Neutron imaging reveals internal plant water dynamics

Background and aims

Knowledge of plant water fluxes is critical for assessing mechanistic processes linked to biogeochemical cycles, yet resolving root water transport dynamics has been a particularly daunting task. Our objectives were to demonstrate the ability to non-invasively monitor individual root functionality and water fluxes within Zea mays L. (maize) and Panicum virgatum L. (switchgrass) seedlings using neutron imaging.

Methods

Seedlings were propagated for 1–3 weeks in aluminum chambers containing sand. Pulses of water or deuterium oxide were then tracked through the root systems by collecting consecutive radiographs during exposure to a cold-neutron source. Water flux was manipulated by cycling on a growth lamp to alter foliar demand for water.

Results

Neutron radiography readily illuminated root structure, root growth, and relative plant and soil water content. After irrigation there was rapid root water uptake from the newly wetted soil, followed by hydraulic redistribution of water through the root system to roots terminating in dry soil. Water flux within individual roots responded differentially to foliar illumination based on supply and demand of water within the root system.

Conclusions

Sub-millimeter scale image resolution revealed timing and magnitudes of root water uptake, redistribution within the roots, and root-shoot hydraulic linkages—relationships not well characterized by other techniques.     

Growth of tree roots in hostile soil: A comparison of root growth pressures of tree seedlings with peas

Background and Aims

As part of a study on growth of tree roots in hostile soil, we envisaged that establishment and survival of trees on hard, dry soil may depend on their ability to exert axial root growth pressures of similar magnitude to those of the roots of agricultural plants (with significant root thickening when roots grow across an air gap or cracks and biopores). We selected tree species originating from a range of different soil and climatic conditions to evaluate whether their relative success on harsh soil (in an evolutionary sense) might be related to the magnitude of root growth pressures they could exert, or how they performed in the very early stages of growth after germination.

Methods

We measured the maximum axial root growth force (F_max) on single lateral root axes of 3- to 4- month old seedlings of 6 small-seeded eucalypts from 2 different habitats and 2 contrasting soil types. Root growth rate, root diameter and F_max were also measured on the primary root axes of a large-seeded acacia and a domesticated annual (Pisum sativum) seedling for up to 10 days following germination.

Results

The lateral roots of the 6 eucalypts and the primary roots of the acacia were considerably smaller than the primary roots of P. sativum and they exerted average forces of similar magnitude to one another (0.198 to 0.312 N). The maximum axial root growth pressures were all in the range 150 to 250 kPa but E. leucoxylon, E. loxophleba and A. salicina exerted the greatest pressures among the trees, and comparable pressures to those exerted by the primary roots of 2-day-old P. sativum (211-252 kPa). Although the primary roots of acacia seedlings exerted increasing axial root growth pressures over a 10-day period following germination, the pressures were still only slightly greater than those of the domesticated plant, P. sativum.

Conclusions

The lack of any very large differences in axial root growth pressures between trees and domesticated plants suggests that trees that grow well in harsh soil don’t do so by exerting higher root growth pressures alone but by also exploring the network of cracks and pores more effectively than do other plants that are less successful.     

Changes in root size and distribution in relation to nitrogen accumulation during maize breeding in China

Background and aims

Modern maize breeding has increased maize yields worldwide. The changes in above-ground traits accompanying yield improvement are well-known, but less information is available as to the effect of modern plant breeding on changes in maize root traits.

Methods

Root growth, nitrogen uptake, dry matter accumulation and yield formation of six maize hybrids released from 1973 to 2000 in China were compared. Experiments were conducted under low and high nitrogen supply in a black soil in Northeast China in 2010 and 2011.

Results

While nitrogen accumulation, dry matter production and yield formation have been increased, modern maize breeding in China since 1990 has reduced root length density in the topsoil without much effect on root growth in the deeper soil. The efficiency of roots in acquiring N has increased so as to match the requirement of N accumulation for plant growth and yield formation. The responses of root growth, nitrogen and dry matter accumulation, and grain yield to low-N stress were similar in the more modern hybrids as in the older ones.

Conclusions

Modern maize breeding has constitutively changed root and shoot growth and plant productivity without producing any specific enhancement in root responsiveness to soil N availability.     

Maize (Zea mays L.) endogenous seed phosphorus remobilization is not influenced by exogenous phosphorus availability during germination and early growth stages

Background and aims

Phosphorus (P) nutrition is very important during early maize seedling growth. Remobilization of endogenous seed P and uptake of exogenous P are therefore of prime importance during this period. Our objectives were to study the effect of the availability of endogenous and exogenous P on i) remobilization of endogenous seed P, ii) the beginning of exogenous P uptake and its intensity, iii) their interaction and effect on seedling development.

Methods

Seeds with high and low reserves of endogenous seed P were cultivated at three rates of availability of exogenous P (0, 100, 1,000?μM) over a growth period of 530 cumulated degree days after sowing. Exogenous P was labeled with radioactive P (³²P) to distinguish the two fluxes of P in seedlings, one due to remobilization of seed P and the other to uptake of exogenous P.

Results

Initially, 86% of endogenous seed P was localized in the scutellum, mainly in the form of phytate, regardless of initial endogenous seed P. At 89 cumulated degree days after sowing (base temperature: 10°C), 98% of seed phytate was hydrolyzed in all treatments. In treatments with available exogenous P, significant uptake of exogenous P started at 71 cumulated degree days after sowing. Efficient uptake of exogenous P depended on its availability, but was independent of phytate hydrolysis and seedling P status. Significant loss of P from germinating seeds due to efflux was observed and was also independent of the availability of exogenous P.

Conclusions

Our results show that hydrolysis of seed P was not influenced by the availability of exogenous P, and conversely, that uptake of exogenous P was not influenced by endogenous P in the seed. This suggests that remobilization of endogenous seed P and uptake of exogenous P by seedling roots are controlled independently.     

A soil-free root observation system for the study of root-microorganism interactions in maize

Background and aims

The root surface of a plant usually exceeds the leaf area and is constantly exposed to a variety of soil-borne microorganisms. Root pathogens and pests, as well as belowground interactions with beneficial microbes, can significantly influence a plants' performance. Unfortunately, the analysis of these interactions is often limited because of the arduous task of accessing roots growing in soil. Here, we present a soil-free root observation system (SF-ROBS) designed to grow maize (Zea mays) plants and to study root interactions with either beneficial or pathogenic microbes.

Methods

The SF-ROBS consists of pouches lined with wet filter paper supplying nutrient solution.

Results

The aspect of maize grown in the SF-ROBS was similar to soil-grown maize; the plant growth was similar for the shoot but different for the roots (biomass and length increased in the SF-ROBS). SF-ROBS-grown roots were successfully inoculated with the hemi-biotrophic maize fungal pathogen Colletotrichum graminicola and the beneficial rhizobacteria Pseudomonas putida KT2440. Thus, the SF-ROBS is a system suitable to study two major belowground phenomena, namely root fungal defense reactions and interactions of roots with beneficial soil-borne bacteria.

Conclusions

This system contributes to a better understanding of belowground plant microbe interactions in maize and most likely also in other crops.     

Net root growth and nutrient acquisition in response to predicted climate change in two contrasting heathland species

Background and aims

Accurate predictions of nutrient acquisition by plant roots and mycorrhizas are critical in modelling plant responses to climate change.

Methods

We conducted a field experiment with the aim to investigate root nutrient uptake in a future climate and studied root production by ingrowth cores, mycorrhizal colonization, and fine root N and P uptake by root assay of Deschampsia flexuosa and Calluna vulgaris.

Results

Net root growth increased under elevated CO₂, warming and drought, with additive effects among the factors. Arbuscular mycorrhizal colonization increased in response to elevated CO₂, while ericoid mycorrhizal colonization was unchanged. The uptake of N and P was not increased proportionally with root growth after 5 years of treatment.

Conclusions

While aboveground biomass was unchanged, the root growth was increased under elevated CO₂. The results suggest that plant production may be limited by N (but not P) when exposed to elevated CO₂. The species-specific response to the treatments suggests different sensitivity to global change factors, which could result in changed plant competitive interactions and belowground nutrient pool sizes in response to future climate change.     

A split-root experiment shows that translocated phosphorus does not alleviate aluminium toxicity within plant tissue

Background and aims

My previous experimental findings suggested that phosphorus (P) enhances aluminium (Al) tolerance in both Al-tolerant and Al-sensitive wheat seedlings. However, the role of P in the amelioration of Al toxicity within plant tissue is still unclear. Therefore, a soil culture horizontal split-root system was used to quantify whether or not translocated P alleviates Al toxicity within the plant tissue.

Methods

Different level of Al and P were added in two compartments in various combinations for separate root halves. Constrasting Al-tolerant (ET8) and Al-sensitive (ES8) wheat genotypes were used as a testing plant.

Results

The limitation of root growth was independent to Al-toxicity in one root half. However, root proliferation occurred as a compensatory growth on the other root half that has no Al-toxicity. Where half of the roots were given 60 mg P/kg, plant did not translocated P in the other part of the root system that grown in Al toxic soil. When 40 mg P/kg were mixed with 60 mg AlCl₃/kg within one root half combinations, root dry weight of both ET8 and ES8 increased markedly in that root half. In contrast, root dry weight of both ET8 and ES8 decreased noticeably only 60 mg AlCl₃/kg treated root half. The shoot P and Al uptake in both ET8 and ES8 was lower in combined 40 mg P/kg and 60 mg AlCl₃/kg addition as compared to other combination with same P and Al level.

Conclusions

Result from this study confirm that addition of P to Al toxic acid soil played dual role like amelioration of Al-toxicity in soil and utilize P as nutrition for plant growth and development. Findings also attributed that added P was reduced by precipitation with added Al. However, evidence found that translocated P was not able to alleviate Al toxicity within plant tissue of both ES8 and ET8.     

Distribution of cutin and suberin biomarkers under forest trees with different root systems

Background and aims

The rhizosphere, the soil immediately surrounding roots, provides a critical bridge for water and nutrient uptake. The rhizosphere is influenced by various forms of root–soil interactions of which mechanical deformation due to root growth and its effects on the hydraulics of the rhizosphere are the least studied. In this work, we focus on developing new experimental and numerical tools to assess these changes.

Methods

This study combines X-ray micro-tomography (XMT) with coupled numerical simulation of fluid and soil deformation in the rhizosphere. The study provides a new set of tools to mechanistically investigate root-induced rhizosphere compaction and its effect on root water uptake. The numerical simulator was tested on highly deformable soil to document its ability to handle a large degree of strain.

Results

Our experimental results indicate that measured rhizosphere compaction by roots via localized soil compaction increased the simulated water flow to the roots by 27 % as compared to an uncompacted fine-textured soil of low bulk density characteristic of seed beds or forest topsoils. This increased water flow primarily occurred due to local deformation of the soil aggregates as seen in the XMT images, which increased hydraulic conductivity of the soil. Further simulated root growth and deformation beyond that observed in the XMT images led to water uptake enhancement of ~50 % beyond that due to root diameter increase alone and demonstrated the positive benefits of root compaction in low density soils.

Conclusions

The development of numerical models to quantify the coupling of root driven compaction and fluid flow provides new tools to improve the understanding of plant water uptake, nutrient availability and agricultural efficiency. This study demonstrated that plants, particularly during early growth in highly deformable low density soils, are involved in active mechanical management of their surroundings. These modeling approaches may now be used to quantify compaction and root growth impacts in a wide range of soils.     

Isolation of ACC deaminase producing PGPR from rice rhizosphere and evaluating their plant growth promoting activity under salt stress

Aims

Bacteria possessing ACC deaminase activity reduce the level of stress ethylene conferring resistance and stimulating growth of plants under various biotic and abiotic stresses. The present study aims at isolating efficient ACC deaminase producing PGPR strains from the rhizosphere of rice plants grown in coastal saline soils and quantifying the effect of potent PGPR isolates on rice seed germination and seedling growth under salinity stress and ethylene production from rice seedlings inoculated with ACC deaminase containing PGPR.

Methods

Soils from root region of rice growing in coastal soils of varying salinity were used for isolating ACC deaminase producing bacteria and three bacterial isolates were identified following polyphasic taxonomy. Seed germination, root growth and stress ethylene production in rice seedlings following inoculation with selected PGPR under salt stress were quantified.

Results

Inoculation with selected PGPR isolates had considerable positive impacts on different growth parameters of rice including germination percentage, shoot and root growth and chlorophyll content as compared to uninoculated control. Inoculation with the ACC deaminase producing strains reduced ethylene production under salinity stress.

Conclusions

This study demonstrates the effectiveness of rhizobacteria containing ACC deaminase for enhancing salt tolerance and consequently improving the growth of rice plants under salt-stress conditions.     

Response characteristics of seed germination and seedling growth of Acorus tatarinowii under diesel stress

Aims

Responses of typical wetland plant Acorus tatarinowii to diesel stress were investigated to provide basis of ecological monitoring system and phytoremediation for diesel-contaminated wetland.

Methods

Greenhouse experiments were established to determine the germinability of seedlings, hydrogen peroxide in leaves, and DNA damage in roots exposed to a range of potentially phytotoxic diesel.

Results

The presence of diesel did not benefit the growth of A. tatarinowii. The germination ratio and germination rate decreased with the increase of diesel concentration, both the lowest value appeared when the concentration of diesel was 10,000 mg?kg^?1. The lowest diesel concentration (2,000 mg?kg^?1) in the soil significantly reduced the length, average diameter, and projected area of root, especially on the stress of the higher diesel concentration (4,000, 8,000, and 10,000 mg?kg^?1). Furthermore, H₂O₂ concentration in leaves rose with the increasing concentration of diesel. However, no DNA oxidative damage to root was observed in our experiment.

Conclusions

Diesel exposure significantly inhabited the seed germination, root elongation, and seedlings growth of A. tatarinowii. Diesel stress caused the accumulation of H₂O₂ in the leaves of A. tatarinowii.     

Capacity of biological soil crusts colonized by the lichen <Emphasis Type="Italic">Diploschistes</Emphasis> to metabolize simple phenols

Background and aims

Soil compaction strongly affects water uptake by roots. The aim of the work was to examine soil—plant interactions with focus on the impact of distribution of compacted soil layers on growth and water uptake by wheat roots.

Methods

The growth-chamber experiment was conducted on wheat growth in soil with compacted soil layers. The system for maintaining constant soil water potential and measurement of daily water uptake from variously compacted soil layers was used.

Results

Layered soil compaction differentiated vertical root distribution to higher extent for root length than root mass. The propagation rate of a water extraction front was the highest through layers of moderately compacted soil. The root water uptake rate was on average 67 % higher from moderately than heavily compacted soil layers. Correlations between water uptake and the length of thick roots were increasing with increasing level of soil compaction.

Conclusions

The study shows that root amount, water uptake, propagation of water extraction and shoot growth strongly depend on the existence of compacted layers within soil profile. The negative effects of heavily compacted subsoil layer on water uptake were partly compensated by increased uptake from looser top soil layers and significant contribution of thicker roots in water uptake.     

Effects of copper on growth, radial oxygen loss and root permeability of seedlings of the mangroves Bruguiera gymnorrhiza and Rhizophora stylosa

Purpose

Mangrove wetlands have experienced significant contaminant input such as copper (Cu), aggravated by rapid urban development. This study aimed to investigate the possible function of root permeability in metal detoxification.

Methods

Pot trials were conducted to evaluate the responses of root permeability in relation to metal (Cu) exposure in seedlings of two mangroves: Bruguiera gymnorrhiza and Rhizophora stylosa.

Results

Copper inhibited plant growth and root permeability of the two species significantly (due to decreases in root porosity, thickening of exodermis and increases in lignification), leading to a significant reduction in radial oxygen loss (ROL). A negative correlation between soil Cu and ROL from root tip was also observed. The observed metal uptake by excised roots further indicated that increased lignification would directly prevent excessive Cu from further entering into the roots.

Conclusions

In summary, the two mangroves reacted to Cu by producing an impermeable barrier in roots. Such an inducible barrier to ROL is likely to be an adaptive strategy against Cu toxicity. This study reveals new evidence of a structural adaptive strategy for metal tolerance by mangrove plants.     

Silicon modifies root anatomy, and uptake and subcellular distribution of cadmium in young maize plants

Background and Aims

Silicon (Si) has been shown to ameliorate the negative influence of cadmium (Cd) on plant growth and development. However, the mechanism of this phenomenon is not fully understood. Here we describe the effect of Si on growth, and uptake and subcellular distribution of Cd in maize plants in relation to the development of root tissues.

Methods

Young maize plants (Zea mays) were cultivated for 10 d hydroponically with 5 or 50 µm Cd and/or 5 mm Si. Growth parameters and the concentrations of Cd and Si were determined in root and shoot by atomic absorption spectrometry or inductively coupled plasma mass spectroscopy. The development of apoplasmic barriers (Casparian bands and suberin lamellae) and vascular tissues in roots were analysed, and the influence of Si on apoplasmic and symplasmic distribution of ¹⁰⁹Cd applied at 34 nm was investigated between root and shoot.

Key Results

Si stimulated the growth of young maize plants exposed to Cd and influenced the development of Casparian bands and suberin lamellae as well as vascular tissues in root. Si did not affect the distribution of apoplasmic and symplasmic Cd in maize roots, but considerably decreased symplasmic and increased apoplasmic concentration of Cd in maize shoots.

Conclusions

Differences in Cd uptake of roots and shoots are probably related to the development of apoplasmic barriers and maturation of vascular tissues in roots. Alleviation of Cd toxicity by Si might be attributed to enhanced binding of Cd to the apoplasmic fraction in maize shoots.     

The effect of phosphorus on cluster-root formation and functioning of Embothrium coccineum (R. et J. Forst.)

Background and aims

Embothrium coccineum (R. et J. Forst.) is a Proteaceae species from the southern part of South America. South-central Chilean soils are younger and contain more phosphorus (P) than soils in Australia and South Africa, where Proteaceae are common. Phosphorus deficiency is the main factor promoting cluster-root formation in Proteaceae. It is not known, however, whether this also applies to E. coccineum, which grows on soils with higher P content.

Methods

Four-month-old seedlings were grown for 4 weeks in hydroponic cultures with 1 μM P or 50 μM P. The number of cluster roots, relative height increment, biomass distribution, cluster root/total root biomass ratio, foliar P concentration, root acid phosphatase activity and root carboxylate-exudation rates were determined.

Results

Seedlings growing at 50 μM P showed a 10?, 1.3? and 3.3-fold greater increase in relative height, total dry mass and foliar P concentration, respectively, compared with those grown at1 μM P. However, seedlings grown at 1 μM P showed a 5?, 16?, 1.7? and 1.3-fold greater number of cluster roots, cluster root/total root biomass ratio, phosphatase activity and total carboxylate exudation, respectively, as compared with those grown at 50 μM P.

Conclusions

A low P supply promotes the initiation, growth and metabolic activity of cluster roots which is in accordance with reports on Proteaceae species occurring in ancient and highly weathered soils.     

Overexpression of a novel salt stress-induced glycine-rich protein gene from alfalfa causes salt and ABA sensitivity in Arabidopsis

Key message

We cloned a novel salt stress-induced glycine-rich protein gene ( MsGRP ) from alfalfa. Its overexpression retards seed germination and seedling growth of transgenic Arabidopsis after salt and ABA treatments.

Abstract

Since soil salinity is one of the most significant abiotic stresses, salt tolerance is required to overcome salinity-induced reductions in crop productivity. Many glycine-rich proteins (GRPs) have been implicated in plant responses to environmental stresses, but the function and importance of some GRPs in stress responses remain largely unknown. Here, we report on a novel salt stress-induced GRP gene (MsGRP) that we isolated from alfalfa. Compared with some glycine-rich RNA-binding proteins, MsGRP contains no RNA recognition motifs and localizes in the cell membrane or cell wall according to the subcellular localization result. MsGRP mRNA is induced by salt, abscisic acid (ABA), and drought stresses in alfalfa seedlings, and its overexpression driven by a constitutive cauliflower mosaic virus-35S promoter in Arabidopsis plants confers salinity and ABA sensitivity compared with WT plants. MsGRP retards seed germination and seedling growth of transgenic Arabidopsis plants after salt and ABA treatments, which implies that MsGRP may affect germination and growth through an ABA-dependent regulation pathway. These results provide indirect evidence that MsGRP plays important roles in seed germination and seedling growth of alfalfa under some abiotic stress conditions.     

Modelling root plasticity and response of narrow-leafed lupin to heterogeneous phosphorus supply

Background & Aims

Searching for root traits underpinning efficient nutrient acquisition has received increased attention in modern breeding programs aimed at improved crop productivity. Root models provide an opportunity to investigate root-soil interactions through representing the relationships between rooting traits and the non-uniform supply of soil resources. This study used simulation modelling to predict and identify phenotypic plasticity, root growth responses and phosphorus (P) use efficiency of contrasting Lupinus angustifolius genotypes to localised soil P in a glasshouse.

Methods

Two L. angustifolius genotypes with contrasting root systems were grown in cylindrical columns containing uniform soil with three P treatments (nil and 20 mg P kg^?1 either top-dressed or banded) in the glasshouse. Computer simulations were carried out with root architecture model ROOTMAP which was parameterized with root architectural data from an earlier published hydroponic phenotyping study.

Results

The experimental and simulated results showed that plants supplied with banded P had the largest root system and the greatest P-uptake efficiency. The P addition significantly stimulated root branching in the topsoil, whereas plants with nil P had relatively deeper roots. Genotype-dependent root growth plasticity in response to P supply was shown, with the greatest response to banded P.

Conclusions

Both experimental and simulation outcomes demonstrated that 1) root hairs and root proliferation increased plant P acquisition and were more beneficial in the localised P fertilisation scenario, 2) placing P deeper in the soil might be a more effective fertilisation method with greater P uptake than top dressing, and 3) the combination of P foraging strategies (including root architecture, root hairs and root growth plasticity) is important for efficient P acquisition from a localised source of fertiliser P.     

Uptake and cellular distribution,in four plant species,of fluorescently labeled mesoporous silica nanoparticles

Key message

We report the uptake of MSNs into the roots and their movement to the aerial parts of four plant species and their quantification using fluorescence, TEM and proton-induced x - ray emission (micro - PIXE) elemental analysis.

Abstract

Monodispersed mesoporous silica nanoparticles (MSNs) of optimal size and configuration were synthesized for uptake by plant organs, tissues and cells. These monodispersed nanoparticles have a size of 20 nm with interconnected pores with an approximate diameter of 2.58 nm. There were no negative effects of MSNs on seed germination or when transported to different organs of the four plant species tested in this study. Most importantly, for the first time, a combination of confocal laser scanning microscopy, transmission electron microscopy and proton-induced X-ray emission (micro-PIXE) elemental analysis allowed the location and quantification MSNs in tissues and in cellular and sub-cellular locations. Our results show that MSNs penetrated into the roots via symplastic and apoplastic pathways and then via the conducting tissues of the xylem to the aerial parts of the plants including the stems and leaves. The translocation and widescale distribution of MSNs in plants will enable them to be used as a new delivery means for the transport of different sized biomolecules into plants.     

Growth and survival of cork oak (Quercus suber) seedlings after simulated partial cotyledon consumption under different soil nutrient contents

Aims

We examined the importance of partial seed consumption (cotyledon loss) by rabbits in the early establishment of seedlings of cork oaks restricted to nutrient-impoverished soils.

Methods

To determine the importance of cotyledons in the growth and development of seedlings, we simulated two levels of predation [light (30 % cotyledon loss) and heavy (60 % loss) partial consumption] and two soil nutrient contents (nutrient-poor soil, nutrient-rich soil). Seedlings height, root length, dry root and shoot biomass, specific leaf mass, leaf density, gas exchange, chlorophyll fluorescence parameters and photosynthetic pigment concentrations were determined.

Results

Results indicated that effect of nutrient level on the growth of the oak seedlings was more important than that of cotyledon biomass. However, in nutrient–poor soils, cotyledon biomass played a major role in the early performance of cork oaks. Acorns grown in nutrient-rich substrate, despite having greater aerial vigor, were slower to develop a vertical root, and hence less likely to reach permanent moisture. Cotyledon loss caused a decrease in the biomass of roots and shoots when acorns were heavily consumed, and as a result experienced a reduction in net photosynthetic rate, stomatal conductance and chlorophyll concentration. Survival of seedlings was unaffected by either soil type or cotyledon loss.

Conclusions

Our results show that effects of soil type on the survival of oak seedlings were more important than those of cotyledon biomass. However, in a competitive situation, cotyledon biomass, as an indicative of growth nutrient support rather than an energy source, could be vital in a nutrient-poor environment, particularly in Mediterranean climate regions and for species with little inherent drought tolerance (as is the case of Quercus spp.), where rapid root growth is required to ensure that contact with soil moisture is maintained over the first summer.