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See detailRoot-emitted volatile organic compounds: can they mediate belowground plant-plant interactions?
Delory, Benjamin ULg; Delaplace, Pierre ULg; Fauconnier, Marie-Laure ULg et al

in Plant and Soil (in press)

Background Aboveground, plants release volatile organic compounds (VOCs) that act as chemical signals between neighbouring plants. It is now well documented that VOCs emitted by the roots in the plant ... [more ▼]

Background Aboveground, plants release volatile organic compounds (VOCs) that act as chemical signals between neighbouring plants. It is now well documented that VOCs emitted by the roots in the plant rhizosphere also play important ecological roles in the soil ecosystem, notably in plant defence because they are involved in interactions between plants, phytophagous pests and organisms of the third trophic level. The roles played by root-emitted VOCs in between- and within-plant signalling, however, are still poorly documented in the scientific literature. Scope Given that (1) plants release volatile cues mediating plant-plant interactions aboveground, (2) roots can detect the chemical signals originating from their neighbours, and (3) roots release VOCs involved in biotic interactions belowground, the aim of this paper is to discuss the roles of VOCs in between- and within-plant signalling belowground. We also highlight the technical challenges associated with the analysis of root-emitted VOCs and the design of experiments targeting volatile-mediated root-root interactions. Conclusions We conclude that root-root interactions mediated by volatile cues deserve more research attention and that both the analytical tools and methods developed to study the ecological roles played by VOCs in interplant signalling aboveground can be adapted to focus on the roles played by root-emitted VOCs in between- and within-plant signalling. [less ▲]

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See detailHow do Rhizobacterial Volatiles Influence Root System Architecture, Biomass Production and Allocation of the Model Grass Brachypodium distachyon?
Delaplace, Pierre ULg; Ormeño - Lafuente, Elena; Nguyen, Minh ULg et al

Conference (2016, January 12)

Plant growth-promoting rhizobacteria are increasingly considered as a complement of conventional inputs in agricultural systems. Their effects on their host plants are diverse and include volatile ... [more ▼]

Plant growth-promoting rhizobacteria are increasingly considered as a complement of conventional inputs in agricultural systems. Their effects on their host plants are diverse and include volatile-mediated growth enhancement. The present study aims at assessing the effects of bacterial volatile production on the biomass production and the root system architecture of Brachypodium distachyon (L.) Beauv. (line Bd-21). An in vitro experimental set-up allowing plant-bacteria interaction through the gaseous phase without any physical contact was used to screen 19 bacterial strains for their growth promotion ability over a 10-day cocultivation period. Using principal component analysis followed by hierarchical clustering and two-way analysis of variance, five groups of bacteria were defined and characterized based on their combined influence on biomass production and root system architecture. The observed effects range from unchanged to highly increased biomass production coupled with increased root length and branching. Primary root length was only increased by the volatile compounds emitted by Enterobacter cloacae JM22 and Bacillus pumilus T4. Overall, the most significant results were obtained with Bacillus subtilis GB03 which induced a 81% increase in total biomass and enhanced total root length, total secondary root length and total adventitious root length by 88, 196 and 473% respectively. The analysis of the emission kinetics of bacterial volatile organic compounds is underway and should lead to the identification of volatile compounds candidates responsible for the observed growth promotion effects. Taking into account the inherent characteristics of our in vitro system, the next experimental steps are identified and discussed from a fundamental and applied viewpoint. [less ▲]

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See detailarchiDART: an R package for the automated computation of plant root architectural traits
Delory, Benjamin ULg; Baudson, Caroline ULg; Brostaux, Yves ULg et al

in Plant and Soil (2016), 398(1), 351-365

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 ... [more ▼]

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. [less ▲]

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See detailarchiDART: Plant Root System Architecture Analysis Using DART and RSML Files
Delory, Benjamin ULg; Baudson, Caroline ULg; Brostaux, Yves ULg et al

Computer development (2015)

Analysis of complex plant root system architectures (RSA) using the output files created by Data Analysis of Root Tracings (DART), an open-access software dedicated to the study of plant root architecture ... [more ▼]

Analysis of complex plant root system architectures (RSA) using the output files created by Data Analysis of Root Tracings (DART), an open-access software dedicated to the study of plant root architecture and development across time series (Le Bot et al (2010) DART: a software to analyse root system architecture and development from captured images, Plant and Soil, DOI: 10.1007/s11104-009-0005-2), and RSA data encoded with the Root System Markup Language (RSML) (Lobet et al (2015) Root System Markup Language: toward a unified root architecture description language, Plant Physiology, DOI: 10.1104/pp.114.253625). More information can be found in Delory et al (2015) archiDART: an R package for the automated computation of plant root architectural traits, Plant and Soil, DOI: 10.1007/s11104-015-2673-4. [less ▲]

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See detailRoot-emitted volatile organic compounds in belowground plant-plant interactions
Delory, Benjamin ULg

Doctoral thesis (2015)

Plants are able to synthesise and release volatile organic compounds (VOCs) aboveground (leaves, stems, flowers and fruits) and belowground (roots). Once emitted, these molecules are key mediators in ... [more ▼]

Plants are able to synthesise and release volatile organic compounds (VOCs) aboveground (leaves, stems, flowers and fruits) and belowground (roots). Once emitted, these molecules are key mediators in biotic interactions as they can be perceived by plant neighbours (first trophic level) and are able to attract/repel organisms of the second (insect herbivores, plant parasitic nematodes) or the third trophic level (entomopathogenic nematodes, parasitoids, etc.). Although many laboratory and field experiments have focused on VOC-mediated plant-plant interactions aboveground, less is known regarding the roles played by root-emitted VOCs in between- and within-plant signalling. In this context, the main goals of this PhD thesis were to (1) identify and quantify the VOCs emitted by barley and chamomile roots and (2) study the influence of chamomile root volatiles on the growth (biomass production and allocation) and the root system architecture (RSA) of barley (interspecific model). Root-emitted VOCs were analysed without extracting the roots from the soil (in situ) using a three-step gas chromatography-mass spectrometry methodology. Plant-plant interaction bioassays were performed using an original experimental device allowing the controlled exposition of growing barley roots to the volatile compounds emitted by chamomile roots for 15 days. In order to speed up the RSA analysis of recipient barley plants, we developed an R package (archiDART) allowing (1) the batch processing of the raw data exported by Data Analysis of Root Tracings (DART) and root image analysis software tools supporting the Root System Markup Language (RSML) format, and (2) the automated computation of RSA traits. Our results showed that crushed barley roots produced mainly hexanal, (E)-hex-2-enal, (E)-non-2-enal and (E,Z)-nona-2,6-dienal. Three-day-old seminal roots were characterised by higher total and individual VOC concentrations compared with older phenological stages. Our experiments also showed that enzymatic activities were required for volatile production. For each developmental stage, the lipoxygenase (LOX) specificity was greater for linoleic acid than for α-linolenic acid. The greatest LOX activities using linoleic and α-linolenic acids as substrates were measured in 7- and 3-day-old roots, respectively. Although undamaged barley roots did not release detectable amounts of VOCs, the analysis of VOCs emitted by mechanically injured roots showed that (E)-non-2-enal (13.8 ± 4.9 ng/g dry wt/h) and (E,Z)-nona-2,6-dienal (4.7 ± 1.8 ng/g dry wt/h) were the only VOCs detected in the plant rhizosphere. Contrasting with these results, the undamaged roots of 61- to 78-day-old chamomile plantlets released mainly one trinorsesquiterpene (albene) and four tricyclic sesquiterpene hydrocarbons (silphinene, modheph-2-ene, α-isocomene and β-isocomene) associated with the Asteraceae family. For each sesquiterpene hydrocarbon, the emission rate was positively correlated with plant age. Based on these results, we performed plant-plant interaction bioassays to investigate the roles played by chamomile root volatiles on the growth and RSA of barley. After 15 days of exposure, plants exposed to the volatiles emitted by the soil and chamomile roots or by the soil alone (control) were morphologically similar. Although not statistically significant (P < 0.09), the leaf area and the total seminal root length were the only parameters that tended to be greater in plants that received the volatile compounds emitted by chamomile roots compared with control plantlets. All these results are discussed in the context of belowground chemical ecology. In addition, some improvements of the experimental devices developed in this research project are also suggested at the end of this PhD thesis. [less ▲]

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See detailInfluence of rhizobacterial volatiles on the root system architecture and the production and allocation of biomass in the model grass Brachypodium distachyon (L.) P. Beauv.
Delaplace, Pierre ULg; Delory, Benjamin ULg; Baudson, Caroline ULg et al

in BMC Plant Biology (2015), 15(195),

Background Plant growth-promoting rhizobacteria are increasingly being seen as a way of complementing conventional inputs in agricultural systems. The effects on their host plants are diverse and include ... [more ▼]

Background Plant growth-promoting rhizobacteria are increasingly being seen as a way of complementing conventional inputs in agricultural systems. The effects on their host plants are diverse and include volatile-mediated growth enhancement. This study sought to assess the effects of bacterial volatiles on the biomass production and root system architecture of the model grass Brachypodium distachyon (L.) Beauv. Results An in vitro experiment allowing plant-bacteria interaction throughout the gaseous phase without any physical contact was used to screen 19 bacterial strains for their growth-promotion ability over a 10-day co-cultivation period. Five groups of bacteria were defined and characterised based on their combined influence on biomass production and root system architecture. The observed effects ranged from unchanged to greatly increased biomass production coupled with increased root length and branching. Primary root length was increased only by the volatile compounds emitted by Enterobacter cloacae JM22 and Bacillus pumilus T4. Overall, the most significant results were obtained with Bacillus subtilis GB03, which induced an 81% increase in total biomass, as well as enhancing total root length, total secondary root length and total adventitious root length by 88.5, 201.5 and 474.5%, respectively. Conclusions This study is the first report on bacterial volatile-mediated growth promotion of a grass plant. Contrasting modulations of biomass production coupled with changes in root system architecture were observed. Most of the strains that increased total plant biomass also modulated adventitious root growth. Under our screening conditions, total biomass production was strongly correlated with the length and branching of the root system components, except for primary root length. An analysis of the emission kinetics of the bacterial volatile compounds is being undertaken and should lead to the identification of the compounds responsible for the observed growth-promotion effects. Within the context of the inherent characteristics of our in vitro system, this paper identifies the next critical experimental steps and discusses them from both a fundamental and an applied perspective. [less ▲]

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See detailRhizobacterial volatiles influence root system architecture, biomass production and allocation of the model grass Brachypodium distachyon (L.) P. Beauv.
Delaplace, Pierre ULg; Ormeño-Lafuente, Elena; Delory, Benjamin ULg et al

Conference (2015, June 18)

Plant growth-promoting rhizobacteria are increasingly considered as a complement of conventional inputs in agricultural systems. Their effects on their host plants are diverse and include volatile ... [more ▼]

Plant growth-promoting rhizobacteria are increasingly considered as a complement of conventional inputs in agricultural systems. Their effects on their host plants are diverse and include volatile-mediated growth enhancement. The present study aims at assessing the effects of bacterial volatile production on the biomass production and the root system architecture of Brachypodium distachyon (L.) Beauv. (line Bd-21). An in vitro experimental set-up allowing plant-bacteria interaction through the gaseous phase without any physical contact was used to screen 19 bacterial strains for their growth promotion ability over a 10-day cocultivation period. Using principal component analysis followed by hierarchical clustering and two-way analysis of variance, five groups of bacteria were defined and characterized based on their combined influence on biomass production and root system architecture. The observed effects range from unchanged to highly increased biomass production coupled with increased root length and branching. Primary root length was only increased by the volatile compounds emitted by Enterobacter cloacae JM22 and Bacillus pumilus T4. Overall, the most significant results were obtained with Bacillus subtilis GB03 which induced a 81% increase in total biomass and enhanced total root length, total secondary root length and total adventitious root length by 88, 196 and 473% respectively. The analysis of the emission kinetics of bacterial volatile organic compounds is underway and should lead to the identification of volatile compounds candidates responsible for the observed growth promotion effects. Taking into account the inherent characteristics of our in vitro system, the next experimental steps are identified and discussed from a fundamental and applied viewpoint. [less ▲]

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See detailBelowground Chemical Ecology: The Case of Wireworms
Barsics, Fanny ULg; Delory, Benjamin ULg; Delaplace, Pierre ULg et al

Conference (2014, December 13)

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See detailPlutella xylostella (L.) infestations at varying temperatures induce the emission of specific volatile blends by Arabidopsis thaliana (L.) Heynh
Hien, Truong Thi Dieu ULg; Delory, Benjamin ULg; Brostaux, Yves ULg et al

in Plant Signaling & Behavior (2014)

The effect of combined abiotic and biotic factors on plant volatile organic compound (VOC) emissions is poorly understood. This study evaluated the VOC emissions produced by Arabidopsis thaliana (L.) Col ... [more ▼]

The effect of combined abiotic and biotic factors on plant volatile organic compound (VOC) emissions is poorly understood. This study evaluated the VOC emissions produced by Arabidopsis thaliana (L.) Col-0 subjected to three temperature regimes (17, 22, and 27 °C) in the presence and absence of Plutella xylostella larvae over two time intervals (0–4 and 4–8 h), in comparison to control plants. The analyses of VOCs emitted by Arabidopsis plants were made by headspace solid phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS). It was found that certain volatile groups (e.g., alcohols, ketones, aldehydes, and terpenes) are induced by both single factors (temperature or larval infestation) and combined factors (temperature and larvae interactions), whereas other volatile groups (e.g., isothiocyanates [ITCs] and nitrile) were specific to the experimental conditions. ITCs (mainly 4-methylpentyl isothiocyanate) were emitted from plants subjected to larval infestation at 17 and 27 °C after the two time intervals. The proportions of sulfides (mainly dimethyl disulfide) and 4-(methylthio) butanenitrile were significantly higher on herbivore-infested plants at 22 °C compared to the other treatments. Overall, our findings indicate that changes in all experimental conditions caused significant changes to the VOC emissions of Arabidopsis plants. Therefore, the interaction between temperature and larval feeding may represent an important factor determining the variability of volatile emissions by plants subjected to multiple simultaneous factors. [less ▲]

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See detailBarley (Hordeum distichon L.) roots produce volatile aldehydes via the lipoxygenase/hydroperoxide lyase pathway with a strong age-dependent pattern
Delory, Benjamin ULg; Delaplace, Pierre ULg; du Jardin, Patrick ULg et al

Conference (2014, August 13)

In chemical ecology, the roles played by root-emitted volatile organic compounds (VOCs) in biotic interactions and the quantitative analysis of such chemicals in root tissues remain poorly documented. In ... [more ▼]

In chemical ecology, the roles played by root-emitted volatile organic compounds (VOCs) in biotic interactions and the quantitative analysis of such chemicals in root tissues remain poorly documented. In this context, this study aims at using a fully automated gas chromatography – mass spectrometry methodology allowing both identification and accurate quantification of VOCs produced by roots of a monocotyledonous plant species at five selected developmental stages from germination to the end of tillering. Results show that barley roots mainly produce four volatile aldehydes, namely hexanal, (E)-hex-2-enal, (E)-non-2-enal and (E,Z)-nona-2,6-dienal. These molecules are well-known linoleic and linolenic acid derivatives produced via the lipoxygenase/hydroperoxide lyase pathway of higher plants. Our findings contrast with analyses documented on aboveground barley tissues that mainly emit C6 aldehydes, alcohols and their corresponding esters. Multivariate statistical analyses performed on individual VOC concentrations indicate quantitative changes in the volatile profile produced by barley roots according to plant age. Barley roots produced higher total and individual VOC concentrations when young seminal roots emerged from the coleorhizae compared to older phenological stages. Moreover, results also show that the C6/C9 volatile aldehyde ratio was the lowest at the end of tillering while the maximum mean value of this ratio was reached in seven day-old barley roots. [less ▲]

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See detailBelowground Chemical Ecology: The Case of Wireworms
Barsics, Fanny ULg; Delory, Benjamin ULg; Delaplace, Pierre ULg et al

Poster (2014, June)

Wireworms, clock-beetles' larvae (Coleoptera, Elateridae), are below ground pests of many crops. They cause dramatic yield losses worldwide. Research on their ecology is crucial to undertake innovative ... [more ▼]

Wireworms, clock-beetles' larvae (Coleoptera, Elateridae), are below ground pests of many crops. They cause dramatic yield losses worldwide. Research on their ecology is crucial to undertake innovative management strategies. In the field of chemical ecology, multitrophic interactions occurring in the rhizosphere are gaining increasing attention from entomologists and agronomists. Our research aims at unveiling the role of volatile organic compounds (VOC) involved in wireworms' foraging behavior, putatively leading to host localization and/or host recognition. [less ▲]

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See detailTemperature regimes and aphid density interactions differentially influence VOC emissions in Arabidopsis
Hien, Truong Thi Dieu ULg; Delory, Benjamin ULg; Vanderplanck, Maryse et al

in Arthropod-Plant Interactions (2014), 8(4), 317-327

The effects of volatile emissions from plants exposed to individual abiotic and biotic stresses are well documented. However, the influence of multiple stresses on plant photosynthesis and defense ... [more ▼]

The effects of volatile emissions from plants exposed to individual abiotic and biotic stresses are well documented. However, the influence of multiple stresses on plant photosynthesis and defense responses, resulting in a variety of volatile profiles has received little attention. In this study, we investigated how temperature regimes in the presence and absence of the sucking insect Myzus persicae affected volatile organic compound emissions in Arabidopsis over three time periods (0-24 h, 24-48 h, and 48-72 h). Headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry was used to evaluate Arabidopsis volatile organic compounds. The results showed that under laboratory conditions, eight volatile classes [alcohols (mainly 2-ethyl-hexan-1-ol), ketone (6-methyl hept-5-en-2-one), esters (mainly (Z)-3-hexenyl acetate), aldehydes (mainly phenylacetaldehyde), isothiocyanates (mainly 4-methylpentyl isothiocyanate), terpenes (mainly (E,E)-α-farnesene), nitrile (5-(methylthio) pentanenitrile), and sulfide (dimethyl trisulfide)] were observed on plants exposed to stress combinations, whereas emissions of six volatile classes were observed during temperature stress treatments alone (with the exception of nitriles and sulfides). Aphid density at high temperature combinations resulted in significantly higher isothiocyanate, ester, nitrile and sulfide proportions. The results of the present study provide an insight into the effects of temperature - aphid interactions on plant volatile emissions. [less ▲]

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See detailDevelopment of an ex-vitro system allowing plant-bacteria interactions through VOCs in the context of water stress
Mendaluk, Magdalena ULg; Baudson, Caroline ULg; Delory, Benjamin ULg et al

Poster (2014, February 07)

Water stress is one of the major environmental factors limiting the crop productivity . Plant stress responses are very complex and drought tolerance may be linked to the presence of specific ... [more ▼]

Water stress is one of the major environmental factors limiting the crop productivity . Plant stress responses are very complex and drought tolerance may be linked to the presence of specific microorganisms in the rhizosphere. Indeed, some plant growth promoting bacteria (PGPR) strains have been found to improve plant growth under abiotic stresses. Among the many mechanisms by which those PGPR can support plant growth, the emission of volatile organic compounds (VOCs) and their biological impact are still under study. The aim of this work is to evaluate the interaction between the model grass Brachypodium distachyon (Bd21) and two strains of PGPR. The impact of volatile emission on Bd21 growth was studied using an ex-vitro cocultivation system without physical contact between plant and bacteria during 10 days. This peculiar system was developed to assess bacterial VOCs impacts on plants under realistic growth and stress conditions. In parallel, the response of Bd21 seedlings to water deficit induced by polyethylene glycol 6000 (PEG 6000) was studied to establish contrasted growth conditions regarding water availability [less ▲]

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See detailQuantitative gas chromatography - mass spectrometry profiling of volatile organic compounds produced by barley (Hordeum distichon L.) roots according to plant age
Delory, Benjamin ULg; Delaplace, Pierre ULg; du Jardin, Patrick ULg et al

Poster (2014, February 07)

In chemical ecology, the roles played by root-emitted volatile organic compounds (VOCs) in biotic interactions and the quantitative analysis of such chemicals in root tissues remain poorly documented. In ... [more ▼]

In chemical ecology, the roles played by root-emitted volatile organic compounds (VOCs) in biotic interactions and the quantitative analysis of such chemicals in root tissues remain poorly documented. In this context, this study aims at developing a fully automated analytical methodology allowing both identification and accurate quantification of VOCs produced by roots of a monocotyledon plant species. Briefly, VOC emitted by crushed barley roots are successively trapped by dynamic headspace sampling on Tenax TA adsorbents, thermally desorbed and cryofocused, separated by gas chromatography (GC) and finally analysed by mass spectrometry (MS) in both SCAN and selected ion monitoring modes. Results show that barley roots mainly produce four volatile aldehydes, namely hexanal, (E)-hex-2-enal, (E)-non-2-enal and (E,Z)-nona-2,6-dienal. These molecules are well-known linoleic (C18:2) and linolenic (C18:3) acid derivatives produced via the lipoxygenase and the hydroperoxide lyase pathways of higher plants. Our findings contrast with analyses documented on aboveground barley tissues that mainly emit C6 aldehydes, alcohols and their derivative esters. Moreover, preliminary results indicate quantitative changes in the volatile profile contained in barley roots according to plant age. Multivariate statistical analyses are currently underway to quantitatively assess these changes using plants at five selected developmental stages ranging from germination to the end of tillering. [less ▲]

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See detailarchiDART: a R package allowing root system architecture analysis using Data Analysis of Root Tracings (DART) output files
Delory, Benjamin ULg; Baudson, Caroline ULg; Brostaux, Yves ULg et al

Poster (2014, February 07)

In 2010, Le Bot et al presented a free and open-access software (Data Analysis of Root Tracings - DART) allowing the analysis of complex root system architectures from captured images, particularly across ... [more ▼]

In 2010, Le Bot et al presented a free and open-access software (Data Analysis of Root Tracings - DART) allowing the analysis of complex root system architectures from captured images, particularly across time series. Using this software, a user has to manually identify roots as a set of links. After vectorization of a root system, three final data sets (RAC, TPS and LIE) can be exported as table files containing several attributes for (a) each individual root (e.g. root length), (b) each observation day or (c) each point used to construct the vectorized root system respectively. These data sets can finally be used either to calculate derived root system architecture (RSA) parameters or to draw the root system architecture at selected observation dates. However when an experiment involves the analysis and comparison of many root systems, the calculation of RSA parameters for each data set and the drawing of the corresponding vectorized root systems become time-consuming. In this context, we developed a R package, called archiDART, allowing both the automatic calculation of common root architecture parameters and the X-Y plotting of vectorized root systems for selected observation dates. [less ▲]

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