Improved scaling of minirhizotron data using an empirically-derived depth of field and correcting for the underestimation of root diameters

Background and aims

Accurate data on the standing crop, production, and turnover of fine roots is essential to our understanding of major terrestrial ecological processes. Minirhizotrons offer a unique opportunity to study the dynamic processes of root systems, but are susceptible to several measurement biases.

Methods

We use roots extracted from minirhizotron tube surfaces to calculate the depth of field of a minirhizotron image and present a model to correct for the underestimation of root diameters obscured by soil in minirhizotron images.

Results

Non-linear regression analysis resulted in an estimated depth of field of 0.78 mm for minirhizotron images. Unadjusted minirhizotron data underestimated root net primary production and fine root standing crop by 61 % when compared to adjusted data using our depth of field and root diameter corrections. Changes in depth of field accounted for >99 % of standing crop adjustments with root diameter corrections accounting for <1 %.

Conclusions

Our results represent the first effort to empirically derive depth of field for minirhizotron images. This work may explain the commonly reported underestimation of fine roots using minirhizotrons, and stands to improve the ability of researchers to accurately scale minirhizotron data to large soil volumes.     

Measuring Fine Root Turnover in Forest Ecosystems

Development of direct and indirect methods for measuring root turnover and the status of knowledge on fine root turnover in forest ecosystems are discussed. While soil and ingrowth cores give estimates of standing root biomass and relative growth, respectively, minirhizotrons provide estimates of median root longevity (turnover time) i.e., the time by which 50% of the roots are dead. Advanced minirhizotron and carbon tracer studies combined with demographic statistical methods and new models hold the promise of improving our fundamental understanding of the factors controlling root turnover. Using minirhizotron data, fine root turnover (y⁻¹) can be estimated in two ways: as the ratio of annual root length production to average live root length observed and as the inverse of median root longevity. Fine root production and mortality can be estimated by combining data from minirhizotrons and soil cores, provided that these data are based on roots of the same diameter class (e.g., < 1 mm in diameter) and changes in the same time steps. Fluxes of carbon and nutrients via fine root mortality can then be estimated by multiplying the amount of carbon and nutrients in fine root biomass by fine root turnover. It is suggested that the minirhizotron method is suitable for estimating median fine root longevity. In comparison to the minirhizotron method, the radio carbon technique favor larger fine roots that are less dynamics. We need to reconcile and improve both methods to develop a more complete understanding of root turnover.     

A comparison of soil core sampling and minirhizotrons to quantify root development of field-grown potatoes

Root growth of potato (Solanum tuberosum L.) is sensitive to soil conditions. A reduced root system size can result in reduced uptake of water and/or nutrients, leading to impaired crop growth. To understand the mechanisms by which soil conditions affect crop growth, study of temporal and spatial development of roots is required.In field experiments, effects of soil temperature, soil compaction and potato cyst nematodes (Globodera pallida) on root growth of potato cultivars were studied using two methods: core sampling and vertically oriented minirhizotrons.Minirhizotrons showed relatively more roots in deeper soil layers than core sampling, probably because of preferential root growth along the tube. Spatial distribution of roots should therefore be analysed by core sampling.To eliminate differences in spatial distribution, total root systems as measured by both methods were compared. Nematodes, cultivars and time did not affect the relationship between both methods. Soil compaction, however, affected it because of a strong response of root length in bulk soil and small differences in root number against the minirhizotron, suggesting that soil coring has to be used to study effects of different bulk densities.With both methods, sequential measurements of roots give the net effect of root growth and decay. Data on root turnover can only be obtained with minirhizotrons by comparing video recordings of different dates. Other information obtained with minirhizotrons is the average orientation of roots. Moreover, the minirhizotron method has the advantage of demanding less labour.     

Applications of minirhizotrons to understand root function in forests and other natural ecosystems

Minirhizotrons have proved useful to understand the dynamics and function of fine roots. However, they have been used comparatively infrequently in forests and other natural plant communities. Several factors have contributed to this situation, including anomalous root distributions along the minirhizotron surface and the difficulty of extracting data from minirhizotron images. Technical and methodological advances have ameliorated some of these difficulties, and minirhizotrons have considerable potential to address some questions of long standing interest. These questions include more fully understanding the role of roots in carbon and nutrient cycling, rates of root decomposition, responses to resource availability and the functional significance of interactions between plant roots and soil organisms. Maximizing the potential for minirhizotrons to help us better understand the functional importance of fine roots in natural plant communities depends upon using them to answer only those questions appropriate to both their inherent strengths and limitations.     

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.     

Soil temperature effects from minirhizotron lighting systems

Observing root dynamics or soil fauna with minirhizotrons requires the use of incandescent or ultraviolet (UV) lighting systems. These light sources can generate heat which would be transferred to the surrounding soil adjacent to the minirhizotron observation tubes and thus may influence root growth and development or fauna activity. The objective of this study was to determine the effect of incandescent and UV light from a minirhizotron camera system on soil temperatures next to minirhizotron tubes. Temperature probes were attached next to and at 0.5 cm from the tube surface and the tubes were then placed in boxes with either a fine sand or a loamy clay soil. Incandescent light was operated stationary for 5 min or moved at 1 cm increments every 10 s down the tube for both dry and wet soils. The UV light was used in a stationary position for 10 minutes in both dry soils. Maximum temperature increases were 3.41–3.52 °C and 1.69–2.14 °C next to the tube for the dry and wet soils, respectively with 5 min of stationary incandescent light. Ultraviolet lights increased soil temperatures to a maximum of approximately 2.5 °C in the dry soil. Probes placed 0.5 cm from the tube surface also showed temperature increases up to 2.15 °C. Moving the light source every 10 s, however, resulted in lower temperature increases (<0.8 °C). Therefore short durations of light resulted in small temperature increases suggesting minimal impact on root development. Increased soil temperatures from longer durations of light, however, may alter root growth and development as well as soil fauna activity and warrants further study.     

氮肥对水曲柳和落叶松细根寿命的影响

采用微根管技术研究了氮肥对水曲柳和落叶松细根生长、衰老和死亡的影响,探讨两树种细根寿命与氮有效性之间的相关关系.结果表明：林地施氮肥后,两树种细根数量都呈减少趋势,细根总体直径增加,分枝程度降低;氮肥使水曲柳细根存活率提高,细根中位值寿命延长105 d,而落叶松细根存活率对氮肥反应不敏感;施氮肥对细根寿命的延长效应主要体现在直径较小的一级根、表层（0～15 cm)根系和春夏季新生的细根,表明氮肥对高生理活性的细根
影响较强.     

An inflatable minirhizotron system for root observations with improved soil/tube contact

Commonly used minirhizotrons consisting of a transparent tube inserted into the soil seldom attain good contact between the tube and the soil, which leads to root growth occurring in a gap rather than in the soil. A new system is described involving an inflatable flexible rubber wall, made from a modified motorcycle tube. Pressure ensures a proper tube/soil contact so that the environmental circumstances for root growth along the tube more closely correspond to those in the undisturbed soil. Before the endoscope slide is introduced into the minirhizotron for taking pictures, the inflatable tube is removed, so that there is no-often opaque-wall between the endoscope and the roots. This improves the picture quality and facilitates the analysis of root images.     

用Minirhizotrons观测柠条根系生长动态

 Minirhizotrons是一种非破坏性、定点直接观察和研究植物根系的新方法。该文介绍了用Minirhizotrons测定植物根系的方法,并同根钻取原状土样法进行了比较;探讨了根系生长动态同土壤含水量间的关系。试验于2004年植物生长季在沙坡头沙漠试验研究站的水分平衡观测场的人工柠条(Caragana korshinskii)林进行,结果表明：Minirhizotrons 管埋入土壤后需要10个月时间允许柠条根系在其周围定居,其观测图片中的根系代表了管子周围2.6 mm土层的根系。柠条根系生长动态和土壤水分变化相关,含水量的升高导致根系的大量繁殖,而根系吸水及蒸发散又导致含水量的减少;在2004年植物生长季,土壤水分和根系的这种相互作用出现了两次,但根系生长高峰比土壤含水量高峰滞后20 d左右。     

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.     

四种不同生活型树种细根寿命及影响因素

以4种不同生活型树种(常绿阔叶和针叶树种、落叶阔叶和针叶树种)为研究对象,通过微根管法现地观测细根的生长动态,比较不同生活型树种细根寿命在种内和种间的差异,探讨影响细根寿命的主要因子,研究结果对理解和预测森林生态系统碳及养分循环过程具有重要的理论意义。结果表明:(1)细根形态特征(分枝结构和直径)显著影响种内细根寿命,分枝等级越低、直径越小,细根的寿命越短;(2)4个树种的细根寿命表现出明显的土层效应和季节效应,即随土壤深度增加,细根的累积存活率逐渐增加,寿命延长;而不同季节出生的细根其寿命长短模式在树种间不一致,春季或夏季出生的细根寿命要长于秋冬季;(3)常绿树种(柳杉、石栎)的细根寿命要长于落叶树种(池杉、麻栎),同时,针叶树种(池杉、柳杉)的细根寿命要长于阔叶树种(麻栎、石栎)。在同一树种内,细根寿命受细根直径、根系分枝结构、土壤环境因子(土层)等因素显著影响,但在不同树种间,细根寿命可能更依赖于树木生长速率、碳分配模式等树木整体的功能性状差异。     

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.     

Root Dynamics in Restored and Naturally Regenerated Atlantic White Cedar Wetlands

This work addressed the seasonal and successional factors of root dynamics in natural and restoration Atlantic white cedar (AWC) wetlands. Using minirhizotrons and soil root cores, fine root dynamics were measured in a chronosequence of reference and restoration AWC wetlands to compare trends in ecosystem development after canopy harvest. Seasonal fine root abundance, production, and mortality were sampled during a 439‐day period in one restoration and three reference AWC wetlands. Soil cores were collected to measure fine root biomass and to determine allometric relationships between root length and biomass. Significant seasonal variation of root dynamics was observed in the young reference and restoration sites. The mature and intermediate‐aged sites exhibited little seasonal variability in root abundance and mortality. Root production was variable but not seasonally consistent. Results suggest that root dynamics become less seasonal as AWC communities shift from herbaceous to woody vegetation dominance. No trend in fine root abundance along the chronosequence was observed, suggesting that roots rapidly reestablish following tree harvest. Measurements of annual root length production suggest increasing annual production with decreasing stand age. However, a reversal of this trend was observed when using production estimates calculated from minirhizotron measurements and root length–mass relationships. These findings underscore the importance of supplementing minirhizotron data with root allometric relationships when analyzing vegetation gradients. Overall, results indicate substantial differences in the form and quantity of root contributions to soil organic matter in the restoration site compared to that in the reference chronosequence. Higher initial planting densities of AWC are recommended to achieve similar contributions of roots to soil organic matter accumulation in the restoration site.     

落叶松和水曲柳人工林细根生长、死亡和周转

 细根周转是陆地生态系统碳分配格局与过程的核心环节,而细根周转估计的关键是了解细根的生长和死亡动态。该研究以18年生落叶松(Larix gmelinii)和水曲柳(Fraxi nus mandshurica)人工林为对象,采用微根管（Minirhizotron）技术对两树种0～40 cm深度的细根生长和死亡动态进行了为期1年的观测,研究了两树种细根在不同土层深度的生长与死亡动态、细根周转以及与土壤有效氮含量、土壤温度、大气温度和降水的关系。结果表明：1） 落叶松平均细根生长（Root length density production, RLDP）0.0045 mm•cm^－2•d^－1）明显低于水曲柳RLDP（0.0077 mm•cm^－2•d^－1）。两个树种细根平均RLDP在表层（0～10 cm）最大,而底层（30～40 cm）最小 ,两树种平均细根死亡（Root length density mortality, RLDM）也表现同样规律 。水曲柳春季生长的细根占41.7%,夏季占39.7%,而落叶松细根生长分别是24.0%和51.2%,水曲柳细根死亡主要发生在春季（34.3%） 和夏季（34.0%）,而落叶松细根死亡主要发生在夏季和秋季（分别占28.5%和32.3%）,两 树种细根生长与死亡在冬季均较小;2）落叶松细根年生长量（0.94 mm•cm^－2•a^－1）和年死亡量（0.72 mm•cm^－2•a^－1）明显低于水曲柳（1.52和1.21 mm•cm^－2•a^－1）,两树种细根表层年生长量和年死亡量均最高,底层最低。落叶松细根年周转为3.1次•a^－1（按年生长量计算）和2.4次•a^－1（按年死亡量计算）,相比较,水曲柳细根年周转分别为2.7次•a^－1和2.2次•a^－1;3）土壤有效氮含量、土壤温度、大气温度和降水综合作用影响细根生长和死亡动态,可以解释细根生长80%的变异和细根死亡95%以上的变异。     

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.     

Short sampling intervals reveal very rapid root turnover in a temperate grassland

Although root growth and mortality play critical regulatory roles in terrestrial ecosystems, little is known about the temporal scale of these dynamics. In temperate grasslands, root dynamics may be particularly rapid because of the high proportion of production allocated to very fine root biomass. In this study, we used minirhizotron tubes to estimate root growth and mortality in an upland grassland in Yellowstone National Park that was grazed by migratory herds of ungulates. Monthly rates of root growth and mortality were estimated from May to September 2005, by measuring the elongation (growth) and disappearance (mortality) of roots at 3-day intervals. Average daily growth (millimeters of root length) was approximately 5 times greater in May and June than in July, August, and September. Average daily mortality (millimeters of root length) did not differ among months. A comparison of the June-September rates of root growth and mortality derived from sampling at short (3-day) and long (1-month) time intervals indicated that the long sampling intervals underestimated both growth and mortality by approximately 60% relative to the short intervals. These results suggest that estimates of grassland root dynamics from minirhizotrons are influenced significantly by sampling interval length, and that rapid root turnover may play a more critical role in regulating energy and nutrient fluxes in temperate grasslands than has previously been recognized.     

两种生境条件下差不嘎蒿细根寿命

细根寿命对细根周转具有重要影响, 是生态系统C分配格局和养分循环研究的重要内容。该文利用微管法研究了流动沙地和固定沙地生长的差不嘎蒿(Artemisia halodendron)灌丛细根生长的动态过程, 通过Kaplan-Meier方法估计了细根存活率和中位值寿命, 并做存活曲线, 用对数轶检验比较了不同生境、不同土壤层次和不同月出生细根寿命的差异程度, 同时分析了不同样地细根寿命同土壤全氮、有机质、体积含水量和容重的相关关系。结果表明, 流动沙地和固定沙地差不嘎蒿细根具有相似的存活曲线, 但在各观测点, 流动沙地的细根累积存活率均高于固定沙地, 流动沙地细根中位值寿命(47 d)显著高于固定沙地(35 d)。细根寿命同各样地的土壤全氮和土壤容重呈显著的负相关关系, 同土壤水分呈显著的正相关关系, 但多元回归分析表明, 土壤水分是引起细根寿命变异的关键因素。土层深度对流动沙地细根寿命没有显著影响, 但两生境深层30~50 cm的细根寿命均显著高于上层(10~30 cm)。不同出生月的细根寿命显著不同, 流动沙地和固定沙地细根寿命具有相似的季节变化规律, 春季(4、5月)细根的寿命最长(71 d), 秋季(8、9月)次之(61 d), 夏季(6、7月)最短(39 d)。     

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。     

连作杨树人工林细根寿命的代际差异及其影响因素

细根寿命是调控森林生产力形成的关键。通过在连作Ⅰ、Ⅱ代杨树人工林固定样地内埋设微根管,对杨树不同根序细根年度生长动态开展连续观测并进行生存分析。结果表明,杨树不同根序细根累积生存率存在显著差异,高级根(3—5级)寿命较长,其累积生存率显著高于1级和2级细根。杨树细根寿命存在显著的代际差异,连作Ⅱ代人工林活根量、死根量和细根总量均高于Ⅰ代林。连作Ⅱ代人工林细根中位值寿命为(90±16)d,显著低于Ⅰ代人工林((102±22)d)。连作Ⅱ代林各根序细根数量、分布比例均高于Ⅰ代林,低级细根累积生存率低于Ⅰ代林而高级细根累积生存率显著高于Ⅰ代林。连作杨树人工林细根寿命显著受制于土壤环境,1级细根寿命与土壤速效氮相关性极显著(r=-0.861),2级细根寿命与土壤物理性状相关性较强且与土壤酚酸含量呈现极显著相关(r=0.870),高级根序细根寿命与土壤物理性质和养分状况等也具有一定相关性。连作杨树人工林土壤酚酸累积和养分有效性下降影响了细根寿命和周转,并进而造成净初级生产力损耗,相关结论为连作杨树人工林生产力衰退机理模型的建立提供了科学依据。     

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.