Phytoextraction of selenium from soils irrigated with selenium-laden effluent

This two-part study compared the efficacy of different plant species to extract Se from soils irrigated with Se-laden effluent. The species used were: Brassica napus L. (canola), Brassica juncea Czern L. and Coss (Indian mustard), and Hordeum vulgare L. (barley). In Study 1 we irrigated the plants with a saline effluent containing 0.150 mg Se L^–1, while in Study 2, the same species were planted in a saline soil selenized with 2 mg Se L^–1. Plants were simultaneously harvested 120 days after planting. In Study 1, there were only slight effects of treatment on dry matter (DM) yield. Plant Se concentrations averaged 21 g Se g^–1DM for the Brassica species, and 4.0 g Se g^–1 DM for barley. Total Se added to soils via effluent decreased by 40% for Brassica species and by 20% for barley. In Study 2, total DM decreased for all species grown in saline soils containing Se. Plant Se concentrations averaged 75 g g^–1 DM for Brassica species and 12 g Se g^–1 DM for barley. Total Se added to soils prior to planting decreased by 40% for Brassica species and up to 12% for barley. In both studies, plant accumulation of Se accounted for at least 50% of the Se removed in soils planted to Brassica and up to 20% in soils planted to barley. Results show that although the tested Brassica species led to a significant reduction in Se added to soil via use of Se-laden effluent, additional plantings are necessary to further decrease Se content in the soil.     

Accumulation of selenium by different plant species grown under increasing sodium and calcium chloride salinity

High levels of naturally occurring selenium (Se) are often found in conjunction with different forms of salinity in central California. Plants considered for use in phytoremediation of high Se levels must therefore be salt tolerant. Selenium accumulation was evaluated for the following species under increasing salt (NaCl and CaCl) conditions:Brassica napus L. (canola),Hibiscus cannibinus L. (kenaf),Festuca arundinacea L. (tall fescue), andLotus tenuis L. (birdsfoot trefoil). The experimental design was a complete randomized block with four salt treatments of <1, 5, 10, and 20 dS m^-1, four plant species, three blocks, and six replicates per treatment. Ninety days after growing in the respective salt treated soil with a Se concentration of 2 mg Se kg^-1 soil, added as Na₂SeO₄, all plant species were completely harvested. Among the species tested, shoot and root dry matter yield of kenaf was most significantly (p<0.001) affected by the highest salt treatment and tall fescue and canola were the least affected species. Generally there was a decrease in tissue accumulation of Se with increasing salt levels, except that low levels of salinity stimulated Se accumulation in canola. Canola leaf and root tissue accumulated the highest concentrations of Se (315 and 80 mg Se kg^-1 DM) and tall fescue the least (35 and 7 mg Se kg^-1 DM). Total soil Se concentrations all harvest were significantly (p<0.05) lower for all species at all salt treatments. Removal of Se from soil was greatest by canola followed by birdsfoot trefoil, kenaf and tall fescue. Among the four species, canola was the best candidate for removing Se under the tested salinity conditions. Kenaf may be effective because of its large biomass production, while tall fescue and birdsfoot trefoil may be effective because they can be repeatedly clipped as perennial crops.     

Induced responses in three alkaloid-containing plant species

In this paper we test three plant species for the inducibility of their alkaloid production. The plants were heavily damaged by cutting off 50% of their leaf surface using a pair of scissors. The cut-off leaf tips were used as controls for possible diurnal fluctuations. After 3, 6, 12, 24 and 48 h, respectively, the leaf bases of the damaged plants were harvested and the alkaloid concentration was measured. In Senecio jacobaea the pyrrolizidine alkaloid (Pa) concentration in damaged plants decreased within 6–12 h after damage. Within 24 h after damage the Pa concentration of Cynoglossum officinale doubled compared to control values. Indole alkaloid production in Catharanthus roseus was found not to be induced in this experiment. The responses are discussed in a functional context. We hypothesize that the nature of the response is not a feature of the type of secondary metabolite, but is related to whether the plants are damaged mainly by generalist or by specialist herbivores.     

Inheritance of isozymes in Brassica campestris L. and genetic divergence among different species of Brassiceae

Electrophoretic investigations of Brassica campestris revealed 8 polymorphic loci that had not been described earlier. The inheritance of 7 of them was determined by crosses between different accessions of B. campestris. A phylogeny of B. campestris, B. nigra, B. alboglabra, Sinapis arvensis and S. alba was estimated from the analysis of 15 isozyme loci. The phylogeny showed a close relationship between B. nigra and S. arvensis and a less close relationship between B. campestris and B. oleracea. S. alba was remotely-related to all of the other species.     

Mycorrhizal fungi and the nutrient ecology of three oldfield annual plant species

Summary Three oldfield annual species (Abutilon theophrasti Medic., Ambrosia artemisiifolia L. and Setaria lutescens (Weigel) Hubb.) were investigated. All three developed substantial mycorrhizal infections when inoculated with Glomus etunicatum Becker & Gerd. Mycorrhizal infection dramatically increased phosphorus content and dry weight of both Abutilon and Ambrosia, but did not significantly affect dry weight and only modestly increased phosphorus content of Setaria. These results were consistent with a lower level of infection and much greater root density in Setaria than in the other species. When Abutilon was grown in the presence of Setaria, mycorrhizal infection had no effect on Abutilon phosphorus content or dry weight. The depressive effect of Setaria on the response to inoculation in Abutilon was probably not caused by water soluble allelopathic chemicals from Setaria roots, but soil leachate from Abutilon plants did inhibit infection in other Abutilon plants. The data were consistent with the hypothesis that the very high root density and effective soil exploitation of Setaria reduced the benefit from mycorrhizal infection in Abutilon via phosphorus depletion in a large proportion of the available soil volume. Furthermore, even if mycorrhizal infection were capable of increasing phosphorus content of Abutilon in the presence of Setaria, the very high competitive ability of Setaria for nitrogen in the soil could have reduced the benefit of an enhanced phosphorus content. Carbon isotope ratios were reduced in Abutilon by mycorrhizal infection, indicating a possible reduction in water use efficiency.     

Ant-seed interactions: combined effects of ant and plant species on seed removal patterns

Seed dispersal by ants (i.e. myrmecochory) is usually considered as a mutualism: ants feed on nutritive bodies, called elaiosomes, before rejecting and dispersing seeds in their nest surroundings. While mechanisms of plant dispersal in the field are well documented, the behaviour of the ant partner was rarely investigated in details. Here, we compared in laboratory conditions the foraging behaviour of two ant species, the omnivorous Lasius niger and the insectivorous Myrmica rubra to which seeds of two European myrmecochorous plants (Chelidonium majus and Viola odorata) were given. Ant colonies were simultaneously presented three types of items: entire seeds with elaiosome (SE), seeds without elaiosome (S) and detached elaiosomes (E). The presence of elaiosomes on seeds did not attract workers from a distance since ants first contact equally each type of items. Although ants are mass-recruiting species, we never observed any recruitment nor trail-laying behaviour towards seeds. For ants having contacted seed items, their antennation, manipulation and seed retrieval behaviour strongly varied depending on the species of each partner. Antennation behaviour, followed by a loss of contact, was the most frequent ant-seed interaction and can be considered as a “hesitation” clue. For both plant species, insectivorous Myrmica ants removed items in larger number and at higher speed than Lasius. This fits with the hypothesis of a convergence between odours of elaiosomes and insect preys. For both ant species, the small Chelidonium seeds were retrieved in higher proportion than Viola ones, confirming the hypothesis that ants prefer the higher elaiosome/diaspore-ratio. Thus, in these crossed experiments, the ant-plant pair Myrmica/Chelidonium was the most effective as ants removed quickly almost all items after a few antennations. The presence of an elaiosome body increased the seed removal by ants excepting for Myrmica which retrieved all Chelidonium seeds, even those deprived of their elaiosome. After 24 h, all the retrieved seeds were rejected out of the nest to the refuse piles. In at least half of these rejected items, the elaiosome was discarded by ants. Species-specific patterns and behavioural differences in the dynamics of myrmecochory are discussed at the light of ant ecology. Received 10 September 2007; revised 5 February 2008; accepted 5 March 2008.     

Selenium and sulfur accumulation and soil selenium dissipation in planting of four herbaceous plant species in soil contaminated with drainage sediment rich in both selenium and sulfur

Four selenium (Se) nonaccumulator plant species, including a forage grass species, Tall Fescue (Festuca arundinacea Schreb.), a forage legume species, Alfalfa (Medicago sativa L.), a wetland species, Rush (Juncus tenuis Wild.), and a dry-land alkaline soil species, Saltgrass (Distichlis spicata L.), were grown in soil contaminated by agricultural drainage sediment having elevated levels of Se and sulfur (S). The above-ground plant tissues were consecutively harvested five times and examined for Se and S accumulation. Plant tissue Se concentrations ranged from 23.0 mg kg-1 to 8.3 mg kg-1. Tissue S concentrations ranged from 3239 mg kg-1 to 7034 mg kg-1. Both tissue Se and S concentrations were significantly different between harvests, species, and species/harvest interactions. Total Se accumulation by the plant biomass harvested ranged from 0.3 to 1.3 mg per soil column and total S accumulations ranged from 87.5 to 321.1 mg per soil column. The reduction in the percentage of total soil Se after 24 weeks growth of the plant species ranged from 12.0% in the Tall Fescue planting to 17.3% in the Rush planting. Over 90% of the soil Se losses were unidentified losses and leaching of Se was prevented. The accumulations of Se and S in the plant biomass were very small compared with the total soil Se and S losses, but substantial amounts of total soil Se (12.0 to 15.0%) and S (28.0 to 50.9%) inventories were dissipated by the growing and harvesting of the plants. The soil S concentration was several hundred times higher than the soil Se concentration, but Se accumulation by the plants and Se dissipation from the soil were not impaired by the high level of soil sulfur. For natural grassland habitat restoration, such as at the Kesterson Wildlife Refuge in the Central Valley of California, or for restoration of large-scale Se contaminated agricultural lands, Se nonaccumulator plant species are favorable candidates, because the possibility of introducing Se toxicity into the food chain can be minimized.     

Tolerance and phytoaccumulation of Chromium by three Azolla species

Azolla, an aquatic fern is ideal candidate for exploitation in constructed wetlands for treating metal-contaminated wastewaters. This study demonstrates the potential of Azolla spp. namely A. microphylla, A. pinnata and A.␣filiculoides to tolerate Cr ions in the growth environment and bioconcentrate them. These species could grow in presence of up to 10 μg ml⁻¹ Cr and showed biomass production 30–70% as compared to controls. Nitrogenase activity was not affected at 1–5 μg ml⁻¹ but at higher concentrations it diminished. There was no growth at higher concentrations of chromium. However, the necrosed biomass harvested from treatments containing higher concentrations of chromium, accumulated Cr to the levels 5000–15,000 μg g⁻¹. At increased levels of Cr, the metal was accumulated in higher amount in dry biomass. Bioconcentration Factor (BCF) ranged between 243 and 4617 for the three species. A. microphylla showed highest bioconcentration potential. Thus, these Azolla spp. can be exploited for treatment of tannery and other Cr contaminated wastewaters.     

Nitrogen resorption from senescing leaves of three salt marsh plant species

Seasonalvariation in leaf nitrogen of mature green and senescent leaves and nitrogenresorption efficiency in three plants (Spartina maritima, Halimioneportulacoides and Arthrocnemum perenne) of aTagus estuary salt marsh are reported. Total nitrogen concentrations in greenand senescent leaves were higher during winter (December and March). Soilinorganic nitrogen availability showed an opposite pattern with higherconcentrations during summer (June and September) when total leaf biomass washigher. Nitrogen resorption efficiency ranged between 31 and 76% andH. portulacoides was the plant that better minimizednitrogen loss by this process. Nitrogen resorption occurred mainly from thesoluble protein pool, although other fractions must have been broken down duringthe resorption process. No significant seasonal variation in nitrogen resorptionefficiency and no relation to leaf total nitrogen or soil nitrogen availabilitywere found. This suggests that the efficiency of the resorption process is notdetermined by the plant nitrogen status nor by the availability of the nutrientin the soil. Nevertheless, resorption from senescing leaves may play animportant role in the nitrogen dynamics of salt marsh plants and reduce thenitrogen requirements for plant growth.     

The mobility of boron in plants

In order to investigate a possible remobilization of boron, tomato plants were first exposed to excessive supplies of B and thereafter transferred to B-free nutrient solutions. Almost all the gain of top biomass after this transfer consisted of the growth of pre-existing immature leaves. Little new growth occurred before severe B deficiency became apparent. Boron was moved out of stems, but there was no evidence for a transfer of B from leaf blades into new tissues at the top.Although the B concentration in roots was low after the transfer to the B-free solution, there was a manyfold increase in root biomass. Consequently, the B content of the entire root system increased.It is concluded that little B was remobilized and transported into plant tops, whereas a small but adequate amount was remobilized and transported to the roots.     

Effect of excess boron on summer and winter squash

The boron tolerance of two summer squash cultivars (Cucurbita pepo L. Aristocrat Zucchini and Peter Pan Scallop) and one winter squash cultivar (Cucurbita moschata Poir. Butter boy) was determined in large, outdoor sand cultures. Boron treatments were imposed by irrigation with culture solutions that contained 1.0, 3.0, 6.0, 9.0, 12.0, or 15.0 mg B L^-1. Relative fruit yields of Zucchini, Scallop, and Butter boy were reduced 5.2%, 9.8%, and 4.3% with each unit (mg L^-1) increase in soil solution B (B_sw)>2.7, 4.9, and 1.0 mg B L^-1, respectively. Reduced yields of all cultivars were attributed to a reduction in fruit number and not fruit size. Boron concentrations in leaves and fruit were directly correlated to B_sw.     

The accumulation and assimilation of dimethylselenide by four plant species

Plants of Agrostis tenuis Sibth., Hordeum vulgare L., Lycopersicon esculentum Mill. and Raphanus sativus L. were grown hydroponically in sealed systems and fumigated with 8 g m^-3 [⁷⁵Se]-dimethylselenide. The accumulation of ⁷⁵Se was measured and the shoot tissues were extracted to examine the products of the ⁷⁵Se assimilation. Characteristic differences were observed between species in the accumulation of ⁷⁵Se and the transport from shoots to roots. High-voltage electrophoresis and chromatography of extracts made with 80% aqueous ethanol revealed the presence of inorganic selenite as an assimilation product as well as the selenium analogues of glutathione and methionine. Extensive incorporation of ⁷⁵Se into protein-bound selenomethionine was observed in all plant species.Abbreviation DMSe dimethylselenide     

Evaluation of three herbaceous index plant species for bioavailability of soil cadmium, chromium, nickel and vanadium

Uncultivated plants growing on disturbed sites may be useful for assessing the bioavailability of some metals in soils, and thus the potential for metal mobilization up the terrestrial food chain, an important element in ecological risk assessment. A planted chicory cultivar (Cichorium intybus L. var. foliosum Hegi.) and the uncultivated plants horseweed (Canada fleabane) (Erigeron canadensis L.) and dogfennel (Eupatorium capillifolium (Lam.) Small) were evaluated for their ability to act as index plant species for soil Cd, Cr, Ni, and V at two field sites where these metals had been applied five yr previously to two highly weathered sandy Ultisols. Soil Cd was available to all analyzed plant tissues of all three plant species at both sites, particularly on the sandier Blanton soil. Chicory was an effective index plant for Cd on the finer textured Orangeburg soil but functioned as an indicator plant (toxicity symptoms were observed) on the sandier Blanton soil. Horseweed and dogfennel were effective index plants for Cd in both contaminated soils. Soil Cr, Ni, and V were less bioavailable than soil Cd and plant metal uptake was more sensitive to residual soil Cr, Ni, and V than was soil extraction with double acid. Horseweed and chicory may have potential as index plants for soil Cr. Chicory may have potential as a Ni index plant. Chicory and dogfennel may have potential as V index plants.     

Callus initiation and regeneration capacities in Brassica species

In order to know the genetic differences of de- and redifferentiation capacities, seven Brassica species (B. campestris, B. nigra, B. oleracea, B. hirta, B. carinata, B. juncea and B. napus) were cultured in vitro, and their response to the medium supplemented with various combinations of auxin and cytokinin hormones was compared. Important factors for callus initiation were shown to be auxin and species. Calli were induced most frequently in Murashige and Skoog (MS) medium with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), whereas -naphthaleneacetic acid (NAA) induced preferentially roots at a concentration of 2 to 5 mg/l. Callus-, root- and shoot-forming capacities from explanted cotyledon tissues were significantly different among the seven Brassica species. Calli derived from cotyledons and hypocotyls of seven species were transferred to MS media with 20g/l sucrose, 0 to 0.1 mg/l NAA and 0 to 4 mg/l kinetin to compare their regeneration capacities. Among the seven species tested, B. napus (2n=4x=38, genome AACC) had the highest shoot forming capacity (20.0%). Other amphiploid species, B. carinata (2n=4x=34, BBCC) and B. juncea (2n=4x=36, AABB) formed shoots at low frequencies (2.8% and 1.2%, respectively). A diploid species, B. oleracea (2n=2x 18, CC) also showed high shoot formation (10.2% on average). This result suggests that the gene(s) controlling shoot formation may be localized in the C genome. Differences were also found among varieties and cultivars within a species. One of the cultivars, Siberian kale (B. oleracea var. acephala) gave about 50% shoot formation. This kale was shown cytologically to be an autotetraploid (2n=4x=36, CCCC), thus probably possessing a double set of the shoot-forming gene(s).     

三种苦苣苔对石灰土和红壤的适应性分析

为探讨苦苣苔科植物对其岩溶生境的适应性,该研究选取黄花牛耳朵(Primulina lutea)、紫花报春苣苔(Pri.purpurea)和桂林蛛毛苣苔(Paraboea guilinensis)三种苦苣苔科植物,将其栽种在石灰土及红壤两种不同类型的土壤中,观测记录其生长性状并对其叶片元素含量进行测定和比较。植株采集过程中,同时采集自然生境中三种苦苣苔科植物叶片及取样植物基部土壤,并对叶片及土壤元素的含量进行测定,作为今后苗圃试验的参照。结果表明:三种苦苣苔科植物在两种土壤上的生长状况及适应性具有差异,其在石灰土上生长良好,在红壤上生长较差;在两种不同土壤中,除N外,桂林蛛毛苣苔的叶片其他元素(P、K、Mn、Mg、Ca、Zn、Cu)差异极显著(P0.01);除P外,紫花报春苣苔的叶片其他元素(N、K、Mn、Mg、Ca、Zn、Cu)差异极显著(P0.01);除N、Cu、Ca外,黄花牛耳朵的叶片元素(P、K、Mn、Mg、Zn)差异极显著(P0.01);三种植物的叶片元素比值,除少数值没有差异外,大部分指标差异都极显著;对叶片元素与栽培土壤元素的相关性分析,发现植物叶片Mn元素与土壤中N、Ca、Mg、Zn、Mn、有机质含量等呈正相关,土壤P元素与叶片中N、P元素呈正相关,而与叶片中Zn元素呈负相关关系。在其他栽培条件一致的条件下,土壤因素及物种差别是造成黄花牛耳朵、紫花报春苣苔和桂林蛛毛苣苔适应性产生差异的主要原因。     

High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species

The plant regeneration ability of zygotic embryo-derived callus cultures was studied for 12 A. cepa varieties and accessions, two A. fistulosum varieties, one A. fistulosum x A. cepa interspecific hybrid and two A. porrum varieties. Compact embryogenic callus was induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid. The embryogenic calluses of all three Allium species were similar in appearance. For all accessions tested plants could be regenerated at a high frequency from this compact callus through somatic embryogenesis, when using kinetin supplemented MS medium (regeneration medium). Addition of abscisic acid to the regeneration medium stimulated the formation of both somatic embryos and shoots for a number of varieties. Concerning shoot regeneration from callus cultures, significant differences existed between genotypes of all accessions except one.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - VDH Van Der Have Seed company     

Production and genetic analysis of partial hybrids in intertribal crosses between Brassica species (B. rapa, B. napus) and Capsella bursa-pastoris

Capsella bursa-pastoris (L.) Medic (2n = 4x = 32) is a natural double-low (erucic acid < 1%, glucosinolates < 30 micromol/g) germplasm and shows high degree of resistance to Sclerotinia sclerotiorum. Hybridizations were carried out between two Brassica species viz. B. rapa (2n = 20) and B. napus (2n = 38) as female and C. bursa-pastoris as male parent to introduce these desirable traits into cultivated Brassica species. Majority of F(1) plants resembled female parents in morphology and only a few expressed some characters of male parent, including the white petals. Based on cytological observation of somatic cells, the F(1) plants were classified into five types: two types from the cross with B. rapa, type I had 2n = 27-29; type II had 2n = 20; three types from the crosses with B. napus, type III was haploids with 2n = 19; type IV had 2n = 29; type V had 2n = 38. One to two chromosomes of C. bursa-pastoris were detected in pollen mother cells (PMCs) of type I plant by genomic in situ hybridization (GISH), together with chromosomal segments in ovary cells and PMCs of some F1 plants. Amplified fragment length polymorphism (AFLP) bands specific for the male parent, novel for two parents and absent bands in Brassica parents were generated in majority of F1 plants, even in Brassica-types and haploids, indicating the introgressions at various levels from C. bursa-pastoris and genomic alterations following hybridization. Some Brassica-type progeny plants had reduced contents of erucic acid and glucosinolates associated with improved resistance to S. sclerotiorum. The cytological and molecular mechanisms behind these results are discussed.     

Patterns of foraging and range use by three species of neotropical primates

This paper describes the diet and range use patterns of the three species of primates in Santa Rosa National Park, Costa Rica (Ateles geoffroyi, Alouatta palliata, andCebus capucinus) and examines the variation in these variables as they relate to seasonal changes and concomitant changes in food availability. These three primate species were studied over a four-year period for a total of 24 months in the field. Santa Rosa National Park is in an area that experiences a long severe dry season in which little if any rain falls and the majority of the non-riparian trees lose their leaves. However, even though the three species were very flexible in terms of diet and range use, the behavioural variability did not correspond to changes in food availability or season.     

Effects of liming and boron fertilization on boron uptake of Picea abies

The effects of liming on concentrations of boron and other elements in Norway spruce [Picea abies (L) Karst.] needles and in the mor humus layer were studied in long-term field experiments with and without B fertilizer on podzolic soils in Finland. Liming (2000+4000 kg ha^-1 last applied 12 years before sampling) decreased needle B concentrations in the four youngest needle age classes from 6–10 mg kg^-1 to 5 mg kg^-1. In boron fertilized plots the corresponding concentrations were 23–35 mg kg^-1 in control plots and 21–29 mg kg^-1 in limed plots. Both liming and B fertilizer decreased the Mn concentrations of needles. In the humus layer, total B concentration was increased by both lime and B fertilizer, and Ca and Mg concentrations and pH were still considerably higher in the limed plots than controls. Liming decreased the organic matter concentration in humus layer, whilst B fertilizer increased it.The results about B uptake were confirmed in a pot experiment, in which additionally the roles of increased soil pH and increased soil Ca concentration were separated by means of comparing the effects of CaCO₃ and CaSO₄. Two-year-old bare-rooted Norway spruce seedlings were grown in mor humus during the extension growth of the new shoot. The two doses of lime increased the pH of soil from 4.1 to 5.6 to 6.1, and correspondingly decreased the B concentrations in new needles from 22 to 12 to 9 mg kg^-1. However, CaSO₄ did not affect the pH of the soil or needle B concentrations. Hence the liming effect on boron availability in these soils appeared to be caused by the increased pH rather than increased calcium concentration.     

Dinitrogen-fixation by three neotropical agroforestry tree species under semi-controlled field conditions

Cultivating dinitrogen-fixing legume trees with crops in agroforestry is a relatively common N management practice in the Neotropics. The objective of this study was to assess the N₂ fixation potential of three important Neotropical agroforestry tree species, Erythrina poeppigiana, Erythrina fusca, and Inga edulis, under semi-controlled field conditions. The study was conducted in the humid tropical climate of the Caribbean coastal plain of Costa Rica. In 2002, seedlings of I. edulis and Vochysia guatemalensis were planted in one-meter-deep open-ended plastic cylinders buried in soil within hedgerows of the same species. Overall tree spacing was 1 × 4 m to simulate a typical alley-cropping design. The ¹⁵N was applied as (NH₄)₂SO₄ at 10% ¹⁵N atom excess 15 days after planting at the rate of 20 kg [N] ha⁻¹. In 2003, seedlings of E. poeppigiana, E. fusca, and V. guatemalensis were planted in the same field using the existing cylinders. The ¹⁵N application was repeated at the rate of 20 kg [N] ha⁻¹ 15 days after planting and 10 kg [N] ha⁻¹ was added three months after planting. Trees were harvested 9 months after planting in both years. The ¹⁵N content of leaves, branches, stems, and roots was determined by mass spectrometry. The percentage of atmospheric N fixed out of total N (%N_f) was calculated based on ¹⁵N atom excess in leaves or total biomass. The difference between the two calculation methods was insignificant for all species. Sixty percent of I. edulis trees fixed N₂; %N_f was 57% for the N₂-fixing trees. Biomass production and N yield were similar in N₂-fixing and non-N₂-fixing I. edulis. No obvious cause was found for why not all I. edulis trees fixed N₂. All E. poeppigiana and E. fusca trees fixed N₂; %N_f was ca. 59% and 64%, respectively. These data were extrapolated to typical agroforestry systems using published data on N recycling by the studied species. Inga edulis may recycle ca. 100 kg ha⁻¹ a⁻¹ of N fixed from atmosphere to soil if only 60% of trees fix N₂, E. poeppigiana 60–160 kg ha⁻¹ a⁻¹, and E. fusca ca. 80 kg ha⁻¹ a⁻¹.