X-ray absorption and phase contrast imaging to study the interplay between plant roots and soil structure

Plant performance is, at least partly, linked to the location of roots with respect to soil structure features and the micro-environment surrounding roots. Measurements of root distributions from intact samples, using optical microscopy and field tracings have been partially successful but are imprecise and labour-intensive. Theoretically, X-ray computed micro-tomography represents an ideal solution for non-invasive imaging of plant roots and soil structure. However, before it becomes fast enough and affordable or easily accessible, there is still a need for a diagnostic tool to investigate root/soil interplay. Here, a method for detection of undisturbed plant roots and their immediate physical environment is presented. X-ray absorption and phase contrast imaging are combined to produce projection images of soil sections from which root distributions and soil structure can be analyzed. The clarity of roots on the X-ray film is sufficient to allow manual tracing on an acetate sheet fixed over the film. In its current version, the method suffers limitations mainly related to (i) the degree of subjectivity associated with manual tracing and (ii) the difficulty of separating live and dead roots. The method represents a simple and relatively inexpensive way to detect and quantify roots from intact samples and has scope for further improvements. In this paper, the main steps of the method, sampling, image acquisition and image processing are documented. The potential use of the method in an agronomic perspective is illustrated using surface and sub-surface soil samples from a controlled wheat trial. Quantitative characterization of root attributes, e.g. radius, length density, branching intensity and the complex interplay between roots and soil structure, is presented and discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Regulation and function of root exudates

Root-secreted chemicals mediate multi-partite interactions in the rhizosphere, where plant roots continually respond to and alter their immediate environment. Increasing evidence suggests that root exudates initiate and modulate dialogue between roots and soil microbes. For example, root exudates serve as signals that initiate symbiosis with rhizobia and mycorrhizal fungi. In addition, root exudates maintain and support a highly specific diversity of microbes in the rhizosphere of a given particular plant species, thus suggesting a close evolutionary link. In this review, we focus mainly on compiling the information available on the regulation and mechanisms of root exudation processes, and provide some ideas related to the evolutionary role of root exudates in shaping soil microbial communities.     

A method to separate plant roots from soil and analyze root surface area

Analysis of the effects of soil management practices on crop production requires knowledge of these effects on plant roots. Much time is required to wash plant roots from soil and separate the living plant roots from organic debris and previous years’ roots. We developed a root washer that can accommodate relatively large soil samples for washing. The root washer has a rotary design and will accommodate up to 24 samples (100 mm diam. by 240 mm long) at one time. We used a flat-bed scanner to digitize an image of the roots from each sample and used a grid system with commercially-available image analysis software to analyze each sample for root surface area. Sensitivity analysis and subsequent comparisons of ‘dirty’ samples containing the roots and all the organic debris contained in the sample and ‘clean’ samples where the organic debris was manually removed from each sample showed that up to 15% of the projected image could be coveredwith debris without affecting accuracy and precision of root surface area measurements. Samples containing a large amount of debris may need to be partitioned into more than one scanning tray to allow accurate measurements of the root surface area. Sample processing time was reduced from 20 h, when hand separation of roots from debris was used, to about 0.5 h, when analyzing the image from an uncleaned sample. The method minimizes the need for preprocessing steps such as dying the roots to get better image contrast for image analysis. Some information, such as root length, root diameter classes and root weights, is not obtained when using this technique. Root length measurements, if needed, could be made by hand on the digital images. Root weight measurement would require sample cleaning and the advantage of less processing time per sample with this method would be lost. The significance of the tradeoff between information not obtained using this technique and the ability to process a greater number of samples with the time and personnel resources available must be determined by the individual researcher and research objectives.     

Root decisions

Root systems have recognizable developmental plans when grown in solution or agar; however, these plans often must be modified to cope with the prevailing conditions in the soil environment such as the avoidance of obstacles and the exploitation of nutrient-rich patches or water zones. The modular structure of roots enables them to respond to their environment, and roots are very adaptive at modifying growth throughout the root system to concentrate their efforts in the areas that are the most profitable. Roots also form associations with microorganisms as a strategy to enhance resource capture. However, while the responses of roots in nutrient patches are well-recognized, overall 'rules of response' and variation in strategy among plant species that can be applied in a number of different environments are still lacking. Finally, there is increasing evidence that root–root interactions are much more sophisticated than previously thought, and the evidence for roots to identify self from non-self roots will be briefly discussed.     

The dynamics of root meristem distribution in the soil

Plants must develop efficient root architectures to secure access to nutrients and water in soil. This is achieved during plant development through a series of expansion and branching processes, mostly in the proximity of root apical meristems, where the plant senses the environment and explores immediate regions of the soil. We have developed a new approach to study the dynamics of root meristem distribution in soil, using the relationship between the increase in root length density and the root meristem density. Initiated at the seed, the location of root meristems in barley seedlings was shown to propagate, wave‐like, through the soil, leaving behind a permanent network of roots for the plant to acquire water and nutrients. Data from observations on barley roots were used to construct mathematical models to describe the density of root meristems in space. These models suggested that the morphology of the waves of meristems was a function of specific root developmental processes. The waves of meristems observed in root systems of barley seedlings exploring the soil might represent a more general and fundamental aspect of plant rooting strategies for securing soil resources.     

Effects of short-term nitrogen addition on fine root biomass,lifespan and morphology of Castanopsis platyacantha in a subtropical secondary evergreen broad-leaved forest

Aims Fine roots are the principal parts for plant nutrients acquisition and play an important role in the underground ecosystem. Increased nitrogen (N) deposition has changed the soil environment and thus has a potential influence on fine roots. The purpose of this study is to reveal the effect of N deposition on biomass, lifespan and morphology of fine root.Methods A field N addition experiment was conducted in a secondary broad-leaved forest in subtropical China from May 2013 to September 2015. Three levels of N treatments: CK (no N added), LN (5 g·m^-2·a^-1), and HN (15 g·m^-2·a^-1) were applied monthly. Responses of fine root biomass, lifespan, and morphology of Castanopsis platyacantha to N addition were analyzed by using a minirhizotron image system from April 2014 to September 2015. Surface soil sample (0-10 cm) was collected in November 2014 and soil pH value, and concentrations of NH₄⁺-N and NO₃^--N were measured.Important findings The biomass and average lifespan of the fine roots of C. platyacantha were 128.30 g·m^-3 and 113-186 days, respectively, in 0-45 cm soil layer. Nitrogen addition had no significant effect on either fine root biomass or lifespan in 0-45 cm soil layer. However, LN treatment significantly decreased C. platyacantha root superficial area in 0-15 cm soil layer. HN treatment significantly decreased soil pH value. Our study indicated that short-term N addition influences soil inorganic N concentration and thus decreased pH value in surface soil, and thereafter affect fine root morphology. Short-term N addition, however, did not affect the fine root biomass, lifespan and morphology in subsoil.     

隔沟交替灌溉条件下玉米根系形态性状及结构分布

为揭示根系对土壤环境的适应机制,研究了隔沟交替灌溉条件下玉米根系形态性状及结构分布。以垄位和坡位的玉米根系为研究对象,利用Minirhizotrons法研究了根系（活/死根）的长度、直径、体积、表面积、根尖数和径级变化及其与土壤水分、土温和水分利用效率（WUE）的相关关系。结果表明,对于活根,在坡位非灌水区域复水后根系平均直径减小,而根系日均生长速率、单位面积土壤根系体积密度、根尖数和表面积均增大,并随灌水区域土壤水分的消退逐渐减小;对于死根,在坡位非灌水区域复水后根系日均死亡速率、根系体积密度、根尖数和表面积变化均减小,其中根系死亡速率和死根直径随土壤水分的消退逐渐降低,而死根体积密度、根尖数和表面积分布随土壤水分降低呈增大趋势;在垄位,根系形态分布趋势与坡位一致,除根系直径与与坡位比较接近外,其他根系形态值均小于坡位。将根系分成4个径级区间分析根系的形态特征,结果表明在根系长度和体积密度分布中以2.5－4.5 mm径级的根系所占比例最大,在根尖数和根系表面积分布中以0.0－2.5 mm径级的根系为主。通过显著性相关分析,死根直径、体积密度、活根表面积等根系形态与土壤含水率、土壤温度和WUE间均存在显著或极显著的正相关关系,部分根系形态指标（如根系的生长速率、活根体积密度）只与坡位土壤含水量、土壤温度具有明显的相关性,表明隔沟交替灌溉对坡位根系形态的调控作用比垄位显著。     

亚热带典型树种根毛特征及其与共生真菌的关系

根毛和共生真菌增加了吸收面积,提高了植物获取磷等土壤资源的能力。由于野外原位观测根表微观结构较为困难,吸收细根、根毛、共生真菌如何相互作用并适应土壤资源供应,缺乏相应的数据和理论。该研究以受磷限制的亚热带森林为对象,选取了21种典型树种,定量了根毛存在情况、属性变异,分析了根毛形态特征与共生真菌侵染率、吸收细根功能属性之间的关系,探讨了根表结构对低磷土壤的响应和适应格局。结果表明:1)在亚热带森林根毛不是普遍存在的, 21个树种中仅发现7个树种存有根毛, 4个为丛枝菌根(AM)树种, 3个为外生菌根(ECM)树种。其中,马尾松(Pinus massoniana)根毛出现率最高,为86%;2)菌根类型是理解根-根毛-共生真菌关系的关键,AM树种根毛密度与共生真菌侵染率正相关,但ECM树种根毛直径与共生真菌侵染率负相关; 3) AM树种根毛长度和根毛直径、ECM树种根毛出现率与土壤有效磷含量呈负相关关系。该研究揭示了不同菌根类型树种根毛-共生真菌-根属性的格局及相互作用,为精细理解养分获取策略奠定了基础。     

施用氮肥对落叶松人工林一级根外生菌根侵染及形态的影响

以落叶松人工林为研究对象,通过施N肥试验,对不同季节、不同土壤深度根系进行取样,研究了1级根外生菌根真菌侵染率和形态,及其与不同季节、土壤深度和土壤N有效性的关系.结果表明:外生菌根真菌对落叶松人工林1级根的侵染率显著受不同季节和土壤深度土壤N有效性的影响.在不同季节和土层之间,施N肥导致菌根真菌侵染率下降.与未侵染菌根真菌相比,菌根真菌侵染导致1级根形态发生明显改变,平均直径增加18.7%,平均根长缩短23.7%,比根长降低16.3%.这种根系形态变化在不同季节、不同土壤深度处理中表现明显.菌根真菌侵染改变了1级根形态,影响根系的生理生态过程.     

Growth and morphology of eucalypt seedling-roots, in relation to soil strength arising from compaction

Information on the response of root growth and morphology to soil strength is useful for testing suitability of existing and new tillage methods and/or for selecting plants suitable for a specific site with or without tillage. Although there is extensive published information on the root growth-soil strength relationships for annual agricultural plants, such information is scarce for woody, perennial tree species. The purpose of this study is to examine growth and morphology of the root systems of 17-day-old eucalypt seedlings with respect to variation in soil strength. Soil strength in this study was varied by compaction of a well-aggregated clay soil to bulk densities of 0.7–1.0 Mg m^-3 whilst maintaining adequate water availability and aeration for plant growth. Lengths and tip-diameters of primary and lateral roots were measured on the excavated root systems of seedlings.With increase in bulk density and also soil strength (expressed as penetrometer resistance), total length of primary and lateral roots decreased. There were 71 and 31% reduction in the lengths of primary and lateral roots respectively with an increase in penetrometer resistance from 0.4 to 4.2 MPa. This indicated primary roots to be more sensitive to high soil strength than the lateral roots. Average length of lateral roots and diameters of both primary and lateral root tips increased with an increase in soil strength as well. There was greater abundance of lateral roots (no. of lateral roots per unit length of primary root) and root hairs with increased soil strength. The observed root behaviour to variable soil strength is discussed in the context of compensatory growth of roots and overall growth of plants.     

An extended root phenotype: the rhizosphere,its formation and impacts on plant fitness

Plants forage soil for water and nutrients, whose distribution is patchy and often dynamic. To improve their foraging activities, plants have evolved mechanisms to modify the physicochemical properties and microbial communities of the rhizosphere, i.e. the soil compartment under the influence of the roots. This dynamic interplay in root?soil?microbiome interactions creates emerging properties that impact plant nutrition and health. As a consequence, the rhizosphere can be considered an extended root phenotype, a manifestation of the effects of plant genes on their environment inside and/or outside of the organism. Here, we review current understanding of how plants shape the rhizosphere and the benefits it confers to plant fitness. We discuss future research challenges and how applying their solutions in crops will enable us to harvest the benefits of the extended root phenotype.     

林木细根寿命及其影响因子研究进展

 细根周转要消耗大量的C,它影响森林生态系统C分配格局与过程和养分循环,对生态系统生产力具有重要意义。细根的周转取决于细根的寿命,细根寿命越短,周转越快,根系对C的消耗也越多。大量研究表明,细根的寿命与地上部分C向根系供应的多少有密切关系,同时也与细根直径大小、土壤中N和水分的有效性、土壤温度以及根际周围的土壤动物和微生物的活动有关。本文综述了国外近年来在该领域里的研究进展,特别是对控制细根寿命的机理和主要影响因子进行了评述,目的是引起国内研究者的关注,促进我国根系生态学的研究与发展。     

种间互作和施氮对蚕豆/玉米间作生态系统地上部和地下部生长的影响

为河西走廊绿洲灌区豆科/禾本科间作体系的养分管理提供科学依据,于2007年在武威绿洲农业试验站应用田间原位根系行分隔技术研究了蚕豆/玉米种间互作和施氮对玉米抽雄期的根系空间分布、根系形态和作物地上部生长的影响。研究结果表明:种间互作和施氮均增加了玉米和蚕豆在纵向和横向两个尺度上的根重密度、根长密度、根表面积、根系体积。根长密度和根表面积与两种作物产量和氮素吸收均呈正相关,而与蚕豆的根瘤重呈负相关;抽雄期的土壤含水量与玉米产量和养分吸收呈显著的负相关。玉米根系可以占据蚕豆地下部空间,但蚕豆的根却较少到间作玉米的地下部空间,也就是间作后增加了玉米根系水平尺度的生态位。蚕豆和玉米根系主要分布分别在0-40 cm浅土层和0-60 cm 土层,且间作玉米根系在60-120 cm比单作和分隔的多。因此,种间互作和施氮扩大了两作物根系纵向和横向的空间生态位,改变了作物根系形态,即扩展了两者水分和养分吸收的生态位,增加了作物吸收养分的有效空间,从而提高了间作生态系统的生产力。     

森林生态系统林木根系对优先流的影响

土壤水和溶质运移是土壤学和环境科学研究的难点和热点,优先流是一种常见的土壤溶质运移形式,绕过土壤基质而优先运移至地下水源,造成土壤养分的流失和水质的恶化。林木根系是土壤层的重要部分,其结构形态影响着优先流过程,为量化林木根系结构对土壤优先流的影响,以首都圈森林生态系统鹫峰定位监测站为研究区域,利用野外染色示踪与室内分析相结合的方法,定量分析根长密度和根系生物量在优先流区和基质流区的变化。结果表明:1)随着土层深度的增加,根长密度表现为减小的趋势,对径级d1 mm,1d3 mm和3d5 mm根系而言,根长密度在优先流区大于基质流区发生概率分别为66.7%,88.9%和83.3%;2)根系d1 mm对优先流贡献度最大,均值为94.8%,1d3 mm和3d5 mm根系对优先流贡献度较小,均值分别为4.3%和0.9%;3)研究点根系生物量进行统计,66.7%优先流区根系生物量大于基质流区根系生物量。开展根系对优先流的影响研究,有助于探明土壤水分运移规律,分析地表地下水质恶化根源,为生态环境安全提供理论指导和技术支持。     

Using a fine root extraction device to quantify small diameter corn roots (≥0.025 mm) in field soils

The study of fine roots growing under field conditions is limited by the techniques currently available for separating these roots from soil. This study had two objectives: to measure the total root length of field grown corn (Zea mays L.) by root diameter class, and to develop an inexpensive and efficient root washing device that would effectively capture all of the roots in a field soil sample. An inexpensive Fine Root Extraction Device (FRED) was constructed from readily available materials and was successful at extracting all roots, including very fine diameter roots (0.025 mm), from field soil samples. Greater than 99.7% of marked roots introduced to the FRED were recaptured by the device. Soil samples from three depths, and on three dates, from field grown corn were placed in the FRED. We found that more than 56% of total root length occurred in roots whose diameters were smaller than 0.175 mm, and more than 35% of root length occurred in roots smaller than 0.125 mm in diameter. Corn roots of the diameters described here have not been reported in field soils prior to this study. Root researchers who fail to measure these very fine roots will significantly underestimate root length density. Widespread use of the FRED should improve our understanding of root distribution in field soils.     

Root interactions and tomato growth in tomato/potato onion companion-cropping system under different phosphorus levels

When two plants interact, changes in plant growth are usually related to variations in root distribution and phosphorus (P) levels. However, root distributions and root tendencies are difficult to study because root systems grow beneath the soil surface. In this study, a transparent root box was used to observe interactions between root systems in situ, and the relation between tomato growth and root proliferation at different depths and distance from the rows at no P added and 120?mg kg^-1 P added levels were also tested. We found that tomato shoot and total biomass increased and roots grew deeper when companion cropped with potato onion under both P levels. Moreover, tomato roots tended to grow away from the potato onion roots. Our results suggest that a deeper and more evasive root distribution may be related to the increased plant growth of tomato when companion cropped with potato onion.     

Quantitative imaging of rhizosphere pH and CO2 dynamics with planar optodes

Background and Aims

Live imaging methods have become extremely important for the exploration of biological processes. In particular, non-invasive measurement techniques are key to unravelling organism–environment interactions in close-to-natural set-ups, e.g. in the highly heterogeneous and difficult-to-probe environment of plant roots: the rhizosphere. pH and CO₂ concentration are the main drivers of rhizosphere processes. Being able to monitor these parameters at high spatio-temporal resolution is of utmost importance for relevant interpretation of the underlying processes, especially in the complex environment of non-sterile plant–soil systems. This study introduces the application of easy-to-use planar optode systems in different set-ups to quantify plant root–soil interactions.

Methods

pH- and recently developed CO₂-sensors were applied to rhizobox systems to investigate roots with different functional traits, highlighting the potential of these tools. Continuous and highly resolved real-time measurements were made of the pH dynamics around Triticum turgidum durum (durum wheat) roots, Cicer arietinum (chickpea) roots and nodules, and CO₂ dynamics in the rhizosphere of Viminaria juncea.

Key Results

Wheat root tips acidified slightly, while their root hair zone alkalized their rhizosphere by more than 1 pH unit and the effect of irrigation on soil pH could be visualized as well. Chickpea roots and nodules acidified the surrounding soil during N₂ fixation and showed diurnal changes in acidification activity. A growing root of V. juncea exhibited a large zone of influence (mm) on soil CO₂ content and therefore on its biogeochemical surrounding, all contributing to the extreme complexity of the root–soil interactions.

Conclusions

This technique provides a unique tool for future root research applications and overcomes limitations of previous systems by creating quantitative maps without, for example, interpolation and time delays between single data points.     

Root:soil adhesion in the maize rhizosphere: the rheological approach

This study was designed to investigate the strength of attachment of plant seedling roots to the soil in which they were grown. The study also assessed the effects of differing soil textures and differing soil matric potentials upon the strength of the root:soil attachment. A device for growing roots upon a soil surface was designed, and was used to produce roots which were attached to the soil. In order to quantify root:soil adhesion, roots of maize seedlings, grown on the soil surface, were subsequently peeled off using a universal test machine, in conjunction with simultaneous time-lapse video observation. To clarify the partitioning of energy in the root:soil peeling test, separate mechanical tests on roots, and on two adherent remoulded topsoil balls were also carried out. The seedling root was characterised by a low bending stiffness. The energy stored in bending was negligible, compared to the root:soil adhesion energy. The mechanical properties of two adherent remoulded topsoil balls were a decrease of the soil:soil adhesion energy as the soil:soil plastic energy increased. These two parameters were therefore interdependent. Using a video-camera system, it was possible to separate the different processes occurring during the root:soil peeling test, in particular, the seed:soil adhesion and the root:soil soil adhesion. An interpretation of the complex and variable force:displacement curves was thus possible, enabling calculation of the root:soil interfacial rupture energy. At a given suction (10 kPa), the results of the peeling test showed a clear soil texture effect on the value of the root:soil interfacial rupture energy. In contrast, for the same silty topsoil, the effect of the soil water suction on the value of the interfacial rupture energy was very moderate. The root:soil interfacial rupture energy was controlled mainly by a product of microscopic soil specific surface area and the macroscopic contact surface area between the root and the soil. Biological and physical interactions contributing to root:soil adhesion such as root:soil interlocking mechanics were also analysed and discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

小麦根系与土壤水分胁迫关系的研究进展

几十年来,大量科学工作者为拓宽小麦根系对土壤水分的吸收能力和调控根系对干旱的适应能力,挖掘干旱地区的生产潜力,实现高产做了大量细致的研究工作,取得了许多重要研究成果,综述了土壤水分胁迫对小麦根系形态、构型建成和生理指标影响的影响。过去进行的研究表明,干旱胁迫条件下,不仅表达小麦根系形态和构型建成指标的根系数量、根系比表面积、根冠比、根生长势、根水势,导管直径等发生显著变化,而且表达根系生理指标的伤流流、根呼吸速率、根系质膜透性、膜脂过氧化水平、保护酶及其同工酶等也发生相应改变,虽然不同的研究者所获得的研究结果不同,有的甚至相互矛盾,但从总体看,各种变要是对干旱胁迫的一种适应性反应,有利于提高小麦的抗旱能力,对干旱条件下产量的形成具有重要作用。