Root sampling methods - applications and limitations of the minirhizotron technique

Applications and limitations of the minirhizotron technique (non-destructive) in relation to two frequently used destructive methods (soil coreing and ingrowth cores) is discussed. Sequential coreing provides data on standing crop but it is difficult to obtain data on root biomass production. Ingrowth cores can provide a quick estimate of relative fine-root growth when root growth is rapid. One limitation of the ingrowth core is that no information on the time of ingrowth and mortality is obtained.The minirhizotron method, in contrast to the destructive methods permits simultaneous calculation of fine-root length production and mortality and turnover. The same fine-root segment in the same soil space can be monitored for its life time, and stored in a database for processing. The methodological difficulties of separating excavated fine roots into living and dead vitality classes are avoided, since it is possible to judge directly the successive ageing of individual roots from the images. It is concluded that the minirhizotron technique is capable of quantifying root dynamics (root-length production, mortality and longevity) and fine-root decomposition. Additionally, by combining soil core data (biomass, root length and nutrient content) and minirhizotron data (length production and mortality), biomass production and nutrient input into the soil via root mortality and decomposition can be estimated.     

A comparison between minirhizotron and monolith sampling methods for measuring root growth of maize (Zea mays L.)

Transparent plastic minirhizotron tubes have been used to evaluate spatial and temporal growth activities of plant root systems. Root number was estimated from video recordings of roots intersecting minirhizotron tubes and of washed roots extracted from monoliths of the same soil profiles at the physiological maturity stage of a maize (Zea mays L.) crop. Root length was measured by the line intercept (LI) and computer image processing (CIP) methods from the monolith samples.There was a slight significant correlation (r=0.28, p<0.005) between the number of roots measured by minirhizotron and root lengths measured by the LI method, however, no correlation was found with the CIP method. Using a single regression line, root number was underestimated by the minirhizotron method at depths between 0–7.6 cm. A correlation was found between root length estimated by LI and CIP. The slope of estimated RLD was significant with depth for these two methods. Root length density (RLD) measured by CIP showed a more erratic decline with distance from the plant row and soil surface than the LI method.     

Length densities,occupancies and weights of apple root systems

The root systems of apple trees from five orchards ranging in age from 1.5-y to 14-y were sampled to depths of between one and two metres using soil cores. Although trees came from orchards which differed in soil-type, tree spacings and management, consistent patterns were found in root systems. In orchards of 4-y and older, roots of adjacent trees met so that soil volumes within the planting grids (i.e, tree spacings of approximately 5 m inter-row×4 m intra-row distances) were completely explored, although not completely occupied by roots. Mean root-length densities declined with depth for these orchards. In the 1.5-y orchard, roots from adjacent trees did not meet and root-length densities declined with radial distance from the stem as well as with depth.Root-length densities in the top 1 m ranged from zero to about 1.0 cm.cm^–3 in all orchards and were highly variable. The proportions of core samples having zero values for root-length density were used to subdivide the root zone into volumes in which all samples contained roots, and volumes in which some samples had no roots.Results suggest that roots in an average tree penetrate to at least one metre depth in all but very young orchards so that soil in this volume is fully explored. Volumes filled by roots and volumes occupied at any particular root-length density appear to reach a maximum at about 4 years. Volumes of soil occupied at any particular root-length density were equal in all orchards older than 4 years. This suggests that root growth was balanced by root death. In contrast woody roots continue to accumulate with time.     

A comparison of minirhizotron techniques for estimating root length density in soils of different bulk densities

Flexible- and rigid-walled minirhizotron techniques were compared for estimating root length density of 14- to 28-day-old Pinto bean (Phaseolus vulgaris L.) and spring whet (Triticum aestivum L.) plants in soil boxes under controlled environment conditions at three soil bulk densities (1.3, 1.5 and 1.7 g cm^–3). The flexible-tube system consisted of bicycle inner tubes inflated inside augered access holes and removed only when measurements were taken. Rigid tubes were constructed of extruded polybutyrate plastic. In both cases tubes were oriented horizontally. Despite similar root densities for wheat and beans based on measurements obtained from soil cores, root densities estimated from both types of minirhizotron were higher in bean than in wheat in uncompacted soil. Estimates of root density by the flexible tube minirhizotron were more closely correlated with soil core image analysis estimates than were those by the rigid minirhizotron system. At high soil bulk density, rigid tube measurements consistently overestimated actual rooting density of both wheat and bean. The relationship between estimated and actual rooting densities in the case of flexible tube measurements was not significantly influenced by soil bulk density. These findings were consistent with the theory that preferential root growth is induced by gaps at the soil-observation tube interface, inherent in the rigid tube technique, and was accentuated under conditions of high soil strength.     

A color composite technique for detecting root dynamics of barley (Hordeum vulgare L.) from minirhizotron images

Quantification of root dynamics by destructive methods is confounded by high coefficients of variation and loss of fine roots. The minirhizotron technique is non-destructive and allows for sequential root observations to be made at the same depth in situ. Observations can be stored on video tape which facilitates data handling and computer-aided image processing. A color composite technique using digital image analyses was adapted in this study to detect barley root dynamics from sequential minirhizotron images. Plants were grown in the greenhouse in boxes (80 × 80 × 75 cm) containing soil from a surface horizon of a Typic Cryoboroll. A minirhizotron was installed at a 45°C angle in each box. Roots intersecting the minirhizotron were observed and video-recorded at tillering, stem extension, heading, dough and ripening growth stages. The images from a particular depth were digitized from the analog video then registered to each other. Discrimination of roots from the soil matrix gave quantitative estimates of root appearance and disappearance. Changes in root appearance and disappearance were detected by assigning a separate primary color (red, green, blue) to selected growth stages, then overlaying the images to create red-green and red-green-blue color composites. The resulting composites allowed for a visual interpretation and quantification of barley root dynamics in situ.     

A comparison of minirhizotron,core and monolith methods for quantifying barley (Hordeum vulgare L.) and fababean (Vicia faba L.) root distribution

Root research has been hampered by a lack of good methods and by the amount of time involved in making measurements. The use of the minirhizotron as a quantitative tool requires comparison with conventional destructive methods. This study was conducted in the greenhouse to compare the minirhizotron technique with core and monolith methods in quantifying barley (Hordeum vulgare L.) and fababean (Vicia faba L.) root distribution. Plants were grown in boxes (80 cm long × 80 cm wide × 75 cm deep) in a hexagonal arrangement to minimize the effects of rooting anistrophy. Minirhizotron observations and destructive sampling to a depth of 70 cm using core and monolith methods were performed at the ripening growth stage. Total root length for the entire depth interval was generally higher in barley (159–309 m) than fababean (110–226 m). Significant correlation coefficients between monolith and core methods for root length density (RLD, cm cm^–3) was observed in both crops (p 0.01). A method and depth interaction showed no significant differences in fababean RLD distribution measured by core and monolith methods. However, the RLD was different for the uppermost 40 cm depth in barley. The relationship for RLD between minirhizotron and core methods was significant only in barley (r=0.77^*). For both crops, estimates of RLD in the top 10-cm layer by the minirhizotron technique were lower than those by core and monolith techniques. In contrast, estimates of RLD were higher in fababean at a depth >30 cm. Destructive sampling still remains the method to quantify root growth in the 0–10 cm soil layer. ei]B E Clothier     

The Wageningen Rhizolab — a facility to study soil-root-shoot-atmosphere interactions in crops

Roots in the Wageningen Rhizolab are observed using two methods: (i) non-destructively, using horizontal, glass minirhizotrons at intervals of 14 days between observations; (ii) with destructive sampling using augers on three dates in the season. This paper reports changes with depth and time in root numbers per unit interface area of the minirhizotron tube (number of intersections) of four crop species (wheat, Brussels sprouts, leek and potato). The number of root intersections of Brussels sprouts, wheat and potato declined with depth at any time, whereas leek showed a different pattern because maximum root growth was observed at a depth of 10–20 cm. Root density generally decreased in the following order: Brussels sprouts, wheat, potato and leek. Plots of root length densities, L_rv(cm. cm^-3), obtained by auger sampling, versus the number of intersections showed considerable variation in slope with species, time in the season and year, implying that a single, universal equation to convert minirhizotron observations into volumetric root densities does not exist. Causes of variation in the slopes are discussed. It is concluded that limited auger sampling combined with minirhizotron observations yield adequate quantitative estimates of relevant root properties.     

Root dynamics in plant and ratoon crops of sugar cane

The root system of a sugar cane crop on an Ultisol in northeastern Brazil was examined throughout the plant and first ratoon crop cycles, using both coring and minirhizotron methods. Total root masses (living plus dead, 0.9–1.1 kg m^-2) and live root lengths (14.0–17.5 km m^-2) were greater during the ratoon cycle than at the end of the plant cane cycle (0.75 kg m^-2 and 13.8 km m^-2, respectively). Root die-back during the two weeks following ratoon harvest was estimated to be 0.15 kg m^-2, about 17% of the total root mass. Root die-back after the plant cane harvest was lower because fire was not used at this harvest and soil humidity was higher under the accumulated litter. A small amount of fine roots proliferated in the litter layer, amounting to 1% of the total mass and 3% of the total length. Root turnover could not be accurately assessed from minirhizotron observations due to variation in the relationship between coring data and the minirhizotron data with both time and soil depth.     

Advancing fine root research with minirhizotrons

Minirhizotrons provide a nondestructive, in situ method for directly viewing and studying fine roots. Although many insights into fine roots have been gained using minirhizotrons, a review of the literature indicates a wide variation in how minirhizotrons and minirhizotron data are used. Tube installation is critical, and steps must be taken to insure good soil/tube contact without compacting the soil. Ideally, soil adjacent to minirhizotrons will mimic bulk soil. Tube installation causes some degree of soil disturbance and has the potential to create artifacts in subsequent root data and analysis. We therefore recommend a waiting period between tube installation and image collection of 6-12 months to allow roots to recolonize the space around the tubes and to permit nutrients to return to pre-disturbance levels. To make repeated observations of individual roots for the purposes of quantifying their dynamic properties (e.g. root production, turnover or lifespan), tubes should be secured to prevent movement. The frequency of image collection depends upon the root parameters being measured or calculated and the time and resources available for collecting images and extracting data. However, long sampling intervals of 8 weeks or more can result in large underestimates of root dynamic properties because more fine roots will be born and die unobserved between sampling events. A sampling interval of 2 weeks or less reduces these underestimates to acceptable levels. While short sample intervals are desirable, they can lead to a potential trade-off between the number of minirhizotron tubes used and the number of frames analyzed per tube. Analyzing fewer frames per minirhizotron tube is one way to reduce costs with only minor effects on data variation. The quality of minirhizotron data should be assessed and reported; procedures for quantifying the quality of minirhizotron data are presented here. Root length is a more sensitive metric for dynamic root properties than the root number. To make minirhizotron data from separate experiments more easily comparable, idiosyncratic units should be avoided. Volumetric units compatible with aboveground plant measures make minirhizotron-based estimates of root standing crop, production and turnover more useful. Methods for calculating the volumetric root data are discussed and an example presented. Procedures for estimating fine root lifespan are discussed.     

The change of soil carbon stocks and fine root dynamics after land use change from a native pasture to a pine plantation

A published meta-analysis of worldwide data showed soil carbon decreasing following land use change from pasture to conifer plantation. A paired site (a native pasture with Themeda triandra dominant, and an adjacent Pinus radiata plantation planted onto the pasture 16 years ago) was set up as a case study to assess the soil carbon reduction and the possible reason for the reduction under pine, including the change in fine root (diameter <2 mm) dynamics (production and mortality). Soil analysis confirmed that soil carbon and nitrogen stocks to 100 cm under the plantation were significantly less than under the pasture by 20 and 15%, respectively. A 36% greater mass of fine root was found in the soil under the pasture than under the plantation and the length of fine root was about nine times greater in the pasture. Much less fine root length was produced and roots died more slowly under the plantation than under the pasture based on observations of fine root dynamics in minirhizotrons. The annual inputs of fine root litter to the top 100 cm soil, estimated from soil coring and minirhizotron observations, were 6.3 Mg dry matter ha⁻¹ year⁻¹ (containing 2.7 Mg C and 38.9 kg N) under the plantation, and 9.7 Mg ha⁻¹ year⁻¹ (containing 3.6 Mg C and 81.4 kg N) under the pasture. The reduced amount of carbon, following afforestation of the pasture, in each depth-layer of the soil profile correlated with the lower length of dead fine roots in the layer under the plantation compared with the pasture. This correlation was consistent with the hypothesis that the soil carbon reduction after land use change from pasture to conifer plantation might be related to change of fine root dynamics, at least in part.     

Evaluating minirhizotron estimates of fine root longevity and production in the forest floor of a temperate broadleaf forest

The minirhizotron technique (MR) for in situ measurement of fine root dynamics offers the opportunity to obtain accurate and unbiased estimates of root production in perennial vegetation only if MR tubes do not affect the longevity of fine roots. Assuming fine root biomass is near steady-state, fine root production (g m^–2 yr^–1) can be estimated as the ratio of fine root biomass (g m^–2) to median fine root longevity (yr). This study evaluates the critical question of whether MR access tubes affect the longevity of fine roots, by comparing fine root survivorship obtained using MR with those from a non-intrusive in situ screen method in the forest floor horizons of a northern hardwood forest in New Hampshire, USA. Fine root survivorship was measured in 380 root screens during 1993–1997 and in six horizontal minirhizotron tubes during 1996–1997. No statistically significant difference was found between estimates of survivorship of fine roots (<1 mm dia.) at this site from MR versus from in situ screens, suggesting that MR tubes do not substantially affect fine root longevity in the forest floor of this northern hardwood forest and providing greater confidence in measurements of fine root production using the MR technique. Furthermore, the methodology for estimating fine root production from MR longevity data was evaluated by comparison of fine root longevity and production estimates made using single vs. multiple root cohorts, and using root-number, root-length, and root-mass weighted methods. Our results indicate that fine root-length longevity estimates based on multiple root cohorts throughout the year can be used to approximate fine root biomass production. Using this method, we estimated fine root longevity and production in the forest floor at this site to be 314 days (or 0.86 yr) and 303 g m^–2 yr^–1, respectively. Fine root production in this northern hardwood forest is approximately equivalent to standing biomass and was previously underestimated by root in-growth cores. We conclude that the use of MR to estimate fine root longevity and production as outlined here may result in improved estimates of fine root production in perennial vegetation.     

弓杠岭不同海拔云杉细根生物量及形态特征

海拔变化是多环境因子的梯度效应,细根作为植物吸收水分与养分的重要器官,其性状特征在指示植物的生长和分布等方面意义重大.该研究以弓杠岭2500～3300 m海拔地的云杉(Picea asperata)细根为研究对象,采用根序分级法对云杉1～5级根序的生物量及细根形态(平均直径、比根长、根长密度、比表面积)进行测定,以明确...     

植物根系研究新技术Minirhizotron的起源、发展和应用

根系是土壤和植物的动态界面,对植物和土壤均具有重要意义。但由于根系深处地下,观测研究十分不便,导致根系研究在广度、深度上均落后于地上部分。随着对根系在生态系统以及全球碳平衡中重要作用的认识,根系渐渐成为国际相关领域的研究热点之一。Minirhizotron(微根区管或小观察窗)技术的诞生和应用,使根系研究手段得到了进一步发展,成为根系研究技术发展的重要里程碑。Minirhizotron技术主要由透明观察管、观测设备和记录设备组成,观测设备曾先后使用了普通镜子、观察镜和相机(或摄像机),记录设备也相应地经历了手工绘制、传统黑白、彩色相片或录像带以及高清晰数字图像。同时,还开发了多种图像自动分析系统,使该项技术日臻完善。Minirhizotron技术可以以非破坏方式,定期对同一根系的出现、生长、衰老、死亡和消失进行连续观察,对根系伸长、根系密度、扎根深度、侧根伸展、分枝特性、菌根特性以及细根动态、根系生命周期和分解等进行观测研究,同时,也可开展根系对不同处理响应的研究。因此,Minirhizotron技术必将在农业、林业和环境等科学领域得到越来越广泛的应用。     

Fine root biomass estimates from minirhizotron imagery in a shrub ecosystem exposed to elevated CO2

Fine root biomass and C content are critical components in ecosystem C models, but they cannot be directly determined by minirhizotron techniques, and indirect methods involve estimating 3-dimensional values (biomass/ soil volume) from 2-dimensional measurements. To estimate biomass from minirhizotron data, a conversion factor for length to biomass must be developed, and assumptions regarding depth of view must be made. In a scrub-oak ecosystem in central Florida, USA, root length density (RLD) was monitored for 10 years in a CO₂ manipulation experiment using minirhizotron tubes. In the seventh year of the study, soil cores were removed from both ambient and elevated CO₂ chambers. Roots from those cores were used to determine specific root length values (m/g) that were applied to the long-term RLD data for an estimation of root biomass over 10 years of CO₂ manipulation. Root length and biomass estimated from minirhizotron data were comparable to determinations from soil cores, suggesting that the minirhizotron biomass model is valid. Biomass estimates from minirhizotrons indicate the <0.25 mm diameter roots accounted for nearly 95% of the total root length in 2002. The long-term trends for this smallest size class (<0.25 mm diameter) mirrored the RLD trends closely, particularly in relation to suspected root closure in this system. Elevated CO₂ did not significantly affect specific root length as determined by the soil cores. A significant treatment effect indicated smallest diameter fine roots (<0.25 mm) were greater under elevated CO₂ during the early years of the study and the largest (2–10 mm) had greater biomass under elevated CO₂ during the later years of the study. Overall, this method permits long-term analysis of the effects of elevated CO₂ on fine root biomass accumulation and provides essential information for carbon models.     

应用微根管法测定细根指标方法评述

树木细根(直径<2mm)在森林生态系统能量流动和物质循环中起着重要的作用。原有的细根生产周转研究中常采用的土钻法、内生长法、挖掘法、根室法和土柱法等,均不能直接观察到细根的动态变化。微根管法是一种非破坏性、可定点直接观察和研究植物根系的方法,为研究细根的生长、衰老、死亡、分解和再生长的过程提供了有效的工具,尤其适用于细根周转、寿命和分解等方面的研究。但该技术不能直接测定单位面积的细根生物量、细根化学组成及细根周转对土壤碳和养分循环的影响,需要与土钻法结合。本文就运用微根管法对细根生物量、生产、周转和寿命等指标的研究方法进行了评述。     

森林生态系统根系生物量研究进展

在森林生态系统功能过程研究中,特别是在生产力和生物地化学循环方面,根系的作用不容忽视,但是,由于研究方法和研究者的观念等方面的限制,对于根系的研究还远不及地上部分受到重视。而关于细根生物量,周转率和生产力等方面的是很少有人问津。为推动我国根系生物学研究的发展,本一面地介绍了９种典型的测度细根生物量的方法。包括：收获法、钻土芯法、内生长土芯法、平衡法、根观测实验室法、土壤碳平衡法、挖土块法、间接法和     

Screening Plant Species for Growth on Weathered,Petroleum Hydrocarbon-Contaminated Sediments

Rapid and cost-effective techniques are needed to select plant species and genotypes for use in phytoremediation, vegetative capping, or revegetation at hazardous waste sites. A greenhouse screening procedure to aid the selection of plant materials would help increase success and decrease the cost. Twenty-nine vascular plant species were compared for growth in weathered sediments contaminated with petroleum hydrocarbons. An uncontaminated reference soil was used to estimate relative seedling growth in stressed and unstressed conditions. Plants were grown in a greenhouse and harvested at 60 and 180 days after planting to estimate variation in seedling growth and full-season growth. Plant growth characteristics measured included height, aboveground biomass, root biomass, root diameter, root-length density, and root surface area density. Concentration of total petroleum hydrocarbons (TPH) was estimated at the final harvest. Considerable variation existed among species for all characteristics except TPH concentration. Under the conditions and length of this trial, no variations in rates of TPH degradation were detected. In general, plant growth was stunted in the contaminated soil compared with the uncontaminated soil; however, differences among plant species for relative seedling growth indicated that they varied in their tolerance to the petroleum hydrocarbon-contaminated soil. For example, tall fescue, Festuca arundinacea, seemed tolerant to the contaminated soil, whereas barley, Hordeum vulgare, seemed sensitive. Comparison of results from the 60- and 180-day harvests suggested that a short-season greenhouse screening could aid selection of species for planting in contaminated soil, if plant growth results are interpreted along with information on the life history characteristics of the species under consideration.     

幼龄柠条细根现存量与环境因子的关系

以晋西北黄土高原区柠条(Caragana korshinskii)幼龄人工林为研究对象, 应用微根管技术(Minirhizotron technique)对林地100 cm土层范围的柠条细根生长动态进行了观测。以2007年生长季(5~9月)的根长密度(RLD, mm·cm^-3)数据为基础, 对柠条细根现存量(RLDst, mm·cm^-3)及其与环境因子(≥10 ℃积温、同期土壤积温、积降雨量和土壤水分等)的关系作了研究。结果表明, 40~90 cm土层是柠条细根的主要分布区和生长活跃区, 其细根占细根总量的59.7%。柠条细根现存量的季节变化特征为: 5月至9月上旬RLDst持续增加, 9月下旬RLDst略有降低。柠条细根现存量季节变化与≥10 ℃积温、同期土壤积温和积降雨量均存在极显著正相关关系。     

微根管在细根研究中的应用

细根（直径≤2 mm）的周转在植物生态系统碳分配过程中具有重要意义.已往细根周转研究主要采用根钻法、分室模型法和内生长法等.这些方法由于不能直接观测到细根生长动态,导致细根周转估计不准确.微根管法是一种非破坏性野外观察细根动态的方法.本文从微根管的发展、功能、安装步骤、图像采集、参数计算、影响观测因素和存在问题等方面逐一进行介绍,并通过水曲柳和落叶松微根管细根观测实例介绍在细根周转过程研究中的应用. 结果表明,微根管可以比较精确地估计出细根长度、单位面积上根长密度、单位体积上根长密度、细根生长量、细根死亡量和细根周转等.微根管是一个观察细根生长、衰老、死亡和分解过程的有效工具.微根管观测精度主要取决于微根管安装的质量和数量、微根管取样间隔期和取样数量、微根管图像分析技术等.此外,土壤质地、石砾多少、微根管材料选择、减少光系统对根系的干扰等也是影响微根管测定精度的因素.如何提高微根管测定精度将成为今后微根管在细根研究中的主要问题.     

Comparison of tomato root distributions by minirhizotron and destructive sampling

Calibration of minirhizotron data against root length density (RLD) was carried out in a field trial where three drip irrigation depths: surface (R0) and subsurface, 0.20 m (RI) and 0.40 m depth (RII) and two processing tomato cultivars: `Brigade' (CI) and `H3044' (CII) were imposed. For each treatment three minirhizotron tubes were located at 10, 37.5 and 75 cm of the way from one plant row to the next. Roots intersecting the minirizotrons walls were expressed as root length intensity (L _a) and number of roots per unit of minirhizotron wall area (N _ra). Root length density (RLD) was calculated from core samples taken for each minirhizotron tube at two locations: near the top of the minirhizotron (BI) and 15 cm apart from it, facing the minirhizotron wall opposite the plant row (BII). Minirhizotron data were regressed against RLD obtained at BI and BII and with their respective means. The results show that for all the situations studied, better correlations were obtained when RLD was regressed with L _a than with N _ra. Also was evident that the relationship between L _a and RLD was strongly influenced by the location of soil coring. RLD was correlated with L _a trough linear and cubic equations, having the last ones higher determination coefficients. For instance at 10 cm from the plant row when values from the top layer (0–40 cm) were analysed separately, L _a was significantly regressed with RLD measured at BII and described by the equations: RLD = 0.5448 + 0.0071 L _a (R ² = 0.51) and RLD = 0.4823 + 0.0074L _a + 8×10^–5 L _a ² – 5×10^–7 L _a ³ (R ² = 0.61). Under the 40 cm depth the highest coefficients of determination for the linear and cubic equations were respectively 0.47 and 0.88, found when L _a was regressed with RLD measured at BI. For minirhizotrons located at 75 cm from the plant row and for location BI it was possible to analyse jointly data from all depths with coefficients of determination of 0.45 and 0.59 for the linear and cubic equations respectively.