known temporal patterns. Hence, their distance relative to the apex reflects root elongation during the period corresponding to their development. The method was tested on maize roots grown in a range of conditions and substrates. It was found that distances from markers to apices were proportional, with some variability, to elongation rates. Remarkably, the linear relationships between these distances were neither affected by substrate, nor by growing conditions. Using several markers allows covering time periods ranging from 0.3 day to 3 days as well as cross validation of estimates. Provided further testing, under a wider range of environmental conditions, is conducted, the concepts presented in this paper may serve to define a new measurement technique.     

Dynamics of heterorhizic root systems: protoxylem groups within the fine-root system of Chamaecyparis obtusa

To understand the physiology of fine-root functions in relation to soil organic sources, the heterogeneity of individual root functions within a fine-root system requires investigation. Here the heterogeneous dynamics within fine-root systems are reported. The fine roots of Chamaecyparis obtusa were sampled using a sequential ingrowth core method over 2 yr. After color categorization, roots were classified into protoxylem groups from anatomical observations. The root lengths with diarch and triarch groups fluctuated seasonally, whereas the tetrarch root length increased. The percentage of secondary root mortality to total mortality increased with increasing amounts of protoxylem. The carbon : nitrogen ratio indicated that the decomposability of primary roots might be greater than that of secondary roots. The position of diarch roots was mostly apical, whereas tetrarch roots tended to be distributed in basal positions within the root architecture. We demonstrate the heterogeneous dynamics within a fine-root system of C. obtusa. Fine-root heterogeneity should affect soil C dynamics. This heterogeneity is determined by the branching position within the root architecture.     

Isolation and culture of mesophyll protoplasts from Rosa hybrida

After 1 h plasmolysis in CPW13M solution, highly viable (>75%) protoplasts were isolated from leaves of axenic shoot cultures of Rosa hybrida L. cv. Abraham Darby using an enzyme mixture containing 1.0% (w/v) Hemicellulase, 0.1% (w/v) Macerozyme, 1.0 (w/v) Cellulase RS, 0.05% (w/v) Pectolyase Y23 and 1.0% (w/v) PVP-10 and from cv. Marie Pavié using an identical mixture but with Cellulase RS and Pectolyase Y23 at 0.7% (w/v) and 0.1% (w/v), respectively. With both cvs., sustained protoplast division was achieved after plating in agarose beads with modified KM8p medium containing 1.0% (w/v) polyvinylpyrrolidone (mol. wt. 10 000; PVP-10), 8.91 μM naphthaleneacetic acid (NAA) and 4.44 μM 6-benzyladenine (BA). Protoplast-derived callus gave rise to roots after transfer to SH medium containing 14 μM 2,4-D. This revised version was published online in June 2006 with corrections to the Cover Date.     

Intrinsic and Extrinsic Controls of Fine Root Life Span

Although fine roots play an integral role in biogeochemical cycling and supporting plant function, fundamental understanding of the mechanisms that control fine root life span is limited. Based on literature, we examined how intrinsic plant characteristics including root diameter, root branching order, rooting depth, and mycorrhizal symbiosis affect fine root life span, and how fine root life span differs with plant life form and foliar habit and between early versus late seral species. We also examined how soil nitrogen and water availability, temperature, and atmospheric carbon dioxide concentration influence fine root life span. We focused on evidence from rhizotron and minirhizotron observations which allow for individual roots to be directly monitored in situ. Fine root life span increased with increasing root diameter, was shorter for more distal than proximal roots, and increased with increasing rooting depth, but was not influenced by mycorrhizal symbiosis. Trees had the longest fine root life spans of all the plant life forms, followed by grasses, lianas, shrubs, and forbs. Among trees, deciduous species had shorter fine root life spans than evergreen species. Fine root life span appears to decrease with increasing temperature and increase with soil water availability, whereas the effects of soil nitrogen availability and atmospheric carbon dioxide concentration on fine root life span were highly inconsistent among studies. Our findings indicate that root morphological characteristics and plant traits are useful predictors of fine root life span. However, environmental influences on fine root life span remain poorly understood due to the limited number of respective studies. Future studies of root demographic processes are needed to better understand environmental controls of fine root life span. It is also critical that research continues into developing more direct and less invasive techniques for studying root demographics.     

A role for redox factors in shaping root architecture under phosphorus deficiency

The developmental response of the Arabidopsis root system to low phosphorus (P) availability involves the reduction in primary root elongation accompanied by the formation of numerous lateral roots. We studied the roles of selected redox metabolites, namely, radical oxygen species (ROS) and ascorbic acid (ASC) in the regulation of root system architecture by different P availability. Rapidly growing roots of plants grown on P-sufficient medium synthesize ROS in root elongation zone and quiescent centre. We have demonstrated that the arrest of root elongation at low P medium coincides with the disappearance of ROS from the elongation zone. P-starvation resulted in a decrease in ascorbic acid level in roots. This correlated with a decrease in cell division activity. On the other hand, feeding P-deficient plants with ASC, stimulated mitotic activity in the primary root meristem and partly reversed the inhibition of root growth imposed by low P conditions. In this paper, we discuss the idea of the involvement of redox agents in the regulation of root system architecture under low P availability.Key words: ascorbic acid, phosphate deficiency, primary root, radical oxygen species, root growth, root system architecture     

Periodicity in the development of the root system of young rubber trees (Hevea brasiliensis Müell. Arg.): relationship with shoot development

The growth pattern of the root system of young rubber trees (Hevea brasiliensis) was studied in relation to shoot development over a period of 3 months. Temporal and spatial variations in elongation and branching processes were examined for the different root types, by means of root observation boxes. Shoot growth was typically rhythmic. Root development was periodic and related to leaf expansion. Root elongation was depressed during leaf growth, whereas branching was enhanced. Consequently, highly branched areas with vigorous secondary roots alternated along the taproot with poorly branched areas with shorter roots. Root types were not affected to the same degree by shoot competition: during leaf expansion, taproot growth was just depressed but remained continuous, the emergence and elongation rates of secondary roots were significantly affected and the elongation rates of tertiary roots fell to zero. These results were consistent with the hypothesis that root growth is related to competition for assimilates and to the sink strength of the different root types, whereas root branching appeared to be promoted by leaf development.     

The effect of drought on mycorrhizal production and very fine root system development of Norway spruce under natural and experimental conditions

Mycorrhizal growth rates were measured monthly, using a new method, in two neighouring plots of a natural spruce stand. One of the plots was irrigated while the other suffered from drought during the late summer and autumn months. Drought did not completely stop mycorrhizal growth. It caused a higher rate of root dormancy and a reduced elongation rate of the parent roots but an increased development of new mucorrhizal last order laterals. Thus, the branching density of the very fine root system was increased, even though fewer growing mycorrhizae were found in the non-irrigated plot during the dry period. Similar results were observed in a water-stress experiment with pot-cultures. After rewetting, elongation rate was stimulated and the number of growing mycorrhizae increased rapidly on the non-irrigated plot. Possible relationships between dry weight, distribution and branching density of growing fine root systems are presented.     

Shade avoidance in the clonal herb Trifolium fragiferum/: a field study with experimentally manipulated vegetation height

In herbaceous canopies light availability can show high degrees of spatial variability in a vertical and also in a horizontal direction. Stoloniferous plants are hence likely to encounter differences in light availability during their ontogenetic development. Different mechanisms, such as petiole elongation, plasticity in internode length and branching, and an enhanced allocation to sexual reproduction have been suggested to represent viable shade-avoidance mechanisms for clonal plants.In a field experiment we tested the response of the stoloniferous herb Trifolium fragiferum L. to experimentally manipulated vegetation heights. Naturally occurring clonal fragments were exposed to four different vegetation heights ranging from 0 cm (high light availability created by clipping the surrounding natural vegetation at ground level) to 20 cm (natural shading in closed canopy). The growth and development of individual clones was followed for two months. At the end of the experiment above-ground plant parts were harvested. Growth-related and morphological parameters (e.g., petiole and internode length) as well as patterns of meristem utilization (i.e., flowering, branching) were recorded.Neither primary stolon growth and biomass accumulation nor branching and flowering were significantly affected by treatments. However, increased vegetation height resulted in a reduced number of secondary ramets and also had strong positive effects on petiole length, leading to marked changes in the architecture of plants growing in canopies of different heights. In addition, the average weight of individual ramets on the primary stolon was markedly higher in plants exposed to taller vegetation as compared to shorter vegetation.The results of this study suggest the occurrence of a trade-off between clonal expansion (i.e., secondary ramet production) and the average size of clonal offspring. If grown under higher vegetation plants invested more into the size of individual ramets, especially into elongating petioles, and less into the growth and development of lateral branches. Placing leaf laminae higher up in the canopy results in an enhanced light interception which has apparently buffered negative effects of increased vegetation height on whole-clone biomass. Plants grown under shorter vegetation invested more into lateral spread by producing more, but smaller ramets.     

Changes in carbon allocation and expression of carbon transporter genes in Betula pendula Roth. colonized by the ectomycorrhizal fungus Paxillus involutus (Batsch) Fr.

Comparative analyses of aspects of the carbon (C) physiology and the expression of C transporter genes in birch (Betula pendula Roth.) colonized by the ectomycorrhizal fungus Paxillus involutus (Batsch) Fr. were performed using mycorrhizal (M) and non‐mycorrhizal (NM) plants of similar foliar nutrient status. After six months of growth, the biomass of M plants was significantly lower than that of NM plants. Diurnal C budgets of both sets of plants revealed that M plants exhibited higher rates of photosynthesis and root respiration expressed per unit dry weight. However, the diurnal net C gain of M and NM plants remained similar. Ectomycorrhizal roots contained higher soluble carbohydrate pools and increased activity of cell wall invertase, suggesting that additional C was allocated to these roots and their ectomycorrhizal fungi consistent with an increased sink demand for C due to the presence of the mycobiont. In M roots, the expression of two hexose and one sucrose transporter genes of birch were reduced to less than one‐third of the expression level observed in NM roots. Analysis using a probe against the birch ribosomal internal transcribed spacer region revealed that M roots contained 22% less plant RNA than NM roots. As the expression of birch hexose and sucrose transporter genes was reduced to a much greater extent, this suggests that these specific genes were down‐regulated in response to alterations in C metabolism within M roots.     

Local NO3- or NH4+ supply modifies the root system architecture of Cedrus atlantica seedlings grown in a split-root device

To study the effects of local nitrate or ammonium supply on the architecture of the Cedrus atlantica root system, cedar seedlings were grown in split-root boxes in a growth chamber. In each box-compartment, roots were fertilized with a solution containing nitrogen, either as nitrate [Ca(NO₃)₂] or ammonium (NH₄Cl), supplied at 0.1 or 5.0 mM. For each seedling, the shoot growth was measured twice a week for 3 months. The root system architecture was also recorded twice a week by tracing the root elongation through the transparent face of the root observation boxes. The apical diameter of the tap-root relay and that of a representative sample of lateral roots were recorded once a month using a monocular magnifier.

The increase of ammonium or nitrate concentration in the nutrient solution has significantly enhanced the production of lateral roots on the tap-root relay. After 90 days of culture, percentages of short lateral roots obtained with nitrate were higher than those obtained using ammonium. A preferential carbon allocation to the shoots was also obtained with an increasing nitrogen supply. Until the 40th day of culture, the elongation of lateral roots was similar for all treatments and ranged from 0.25 to 0.5 cm day⁻¹. From the 40th day to the 95th day, significant differences were observed between the compared modes and maximum elongation rates were obtained with 5 mM NH₄⁺ (2.18 cm day⁻¹) and 5 mM NO₃⁻ (1.18 cm day⁻¹). Local applications of nitrate and ammonium at a low or a high concentration had local effects on elongation and branching of the root system in the fertilized compartment. Contrasting effects of ammonium and nitrate were observed on the apical diameter of tap-roots and lateral roots. The root-split culture device confirmed that nitrate had local effects on the architecture of the C. atlantica root system.