Biology of theParasponia-Bradyrhizobium symbiosis

Parasponia remains the only non-legume known to nodulate withRhizobium/Bradyrhizobium. It is a pioneer plant that is capable of rapid growth and fixing large quantities of nitrogen. In addition to its high agronomic potential, the symbiosis offers the scientist the unique opportunity of studying differences at the molecular level of both partners, and to investigate any possible extension of the symbiosis to other non-legumes of importance. Haemoglobin has been found in the nodule tissue ofParasponia and other nodulated non-legumes and the gene for it has been found and expressed in non-nodulating plants such asTrema tomentosa andCeltis australis. Bradyrhizobium strains isolated from species ofParasponia growing in Papua New Guinea form a group that are more specific in their host requirements thanBradyrhizobium strains from tropical legumes from the same area. They do not effectively nodulate (except CP283) tropical legumes, andParasponia is not readily nodulated withRhizobium andBradyrhizobium strains from legumes. The effectiveness of the symbiosis is influenced by host species, theBradyrhizobium strain and the environment.Parasponia andersonii forms a more effective symbiosis than the other species tested. In competition studies with strains from legumes, isolates fromParasponia always dominate in nodules onParasponia.     

Estimation of symbiotic N2 fixation in an Amazon floodplain forest

Sustainable management for existing Amazonian forests requires an extensive knowledge about the limits of ecosystem nutrient cycles. Therefore, symbiotic nitrogen (N₂) fixation of legumes was investigated in a periodically flooded forest of the central Amazon floodplain (Várzea) over two hydrological cycles (20 months) using the ¹⁵N natural abundance method. No seasonal variation in ¹⁵N abundance (δ ¹⁵N values) in trees which would suggest differences in N₂ fixation rates between the terrestrial and the aquatic phase was found. Estimations of the percentage of N derived from atmosphere (%Ndfa) for the nodulated legumes with Neptunia oleracea on the one side and Teramnus volubilis on the other resulted in mean %Ndfa values between 9 and 66%, respectively. More than half of the nodulated legume species had %Ndfa values above 45%. These relatively high N gains are important for the nodulated legumes during the whole hydrological cycle. With a %Ndfa of 4–5% for the entire Várzea forest, N₂ fixation is important for the ecosystem and therefore, has to be taken into consideration for new sustainable land-use strategies in this area.     

The contribution of symbiotic nitrogen fixation to the nitrogen economy of natural ecosystems

Summary Fourty four species of native wild herbaceous legumes belonging to 12 genera and associated with mature fallow lands of the derived savanna were collected from seven random locations encompassing an estimated area of 10,800 km² and containing the seven different geomorphological soil formations in Anambra State of Nigeria.The number of legume species found differed according to the dominant grass in the fallows sampled, more species being associated with Andropogon, Hyperrhenia and Pennisetum than with Imperata and Loudetia. Detailed vegetation analysis of one hectare of fallow land dominated by Loudetia in one of the locations revealed that legumes comprised about 3% of the species encountered.In greenhouse trials, all the 19 species studied nodulated. The correlation (r=0.646) between fresh weight of nodules and dry weight of tops was significant at 0.01 level. Leaf N in these species ranged from 4.27% to 1.88%.The study indicated that a large number of naturally occurring herbaceous leguminous species, some of which appear to have promising potentials for increasing the N economy of the ecosystem, exists in the fallow land of the derived savanna.     

Source of the soybean N credit in maize production

Nitrogen response trials throughout the United States Corn Belt show that economic optimum rates of N fertilization are usually less for maize (Zea mays L.) following soybean (Glycine max L.) than for maize following maize; however, the cause of this rotation effect is not fully understood. The objective of this study was to investigate the source of the apparent N contribution from soybean to maize (soybean N credit) by comparing soil N mineralization rates in field plots of unfertilized maize that had either nodulated soybean, non-nodulated soybean, or maize as the previous crop. Crop yields, plant N accumulation, soil inorganic N, and net soil mineralization were measured. Both grain yield (6.3 vs. 2.8 Mg ha^–1) and above-ground N accumulation (97 vs. 71 kg ha^–1) were greatly increased when maize followed nodulated soybean compared with maize following maize. A partial benefit to yield and N accumulation was also observed for maize following non-nodulated soybean. Cumulative net soil N mineralization following nodulated soybean, non-nodulated soybean, and maize was 112, 92 and 79 kg N ha^–1, respectively. Net mineralization of soil N appeared to be influenced by both quality (C:N ratio) and quantity of residue from the previous crop. In addition to an increase in plant available N from mineralization, the amount of soil inorganic N (especially in soil 5 cm from the row) was greater following nodulated soybean than non-nodulated soybean or maize. Based on these data, the soybean N credit appears to result from a combination of a decrease in net soil mineralization in continuous maize production and an increase in residual soil N from symbiotic fixation.     

Estimation of N2-fixation by sugarcane,Saccharum sp., and soybean,Glycine max,grown in soil with15N-labelled organic matter

Summary Two varieties of sugarcane, and nodulated and non-nodulated soybean isolines, were planted in a soil previously mixed with¹⁵N-labelled plant material. 45 days was allowed to elapse before planting, to permit initiation of organic matter mineralization. Plants were grown for 60 days, then harvested, dried, weighed and analysed for total N. Analysis of soil samples pre-incubated in the laboratory was carried out to evaluate ammonium and nitrate from added organic matter. Dry weights of the soybean isolines were similar, but total N was higher for the nodulated line. Both sugarcane varieties showed similar weight and total N. Nitrogen derived from applied organic matter (NdfOM) was higher in non-nodulated soybean than in all other plants. Although there is the possibility of different¹⁵N availabilities between species, nitrogen derived from fixation (Nfix) was calculated based on the¹⁵N enrichment of the non-nodulating soybean. Nfix was 72% for nodulating soybean and ranged from 19 to 39% for different parts of sugarcane plants, despite high levels of available-N. Nitrogen derived from soil was calculated by difference. NdfOM was lower in roots than in upper parts (leaves+stalks) of plants. Use of¹⁵N labelled organic matter seems a useful approach to the longer term measurement of N₂-fixation.IAEA Project BRA/5/009-CENA.     

From North to South: A latitudinal look at legume nodulation processes

Legumes and some nodulation processes evolved about 55–60 Ma. Since then they have radiated from their origin at either side of the Tethys seaway, to high latitudes in both the northern and southern hemispheres. In many cases this has involved different tribes and genera, and different nodule processes, but with the common feature that almost all legumes in the higher latitudes are potentially nodulated and, with the exception of some herbaceous species of Chamaecrista, nodulated caesalpinioid legumes are rare. This is not true of tropical regions where all three sub-families are found, with many of their species lacking the ability to nodulate. Whether or not this is related to the availability of combined nitrogen is a matter of current discussion. This review will consider the range of nodulation phenotypes (infection, morphology, structure) and show how many of these are confined to one or other hemisphere. How this might relate to the different genera and species of endophytic (nitrogen fixing) rhizobia in relation to soil conditions will also be discussed. Better knowledge of the wide variation in nodulation parameters is important for understanding the ecology of different regions and for management purposes. Nodule characters are of great potential use in defining taxonomic groupings in legumes.     

Enhancing the early performance of the leguminous shrub Retama sphaerocarpa (L.) Boiss.: fertilisation versus Rhizobium inoculation

We have investigated the effect on growth of fertilisation versus biological nitrogen fixation by rhizobial nodules in Retama sphaerocarpa(L.) Boiss, a leafless leguminous shrub native to the Iberian Peninsula and North-West Africa that has generated interest for revegation of dry Mediterranean habitats. Our main objective was to optimise the formation of root nodules under nursery conditions and to evaluate their influence on the first year of seedling growth in comparison with standard fertilisation. Seedlings of R. sphaerocarpa from two Spanish localities were grown under two levels of fertilisation, and half of each were inoculated with rhizobia isolated from adult Retama, Cytisus and Adenocarpusplants in the field. Although some promiscuity was observed, nodulation was significantly successful with specific rhizobia. At the end of the experiment, highly fertilised plants were taller and heavier and exhibited larger photosynthetic rates than either nodulated or non-nodulated plants under low fertilisation. High fertilisation enhanced seedling growth but inhibited both the nodulation and the nitrogenase activity of the nodules. Thus, physiological differences between nodulated and non-nodulated plants were observed in the low but not in the high fertilisation treatment. Nitrogen uptake and use was enhanced by root nodules, which translated into enhanced photosynthesis and growth. Since inoculation is simple, environmentally friendly and cheap, and nodulated plants are more likely to overcome transplant stress than non-nodulated ones, our results suggest that inoculation together with low, background fertilisation (instead of high fertilisation) should be used when producing high quality seedlings of this autochthonous Mediterranean shrub.     

Weather and nodule mediated variations in delta 13C and delta 15N values in field-grown soybean (Glycine max L.) with special interest in the analyses of xylem fluids

The nodulating soybean (Enrei) and its non-nodulating mutant (EN 1282) were grown in outdoor plots for 2 years (1994: extraordinary dry, high temperature, 1995: ordinary year). Carbon and nitrogen accumulation, delta 13C and delta 15N values in plant parts and xylem fluids and delta 15N values in the water-extractable soil N were analysed throughout the growing period. Plant growth in 1994 was rapid during the early growth stages, but no pods were produced. In 1995, plant growth was normal and pods were formed. The delta 13C values of the leaves were less negative in 1994 than in 1995 and the nodulated plants showed less negative delta 13C values than non-nodulated plants in both years. The delta 13C values of the leaves during the vegetative phase were positively correlated to the leaf N concentrations. Leaf N concentrations in their turn were influenced by nodulation and weather conditions and/or soil available N. The delta 15N values in the plants and xylem fluids were lower in the nodulated soybean than in non-nodulated soybean in both years, and estimates of the contribution of N2 fixation in nodulated plants based on plant top delta 15N values were 7-14% in 1994 and 37-63% in 1995. The delta 13C values of xylem fluids did not differ between nodulated and non-nodulated plants. Thus, the expected contribution by phosphopenolpyruvate carboxylase-mediated CO2 fixation in the root nodules to plant C-incorporation could not have been significant.     

Phosphorus addition reduces invasion of a longleaf pine savanna (Southeastern USA) by a non-indigenous grass (Imperata cylindrica)

Imperata cylindrica is an invasive C₄ grass, native to Asia and increasing in frequency throughout the tropics, subtropics, and southeastern USA. Such increases are associated with reduced biodiversity, altered fire regimes, and a more intense competitive environment for commercially important species. We measured rates of clonal spread by I. cylindrica from a roadside edge into the interior of two longleaf pine savannas. In addition, we measured the effects of fertilization with nitrogen and phosphorus on clonal invasion of one of these sites. Clonal invasion occurred at both sites and at similar rates. Older portions of an I. cylindrica sward contained fewer species of native pine-savanna plants. Clonal growth rates and aboveground mass of I. cylindrica were reduced by the addition of phosphorus relative to controls by the second growing season at one site. As a group, native species were not affected much by P-addition, although the height of legumes was increased by P addition, and the percent cover of legumes relative to native non-legumes decreased with increasing expected P limitation (i.e., going from P-fertilized to controls to N-fertilized treatments). Clonal invasion was negatively correlated with the relative abundance of legumes in control plots but not in P-fertilized plots. Species richness and percent cover of native plants (both legumes and non-legumes) were dramatically lower in N-fertilized plots than in controls or P-fertilized plots. Species richness of native plants was negatively correlated with final aboveground mass of I. cylindrica in control and P-fertilized plots, but not in N-fertilized plots. The results suggest that I. cylindrica is a better competitor for phosphorus than are native pine-savanna plants, especially legumes, and that short-lived, high-level pulses of phosphorus addition reduce this competitive advantage without negatively affecting native plant diversity. Ratios of soil P to N or native legume to non-legume plant species may provide indicators of the resistance of pristine pine savannas to clonal invasion by I. cylindrica.     

Belowground mutualists and the invasive ability of Acacia longifolia in coastal dunes of Portugal

The ability to form symbiotic associations with soil microorganisms and the consequences for plant growth were studied for three woody legumes grown in five different soils of a Portuguese coastal dune system. Seedlings of the invasive Acacia longifolia and the natives Ulex europaeus and Cytisus grandiflorus were planted in the five soil types in which at least one of these species appear in the studied coastal dune system. We found significant differences between the three woody legumes in the number of nodules produced, final plant biomass and shoot ¹⁵N content. The number of nodules produced by A. longifolia was more than five times higher than the number of nodules produced by the native legumes. The obtained ¹⁵N values suggest that both A. longifolia and U. europaeus incorporated more biologically-fixed nitrogen than C. grandiflorus which is also the species with the smallest distribution. Finally, differences were also found between the three species in the allocation of biomass in the different studied soils. Acacia longifolia displayed a lower phenotypic plasticity than the two native legumes which resulted in a greater allocation to aboveground biomass in the soils with lower nutrient content. We conclude that the invasive success of A. longifolia in the studied coastal sand dune system is correlated to its capacity to nodulate profusely and to use the biologically-fixed nitrogen to enhance aboveground growth in soils with low N content.     

Carbon isotope composition of N2-fixing and N-fertilized legumes along an elevational gradient

Dinitrogen-fixing legumes are frequently assumed to be less water-use efficient than plants utilizing soil mineral N, because of the high respiratory requirements for driving N₂ fixation. However, since respiration is assumed not to discriminate against ¹³C, any differences in water-use efficiency exclusively due to respiration should not be apparent in carbon isotope discrimination () values. Our objective was to determine if the source of N (N₂ fixation versus soil N) had any effect on of field-grown grain legumes grown at different elevations. Four legume species, Glycine max, Phaseolus lunatus, P. vulgaris, and Vigna unguiculata, were grown on five field sites spanning a 633 m elevational gradient on the island of Maui, Hawaii. The legumes were either inoculated with a mixture of three effective strains of rhizobia or fertilized weekly with urea at 100 kg N ha^-1 in an attempt to completely suppress symbiotic N₂-fixing activity. In 14 of 20 analyses of stover and 12 of 15 analyses of seed values were significantly higher (p=0.10) in the inoculated plants than the N-fertilized plants. Nitrogen concentrations were generally higher in the fertilized treatments than the inoculated treatments. The different values obtained depending on N-source may have implications in using as an indicator of water-use efficiency or yield potential of legumes.     

Soil fertility and herb facilitation mediated by Retama sphaerocarpa

Abstract. Woody legumes growing in dry climates can increase soil nutrient content and facilitate plant growth in their understorey. We investigated differences in soil fertility and herbaceous community in relation to the presence and absence of the legume Retama sphaerocarpa in a shrubland in a mediterranean type climate. The results showed a higher content of limiting nutrients for plant growth, such as phosphorus and nitrogen, under the R. sphaerocarpa canopy. Herbaceous biomass, cover and nitrogen content increased below the canopy. However, species richness and diversity were diminished in the presence of a dense canopy of R. sphaerocarpa. Nitrogen isotopic analysis did not indicate a clear relationship between symbiotic fixation in R. sphaerocarpa and nitrogen content of soil and plants under its canopy. Nevertheless, herbs growing in the understorey showed a linear correlation between foliar N content and ¹⁵N values. The existence of a dense shrub canopy induced a smaller monthly variation in herb richness, diversity and biomass, suggesting that it provides a stable microhabitat facilitating herbaceous vegetation establishment and growth. The ‘shelter’ effect was more important when environmental conditions became stressful for herbaceous plants.     

Amino acid content in xylem sap of regrowing alfalfa (Medicago sativa L.): Relations with N uptake,N2 fixation and N remobilization

During vegetative regrowth of Medicago sativa L., soil N, symbiotically fixed N₂ and N reserves meet the nitrogen requirements for shoot regrowth. Experiments with nodulated or non-nodulated plants were carried out to investigate the changes in N flows originating from the different N sources and in xylem transport of amino acids during regrowth. Exogenous N uptake, N₂ fixation and endogenous N remobilization were estimated by ¹⁵N labelling and amino acids in xylem sap were analysed. Removal of shoots resulted in great declines of exogenous N flows derived either from N₂ or from NH₄NO₃ during the first week of regrowth, thereafter recovery increased linearly. Mineral N uptake as well as N₂ fixation occurred mainly between the 10th and 18th day after removal of shoots while exogenous N assimilation in intact plants remained at a steady level. Nitrogen remobilization rates in defoliated plants increased by at least three to five-fold, especially during the first 10 days following shoot removal. Compared to control plants, contents of amino acids in xylem sap, during the first 10 days of regrowth, were reduced by about 72% and 82% in NH₄NO₃ grown and in N₂ fixing plants, respectively. Asparagine was the main amino acid transported in xylem sap of both treated plants. Its relative contents during this period significantly decreased from 75% to 59% and from 67% to 36% respectively in non-nodulated plants and in nodulated ones. This decline was accompanied by compensatory increase in the relative contents of aspartate and glutamine.     

Natural abundance of 15N in actinorhizal plants and nodules

Substantial enrichment of some plant parts in ¹⁵N relative to the rest of the plant is unusual, but is found in the nitrogen-fixing nodules of many legumes. A range of actinorhizal plants was surveyed to determine whether the nodules of any of them are also substantially enriched in ¹⁵N. The nonlegume Parasponia, nodulated by a rhizobium, was also included. Four of the actinorhizal genera and Parasponia were grown in N-free culture, and three actinorhizal genera were collected from the field. Nodules of Parasponia, Casuarina and Alnus were¹⁵N enriched relative to other plant parts, but only Parasponia approached the degree of enrichment found in some legume nodules. The nodules of Datisca, Myrica, Elaeagnus, Shepherdia, and Coriaria were depleted in ¹⁵N. Thus many actinorhizal nodules are depleted in ¹⁵N compared to other plant parts and enrichment is modest when it does occur. Whole plant ¹⁵N content (¹⁵N) in four actinorhizal plants and Parasponia showed a relatively narrow range of –1.41 to –1.90. Hence regardless of the degree of nodule enrichment or depletion, whole plant ¹⁵N content appears to vary little in plants grown in N-free culture.     

Autumnal changes in tissue nitrogen of autumn olive,black alder and eastern cottonwood

Two experiments were conducted to determine patterns of N change in tissues of autumn olive (Elaeagnus umbellata Thunb.) and black alder (Alnus glutinosa [L.] Gaertn.) during autumn in central Illinois, U.S.A. In the first study leaf nitrogen concentrations of autumn olive decreased 40% at an infertile minespoil site and 39% at a fertile prairie site throughout autumn whereas nitrogen concentrations in respective bark samples increased by 39% and 37%. Salt-extractable protein concentrations increased in bark and decreased in leaves over the sampling period. Free amino acid concentrations of autumn olive leaves decreased over the course of the experiment from peak concentrations in August. Asparagine, glutamic acid and proline were major constituents of the free amino acid pools in leaves. Total phosphorus concentrations of autumn olive leaves declined by 40–46% during autumn while bark concentrations of P did not significantly change.In the second experiment non-nodulated seedlings of alder receiving a low level of N-fertilization did not exhibit net resorption of leaf N during autumn whereas foliar N concentration of contrasting nonactinorhizal cottonwood plants (Populus deltoides Bartr. ex. Marsh) under the same fertilization regime decreased by 27% after the first frost. A gradual but significant decrease of 38% in foliar N concentration of nodulated alder seedlings grown under a low N-fertilization regime was associated with the cessation of nitrogenase activity during autumn in nodules. Compared with the low N fertilization regime, the higher level of N-fertilization resulted in smaller autumnal decreases of foliar N concentration in nodulated alder (17%) and in cottonwood (20%); but there was no decrease in foliar N concentration in non-nodulated alder. The higher level of N-fertilization promoted a greater accumulation of N in the roots than in the bark of both tree species after the first frost.Our results suggest that black alder lackingFrankia symbionts does not exhibit net leaf N resorption and that autumnal decreases in leaf N ofFrankia-nodulated black alder result primarily from declining foliar N import relative to export due to low temperature inhibition of N₂ fixation. In contrast, autumn olive exhibited greater and more precipitous autumnal declines in foliar N concentration than those of alder, and the pattern of N decline was unaffected by site fertility.     

长白山苔原带凋落物生态化学计量特征及其对模拟氮沉降的响应

长白山苔原是我国乃至欧亚大陆东部独有的高山苔原,根据前人调查植被以灌木苔原为主要类型。在全球变暖背景下,近30年来,长白山岳桦林下的草本植物侵入苔原带,原生灌木苔原分化为灌木苔原、灌草苔原和草本苔原,形成了灌木、灌草混合和草本3种不同类型的凋落物,凋落物数量和质量发生显著改变。与此同时长白山苔原氮沉降量也在逐年增加,导致了土壤中氮的累积,势必影响凋落物的分解。凋落物作为连接植物和土壤的纽带,其分解过程中碳（C）、氮（N）、磷（P）等化学组分和化学计量比的变化直接和间接影响着土壤养分有效性和植物养分利用策略。为揭示氮沉降增加对长白山苔原带不同类型凋落物化学组分及生态化学计量特征早期变化的影响,开展了为期8个月的模拟氮沉降室内凋落物分解实验。在苔原带采集灌木优势种牛皮杜鹃和草本优势种小叶章的凋落物带回实验室,模拟灌木牛皮杜鹃群落、灌草混合的牛皮杜鹃-小叶章群落和草本小叶章群落的3种不同类型凋落物,设置三个施氮处理：对照（CK,0 g N m^-2 a^-1）、低氮（LN,10 g N m^-2 a^-1）、高氮（HN,20 g N m^-2 a^-1）。研究表明：（1）不施氮处理时,3种凋落物的C、P均呈释放状态,木质素（Li）呈先累积再略有降解趋势;牛皮杜鹃凋落物的N元素富集而其余两种凋落物N元素呈释放状态;灌草混合和草本凋落物比原生的灌木凋落物C和N元素释放快、Li累积少;而灌木凋落物的P释放略快于灌草和草本凋落物。3种植被类型凋落物的C/N、C/P、Li/N大小表现为：牛皮杜鹃凋落物>牛皮杜鹃-小叶章混生群落凋落物>小叶章凋落物;N/P表现为：小叶章凋落物>牛皮杜鹃凋落物>牛皮杜鹃-小叶章混生群落凋落物。（2）氮沉降促进3种类型凋落物分解过程中C、N和P化学组分的释放,且氮浓度越高促进作用越显著。在牛皮杜鹃凋落物分解过程中,氮素添加到达某一阈值后,其C/N、C/P、N/P、Li/N的降幅最大,后续若再增加氮素,其对化学计量比的影响均会减弱;本实验中的氮素添加量增加促进了小叶章凋落物的C/N、Li/N下降。（3）草本植物入侵引起凋落物类型的变化带来凋落物分解加快,将导致长白山苔原带养分循环的变化;氮沉降增加对小叶章凋落物化学组分的释放及C/N、Li/N的下降更为促进,小叶章凋落物内难分解化合物减少,分解受到促进。高氮沉降加快了小叶章凋落物与土壤、草本植物之间的养分循环。因此,随着未来苔原带氮沉降量的增加,将更有利于小叶章在与牛皮杜鹃的竞争中获胜,使苔原带呈现草甸化趋势。     

模拟N沉降对太岳山油松人工林和天然林草本群落的影响

由于人类活动氮沉降呈逐年增加的趋势,进而增加了陆地生态系统氮的输入,从而影响陆地生态系统多样性、物种组成和功能。为揭示氮沉降增加对油松林草本群落的影响,于2009年7月在太岳山油松人工林和天然林,设计4个施氮水平:对照(CK,0 kg N hm-2a-1),低氮(LN,50 kg N hm-2a-1),中氮(MN,100 kg N hm-2a-1)和高氮(HN,150 kg N hm-2a-1),研究草本群落的生物多样性、生物量以及草本元素含量对模拟N沉降的响应。研究结果表明:模拟N沉降未能显著影响人工林草本群落的生物多样性(P0.05),而中氮、高氮显著降低了天然林草本群落的生物多样性(P0.05);从Jaccard指数和Sorensen指数分析得出人工林不同氮水平之间草本群落差异性较小,而天然林不同氮水平之间草本群落差异性较大。模拟N沉降没有显著改变人工林草本群落生物量(P0.05),而高氮明显促进天然林草本群落生物量的增加(P0.05)。与对照相比,模拟N沉降提高了人工林和天然林羊胡子苔草叶根中的全N含量(P0.05),而降低了全Mg的含量(P0.05),并且根部元素含量变化与土壤养分含量变化较为一致。施氮提高了N/K、N/Ca、N/Mg(P0.05)的比值。说明油松林下草本群落对氮沉降的响应因林分土壤N饱和程度以及林地利用历史的不同而产生差异,其中天然林响应最为敏感。     

Functional diversity in an Amazonian rainforest of French Guyana: a dual isotope approach (δ15N and δ13C)

Functional aspects of biodiversity were investigated in a lowland tropical rainforest in French Guyana (5°2′N, annual precipitation 2200 mm). We assessed leaf δ¹⁵N as a presumptive indicator of symbiotic N₂ fixation, and leaf and wood cellulose δ¹³C as an indicator of leaf intrinsic water-use efficiency (CO₂ assimilation rate/leaf conductance for water vapour) in dominant trees of 21 species selected for their representativeness in the forest cover, their ecological strategy (pioneers or late successional stage species, shade tolerance) or their potential ability for N₂ fixation. Similar measurements were made in trees of native species growing in a nearby plantation after severe perturbation (clear cutting, mechanical soil disturbance). Bulk soil δ¹⁵N was spatially quite uniform in the forest (range 3–5‰), whereas average leaf δ¹⁵N ranged from −0.3‰ to 3.5‰ in the different species. Three species only, Diplotropis purpurea, Recordoxylon speciosum (Fabaceae), and Sclerolobium melinonii (Caesalpiniaceae), had root bacterial nodules, which was also associated with leaf N concentrations higher than 20 mg g⁻¹. Although nodulated trees displayed significantly lower leaf δ¹⁵N values than non-nodulated trees, leaf δ¹⁵N did not prove a straightforward indicator of symbiotic fixation, since there was a clear overlap of δ¹⁵N values for nodulated and non-nodulated species at the lower end of the δ¹⁵N range. Perturbation did not markedly affect the difference δ¹⁵N_soil−δ¹⁵N_leaf, and thus the isotopic data provide no evidence of an alteration in the different N acquisition patterns. Extremely large interspecific differences in sunlit leaf δ¹³C were observed in the forest (average values from −31.4 to −26.7‰), corresponding to intrinsic water-use efficiencies (ratio CO₂ assimilation rate/leaf conductance for water vapour) varying over a threefold range. Wood cellulose δ¹³C was positively related to total leaf δ¹³C, the former values being 2–3‰ higher than the latter ones. Leaf δ¹³C was not related to leaf δ¹⁵N at either intraspecific or interspecific levels. δ¹³C of sunlit leaves was highest in shade hemitolerant emergent species and was lower in heliophilic, but also in shade-tolerant species. For a given species, leaf δ¹³C did not differ between the pristine forest and the disturbed plantation conditions. Our results are not in accord with the concept of existence of functional types of species characterized by common suites of traits underlying niche differentiation; rather, they support the hypothesis that each trait leads to a separate grouping of species. Received: 18 August 1997 / Accepted: 14 April 1998     

Optimising biological N2 fixation by legumes in farming systems

Whether grown as pulses for grain, as green manure, as pastures or as the tree components of agro-forestry systems, the value of leguminous crops lies in their ability to fix atmospheric N₂, so reducing the use of expensive fertiliser-N and enhancing soil fertility. N₂ fixing legumes provide the basis for developing sustainable farming systems that incorporate integrated nutrient management. By exploiting the stable nitrogen isotope ¹⁵N, it has been possible to reliably measure rates of N₂ fixation in a wide range of agro-ecological field situations involving many leguminous species. The accumulated data demonstrate that there is a wealth of genetic diversity among legumes and their Rhizobium symbionts which can be used to enhance N₂ fixation. Practical agronomic and microbiological means to maximise N inputs by legumes have also been identified.     

Symbiotic performance of grain and wild herbaceous legumes in the Okavango Delta and Tswapong region of Botswana

The low inherent soil fertility, especially nitrogen (N) constrains arable agriculture in Botswana. Nitrogen is usually added to soil through inorganic fertilizer application. In this study, biological nitrogen fixation by legumes is explored as an alternative source of N. The objectives of this study were to measure levels of N₂ fixation by grain legumes such as cowpea, Bambara groundnut and groundnut in farmers’ fields as well as to estimated N₂ fixation by indigenous herbaceous legumes growing in the Okavango Delta. Four flowering plants per species were sampled from the panhandle part of the Okavango Delta and Tswapong area. Nitrogen fixation was measured using the ¹⁵N stable isotope natural abundance technique. The δ¹⁵N values of indigenous herbaceous legumes indicated that they fixed N₂ (?1.88 to +1.35 ‰) with the lowest value measured in Chamaecrista absus growing in Ngarange (Okavango Delta). The δ¹⁵N values of grain legumes growing on farmers’ fields ranging from ?1.2 ‰ to +3.3 ‰ indicated that they were fixing N₂. For grain legumes growing at most farms, %Ndfa were above 50% indicating that they largely depended on symbiotic fixation for their N nutrition. With optimal planting density, Bambara groundnuts on farmers’ fields could potentially fix over 90 kg N/ha in some parts of Tswapong area and about 60 kg N/ha in areas around the Okavango Delta. Results from this study have shown that herbaceous indigenous legumes and cultivated legumes play an important role in the cycling of N in the soil. It has also been shown that biological N₂ on farmer’s field could potentially supply the much needed N for the legumes and the subsequent cereal crops if plant densities are optimized with the potential to increase food security and mitigate climate change.