Mobilization of nitrogen reserves during regrowth of defoliated Trifolium repens L. and identification of potential vegetative storage proteins

Although it is well established that carbon reserves contributeto shoot regrowth of leguminous forage species, little informationis available on nitrogen reserves except in Medicaqo sativaL. and Trifolium subterraneum L. In this study, reserves werelabelled with ¹⁵N to demonstrate the mobilization of endogenousnitrogen from roots and stolons to regrowing leaves and newstolons during 24 d of regrowth in white clover (Thfolium repensL.). About 55% and 70%, respectively, of the nitrogen contentsof these organs were mobilized to support the regrowth of leaves.During the first 6 d, nitrogen in regrowing leaves came mainlyfrom N reserves of organs remaining after defoliation. Afterthese first 6 d of regrowth, most of the shoot nitrogen wasderived from exogenous nitrogen taken up while the contributionof nitrogen reserves decreased. After defoliation, the buffer-solubleprotein content of roots and stolons decreased by 32% duringthe first 6 d of regrowth. To identify putative vegetative storageproteins, soluble proteins were separated using SDS-PAGE ortwo-dimensional electrophoresis. One protein of 17.3 kDa instolons and two proteins of 15 kDa in roots seemed to behaveas vegetative storage proteins. These three polypeptides, initiallyfound at high concentrations, decreased in relative abundanceto a large extent during early regrowth and then were accumulatedagain in roots and stolons once normal growth was re-established. Key words: White clover, regrowth, ¹⁵N-labelled, vegetative storage proteins, electrophoresis     

Regrowth dynamics of <Emphasis Type="Italic">Calamagrostis epigejos</Emphasis> after defoliation as affected by nitrogen availability

Young plants of a rhizomatous grass Calamagrostis epigejos (L.) Roth were grown from seed in nutrient solutions containing nitrogen in concentrations 0.1, 1.0, and 10 mM. After six weeks of cultivation the plants were defoliated and changes in growth parameters and in content of storage compounds were measured in the course of regrowth under highly reduced nitrogen availability. Plants grown at higher nitrogen supply before defoliation had higher amount of all types of nitrogen storage compounds (nitrates, free amino acids, soluble proteins), which was beneficial for their regrowth rate, in spite of lower content of storage saccharides. Amino acids and soluble proteins from roots and stubble bases were the most important sources of storage compounds for regrowth of the shoot. Faster growth of plants with higher N content was mediated by greater leaf area expansion and greater number of leaves. In plants with lower contents of N compounds number of green leaves decreased after defoliation significantly and senescing leaves presumably served as N source for other growing organs. Results suggest that internal N reserves can support regrowth of plants after defoliation even under fluctuating external N availability. Faster regrowth of C. epigejos with more reserves was mediated mainly by changes in plant morphogenesis.     

Nitrogen and Carbon Flows Estimated by 15N and 13C Pulse-Chase Labeling during Regrowth of Alfalfa

The flow of 15N and 13C from storage compounds in organs remaining after defoliation (sources) to regrowing tissue (sinks), and 13C losses through root or shoot respiration were assessed by pulse-chase labeling during regrowth of alfalfa (Medicago sativa L.) following shoot removal. A total of 73% of labeled C and 34% of labeled N were mobilized in source organs within 30 d. Although all of the 15N from source organs was recovered in the regrowing tissue, much of the 13C was lost, mainly as CO2 respired from the root (61%) or shoot (8%), and was found to a lesser extent in sink tissue (5%). After 3, 10, or 30 d of regrowth, 87, 66, and 52% of shoot N, respectively, was derived from source tissue storage compounds; the rest resulted from translocation of fixed N2. Overall results suggest that most shoot C was linked to photosynthetic activity rather than being derived from mobilization of stored C in source organs. Furthermore, isotopic analysis of different chemical fractions of plant tissue suggests that between 14 and 58% of the shoot C derived from source tissues was linked to the mobilization of N compounds, not carbohydrates.     

Influence of initial levels of carbohydrates, fructans, nitrogen, and soluble proteins on regrowth of Lolium perenne L. cv. Bravo following defoliation

An experiment was designed to evaluate the role of N and C reserves on regrowth of Lolium perenne cv. Bravo following defoliation. By using two nitrogen fertilization levels together with three photoperiodic conditions, plants with variable contents of water-soluble carbohydrates (43-216 mg g^-1 DW in stubble) and contrasting amounts of nitrogen (7-49 mg g^-1 DW) were obtained. Plants were severely defoliated and regrowth was followed for 28 d under the same environmental conditions. The yield of leaf dry matter at the end of the regrowth period was not related to the initial level of carbohydrate reserves. However, levels of fructan in leaf sheaths and in elongating leaf bases strongly influenced the shoot yield during the first 2 d following defoliation. Fructan exohydrolase activity increased 2-3-fold in sheaths and 3.5-5-fold in elongation leaf bases, suggesting that not only fructans from sheaths but also fructans from immature cells may be used as substrates for growth. In contrast, no direct relationship was found between shoot production and nitrogen or soluble protein accumulation in source organs during early regrowth. A significant correlation existed with the initial amount of soluble proteins in sheaths and in elongating leaf bases after only 6 d of regrowth.     

Partitioning of Nitrogen Derived from N2 Fixation and Reserves in Nodulated Medicago sativa L. During Regrowth

Nodul{macron}ted alfalfa plants were grown hydroponically. Inorder to quantify N₂ fixation and remobilization of N reservesduring regrowth the plants were pulse-chase-labelled with ¹⁵N.Starch and ethanol-soluble sugar contents were analysed to examinechanges associated with those of N compounds. Shoot removalcaused a severe decline in N₂ fixation and starch reserves within6 d after cutting. The tap root was the major storage site formetabolizable carbohydrate compounds used for regrowth; initiallyits starch content decreased and after 14 d started to recoverreaching 50% of the initial value on day 24. Recovery of N₂fixation followed the same pattern as shoot regrowth. Afteran initial decline during the first 10 d following shoot removal,the N₂ fixation, leaf area and shoot dry weight increased sorapidly that their levels on day 24 exceeded initial values.Distribution of ¹⁵N within the plant clearly showed that a significantamount of endogenous nitrogen in the roots was used by regrowingshoots. The greatest use of N reserves (about 80% of N incrementin the regrowing shoot) occurred during the first 10 d and thencompensated for the low N₂ fixation. The distribution of N derivedeither from fixation or from reserves of source organs (taproots and lateral roots) clearly showed that shoots are thestronger sink for nitrogen during regrowth. In non-defoliatedplants, the tap roots and stems were weak sinks for N from reserves.By contrast, relative distribution within the plant of N assimilatedin nodules was unaffected by defoliation treatment. Key words: Medicago sativa L., N₂ fixation, N remobilization, N₂ partitioning, regrowth     

Nitrogen Reserve Mobilization during Regrowth of Medicago sativa L. (Relationships between Availability and Regrowth Yield)

An experiment was designed to study the role of N and C reserves on regrowth of the shoots following defoliation of forage species. Starch and N accumulation in root and crown tissue of nonnodulated Medicago sativa L. were modified during regrowth by applying different levels of N and different cutting heights. Plants were obtained with similar crown and root dry weights, but having either low starch and high tissue N or high starch and low tissue N. The plants were then submitted to a second defoliation and supplied with optimal N nutrition, and N flow from reserve was quantified using pulse-chase 15N labeling. Maximum yields following the second regrowth were obtained from those plants having a high tissue N, despite their low level of nonstructural carbohydrate. When N in the roots and crown exceeded 5 mg N plant-1 at the beginning of regrowth, about 68% was translocated to regrowing shoots. Highly significant correlations were also found between the amounts of N available in roots and crown at the beginning of regrowth and (a) the amount of N that was mobilized to new tissues, (b) the amount of N taken up during the regrowth period, and (c) the final shoot yield after 24 d of regrowth. No similar correlations were found for plants that varied in their initial starch content of roots and crown. It is suggested that N reserves were used mainly during the first 10 d after defoliation, and that the resulting aerial growth during this period should be sufficient to restore N2 fixation and/or N uptake to levels equal to those prior to defoliation. These data emphasize (a) the importance of root N reserves in initiating and sustaining new shoot growth, and (b) the need for a re-evaluation of the contribution of C reserves to shoot regrowth.     

Influence of initial organic N reserves and residual leaf area on growth, N uptake, N partitioning and N storage in alfalfa (Medicago sativa) during post-cutting regrowth

BACKGROUND AND AIMS: The influence of initial residual leaf area and initial N reserves on N uptake, final N distribution, and yield in alfalfa regrowing after cutting, were studied. METHODS: The effects of two levels of initial residual leaf area (plants cut to 15 cm, with (L+) or without (L-) their leaves) and two initial levels of N status [high N (HN) or low N (LN)] on growth, N uptake and N partitioning, allocation and storage after 29 d of post-cutting regrowth were analysed. KEY RESULTS: During most of the regrowth period (8-29 d after the initial harvest), HN and L+ plants had higher net N uptake rates than LN and L- plants, respectively, resulting in a greater final mineral N uptake for these treatments. However, the final partitioning of exogenous N to the regrowing shoots was the same for all treatments (67 % of total exogenous N on average). Final shoot growth, total plant N content, and N allocation to the different taproot N pools were significantly lower in plants with reduced initial leaf area and initial N reserve status. CONCLUSIONS: Although both initial residual leaf area and initial N reserves influenced alfalfa regrowth, the residual leaf area had a greater effect on final forage production and N composition in the taproot, whereas the N uptake rate and final total N content in plant were more affected by the initial N reserve status than by the residual leaf area. Moreover, N storage as proteins (especially as vegetative storage proteins, rather than nitrate or amino acids) in the taproot allowed nitrate uptake to occur at significant rates. This suggests that protein storage is not only a means of sequestering N in a tissue for further mobilization, utilization for growth or tissue maintenance, but may also indirectly influence both N acquisition and reduction capacities.     

Phosphorus reserves increase grass regrowth after defoliation

Accumulation of P above levels that promote growth, a common plant response called “luxury consumption”, can be considered as a form of reserve to support future growth when the nutrient can subsequently be mobilized. However, the effect of P reserves on regrowth following defoliation has not been demonstrated. We tested the hypothesis that P luxury consumption increases plant tolerance to defoliation. We performed two experiments with four grass species from a continuously grazed temperate grassland in the Flooding Pampa (Argentina). The first experiment, aimed at generating P luxury consumption by fertilization, resulted in one species (Sporobolus indicus) showing luxury consumption. In this way, we were able to obtain plants of S. indicus with similar biomass but contrasting amounts of P reserves. The second experiment evaluated the subsequent regrowth following defoliation on a P-free medium of these plants differing in P reserves. Regrowth was larger for plants that had shown P luxury consumption during a previous period than for plants with lower levels of P reserves. During regrowth these plants showed a clear pattern of P remobilization from the stubble, crown, and root compartments to the regrowing tissue, in addition to a likely reutilization of P present in leaf-growth zones. This work is the first showing that high levels of P reserves can confer tolerance to defoliation by promoting compensatory growth under P deficiency.     

Regrowth by Swards of Subterranean Clover after Defoliation. 1. Growth, Non-structural Carbohydrate and Nitrogen Content

Swards of subterranean clover (Trifolium subterraneum L.) atLAl 6 grown in N-free nutrient solution were subjected to threedefoliation treatments which removed 30, 70 and 80% of shootdry weight. Subsequent regrowth and changes in the concentrationsof carbohydrate and nitrogen in plant components were measuredat 0, 1, 5, 9 and 13 d after defoliation and compared with thosein uncut swards. The rate of shoot regrowth declined with increasing severilyof defoliation. In all defoliation treatments, growth was confinedto leaves for up to 5 d. Root growth ceased in all treatmentsfor a longer period. Reestablishment of the leaf area in severely-defoliatedswards was facilitated by the rapid opening of developing leavesand by changes in the allocation of carbon which favoured leafover branch and root, and lamina over petiole growth. Loss of carbohydrate and nitrogen from roots and branches lasting5–9 d was observed in the more severe defoliation treatments.Loss of protein (N x 6.25) exceeded that of total non-structuralcarbohydrate, and could have accounted for the nitrogen contentof new leaf during this period. Branches lost 62% of their initialcarbohydrate content compared with 25% from roots in the 80%cut swards. In contrast, roots, by virtue of their greater mass,were the principle source of mobilized nitrogen. Nitrogen accumulationceased in 80% cut swards for 9 d. However, carbohydrate levelsin the crown nodules were not severely depleted. It was concluded that partitioning of growth to lamina and themobilization of carbohydrates and nitrogen were important forrecovery from defoliation. Carbohydrates, carbon partitioning, defoliation, nitrogen, mobilization, regrowth, subterranean clover, Trifolium subterraneum L     

P effects on N uptake and remobilization during regrowth of Italian ryegrass (Lolium multiflorum)

The influence of P deficiency on the utilization of two sources of N, mineral N (exogenous N) and reserved N (endogenous N), for regrowth of Italian ryegrass (Lolium multiflorum) was studied. P-sufficient (+P) or P-free (−P) nutrition solution was applied from 7 days before defoliation to 24 days of regrowth and the N flows derived from two different N sources within the plant were quantified by ¹⁵N pulse-chase labeling. Shoot regrowth significantly reduced by 12 days of regrowth, while root growth was more in −P plants. Inorganic P (P_i) concentration was highly reduced by P deprivation more in the stubble and regrowing shoots and less in the roots. At defoliation, P deprivation had induced a higher accumulation for all N compounds in the stubble and for amino acids in the roots. The previously incorporated ¹⁵N in stubble and roots as nitrate and amino acids was much decreased in −P plants especially for the first 6 days of regrowth. Total N content in the regrowing leaves was not significantly different between +P and −P plants, but percentage contribution of remobilized N for total leaf N formation was significantly higher in −P plants (78%) than in +P plants (69%) at 6 days of regrowth. From day 12, the utilization of both endogenous and exogenous N was apparently inhibited in −P plants.     

Carbohydrate storage and use in an alpine population of the perennial herb, Oxytropis sericea

I tested hypotheses for ecological roles of storage carbohydrates in perennating organs (roots and branches) of alpine Oxytropis sericea, a leguminous herb. In naturally growing plants, total nonstructural carbohydrates achieved their maximal concentration in the fall, declined during winter, and reached minimal levels immediately after growth initiation in the spring. Experimental manipulation of carbon sink-source relations through shading of leaves of reproductive plants revealed that the normally unused portion of these carbohydrates is largely available for withdrawal. In another experiment, plants subjected to carbohydrate depletion through shading suffered decreased leaf growth after winter dormancy and had a lower probability of flowering and decreased inflorescence biomass. The dependence of reproductive growth on stored carbohydrates, however, was limited to its initial stages, because accumulation of storage carbohydrates occurred simultaneously with inflorescence expansion, flowering, and fruiting. Moreover, the whole-plant photosynthetic rate, estimated from gas exchange measurements also peaked at the time of inflorescence growth. To address whether stored reserves allow compensatory regrowth following defoliation, plants were subjected to experimental removal of leaves and inflorescences. Defoliated O. sericea partly regrew the lost leaves but withdrawal of stored carbohydrates was limited. Similarly, in a second defoliation experiment where infructescences were left intact, the plants used little stored carbohydrate and only partly compensated for fruit growth. However, carbohydrate accumulation was negatively affected by defoliation. While the ecological importance of stored nonstructural carbohydrates cannot be attributed to any function in isolation, winter respiration, leaf regrowth after winter, and early reproductive growth in O. sericea all depend to a significant extent on stored reserves. Maintaining a large storage pool may protect these functions in years when carbon status is less favorable than during this study. Received: 13 May 1998 / Accepted: 24 November 1998     

The use of internal nitrogen stores in the rhizomatous grass Calamagrostis epigejos during regrowth after defoliation

BACKGROUND AND AIMS: The regrowth dynamics after defoliation of the invasive grass Calamagrostis epigejos were studied. As nitrogen (N) reserves have been shown to play an important role during plant regrowth, the identity, location and relative importance for regrowth of N stores were determined in this rhizomatous grass. METHODS: Plant growth, nitrate uptake and root respiration were followed during recovery from defoliation. Water soluble carbohydrates, nitrate, free amino acids and soluble proteins were analysed in the remaining organs. KEY RESULTS: Nitrate uptake and root respiration were severely reduced during the first days of regrowth. Roots were the main net source of mobilized N. The quantitatively dominant N storage compounds were free amino acids. Free amino acids and soluble proteins in the roots decreased by 55 and 50%, respectively, and a substantial (approximately 38%) decrease in stubble protein was also observed. Although the relative abundance of several soluble proteins in roots decreased during the initial recovery from defoliation, no evidence was found for vegetative storage protein (VSP). Furthermore, rhizomes did not act as a N storage compartment. CONCLUSIONS: Production of new leaf area was entirely reliant, during the first week after defoliation, on N stores present in the plant. Mobilized N originated mainly from free amino acids and soluble proteins located in roots, and less so from proteins in stubble. Presence of VSP in the roots was not confirmed. The data suggest that rhizomes played an important role in N transport but not in N storage.     

Seasonal changes in shoot regrowth potential in Calamagrostis canadensis

Summary A series of laboratory experiments was conducted to examine seasonal change in shoot regrowth potential following disturbance in Calamagrostis canadensis. On several dates during the 1988 and 1989 growing seasons, soil cores were collected from field sites dominated by this grass. Shoot regrowth from cores after clipping at the soil surface was monitored under dark or light laboratory conditions at 20°C. seasonal changes in field concentrations of total nonstructural carbohydrate and nitrogen in rhizomes largely accounted for the observed seasonal change in etiolated regrowth potential of shoots in laboratory experiments. In contrast, shoot regrowth potential in the light showed a very different seasonal pattern. The ratio of shoot biomass regrowth 20 d after clipping in the light versus dark treatment showed a gradual seasonal decrease from 12:1 in the early May experiment to near 1:1 in the September experiment. However, the rate of photosynthesis of regrowing shoots in the light was highest in experiments conducted late in the growing season. This may indicate a strong seasonal decrease in the proportion of current photosynthate of regrowing shoots that is allocated to new shoot growth. Alternatively, mobilization of rhizome carbohydrate reserves for shoot regrowth may have been inhibited during the re-establishment of photosynthesis in the light treatment. Either mechanism would explain why shoot regrowth in the light is poorly correlated with levels of belowground carbohydrate reserves, even under controlled laboratory conditions.     

Coping with herbivory: Photosynthetic capacity and resource allocation in two semiarid Agropyron bunchgrasses

Summary Agropyron desertorum, a grazing-tolerant bunchgrass introduced to the western U.S. from Eurasia, and Agropyron spicatum, a grazing-sensitive bunchgrass native to North America, were examined in the field for photosynthetic capacity, growth, resource allocation, and tiller dynamics. These observations allowed identification of physiological characteristics that may contribute to grazing tolerance in semiarid environments. A uniform matrix of sagebrush, Artemisia tridentata, provided an ecologically relevant competitive environment for both bunch-grass species. Physiological activity, growth, and allocation were also followed during recovery from a severe defoliation treatment and were correlated with tiller dynamics.Potential photosynthetic carbon uptake of both species was dominated by stems and leaf sheaths during June, when maximum uptake rates occurred. For both species, water use efficiency of stems and sheaths was similar to that of leaf blades, but nitrogen investment per photosynthetic surface area was less than in blades. In addition, soluble carbohydrates in stems and sheaths of both species constituted the major labile carbon pools in control plants. Contrary to current theory, these findings suggest that culms from which leaf blades have been removed should be of considerable value to defoliated bunchgrasses, and in the case of partial defoliation could provide important supplies of organic nutrients for regrowth. These interpretations, based on total pool sizes, differ markedly from previous interpretations based on carbohydrate concentrations alone, which suggested that crowns contain large carbohydrate reserves. In this study, crowns of both species contained a minor component of the total plant carbohydrate pool.Following defoliation, A. desertorum plants rapidly reestablished a canopy with 3 to 5 times the photosynthetic surface of A. spicatum plants. This difference was primarily due to the greater number of quickly growing new tillers produced following defoliation. Agropyron spicatum produced few new tillers following defoliation despite adequate moisture, and carbohydrate pools that were equivalent to those in A. desertorum.Leaf blades of regrowing tillers had higher photosynthetic capacity than blades on unclipped plants of both species, but the relative increase, considered on a unit mass, area, or nitrogen basis, was greater for A. desertorum than for A. spicatum. Agropyron desertorum also had lower investment of nitrogen and biomass per unit area of photosynthetic tissues, more tillers and leaves per bunch, and shorter lived stems, all of which can contribute to greater tolerance of partial defoliation.Greater flexibility of resource allocation following defoliation was demonstrated by A. desertorum for both nitrogen and carbohydrates. Relatively more allocation to the shoot system and curtailed root growth in A. desertorum resulted in more rapid approach to the preclipping balance between the root and shoot systems, whereas root growth in A. spicatum continued unabated following defoliation. Nitrogen required for regrowth in both species was apparently supplied by uptake rather than reserve depletion. Carbohydrate pools in the shoot system of both species remained very low following severe defoliation and were approximately equivalent to carbon fixed in one day by photosynthesis of the whole canopy.Dedicated to Drs. Michael Evenari and Konrad Springer     

Oxygen Supply Limits Nitrogenase Activity of Clover Nodules After Defoliation

The aim of this study was to test the effect of oxygen partialpressure as a possible limiting factor of nitrogen fixationfollowing defoliation. The response of nitrogenase activity(C₂H₂-reduction) of defoliated and undefoliated white and redclover plants (Trifolium repens L. and Trifolium pratense L.)to either 19 kPa oxygen or 55 kPa oxygen was investigated. Priorto defoliation, white clover plants were grown for five weeksin a growth chamber, and red clover plants for 7 or 11 weeksin a glasshouse. The results included measurements of ¹⁶N₂-uptake. Increasing oxygen partial pressure from 19 to 55 kPa severelyrestricted nitrogenase activity of undefoliated white cloverplants; however, 2 h after complete defoliation, the same treatmentcaused a significant increase. A fivefold increase in nitrogenaseactivity upon exposure to the elevated oxygen partial pressurewas found at the end of a 24 h period. This beneficial effectdecreased gradually from 1 to 5 d after defoliation. The responseof recently defoliated red clover plants to 55 kPa oxygen partialpressure was similar to that of white clover, independentlyof plant age. The gradual recovery of nitrogenase activity duringthree weeks of regrowth was associated with a simultaneous changein the response to increased oxygen partial pressure, leadingagain to the response of undefoliated plants. These data suggested that lack of oxygen at the site of nitrogenfixation, resulting from a dramatic increase in oxygen-diffusionresistance, is the main factor limiting nitrogenase activityfollowing defoliation. Trifolium repens L., Trifolium pratense L., white clover, red clover, defoliation, regrowth, nodules, nitrogen fixation, nitrogenase activity, oxygen limitation     

Potential soil carbon sequestration in a semiarid Mediterranean agroecosystem under climate change: Quantifying management and climate effects

Repeated defoliation and flooding trigger opposite plant morphologies, prostrated and erect ones, respectively; while both induce the consumption of carbohydrate reserves to sustain plant recovery. This study is aimed at evaluating the effects of the combination of defoliation frequency and flooding on plant regrowth and levels of crown reserves of Lotus tenuis Waldst. & Kit., a forage legume of increasing importance in grazing areas prone to soil flooding. Adult plants of L. tenuis were subjected to 40 days of flooding at a water depth of 4 cm in combination with increasing defoliation frequencies by clipping shoot mass above water level. The following plant responses were assessed: tissue porosity, plant height, biomass of the different organs, and utilization of water-soluble carbohydrates (WSCs) and starch in the crown. Flooding consistently increased plant height independently of the defoliation frequency. This response was associated with a preferential location of shoot biomass above water level and a reduction in root biomass accumulation. As a result, a second defoliation in the middle of the flooding period was more intense among plants that are taller due to flooding. These plants lost ca. 90% of their leaf biomass vs. ca. 50% among non-flooded plants. The continuous de-submergence shoot response of frequently defoliated plants was attained in accordance to a decrease of their crown reserves. Consequently, these plants registered only 27.8% of WSCs and 9.1% of starch concentrations with respect to controls. Under such stressful conditions, plants showed a marked reduction in their regrowth as evidenced by the lowest biomass in all plant compartments: shoot, crowns and roots. Increasing defoliation frequency negatively affects the tolerance of the forage legume L. tenuis to flooding stress. Our results reveal a trade-off between the common increase in plant height to emerge from water and the amount of shoot removed to tolerate defoliation. When both factors are combined and defoliation persists, plant regrowth would be constrained by the reduction of crown reserves.     

The Influence of Carbohydrate Reserves on the Response of Nodulated White Clover to Defoliation

A growth-chamber study was carried out to determine whetherthe response of apparent nitrogenase activity (C₂ H₂ reduction)to complete defoliation is influenced by the availability ofcarbohydrate reserves Reserve carbohydrate (TNC) concentrationsof 6-week-old white clover (Trifoliun repens L) plants weremodified by CO₂ pretreatments There was no difference in theresponse of apparent nitrogenase activity to defoliation betweenplants with different TNC concentrations C₂H₂ reduction activitydeclined sharply after defoliation and then recovered similarlyin both high- and low-TNC plants Further experiments were conductedto explain the lack of response of apparent nitrogenase activityto TNC levels Bacteroid degradation was ruled out because invitro nitrogenase activity of crude nodule extracts was stillintact 24 h after defoliation Sufficient carbohydrates appearedto be available to the nodules of defoliated plants becauseadding [¹⁴C]glucose to the nutrient solution did not preventthe decline in apparent nitrogenase activity These conclusionswere supported by the finding that an increase in pO₂ aroundthe nodules of defoliated plants completely restored their C₂H₂reduction activity The comparison of the effects of defoliationand darkness suggested that the decrease in apparent nitrogenaseactivity was not related directly to the interruption of photosynthesisIt appears that lack of photosynthates is not the immediatecause of the decline of nitrogen-fixing activity after defoliation White clover, Trifolium repens L, defoliation, nitrogen fixation, regrowth, reserves, carbohydrates, acetylene reduction, nodule extract     

The Effect of Defoliation on the Nitrogen Economy of White Clover: Regrowth and the Remobilization of Plant Organic Nitrogen

Single plants of white clover (Trifolium repens) were establishedfrom stolon cuttings rooted in acid-washed silver sand. Allplants were inoculated with Rhizobium trifolii, and receivednutrient solution containing 0·5 mg ¹⁵N as either ammoniumor nitrate weekly for 12 weeks (i.e. 6 mg ¹⁵N in total). Plantswere then leniently defoliated or left intact, and the labelledN supply was replaced with unlabelled N. Lenient defoliationremoved fully expanded leaves only, leaving immature leaveswhich accounted for 50–55% of the total; growing pointnumbers were not reduced. Nodules, leaves and growing pointswere counted over the following 21 d period, and d. wts, N contents,and ¹⁵N enrichments of individual plant organs were determined. Defoliated plants had fewer nodules, but numbers of growingpoints were unaffected by defoliation. The rates of both leafemergence and expansion were accelerated in defoliated plants;in consequence the number of young leaves remained less thanin intact plants until day 21. Total dry matter (DM) and N accumulationwere less in defoliated plants, and a greater proportion oftotal plant DM was invested in roots. About 97 % of plant totalN was derived from fixed atmospheric N, but there was incompletemixing of fixed and mineral N within the plant. Relatively moremineral N was incorporated into roots, whereas there was relativelymore fixed N in nodules. There was isotopic evidence that Nwas remobilized from root and stolon tissue for leaf regrowthafter defoliation; approximately 2 % of plant N turned overdaily in the 7-d period after defoliation, and this contributedabout 50% of the N increment in leaf tissue. White clover, Trifolium repens L. cv. SI84, lenient defoliation, N economy, regrowth, N remobilization     

Effects of competitive stress on vegetative growth, storage, and regrowth after defoliation in Phleum pratense

The capacity for resource storage is important to population persistence in nonforaging plants with a consolidation strategy. In a competitive environment, selection may favor genetic variants with a well-developed ability to store resources for future vegetative growth or to regrow following herbivory. To determine the evolutionary potential for changes in vegetative growth, storage and regrowth after defoliation in response to competitive stress, half-sib families of the caespitose grass Phleum pratense were grown in pots in a glasshouse either alone or in competition with four individuals of Lolium perenne . They were defoliated after 16 wk and permitted to regrow for 8 wk. Production of new tillers, leaf area and leaf dry mass were recorded before and after defoliation; resource storage was estimated by the dry mass of stem bases. Tiller numbers, leaf area and mass, regrowth, and stem base mass were significantly reduced by competition; however, there was significant variation among families for many variables. A significant proportion of the variation in clipped and regrowth leaf mass, and in stem base mass, was due to a competition by family interaction. Storage in stem bases was correlated with total accumulated leaf mass in both control and competition groups. Genetic variation for competitive ability and the ability to regrow rapidly after defoliation exist in this P. pratense population. A well-developed ability to regrow following release from competitive or grazing pressures is likely to be a critical adaptation of caespitose grasses.