Issues in using the WinRHIZO system to determine physical characteristics of plant fine roots☆

Image analysis systems have facilitated rapid measurements of fine root length (RL), diameter (RD), volume (RV), etc. The WinRHIZO system is unlike other image analysis systems in that it can detect, and make corrections for, areas of root overlap. It is designed to be capable of using both Regent’s non-statistical method (WinRHIZO method) and Tennant’s statistical method (line-intersect method), and can simultaneously output the root measurements by both methods when they are chosen at the same time. This study tested: (1), the efficacy of the overlap correction function in the WinRHIZO system; and (2), the consistency of fine root measurements between the WinRHIZO and Tennant methods with two sets of root measurement data from winter wheat (Triticum aestivium L.). The results showed that there were significant differences in RL, RD and RV between small root samples with and without stumps. The impact of root stumps outweighed the overlap correction efficacy in WinRHIZO. The values from the Tennant method are significantly different from those using the WinRHIZO method, although both results are statistically closely correlated. This indicated how critical it was to use without-stump root samples when using image analysis systems to measure RL, RD, RV, etc., and to keep in mind that a significant difference in root measurements may be methodologically related when comparing the results of various experiments from these two methods. Our research results bear important implications for the study of root ecology.     

Sample preparation and scanning protocol for computerised analysis of root length and diameter

Root length and diameter distribution are important characteristics to be considered when describing and comparing root systems. Root length and root-diameter distribution may be obtained in two ways: by microscopical measurements, which are laborious, or by computerised analysis, which is fast but sensitive to the scanning protocol. Although scanning protocols vary widely between laboratories, papers rarely report the details. Using two commercially available root analysis software packages (Delta-T Scan and WinRHIZO), we performed a sensitivity analysis of scanning protocols for estimating root length and diameter distribution. The results are shown graphically, rather than numerically, because the interactions between different parameters in the overall scanning protocol are most clearly illustrated by the shape of the curves. The present analysis clearly demonstrated the sensitivity of the two scanning methods with regard to staining period, maximum root density, scanning resolution and transformation threshold. For example, estimating the root-diameter distribution versus measuring root length, puts opposite constraints on the transformation threshold settings. We suggest the following settings for the most sensitive parameters: a staining period of 24 h, a root density of less than 0.5 mm root per mm² surface, a resolution of 400 dpi and the automatic threshold for WinRHIZO and a brightness of 200 for Delta-T Scan. According to this protocol, comparison of computerised analyses with microscopic measurements showed good total root length and diameter distribution agreement for three contrasting root systems. We suggest to always start with the present protocol when studying other species. If validation indicates that the proposed scanning protocol needs to be modified for other species, the present sensitivity analysis may be used as a guideline for changing the most critical parameters. Similarly, the use of another stain than neutral red may also require modifications of the scanning protocol. In general, a long staining period (e.g., 24 h) is recommended for all stains, as small differences in staining period have the least effect when the root tissue is saturated. To enhance comparability of results in the literature, the staining period, stain, (maximum) root density, scanner resolution and threshold should always be listed when root data obtained by scanning are presented. This revised version was published online in August 2006 with corrections to the Cover Date.     

IJ_Rhizo: an open-source software to measure scanned images of root samples

Background and aims

This paper provides an overview of the measuring capabilities of IJ_Rhizo, an ImageJ macro that measures scanned images of washed root samples. IJ_Rhizo is open-source, platform-independent and offers a simple graphic user interface (GUI) for a main audience of non-programmer scientists. Being open-source based, it is also fully modifiable to accommodate the specific needs of the more computer-literate users. A comparison of IJ_Rhizo’s performance with that of the widely used commercial package WinRHIZO? is discussed.

Methods

We compared IJ_Rhizo’s performance with that of the commercial package WinRHIZO? using two sets of images, one comprising test-line images, the second consisting of images of root samples collected in the field. IJ_Rhizo and WinRHIZO? estimates were compared by means of correlation and regression analysis.

Results

IJ_Rhizo “Kimura” and WinRHIZO? “Tennant” were the length estimates that were best linearly correlated with each other. Correlation between average root diameter estimates was weaker, due to the sensitivity of this parameter to thresholding and filtering of image background noise.

Conclusions

Overall, IJ_Rhizo offers new opportunities for researchers who cannot afford the cost of commercial software packages to carry out automated measurement of scanned images of root samples, without sacrificing accuracy.     

干旱对杉木幼苗根系构型及非结构性碳水化合物的影响

通过比较不同时期不同强度干旱胁迫下杉木1年生盆栽苗地上部分生长、根系构型以及根系中非结构性碳水化合物含量（TNC）的变化,并分析各指标之间的相关性,探究杉木根系在干旱胁迫下的适应性策略以及抗旱生理机制,以期为杉木造林生产和水分管理提供科学依据和技术指导。结果表明：随着干旱程度的加强,杉木幼苗地上部分干重（SDW）、根干重（RDW）、根长（RL）、根表面积（SA）、根体积（RV）、根尖数（RT）、根系分支角度（Angle）、分形维数（FD）逐渐减小,根冠比（R/T）逐渐增大,根系拓扑指数（TI）、根系平均直径（RD）先增大后减小,比根长（SRL）先减小后增大。而根系连接长度（LL）、TNC、糖淀比在不同时期表现出不同的趋势。连接长度随着干旱胁迫的加强在30 d和60 d时表现出逐渐增加趋势而在90 d时则表现出先减小后增大的趋势。TNC在30 d和60 d时先增大后减小,但90 d时,呈逐渐下降的趋势。糖淀比随着干旱胁迫的加强在30 d和60 d时表现出先增加后减小趋势,90 d时,表现为先减小后增大。干旱胁迫显著影响根系在不同径级的分布长度,且随着胁迫时间的延长不断变化。杉木地上部分生长与根系生长指标（RL、SA、RV、RT、RDW）以及根系构型指标（Angle、FD）之间存在显著的正相关（P < 0.01）,根系平均直径与TNC存在显著的正相关（P < 0.05）。总之,杉木通过增加根系光合产物的积累、提高根系建成成本,增加有限成本下根系的复杂程度和延伸范围,降低根系分支角度,使根系"更陡更深"来适应不同强度的干旱胁迫。     

Measurement of fine root tissue density: a comparison of three methods reveals the potential of root dry matter content

Aims

Root tissue density (RTD, the ratio of root dry mass to root volume) is a fundamental trait in comparative root ecology, being increasingly used as an indicator of plant species’ resource use strategy. However, the lack of standardized method to measure this trait makes comparisons tricky. This study aims to compare three methods commonly used for determining fine RTD and to test whether root dry matter content (RDMC, the ratio between root dry mass and root fresh mass) could be used as a surrogate of fine root tissue density.

Methods

RTD of 163 fine root samples was determined using (i) Archimedes’ method, (ii) image analysis (WinRHIZO software), and (iii) using the root dry matter content as a proxy. Root samples belonged to different herbaceous species grown in different conditions.

Results

RTD measured with Archimedes’ method was positively correlated with RTD estimated with image analysis and with RDMC. However we demonstrated that RTD measured with Archimedes’ method was better predicted by RDMC (R²?=?0.90) than by RTD measured with image analysis (R²?=?0.56). The performance and limitations of each method were discussed.

Conclusion

RDMC is a quick, cheap and relatively easy measurable root attribute; we thus recommended its measurement as a proxy of fine root tissue density.     

Use of a lux-based procedure to rapidly visualize root colonisation by Pseudomonas fluorescens in the wheat rhizosphere

The bioluminescently marked Pseudomonas fluorescens strain 5RL, has been used previously to follow colonisation of soy bean roots (De Weger et al. [1991] Appl. Environ. Microbiol. 57:36-41). In the present paper the method has been further developed and optimized for wheat roots and it is used to get a quick overview of the colonisation patterns of many different root systems at the same time. Colonisation was followed on wheat plants grown in our gnotobiotic sand system (Simons et al., 1996. Mol Plant Microbe Interact 9: 600–607) and the following results were obtained. (i) A spatio-temporal analysis of the colonisation of wheat roots showed that 4 days after planting the highest bacterial activity was observed at the upper part of the root. After 6 days the high bacterial activity at the upper part was further increased, whereas spot-like activities were observed on the lower root parts, possibly due to micro-colonies. (ii) Bacterial mutations causing lack of motility or auxotrophy for amino acids resulted in impaired colonisation of the lower root parts, indicating that motility and prototrophy for the involved amino acid(s) are important factors for wheat root colonisation by strain 5RL. (iii) Coinoculation of strain 5RL with other wild type Pseudomonas strains on the root influenced the colonisation pattern observed for strain 5RL. Colonisation was not visually affected when the competing strain was a poor root coloniser, but was severely reduced when the competing strain was a good root coloniser. The results show that the spatio-temporal colonisation of wheat root by P. fluorescens strain 5RL and derivatives is similar to that of strain WCS365 on tomato. The advantage of the use of lux-marked strains is that the results are obtained much quicker than when conventional methods are used and that the result is supplied as an image of the colonisation pattern of many different roots.     

不同氮素添加量对大兴安岭冻土区泥炭地植物细根形态的影响模拟研究

为探讨氮素营养环境变化对冻土区泥炭地植物细根形态的影响,在大兴安岭泥炭地开展了不同浓度氮素添加模拟试验,添加量分别为0 g N m^-2 a^-1（CK）、6 g N m^-2 a^-1（N1）、12 g N m^-2 a^-1（N2）和24 g N m^-2 a^-1（N3）。在2020年8月和9月,利用微根管技术观测泥炭地不同深度（0-20 cm、20-40 cm）土壤中的植物细根形态,应用WinRHIZO图像分析软件分析根系特征。结果表明,在表层土壤（0-20 cm）中植物细根的总根长、总表面积、总体积和根长密度随施氮量增加而增加,其中8月份N3处理下细根总根长、总表面积、总体积和根长密度显著高于其他处理（P< 0.05）,N2处理下细根总表面积、总体积显著高于对照组和N1处理;9月份N3处理下细根总根长和根长密度显著高于对照组,总表面积和总体积显著高于对照组和N1处理。说明高浓度氮素添加在一定程度上缓解了植物氮素限制,能够显著促进表层土壤（0-20 cm）中植物细根的生长,但对亚表层土壤（20-40 cm）中细根的影响幅度小于表层土壤。     

Do propagation methods affect the fine root architecture of African plum (Dacryodes edulis)?

Belowground tree growth attributes determine whether associations will be complementary or competitive in an agroforestry context. A study on fine root (d?≤?2?mm) distribution patterns of Dacryodes edulis based on root density (RD), root length density (RLD) and root weight density (RWD) was conducted to evaluate the effect of propagation methods on rooting distribution. Results showed that D. edulis trees of seed origin had greater RD (P?≤?0.001) than trees of vegetative origin (cuttings and marcots) in the upper soil stratum (0–30?cm). Similarly, in the uppermost soil stratum (0–10?cm), RLD and RWDs varied significantly (P?<?0.01). Trees of seed origin had an exponential distribution pattern for fine RD, RLD and RWD with depth to 80?cm. In contrast, the distribution pattern of fine roots of trees of vegetative origin (cuttings and marcots) were quadratic for the same variables which increased in the 20–30?cm soil depth stratum before declining steadily to a depth of 80?cm. The findings of this study suggest that D. edulis trees of vegetative origin (cuttings and marots) are likely to be less competitive than trees of seed origin when intercropped with shallow-rooted annual plants in an agroforestry system for belowground resources.     

增温、施肥与种内竞争的交互作用对云杉根系属性的影响

增温、施肥与种内竞争的交互作用对云杉根系属性的影响 物种竞争、气温和土壤养分是青藏高原东部高寒地区影响树木生长的重要因素。虽然已开展了大量关于物种竞争、气温、施肥单因素对树木生长的影响研究,但关于这三者的交互作用对根系生长的影响还知之甚少。因此,本研究拟通过测量根系属性(细根长、根表面积、比根长、比表面积、根尖数、根系分支数等)、根生物量,以及根系养分吸收,研究施肥和增温对物种竞争的影响,并进一步探讨施肥、增温与物种竞争的交互作用对云杉(Picea asperata)生长的影响机制以及所采取的适应策略。研究结果表明,增温、施肥和竞争均提高了细根的氮、钾浓度,但并未影响细根生物量和根长、根表面积、根尖数和根分支数等根系特征。然而,无论是增温、施肥,或是它们的联合作用,与物种竞争进行交互时,均增加了根长、根表面积、根尖数、根系分支数和养分吸收。此外,施肥降低了根比表面积、比根长和单位面积的根尖数和根分支数,增温和竞争的交互作用使根比表面积、比根长下降,其他参数不受温度和竞争的影响。该结果表明,云杉在物种竞争、气候变暖、施肥及其交互作用下保持着保守的营养策略。该研究加强了对树木应对全球变化的生理和生态适应性的理解。     

Indirect emissions of forest bioenergy: detailed modeling of stump‐root systems

Indirect carbon dioxide emissions from producing bioenergy from tree stumps and roots depend critically on the decomposition rate of these harvest residues if they were left in forest to decay. We developed a method to improve the current estimates of these emissions. First, the 3D structure of uprooted stump‐root systems was modeled based on terrestrial laser‐scanning data. Second, information obtained on the size distribution of the stumps and the roots was used to simulate their decomposition and to estimate the indirect emissions. The method was able to describe the structure of stump‐root systems at a clear‐cut boreal Norway spruce test site. Compared with earlier results based on the diameter of stumps alone, the new estimates of the decomposition rate were slightly higher and, consequently, those of the indirect emissions slightly lower. The method is useful to collect information on the indirect emissions of stump‐root bioenergy quickly in different forests.     

Fine root turnover in forest ecosystems in relation to quantity and form of nitrogen availability: a comparison of two methods

Summary Two methods of estimating fine root production and turnover are compared for 13 forest ecosystems exhibiting a wide range in form (NH₄ ⁺ vs. NO₃ ^-) and quantity of available nitrogen. The two methods are by comparison of seasonal maxima and minima in biomess and by nitrogen budgeting. Both methods give similar results for stands with low rates of nitrification. The budgeting method predicts higher fine root turnover and productivity than the max-min method for systems with significant rates of nitrification.     

A plane intersect method for estimating fine root productivity of trees from minirhizotron images

The advent of minirhizotrons more than a decade ago has made the careful and widespread study of fine root dynamics of trees possible. However, to this day, the estimation of fine root productivity in terms of mass production per unit of ground surface from the minirhizotron data remains hampered by the difficulty in transforming images of roots captured along a two-dimensional plane into estimates of root volume or mass within a soil volume. In this work, we propose that the date of fine root appearance and the diameter of fine roots are the most robust variables that can be obtained from minirhizotron measurements of tree roots and that these two variables should be the basis of productivity estimates. The method proposed for estimating fine root productivity expands the line intersect method of Van Wagner (1968) into a plane intersect method that permits, with the appropriate volumetric transformations and corrections for tube and slope angles, the estimation of fine root productivity per unit ground area for specific periods. Examples of calculations are presented for two datasets obtained within two different forested sites, as well as a comparison with a methodology based on camera depth-of-view. The main weakness of the plane intersect method is the assumption that all fine root segments are independent. The correction for the fraction of coarse particles also creates uncertainty in the final estimate.     

The potential of the decay fungus Chondrostereum purpureum in the biocontrol of broadleaved tree species

The ability of the decay fungus Chondrostereum purpureum to grow within birch (Betula pendula and Betula pubescens), aspen (Populus tremula), rowan (Sorbus aucuparia) and willow (Salix caprea) stumps or root systems is unknown, although such information would be crucial in understanding its impact on the vegetative growth of broadleaved trees, their ecology and potential biocontrol. Saplings of these tree species were cut and inoculated with the fungus. After 3 months, 47, 14, 0 and 0% of birch, aspen, rowan and willow stumps, respectively, were dead, and C. purpureum was frequently present within the stumps of all species. In more than half of the birch stumps investigated, the fungus had penetrated into the roots unlike in the other tree species. Our results indicate that C. purpureum can utilize the woody material of birch better than that of other species, and that penetration into the roots is needed to kill the host.     

Modelling root structure and stability

Summary Seven fully excavated 16 year-old root systems of Sitka spruce were analysed. All roots in excess of 1 cm diameter at their origin on the stumps were analysed, data being collected until root diameter declined to less than 0.5 cm.Root morphology and distribution was identified as a balance between systematic biological mechanisms and their disruption by environmental factors, particularly changes of soil density and soil surface contours. The biological mechanisms have been modelled and the model is capable of simulating root systems in response to a few simple input variablese.g. the number of roots originating at stumps, stem ratius, total number of roots of all ordersetc.Additionally equations have been developed to estimate the distribution of root diameter, and root weight at all distances from tree stems and a similar equation permits the estimation of tree diameter at chosen heights. These latter estimates being utilized to calculate the turning moment of stems when blown by the wind.The influence of the wind on turning moment is explored for simulated root systems of differing strength and gross morphology.     

Accurate root length measurement by image analysis

Algorithms for estimating root length by image analysis should lead to results that have no systematic error (bias), be insensitive to preferential root orientation, valid across a wide range of sample sizes and adjust for overlap between roots in samples, to reduce the effort needed in spreading out root systems. We propose a new algorithm that forms a compromise between small bias and robustness (insensitivity to variation in sample size and preferential root orientation), and provide a simple way of dealing with root overlap. Image analysis was performed on a Macintosh computer using the public domain NIH Image program. The digital image of the root was processed to get the thinned image (skeleton). The numbers of orthogonally and diagonally connected pairs of pixels (N _o and N _d, respectively) in the skeleton were counted separately and used for length (L) calculation. A new length calculation equation was introduced so that the effect of orientation on length calculation was minimized; L=[N d ²+(N _d+N _o/2)²]^1/2+N _o/2. The maximum error due to orientation of a single line was evaluated for an ideal line, and the analysis revealed that the new equation was less affected by orientation than previous equations. Copper wire and rice (Oryza sativa L.) roots containing both primary and fine secondary root were measured manually and with image analysis. The two methods showed good agreement within 1.5%. The proposed image analysis method yielded length estimates with CV from 0.23 to 0.88%, which was lower than the CVs of the line-intersect method. Moreover, the lengths of overlapping samples were calculated correctly because the image analysis method distinguished an overlapping pixel from a thinned image, while the calculation with the line-intersect method showed underestimation because overlaps were not considered in that method. This revised version was published online in June 2006 with corrections to the Cover Date.     

Mapping quantitative trait loci for root development under hypoxia conditions in soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L. Merr.)

Key message

Greatest potential, QTLs for hypoxia and waterlogging tolerance in soybean roots were detected using a new phenotypic evaluation method.

Abstract

Waterlogging is a major environmental stress limiting soybean yield in wet parts of the world. Root development is an important indicator of hypoxia tolerance in soybean. However, little is known about the genetic control of root development under hypoxia. This study was conducted to identify quantitative trait loci (QTLs) responsible for root development under hypoxia. Recombinant inbred lines (RILs) developed from a cross between a hypoxia-sensitive cultivar, Tachinagaha, and a tolerant landrace, Iyodaizu, were used. Seedlings were subjected to hypoxia, and root development was evaluated with the value change in root traits between after and before treatments. We found 230 polymorphic markers spanning 2519.2 cM distributed on all 20 chromosomes (Chrs.). Using these, we found 11 QTLs for root length (RL), root length development (RLD), root surface area (RSA), root surface area development (RSAD), root diameter (RD), and change in average root diameter (CARD) on Chrs. 11, 12, 13 and 14, and 7 QTLs for hypoxia tolerance of these root traits. These included QTLs for RLD and RSAD between markers Satt052 and Satt302 on Chr. 12, which are important markers of hypoxia tolerance in soybean; those QTLs were stable between 2 years. To validate the QTLs, we developed a near-isogenic line with the QTL region derived from Iyodaizu. The line performed well under both hypoxia and waterlogging, suggesting that the region contains one or more genes with large effects on root development. These findings may be useful for fine mapping and positional cloning of gene responsible for root development under hypoxia.

     

Drought affects the coordination of belowground and aboveground resource‐related traits in Solidago canadensis in China

Quantifying patterns of variation and coordination of plant functional traits can help to understand the mechanisms underlying both invasiveness and adaptation of plants. Little is known about the coordinated variations of performance and functional traits of different organs in invasive plants, especially in response to their adaptation to environmental stressors. To identify the responses of the invasive species Solidago canadensis to drought, 180 individuals were randomly collected from 15 populations and 212 ramets were replanted in a greenhouse to investigate both the response and coordination between root and leaf functional traits. Drought significantly decreased plant growth and most of the root and leaf functional traits, that is, root length, surface area, volume and leaf size, number, and mass fraction, except for the root length ratio and root mass fraction. Phenotypic plasticity was higher in root traits than in leaf traits in response to drought, and populations did not differ significantly. The plasticity of most root functional traits, that is, root length (RL), root surface area (RSA), root volume (RV), and root mass fraction (RMF), were significantly positively correlated with biomass between control and drought. However, the opposite was found for leaf functional traits, that is, specific leaf area (SLA), leaf area ratio (LAR), and leaf mass fraction (LMF). Drought enhanced the relationship between root and leaf, that is, 26 pairwise root–leaf traits were significantly correlated under drought, while only 15 pairwise root–leaf traits were significantly correlated under control conditions. Significant correlations were found between biomass and all measured functional traits except for leaf size. RV, root length ratio, RMF, total area of leaves, and LMF responded differently to water availability. These responses enable S. canadensis to cope with drought conditions and may help to explain the reason of the vast ecological amplitude of this species.     

高寒草甸植被细根生产和周转的比较研究

植物根系是陆地生态系统重要的碳汇和养分库,细根周转过程是陆地生态系统地下部分碳氮循环的核心环节,在陆地生态系统如何响应全球变化中起着关键作用。在全球变化敏感地区之一的青藏高原,对该地区的主要植被类型矮嵩草草甸同时采用根钻法、内生长袋法和微根管法3种观测方法研究细根生产和周转速率,并探讨了极差法、积分法、矩阵法和Kaplan-Meier法等数据处理方法对计算值的影响。研究结果显示:在估算细根净初级生产力时,根钻法宜采用积分法,内生长袋法宜选用矩阵法;由此进一步以最大细根生物量为基础,根钻法和内生长袋法估测的细根年周转速率分别为0.36 a-1和0.52 a-1,内生长袋法的估算结果是根钻法的1.44倍。对于微根管法,将其观测得到的细根长度转换为单位面积的生物量值后,采用积分法计算出细根周转速率为0.84 a-1,远高于传统方法的估算结果;若采用Kaplan-Meier生存分析方法,则计算出的细根周转速率更高达3.41 a-1。     

土壤水分和氮磷营养对小麦根系生理特性的调节作用

研究结果表明：在土壤相对含水量(SRWC)为40％～70％范围内,随着水分亏缺加重,小麦根系生长受到限制,根系比表面积(RA)、根水势(RΨw)明显降低,根呼吸(RP)作用加强,根长(RL)变短,根系干物重(RDW)减少;随着土壤水分趋于良好,RΨw明显增加,Rp作用显著降低,SRWC在54％～57％时RA最高,RDW在SRWC为55％～62％之间时最大,而SRWC在55％上下时RL达最长,表明土壤水分趋于轻度干旱有利根系下扎和RA的提高,土壤水分趋于良好有利于根量增长。氮磷营养对根系生理特性具有明显作用。磷营养可显著提高RA,RΨw,RL,RDW,降低Rp,在较为严重干旱条件下,磷对RA,RP,RΨw,RL的调节效果更好。氮素营养对小麦根系生理特性的调节效果与磷相比具有明显的差别。土壤严重干旱,氮引起RΨw的明显下降,为负效应;轻度干旱氮对RΨw无明显作用;在水分良好条件下,氮对RΨw具有显著的正向调节效果。氮营养可使Rp作用明显加强。随着SRWC的提高,增施氮肥可导致RDW的显著增加。小麦RL对氮的反应不敏感。磷除作为一种营养物质促进作物根系生长发育外,在水分胁迫条件下,磷可明显改善植株体内的水分关系,增强对干旱缺水环境的适应能力,提高作物抗旱性。     

Conventional detection methodology is limiting our ability to understand the roles and functions of fine roots

We lack a thorough conceptual and functional understanding of fine roots. Studies that have focused on estimating the quantity of fine roots provide evidence that they dominate overall plant root length. We need a standard procedure to quantify root length/biomass that takes proper account of fine roots. Here we investigated the extent to which root length/biomass may be underestimated using conventional methodology, and examined the technical reasons that could explain such underestimation. Our discussion is based on original X-ray-based measurements and on a literature review spanning more than six decades. We present evidence that root-length recovery depends strongly on the observation scale/spatial resolution at which measurements are carried out; and that observation scales/resolutions adequate for fine root detection have an adverse impact on the processing times required to obtain precise estimates. We conclude that fine roots are the major component of root systems of most (if not all) annual and perennial plants. Hence plant root systems could be much longer, and probably include more biomass, than is widely accepted.