Loss of whole-tree hydraulic conductance during severe drought and multi-year forest die-off

Understanding the pathways through which drought stress kills woody vegetation can improve projections of the impacts of climate change on ecosystems and carbon-cycle feedbacks. Continuous in situ measurements of whole trees during drought and as trees die hold promise to illuminate physiological pathways but are relatively rare. We monitored leaf characteristics, water use efficiency, water potentials, branch hydraulic conductivity, soil moisture, meteorological variables, and sap flux on mature healthy and sudden aspen decline-affected (SAD) trembling aspen (Populus tremuloides) ramets over two growing seasons, including a severe summer drought. We calculated daily estimates of whole-ramet hydraulic conductance and modeled whole-ramet assimilation. Healthy ramets experienced rapid declines of whole-ramet conductance during the severe drought, providing an analog for what likely occurred during the previous drought that induced SAD. Even in wetter periods, SAD-affected ramets exhibited fivefold lower whole-ramet hydraulic conductance and sevenfold lower assimilation than counterpart healthy ramets, mediated by changes in leaf area, water use efficiency, and embolism. Extant differences between healthy and SAD ramets reveal that ongoing multi-year forest die-off is primarily driven by loss of whole-ramet hydraulic capability, which in turn limits assimilation capacity. Branch-level measurements largely captured whole-plant hydraulic limitations during drought and mortality, but whole-plant measurements revealed a potential role of other losses in the hydraulic continuum. Our results highlight the importance of a whole-tree perspective in assessing physiological pathways to tree mortality and indicate that the effects of mortality on these forests’ assimilation and productivity are larger than expected based on canopy leaf area differences.     

干旱低磷胁迫对不同品种小麦根系导水率的影响

控制磷素水平,采用控制灌水量(正常供水、中度及重度干旱胁迫)的盆栽试验法,选择抗旱性小麦品种陕合6号(W1)和水分敏感型品种郑引1号(W2)为供试材料。用压力室法测定了三叶期的两品种小麦根系导水率(LPr)的变化规律。结果表明：陕合6号,在有磷正常供水处理( PH)下具有较高的导水率,干旱胁迫时LPr降低较少,且复水后有较强的恢复能力。郑引1号, PH的LPr值相对较小,干旱导致的根系导水率下降非常突出,复水后的恢复能力也较弱。另外,干旱胁迫对小麦苗期根系导水率的影响大于磷胁迫对其导水率的影响,且两品种小麦无磷止常供水处理(-PH)的LPr分别为 PH的31．9%和53．6％,即磷对前者LPr的影响大于后者。     

The importance of mesophyll conductance in regulating forest ecosystem productivity during drought periods

Water availability is the most limiting factor to global plant productivity, yet photosynthetic responses to seasonal drought cycles are poorly understood, with conflicting reports on which limiting process is the most important during drought. We address the problem using a model‐data synthesis approach to look at canopy level fluxes, integrating twenty years of half hour data gathered by the FLUXNET network across six Mediterranean sites. The measured canopy level, water and carbon fluxes were used, together with an inverse canopy ecophysiological model, to estimate the bulk canopy conductance, bulk mesophyll conductance, and the canopy scale carbon pools in both the intercellular spaces and at the site of carboxylation in the chloroplasts. Thus the roles of stomatal and mesophyll conductance in the regulation of internal carbon pools and photosynthesis could be separated. A quantitative limitation analysis allowed for the relative seasonal responses of stomatal, mesophyll, and biochemical limitations to be gauged. The concentration of carbon in the chloroplast was shown to be a potentially more reliable estimator of assimilation rates than the intercellular carbon concentration. Both stomatal conductance limitations and mesophyll conductance limitations were observed to regulate the response of photosynthesis to water stress in each of the six species studied. The results suggest that mesophyll conductance could bridge the gap between conflicting reports on plant responses to soil water stress, and that the inclusion of mesophyll conductance in biosphere–atmosphere transfer models may improve their performance, in particular their ability to accurately capture the response of terrestrial vegetation productivity to drought.     

Mycorrhizal influence on hydraulic and hormonal factors implicated in the control of stomatal conductance during drought

During drying, mycorrhizal plants often maintain higher stomatalconductance (g_s) than similarly-sized and -nourished non-mycorrhizalplants, but the mechanism of mycorrhizal influence remains unclear.Several hydraulic and non-hydraulic factors previously implicatedin control of stomatal behavior during drought were measured,to learn which are affected when roots of cowpea (Vigna unguiculata[L.] Walp. cv. White Acre) are extensively colonized by Glomusintraradices Schenck and Smith isolate UT143. At low soil watercontents (), mycorrhizal plants maintained higher g_s, transpirationand shoot water potential () than non-mycorrhizal plants. Thesehigher foliar water status characters were associated with lowerxylemsap abscisic acid concentrations ([ABA]) and lower ABAfluxes to leaves in mycorrhizal plants at low soil . Stomatalconductance was most closely correlated with xylem-sap [ABA],ABA flux to leaves and shoot . Stomatal conductance was notcorrelated with xylemsap concentrations of calcium or zeatinriboside equivalents, or with xylem-sap pH, nor were these xylem-sapconstituents affected by mycorrhizal symbiosis. Stomata of mycorrhizaland non-mycorhizal leaves showed similar sensitivities to ABA,whether leaves were intact or detached. It is concluded thatmycorrhizal fungi probably increased the capability of rootsystems to scavenge water in drier soil, resulting in less strainto foliage and hence higher g_s, and shoot at particular soil. Key words: Abscisic acid, cytokinins, Glomus intraradices     

Role of a single aquaporin isoform in root water uptake

Aquaporins are ubiquitous channel proteins that facilitate the transport of water across cell membranes. Aquaporins show a typically high isoform multiplicity in plants, with 35 homologs in Arabidopsis. The integrated function of plant aquaporins and the function of each individual isoform remain poorly understood. Matrix-assisted laser desorption/ionization time-of-flight analyses suggested that Plasma Membrane Intrinsic Protein2;2 (PIP2;2) is one of the abundantly expressed aquaporin isoforms in Arabidopsis root plasma membranes. Two independent Arabidopsis knockout mutants of PIP2;2 were isolated using a PCR-based strategy from a library of plant lines mutagenized by the insertion of Agrobacterium tumefaciens T-DNA. Expression in transgenic Arabidopsis of a PIP2;2 promoter-beta-glucuronidase gene fusion indicated that PIP2;2 is expressed predominantly in roots, with a strong expression in the cortex, endodermis, and stele. The hydraulic conductivity of root cortex cells, as measured with a cell pressure probe, was reduced by 25 to 30% in the two allelic PIP2;2 mutants compared with the wild type. In addition, free exudation measurements revealed a 14% decrease, with respect to wild-type values, in the osmotic hydraulic conductivity of roots excised from the two PIP2;2 mutants. Together, our data provide evidence for the contribution of a single aquaporin gene to root water uptake and identify PIP2;2 as an aquaporin specialized in osmotic fluid transport. PIP2;2 has a close homolog, PIP2;3, showing 96.8% amino acid identity. The phenotype of PIP2;2 mutants demonstrates that, despite their high homology and isoform multiplicity, plant aquaporins have evolved with nonredundant functions.     

Chemical and hydraulic signals regulate stomatal behavior and photosynthetic activity in maize during progressive drought

The objectives of this study were to investigate stomatal regulation in maize seedlings during progressive soil drying and to determine the impact of stomatal movement on photosynthetic activity. In well-watered and drought-stressed plants, leaf water potential (Ψ _leaf), relative water content (RWC), stomatal conductance (g _s), photosynthesis, chlorophyll fluorescence, leaf instantaneous water use efficiency (iWUE_leaf), and abscisic acid (ABA) and zeatin-riboside (ZR) accumulation were measured. Results showed that g _s decreased significantly with progressive drought and stomatal limitations were responsible for inhibiting photosynthesis in the initial stages of short-term drought. However, after 5 days of withholding water, non-stomatal limitations, such as damage to the PSII reaction center, became the main limiting factor. Stomatal behavior was correlated with changes in both hydraulic and chemical signals; however, changes in ABA and ZR occurred prior to any change in leaf water status. ABA in leaf and root tissue increased progressively during soil drying, and further analysis found that leaf ABA was negatively correlated with g _s (R ² = 0.907, p < 0.05). In contrast, leaf and root ZR decreased gradually. ZR in leaf tissue was positively correlated with g _s (R ² = 0.859, p < 0.05). These results indicate that ABA could induce stomatal closure, and ZR works antagonistically against ABA in stomatal behavior. In addition, the ABA/ZR ratio also had a strong correlation with g _s, suggesting that the combined chemical signal (the interaction between ABA and cytokinin) plays a role in coordinating stomatal behavior. In addition, Ψ _leaf and RWC decreased significantly after only 3 days of drought stress, also affecting stomatal behavior.     

Nitrate induction of root hydraulic conductivity in maize is not correlated with aquaporin expression

Some plant species can increase the mass flow of water from the soil to the root surface in response to the appearance of nitrate in the rhizosphere by increasing root hydraulic conductivity. Such behavior can be seen as a powerful strategy to facilitate the uptake of nitrate in the patchy and dynamically changing soil environment. Despite the significance of such behavior, little is known about the dynamics and mechanism of this phenomenon. Here we examine root hydraulic response of nitrate starved Zea mays (L.) plants after a sudden exposure to 5 mM NO₃ ⁻ solution. In all cases the treatment resulted in a significant increase in pressure-induced (pressure gradient ~ 0.2 MPa) flow across the root system by ~50% within 4 h. Changes in osmotic gradient across the root were approximately 0.016 MPa (or 8.5%) and thus the results could only be explained by a true change in root hydraulic conductance. Anoxia treatment significantly reduced the effect of nitrate on xylem root hydraulic conductivity indicating an important role for aquaporins in this process. Despite a 1 h delay in the hydraulic response to nitrate treatment, we did not detect any change in the expression of six ZmPIP1 and seven ZmPIP2 genes, strongly suggesting that NO₃ ⁻ ions regulate root hydraulics at the protein level. Treatments with sodium tungstate (nitrate reductase inhibitor) aimed at resolving the information pathway regulating root hydraulic properties resulted in unexpected findings. Although this treatment blocked nitrate reductase activity and eliminated the nitrate-induced hydraulic response, it also produced changes in gene expression and nitrate uptake levels, precluding us from suggesting that nitrate acts on root hydraulic properties via the products of nitrate reductase.     

Influence of saline stress on root hydraulic conductance and PIP expression in Arabidopsis

Measurements of the root hydraulic conductance (L0) of roots of Arabidopsis thaliana were carried out and the results were compared with the expression of aquaporins present in the plasma membrane of A. thaliana. L0 of plants treated with different NaCl concentrations was progressively reduced as NaCl concentration was increased compared to control plants. Also, L0 of plants treated with 60 mmol/L NaCl for different lengths of time was measured. Variations during the light period were seen, but only for the controls. A good correlation between mRNA expression and L0 was observed in both experiments. Control plants and plants treated with 60 mmol/L NaCl were incubated with Hg and then with DTT. For these plants, L0 and cell-to-cell pathway contributions to root water transport were determined. These results revealed that in control plants most water movement occurs via the cell-to-cell pathway, thus implying aquaporin involvement. But, in NaCl-stressed plants, the Hg-sensitive cell-to-cell pathway could be inhibited already by the effect of NaCl on water channels. Therefore, short periods of NaCl application to Arabidopsis plants are characterised by decreases in the L0 of roots, and are related to down-regulation of the expression of the PIP aquaporins. This finding indicates that the well known effect of salinity on L0 could involve regulation of aquaporin expression.     

Root hydraulic conductance and aquaporin abundance respond rapidly to partial root-zone drying events in a riparian Melaleuca species

? Drying a portion of a root system (partial root-zone drying (PRD)) can induce partial stomatal closure, but this response is not always observed. We hypothesized that some of the variation in PRD response reflects adaptations to the native environment, where plants subjected to frequent PRD events may display a greater degree of root-level compensation. ? Here, we examined PRD responses of Melaleuca argentea, a tree native to intermittent waterways in which PRD events are common. Seedlings were grown with part of their root system in soil and part in an aquatic compartment, mimicking conditions often observed in the field. ? The aquatic roots initially provided two-thirds of total water uptake, but draining the aquatic compartment had no effect on stomatal conductance, so long as soil moisture remained c. 80% of field capacity. Water uptake from the soil compartment increased threefold within 24 h, with a corresponding transient threefold increase in root hydraulic conductance (L(p)), an increase in plasma membrane intrinsic protein 1 (PIP1) aquaporins at 24 h, and a decrease in PIP2 aquaporins by 48 h. ? Our results demonstrate that PRD can induce rapid changes in L(p) and aquaporin expression in roots, which may play a role in short-term water uptake adjustments, particularly in species adapted to heterogeneous water availability.     

Amelioration of chilling-induced water stress by abscisic Acid-induced changes in root hydraulic conductance

Pretreatment of soybean (Glycine max L. var Ransom) root systems with abscisic acid (ABA) ameliorates the deleterious effect of low temperatures on root hydraulic conductance. ABA treatment of root systems subsequently chilled to 10°C with shoots at 25°C resulted in higher leaf water potentials and lower stomatal resistances. If the root systems are left at 25°C, ABA causes stomatal closure. Membrane alterations are suggested as a mechanism for the ABA action in plant response to chilling stress.     

Influence of xylem development on axial hydraulic conductance within Prunus root systems

The effect of secondary growth on the distribution of the axial hydraulic conductance within the Prunus root system was investigated. Secondary growth resulted in a large increase in both the number (from about 10 to several thousand) and diameter of xylem vessels (from a few micrometres to nearly 150 µm). For fine roots (<3 mm), an increase in root diameter was correlated with a slight increase in the number of xylem vessels and a large increase in their diameter. Conversely, for woody roots, an increase in root diameter was associated with a dramatic increase in the number of xylem vessels, but little or no change in vessel diameter. The theoretical axial conductivity (Kh, m⁴.s^-1.MPa^-1) of root segments was calculated with the Poiseuille-Hagen equation from measurements of vessel diameter. Kh measured using the tension-induced technique varies over several orders of magnitude (7.4᎒^-11 to 5.7᎒^-7 m⁴.s^-1.MPa^-1) and shows large discrepancies with theoretical calculated Kh. We concluded that root diameter is a pertinent and useful parameter to predict the axial conductance of a given root, provided the root type is known. Indeed, the relationship between measured Kh and root diameter varies according to the root type (fine or woody), due to differences in the xylem produced by secondary growth. Finally, we show how the combination of branching pattern and axial conductance may limit water flow through root systems. For Prunus, the main roots do not appear to limit water transfer; the axial conductance of the main axes is at least 10% higher than the sum of the axial conductance of the branches.     

Drought resistance of Quercus pubescens as a function of root hydraulic conductance, xylem embolism and hydraulic architecture

Water relations, xylem embolism, root and shoot hydraulic conductance of both young plants in the field and potted seedlings of Quercus pubescens have been studied with the aim of investigating whether these variables may account for the well known adaptation of this oak species to arid habitats. Our data revealed that Q. pubescens is able to maintain high leaf relative water contents under water stress conditions. In fact, relative water contents measured in summer (July) did not differ from those recorded in April. This was apparently achieved by compensating water loss by an equal amount of water uptake. Such a drought avoidance strategy was made possible by the recorded high hydraulic efficiency of stems and roots under water stress. In fact, root hydraulic conductance of field-grown plants was maintained high in summer when the percentage loss of hydraulic conductance of stems was lowest. The hydraulic architecture of young plants of Q. pubescens measured in terms of partitioning of hydraulic resistances along the water pathway revealed that the highest hydraulic resistance was located in stems of the current year's growth. This hydraulic architecture is interpreted as consistent with the adaptation of Q. pubescens to arid habitats as a consequence of the recorded seasonal changes in water relation parameters as well as in root and stem hydraulics.     

Hydraulic responses to extreme drought conditions in three co-dominant tree species in shallow soil over bedrock

An important component of the hydrological niche involves the partitioning of water sources, but in landscapes characterized by shallow soils over fractured bedrock, root growth is highly constrained. We conducted a study to determine how physical constraints in the root zone affected the water use of three tree species that commonly coexist on the Edwards Plateau of central Texas; cedar elm (Ulmus crassifolia), live oak (Quercus fusiformis), and Ashe juniper (Juniperus ashei). The year of the study was unusually dry; minimum predawn water potentials measured in August were ?8 MPa in juniper, less than ?8 MPa in elm, and ?5 MPa in oak. All year long, species used nearly identical water sources, based on stable isotope analysis of stem water. Sap flow velocities began to decline simultaneously in May, but the rate of decline was fastest for oak and slowest for juniper. Thus, species partitioned water by time when they could not partition water by source. Juniper lost 15–30 % of its stem hydraulic conductivity, while percent loss for oak was 70–75 %, and 90 % for elm. There was no tree mortality in the year of the study, but 2 years later, after an even more severe drought in 2011, we recorded 34, 14, 6, and 1 % mortality among oak, elm, juniper, and Texas persimmon (Diospyros texana), respectively. Among the study species, mortality rates ranked in the same order as the rate of sap flow decline in 2009. Among the angiosperms, mortality rates correlated with wood density, lending further support to the hypothesis that species with more cavitation-resistant xylem are more susceptible to catastrophic hydraulic failure under acute drought.     

Measurement of apoplasmic and cell-to-cell components of root hydraulic conductance by a pressure-clamp technique

A pressure-clamp technique was devised for the direct measurement of cell-to-cell and apoplasmic components of root hydraulic conductance; the experimental results were analyzed in terms of a theoretical model of water and solute flow, based on a composite membrane model of the root. When water is forced under a constant pressure into a cut root system, an exponential decay of flow is observed, until a constant value is attained; when pressure is released, a reverse water flow out of the root system is observed which shows a similar exponential behavour. The model assumes that the transient flow occurs through a cell-to-cell pathway and the observed decrease is the result of accumulation of solutes in front of the root semi-permeable membrane, whilst the steady-state component results from the movement of water through the parallel apoplasmic pathway. Root conductance components are estimated by fitting the model to experimental data. The technique was applied to the root systems of potted cherry (Prunus avium L.) seedlings; average apoplasmic conductance was 15.5 × 10^–9m³· s^–1· MPa^–1, with values ranging from 12.0 × 10^–9 to 18.5 × 10^–9m³· s^–1· MPa^–1; average cell-to-cell conductance was 11.7 × 10⁹ m³· s^–1· MPa^–1, with values ranging from 8.5 × 10^–9 to 15.3 × 10^–9 m³ · s^–1·MPa^–1. Cell-to-cell conductance amounted on average to 43% of total root conductance, with values between 41 and 45%. Leaf specific conductance (conductance per unit of leaf area supported) of the root systems ranged from 2.7 × 10^–8 to 5.6 × 10^–8 m· s^–1·MPa^–1, with an average of 3.7 × 10^–8 m · s^–1·MPa^–1. The newly developed technique allows the interaction of mass flow of water and of solutes to be explored in the roots of soil-grown plants.Abbreviations and Symbols A L_p root hydraulic conductance - A^aL _p ^a root apoplasmic conductance - A^ccL _p ^cc root cell-to-cell conductance - C_s(t) concentration of solutes in apical root compartment at time t - J_v flow of water through the root - J _v ^a apoplasmic flow of water - J_v/cc cell-to-cell flow of water - LSC leaf specific conductance of the root system - P root hydrostatic pressure - P_appl applied pressure - _s(t) root osmotic pressure at time t - _m osmotic pressure of rooting medium - reflection coefficient of root membrane - time constant of cell-to-cell flow decay This research was funded within the EC Project Long-term effects of CO₂-increase and climate change on European forests (LTEEF) (EV5V-CT94-0468); F.M. was supported by a Ministero dell' Universitá e della Ricerca Scientifica e Tecnologica — British Council agreement (Project The ecological significance of cavitation in woody plants); M.C. was supported by a Consiglio Nazionale delle Ricerche — British Council agreement. We gratefully thank Prof. P.G. Jarvis (University of Edinburgh, UK) for revising an earlier version of this paper and Prof. E. Steudle (University of Bayreuth, Germany) for helpful comments.     

A method for measuring xylem hydraulic conductance and embolism in entire root and shoot systems

Current methods for determining the influence of xylem cavitationon hydraulic conductance are limited to unbranched stem or rootsegments with hydraulic conductances above c. 2 mmol s^–1MPa^–1. Lower conductances and/or highly branched systemsare encountered in seedlings, arid-land shrubs, herbs, and distalportions of shoot and root systems of trees. In order to quantifythe hydraulic impact of cavitation in such systems, existingtechniques have been modified. Branched shoot or root systemswere prepared for measurement by removal of leaves, or roottips, respectively. The shoot or root system was enclosed ina vacuum chamber with the proximal end protruding and suppliedwith perfusing solution. Flow through the xylem was inducedby chamber vacuum. Hydraulic conductance was determined fromthe slope of the flow rate versus pressure relationship. Xylemembolism was quantified from the increase in hydraulic conductancefollowing high pressure (100 kPa) perfusion of solution throughthe plant. Examples are provided of the application of the methodto cavitation studies in the cold desert shrub Artemisia tridentata. Key words: Hydraulic conductance, xylem cavitation, embolism, whole root/shoot system     

Role of small conductance calcium-activated potassium channels expressed in PVN in regulating sympathetic nerve activity and arterial blood pressure in rats

Small conductance Ca(2+)-activated K(+) (SK) channels regulate membrane properties of rostral ventrolateral medulla (RVLM) projecting hypothalamic paraventricular nucleus (PVN) neurons and inhibition of SK channels increases in vitro excitability. Here, we determined in vivo the role of PVN SK channels in regulating sympathetic nerve activity (SNA) and mean arterial pressure (MAP). In anesthetized rats, bilateral PVN microinjection of SK channel blocker with peptide apamin (0, 0.125, 1.25, 3.75, 12.5, and 25 pmol) increased splanchnic SNA (SSNA), renal SNA (RSNA), MAP, and heart rate (HR) in a dose-dependent manner. Maximum increases in SSNA, RSNA, MAP, and HR elicited by apamin (12.5 pmol, n = 7) were 330 ± 40% (P < 0.01), 271 ± 40% (P < 0.01), 29 ± 4 mmHg (P < 0.01), and 34 ± 9 beats/min (P < 0.01), respectively. PVN injection of the nonpeptide SK channel blocker UCL1684 (250 pmol, n = 7) significantly increased SSNA (P < 0.05), RSNA (P < 0.05), MAP (P < 0.05), and HR (P < 0.05). Neither apamin injected outside the PVN (12.5 pmol, n = 6) nor peripheral administration of the same dose of apamin (12.5 pmol, n = 5) evoked any significant changes in the recorded variables. PVN-injected SK channel enhancer 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazol-2-one (DCEBIO, 5 nmol, n = 4) or N-cyclohexyl-N-[2-(3,5-dimethyl-pyrazol-1-yl)-6-methyl-4-pyrimidin]amine (CyPPA, 5 nmol, n = 6) did not significantly alter the SSNA, RSNA, MAP, and HR. Western blot and RT-PCR analysis of punched PVN tissue showed abundant expression of SK1-3 channels. We conclude that SK channels expressed in the PVN play an important role in the regulation of sympathetic outflow and cardiovascular function.     

Effect of deep and shallow root systems on the dynamics of soil inorganic N during 3-year crop rotations

Unused inorganic nitrogen (N_inorg) left in agricultural soils will typically leach to deeper soil layers. If it moves below the root zone it will be lost from the system, but the depth of the root zone depends on the crop species grown. In this experiment we studied the effect of 3-year crop sequences, with different combinations of deep-rooted and shallow-rooted crops, on soil N_inorg dynamics to 2.5 m soil depth and the possibility of crop utilization of N leached to deep soil layers. We grew ten different crop sequences for 3 years. The crops and catch crops grown were selected to allow different sequences of deep-rooted and shallow-rooted crops. Very different rooting depths were obtained, from only 0.5 m (leek), to ∼1.0 m (ryegrass and barley), 1.5 m (red beet), 2.0 m (fodder radish and white cabbage) and more than 2.5 m by the chicory catch crop. The results showed a significant retention of N_inorg within the 2.5 m soil profile from one year to the next, but the retained N had leached to deeper parts of the profile during the winter season. Only little N_inorg was retained over two winter seasons. The retention in the deeper soil layers allowed N_inorg to be taken up by succeeding deep-rooted main crops or catch crops. The effects of crop rooting depth on N_inorg in the subsoil layers from 1.0 to 2.5 m were striking. White cabbage reduced N_inorg below 1.0 m with up to 113 kg N ha^-1 during its growth. Grown after catch crops, leek and red beet left on average 60 kg N ha⁻¹ less below 1.0 m than leek and red beet grown without a preceding catch crop. We conclude that it is possible to design crop rotations with improved nitrogen use efficiency by using the differences in crop rooting patterns; deep-rooted crops or catch crops can be used to recover N_inorg leached after previous crops, and catch crops can be grown before shallow-rooted crops to lift the deep N_inorg up to layers where these crops have their roots.