Fine root heterogeneity by branch order: exploring the discrepancy in root turnover estimates between minirhizotron and carbon isotopic methods

Fine roots constitute a large and dynamic component of the carbon cycles of terrestrial ecosystems. The reported fivefold discrepancy in turnover estimates between median longevity (ML) from minirhizotrons and mean residence time (MRT) using carbon isotopes may have global consequences. Here, a root branch order-based model and a simulated factorial experiment were used to examine four sources of error. Inherent differences between ML, a number-based measure, and MRT, a mass-based measure, and the inability of the MRT method to account for multiple replacements of rapidly cycling roots were the two sources of error that contributed more to the disparity than did the improper choice of root age distribution models and sampling bias. Sensitivity analysis showed that the rate at which root longevity increases as order increases was the most important factor influencing the disparity between ML and MRT. Assessing root populations for each branch order may substantially reduce the errors in longevity estimates of the fine root guild. Our results point to the need to acquire longevity estimates of different orders, particularly those of higher orders.     

微根管法和同位素法在细根寿命研究中的应用及比较

细根的生产和周转在陆地生态系统的碳和养分循环中起着重要作用,并且对全球环境变化具有一定的指示意义。细根寿命是估计细根周转的关键,其长短决定了养分和碳消耗与循环的速度。由于采用的研究方法不同,导致所得细根寿命估计值存在较大差异,目前最新的同位素和微根管2种方法之间寿命估计值差异可达10倍以上。本文对这2种研究方法的原理和优点进行了阐述,并从细根定义、细根寿命理论分布假设、细根取样误差等方面对导致这2种方法研究结果存在差异的原因进行分析,以期有助于今后根系研究的发展。     

微根窗技术及其在植物根系研究中的应用

植物根系对固定植株和获得水分和养分起重要作用,但是土壤不可观测性的限制,给根系生态学的研究带来一定的困难。因此,找到原位观察根系生长的方法对研究根系生态学就显得尤为重要。目前微根窗技术被认为是研究根系生态学最有前途的方法。从微根窗系统的组成、微根窗管的安装、微根窗图象的收集及微根窗数据的利用等几个方面进行了概述。阐述了在微根窗使用和操作过程中需要注意的几个问题,微根窗管与土壤之间的良好接触是获得高质量微根窗图像数据的前提和基础;图象收集的频率依赖于测定和计算的根系参数,如果想得到根系现存量、生产力、更新和寿命的信息,必须避免采样间隔时间过长。     

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。     

Automatic discrimination of fine roots in minirhizotron images

Minirhizotrons provide detailed information on the production, life history and mortality of fine roots. However, manual processing of minirhizotron images is time-consuming, limiting the number and size of experiments that can reasonably be analysed. Previously, an algorithm was developed to automatically detect and measure individual roots in minirhizotron images. Here, species-specific root classifiers were developed to discriminate detected roots from bright background artifacts. Classifiers were developed from training images of peach (Prunus persica), freeman maple (Acer x freemanii) and sweetbay magnolia (Magnolia virginiana) using the Adaboost algorithm. True- and false-positive rates for classifiers were estimated using receiver operating characteristic curves. Classifiers gave true positive rates of 89-94% and false positive rates of 3-7% when applied to nontraining images of the species for which they were developed. The application of a classifier trained on one species to images from another species resulted in little or no reduction in accuracy. These results suggest that a single root classifier can be used to distinguish roots from background objects across multiple minirhizotron experiments. By incorporating root detection and discrimination algorithms into an open-source minirhizotron image analysis application, many analysis tasks that are currently performed by hand can be automated.     

2004—2008年落叶松人工林细根生产和死亡的季节动态

2004—2008年,采用微根管(minirhizotron)技术,对落叶松人工林细根生产和死亡进行连续动态观测,同时测定了温度(大气温度和土壤10 cm温度)和水分(降雨量和土壤10 cm深处含水量)的变化,研究细根生产、死亡的动态及其与温度和水分的关系.结果表明：落叶松细根年根长生产量在0.20～0.78 mm·cm^-2,死亡量在0.26～0.72 mm·cm^-2;2004—2006年细根年根长平均生产量(0.67 mm·cm^-2)和死亡量(0.59 mm·cm^-2)均高于2007—2008年细根年根长平均生产量和死亡量(0.37和0.39 mm·cm^-2);在生长季内（5—10月）,落叶松春末至夏季(6—7月) 的细根生产量占全年产量的51%～68%,秋末(10月)仅占全年的1%～4%;而夏末(8月)和秋季(9—10月)细根死亡量占全年的59%～70%,早春(5月)占全年的1%～5%.相关分析表明,大气温度变化可以解释细根生产量66%的变异,而土壤10 cm深处温度解释24%,降雨量解释27%.细根的死亡量与土壤10 cm深处温度呈指数正相关.     

细柄阿丁枫和米槠细根寿命影响因素

采用微根管技术对福建建瓯万木林自然保护区细柄阿丁枫(ALG)和米槠(CAC)细根进行了连续2 a的观测。估计细根寿命采用Kaplan-Meier方法,用对数秩检验(log-rank test)比较单一因素(细根直径、序级、出生年份、出生季节、土层以及邻近细根数量)对细根寿命的影响。同时采用Cox比例风险回归分析方法,分析上述因素对细根存活的影响程度。结果表明:细柄阿丁枫细根平均寿命和中值寿命分别为(286±8)d和(184±9)d,而米槠的则分别为(261±10)d和(212±8)d。仅考虑单一因素时,出生季节、径级、序级以及邻近细根数量对细柄阿丁枫和米槠细根寿命皆有极显著影响(P<0.01);出生年份对米槠细根寿命有极显著影响(P<0.01),但对细柄阿丁枫细根寿命的影响无统计学意义(P>0.05);土层深度对细柄阿丁枫细根寿命有极显著影响(P<0.01),而对米槠细根寿命的影响无统计学意义(P>0.05)。Cox比例风险回归分析则表明出生年份对二者细根寿命的影响皆无统计学意义(P>0.05),影响因素按照影响程度大小排列均依次是序级、出生季节、细根直径、邻近细根数量,而土层对细柄阿丁枫细根寿命的影响最弱,对米槠细根寿命的影响无统计学意义(P>0.05)。     

Annual and seasonal changes in fine root biomass of a Quercus ilex L. forest

The biomass, production and mortality of fine roots (roots with diameter <2.5 mm) were studied in a typical Mediterranean holm oak (Quercus ilex L.) forest in NE Spain using the minirhizotron methodology. A total of 1212 roots were monitored between June of 1994 and March of 1997. Mean annual fine root biomass in the holm oak forest of Prades was 71±8 g m^–2 yr^–1. Mean annual production for the period analysed was 260+11 g m^–2 yr^–1. Mortality was similar to production, with a mean value of 253±3 g m^–2 yr^–1. Seasonal fine root biomass presented a cyclic behaviour, with higher values in autumn and winter and lower in spring and summer. Production was highest in winter, and mortality in spring. In summer, production and mortality values were the lowest for the year. Production values in autumn and spring were very similar. The vertical distribution of fine root biomass decreased with increasing depth except for the top 10–20 cm, where values were lower than immediately below. Production and mortality values were similar between 10 and 50 cm depth. In the 0–10 cm and the 50–60 cm depth intervals, both production and mortality were lower.     

Temperature-induced alteration to root longevity in Lolium perenne

The production and mortality of roots is an important factor determining nutrient and carbon fluxes in terrestrial ecosystems. However, the effect of temperature on the longevity of roots is not understood. In this study the impact of changes in temperature on the longevity of Lolium perenne roots was determined.Plants were grown at 15 °C, 21 °C and 27 °C and their roots observed using a minirhizotron system. Major effects of temperature were measured with large reductions in root longevity occurring at higher temperatures. The implications of these data are discussed.     

去叶对水曲柳和落叶松苗木当年生长及细根动态的影响

叶片被取食会导致树木生长发育和生理代谢发生显著的变化。目前对细根动态如何对叶片损失做出响应的了解仍然有限。以生物量分配和高生长策略不同的水曲柳(Fraxinus mandschurica)和落叶松(Larix gmelinii)苗木为研究对象, 进行了不同强度的人为去叶处理(叶面积去除0% (对照)、40%和80%), 采用微根管技术对细根(直径≤2 mm)生产和死亡的季节动态进行了定量观测, 同期测定了地上部分(苗高和地径)的生长。结果表明: 1)去叶降低了两树种苗高(统计上均不显著)和地径的生长, 但是对苗高生长的影响小于地径。随着去叶强度的提高, 苗木地上生长受到的影响加大, 生长季末期水曲柳苗高比对照降低3.3%-12.1%, 地径降低5.7%-23.1%; 而落叶松苗高和地径降低相对较少(< 12%)。2)去叶显著地减少了水曲柳和落叶松细根现存量(p< 0.001), 其相对增长量((去叶后现存量高峰-去叶当日现存量)/去叶当日现存量)随着去叶强度的加大而降低。3)与对照相比, 去叶后两树种细根生产量显著减少(p< 0.05), 而细根死亡量在不同处理间没有显著差异。综合来看, 去叶对水曲柳地上部分(特别是地径)生长影响较大, 而对落叶松地下部分(主要是新根)生长影响较大。研究结果为理解冠层碳供应对根系动态影响的种间差异及其机制提供了必要的理论依据。     

樟子松人工林细根寿命估计及影响因子研究

细根寿命的估计是了解细根生产和死亡的关键, 对了解陆地生态系统碳分配格局和养分循环具有重要意义。该研究采用微根管(minirhizotron)技术, 以23年生樟子松(Pinus sylvestris var. mongolica)人工林为研究对象, 对细根生长和死亡过程进行了连续两年的观测。细根寿命的估计采用Kaplan-Meier方法, 计算细根的平均寿命(mean longevity)、中值寿命(median longevity)和累积存活率(cumulative survival rate), 用对数秩检验(log-rank test)比较单一因素, 包括细根直径、根序、出生季节和土层以及菌根侵染对细根寿命的影响。采用Cox比例风险回归分析方法, 同时分析上述因素对细根存活的影响程度。结果表明, 樟子松细根的生产和死亡具有明显的季节性, 春末和夏季(6月和7月)为生产高峰; 而死亡高峰出现在夏末至秋末, 以及冬季。樟子松细根的平均和中值寿命分别为(322 ± 10)天和(310 ± 15)天, 对数秩检验表明, 仅考虑单一因子时, 细根直径、根序、出生季节和土层以及菌根侵染均对细根寿命有显著影响。Cox回归分析表明, 菌根侵染、细根直径和土层是影响樟子松细根寿命的重要因子。细根直径每增加1 mm, 细根死亡危险率就降低99%, 即相当于寿命延长99%; 细根出生土层每增加1 cm, 其寿命延长5%; 而菌根侵染后, 会导致细根死亡危险率增加175%; 但根序和出生季节的影响不显著。这些发现证实: 林木细根寿命受到内在与外在因素的共同控制, 而多变量回归分析的方法有助于我们全面揭示细根寿命变异的潜在机制。     

中亚热带细柄阿丁枫和米槠群落细根的生产和死亡动态

采用微根管技术与挖掘法相结合的方式对福建省万木林自然保护区细柄阿丁枫和米槠天然林细根生产和死亡动态进行了为期两年多的观测,分析细根生产和死亡的季节变化、垂直分布及径级和序级分配,并估计细根的年生产量和年死亡量.结果表明:细柄阿丁枫细根年生产量和年死亡量分别为(230.1±162.8) g·m-2·a-1和(188.8±75.5)g·m-2· a-1,均略大于米槠的(214.5±185.8) g·m-2·a-1和(178.8±26.5) g·m-2·a-1,但两种森林群落的细根年生产量和年死亡量均无显著差异(P＞0.05).两森林群落细根生产均在春季达到高峰,其中米槠细根生产与月降水量呈极显著相关(P＜0.01,r=0.566);细根死亡则呈现季节性地波动,米槠细根死亡峰值主要发生于夏季和秋季,而细柄阿丁枫则出现在秋季.两森林群落细根生产和死亡皆主要集中于土壤表层0-40 cm中,而且不同径级细根生产和死亡集中于0-1 mm细根中,其中0.3-0.6 mm细根的生产和死亡在两森林群落中均最大.两森林群落一级根的生产和死亡均大于高级根.     

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.     

Arbuscular mycorrhizal fungi induced alteration to root longevity of poplar

Root longevity is an important element determining the fluxes of nutrients and carbon within terrestrial ecosystems. The effect of colonisation by an arbuscular mycorrhizal (AM) fungus on root longevity of poplar was determined using a minirhizotron camera to make direct observation of roots in situ in conjunction with image analysis. significant influences of colonisation on the longevity of roots were observed with most colonised roots surviving for a shorter period than non-colonised roots: only 16% of colonised roots survived for longer than 49 days compared to 49% of non-colonised roots. After 21 days approximately 5 times as many colonised as non-colonised roots had died. Possible mechanisms are discussed.     

Interactive effects of soil warming and nitrogen addition on fine root production of Chinese fir seedlings

Aims There have been a large number of studies on the independent separate responses of fine roots to warming and nitrogen deposition, but with contradictory reporting. Fine root production plays a critical role in ecosystem carbon, nutrient and water cycling, yet how it responds to the interactive warming and nitrogen addition is not well understood. In the present study, we aimed to examine the interactive effects of soil warming and nitrogen addition on fine root growth of 1-year-old Chinese fir (Cunninghamia lanceolata) seedlings in subtropical China.
Methods A mesocosm experiment, with a factorial design of soil warming (ambient, +5 °C) and nitrogen addition (ambient, ambient + 40 kg·hm^-2·a^-1, ambient + 80 kg·hm^-2·a^-1), was carried out in the Chenda State-owned Forest Farm in Sanming City, Fujian Province, China. Fine root production (indexed by the number of fine roots emerged per tube of one year) was measured biweekly using minirhizotrons from March of 2014 to February of 2015.
Important findings (1) The two-way ANOVA showed that soil warming had a significant effect on fine root production, while nitrogen addition and soil warming × nitrogen addition had no effect. (2) The three-way ANOVA (soil warming, nitrogen addition and diameter class) showed that soil warming, diameter class and soil warming × diameter class had significant effects on fine root production, especially for the number of fine roots in 0-1 mm diameter class that had been significantly increased by soil warming. Compared with the 1-2 mm roots, the 0-1 mm roots seemed more flexible. (3) Repeated measures of ANOVA (soil warming, nitrogen addition and season) showed that soil warming, season, soil warming × season, and soil warming × nitrogen addition × season had significant effects on fine root production. In spring, the number of fine roots was significantly increased both by soil warming and soil warming × season, while soil warming, nitrogen addition, soil warming × nitrogen addition significantly decreased fine root production in the summer. (4) Soil warming, soil layer, soil warming × soil layer had significant effects on fine root production. The number of in-growth fine roots was significantly increased by soil warming at the 20-30 cm depth only. It seemed that warming forced fine roots to grow deeper in the soil. In conclusion, soil warming significantly increased fine root production, but they had different responses and were dependent of different diameter classes, seasons and soil layers. Nitrogen addition had no effect on fine root production. Only in spring and summer, soil warming and nitrogen addition had significant interactive effects.     

Soil fertility and fine root dynamics in response to 4 years of nutrient (N,P, K) fertilization in a lowland tropical moist forest,Panama

The question of how tropical trees cope with infertile soils has been challenging to address, in part, because fine root dynamics must be studied in situ. We used annual fertilization with nitrogen (N as urea, 12.5 g N m^?2 year^?1), phosphorus (P as superphosphate, 5 g P m^?2 year^?1) and potassium (K as KCl, 5 g K m^?2 year^?1) within 38 ha of old‐growth lowland tropical moist forest in Panama and examined fine root dynamics with minirhizotron images. We expected that added P, above all, would (i) decrease fine root biomass but, (ii) have no impact on fine root turnover. Soil in the study area was moderately acidic (pH = 5.28), had moderate concentrations of exchangeable base cations (13.4 cmol kg^?1), low concentrations of Bray‐extractable phosphate (PO₄ = 2.2 mg kg^?1), and modest concentrations of KCl‐extractable nitrate (NO₃ = 5.0 mg kg^?1) and KCl‐extractable ammonium (NH₄ = 15.5 mg kg^?1). Added N increased concentrations of KCl‐extractable NO₃ and acidified the soil by one pH unit. Added P increased concentrations of Bray‐extractable PO₄ and P in the labile fraction. Concentrations of exchangeable K were elevated in K addition plots but reduced by N additions. Fine root dynamics responded to added K rather than added P. After 2 years, added K decreased fine root biomass from 330 to 275 g m^?2. The turnover coefficient of fine roots <1 mm diameter ranged from 2.6 to 4.4 per year, and the largest values occurred in plots with added K. This study supported the view that biomass and dynamics of fine roots respond to soil nutrient availability in species‐rich, lowland tropical moist forest. However, K rather than P elicited root responses. Fine roots smaller than 1 mm have a short lifetime (<140 days), and control of fine root production by nutrient availability in tropical forests deserves more study.     

5种温带森林生态系统细根的时间动态及其影响因子

细根(直径≤2 mm)的生长和死亡动态及其影响因子是森林生态系统能量流动和物质循环的重要研究内容,但因受到研究方法的限制而了解甚少.于2010年5-10月采用微根管技术对东北东部山区5种温带森林生态系统的细根生长量(FRP)和死亡量(FRM)进行了动态跟踪测定,并同步测定了土壤温度(Ts)、土壤湿度(Ms)、叶面积指数(LAI)等相关因子.结果表明:不同林型和取样时间的FRP和FRM均差异显著(P＜0.001).杨桦林、硬阔叶林、兴安落叶松林、红松林、蒙古栎林的FRP和FRM分别为:(13.34 ±0.90) μm·cm-2·d-1(平均值±标准误)和(5.02±0.36) μm· cm-2·d-1、(13.04±0.82)μm·cm-2·d-1和(6.85±0.32) μm· cm-2·d-1、(8.74±1.44) μm·cm-2·d-1和(5.05±0.61) μm·cm-2·d-1、(8.02±2.27) μm·cm-2·d-1和(3.88±0.35)μm·em-2·d-1、(7.59±0.82) μm·cm-2·d-1和(3.88±0.61) μm· cm-2·d-1.所有林型生长季期间FRP的时间变化均呈现明显的单峰型,但峰值出现的时间却因林型而异.FRM随生长季的进程而逐渐增加,杨桦林和硬阔叶林FRM在8月初出现峰值,而红松林、兴安落叶松林和蒙古栎林的FRM峰值均出现在生长季末期.Ts、Ms和LAI对FRP和FRM均存在显著的正效应(P＜0.05),3个因子的综合作用对各个林型FRP和FRM变异性的解释率分别达68％和53％以上,表明这些温带森林生态系统细根生长和死亡的时间动态主要受土壤温湿度和叶面积变化的联合影响.     

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.     

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。