Sustainable agriculture in the semi-arid tropics through biological nitrogen fixation in grain legumes

Sustainable agriculture relies greatly on renewable resources like biologically fixed nitrogen. Biological nitrogen fixation plays an important role in maintaining soil fertility. However, as BNF is dependent upon physical, environmental, nutritional and biological factors, mere inclusion of any N₂-fixing plant system does not guarantee increased contributions to the soil N pool. In the SAT where plant stover is also removed to feed animals, most legumes might be expected to deplete soil N. Yet beneficial legume effects in terms of increased yields in succeeding cereal crops have been reported. Such benefits are partly due to N contribution from legumes through BNF and soil N saving effect. In addition, other non-N rotational benefits, for example, improved nutrient availability, improved soil structure, reduced pests and diseases, hormonal effects are also responsible. In this paper we have reviewed the research on the contribution of grain legumes in cropping systems and the factors affecting BNF. Based on the information available, we have suggested ways for exploiting BNF for developing sustainable agriculture in the semi-arid tropics (SAT). A holistic approach involving host-plant, bacteria, environment and proper management practices including need based inoculation for enhancing BNF in the cropping systems in the SAT is suggested.     

Enhancing biological nitrogen fixation: An appraisal of current and alternative technologies for N input into plants

Biological nitrogen fixation (BNF) involves a highly specialized and intricately evolved interactions between soil microorganisms and higher plants for harnessing the atmospheric elemental nitrogen (N). This process has been researched for almost a century for efficient N input into plants. The basic mechanism and biochemical steps involved in BNF have been unraveled. It has become abundantly clear that the host plant (legumes) dominates in regulating the BNF process. Environmental factors as well influence this process. Perturbation or any manipulation of the interactions between the bacteria and the legumes seems to offset the critical balance, usually to the detriment of N fixation efficiency. Not much success has been obtained in either enhancing BNF in legumes or transferring important BNF traits to non-nitrogen fixing organisms. An appraisal is given for the lack of success in making the BNF process a popular and efficient agronomic practice. Alternative physiological approaches are presented for improving mobilization, redistribution and utilization of stored N reserves within the host plant.     

Evidence against associative N2 fixation as a significant N source in long-term wheat plots

In monocropped cereal systems, annual N inputs from non-fertilizer sources may be more than 30 kg ha^-1. We examined the possibility that these inputs are due to biological N₂ fixation (BNF) associated with roots or decomposing residues. Wheat was grown under greenhouse conditions in pots (34 cm long by 10 cm diameter) containing soil from a plot cropped to spring wheat since 1911 without fertilization. The roots and soil were sealed from the atmosphere and exposed to a¹⁵N₂-enriched atmosphere for three to four weeks during vegetative, reproductive or post-reproductive stages. This technique permitted detection of as little as 1 μg fixed N plant^-1 in plant material and 40 μg fixed N plant^-1 in soil. No fixation of¹⁵N₂ occurred during either of the first two labelling periods. In the final labelling period, straw returned to the soil was significantly enriched in¹⁵N, especially in a pot with a higher soil moisture content. Total BNF in this pot was 13 μg N plant^-1, or about 30 g N ha^-1. In a separate experiment with soil from the same plot, we detected BNF only when soil was amended with glucose at a high soil moisture content. Measured associative BNF was insufficient to account for observed N gains under field conditions. Lethbridge Research Centre contribution no. 3879488. Lethbridge Research Centre contribution no. 3879488.     

Nitrogen-15 labeling effectiveness of two tropical legumes

Nitrogen-15 foliar applications for the production of field-labeled plant tissues may achieve more effective labeling of plant shoot and root tissues and minimize directly labeling the soil N fraction as occurs when¹⁵ N is soil applied. Consequently, foliar-labeled plant tissues should be better suited for subsequent ¹⁵N mineralization studies. A field experiment was conducted to determine the effectiveness of ¹⁵N-labeling and the accumulation of ¹⁵N in various plant parts of two tropical legumes. Desmodium ovalifolium Guillemin and Perrottet and Pueraria phaseoloides (Roxb.) Benth., grown in 0.5 m² microplots, were labeled with foliar-applied urea containing 99 atom% ¹⁵N. Plants in each microplot received a total of 0.1698 g ¹⁵N that was applied all at once or split equally into two, three or four applications. Legume shoots and roots and soil were destructively harvested and analyzed for total ¹⁵N content. Averaged over both legumes and foliar application rates, total plant (shoots, flowers, leaf litter, and roots) recovery was approximately 79% of the ¹⁵N applied. The soil contained 3% of the ¹⁵N applied, of which 2.5 and 0.5% were in the inorganic and organic fractions, respectively. Nitrogen-15 recovery in shoots (76%) was sixty-five fold greater than in roots (1%) and about nineteen fold greater than the sum of roots and soil (4.1%), a much greater percent recovery than observed in other foliar labeling studies. Averaged over all four foliar split-application rates, ¹⁵N recovery by Desmodium shoots was greater than Pueraria. Results demonstrate that ¹⁵N foliar application to legumes is an effective method for labeling, resulting in atom% excess ¹⁵N levels and ¹⁵N recoveries comparable to those reported with the more traditional soil-labeling approach. Another advantage of this method is a nondestructive, in situ labeling method that permits separation of shoot and root residual N contribution to subsequent crops in N tracer studies.     

Managing native and legume-fixed nitrogen in lowland rice-based cropping systems

Lowlands comprise 87% of the 145 M ha of world rice area. Lowland rice-based cropping systems are characterized by soil flooding during most of the rice growing season. Rainfall distribution, availability of irrigation water and prevailing temperatures determine when rice or other crops are grown. Nitrogen is the most required nutrient in lowland rice-based cropping systems. Reducing fertilizer N use in these cropping systems, while maintaining or enhancing crop output, is desirable from both environmental and economic perspectives. This may be possible by producing N on the land through legume biological nitrogen fixation (BNF), minimizing soil N losses, and by improved recycling of N through plant residues. At the end of a flooded rice crop, organic- and NH₄-N dominate in the soil, with negligible amounts of NO₃. Subsequent drying of the soil favors aerobic N transformations. Organic N mineralizes to NH₄, which is rapidly nitrified into NO₃. As a result, NO₃ accumulates in soil during the aerobic phase. Recent evidence indicates that large amounts of accumulated soil NO₃ may be lost from rice lowlands upon the flooding of aerobic soil for rice production. Plant uptake during the aerobic phase can conserve soil NO₃ from potential loss. Legumes grown during the aerobic phase additionally capture atmospheric N through BNF. The length of the nonflooded season, water availability, soil properties, and prevailing temperatures determine when and where legumes are, or can be, grown. The amount of N derived by legumes through BNF depends on the interaction of microbial, plant, and environmental determinants. Suitable legumes for lowland rice soils are those that can deplete soil NO₃ while deriving large amounts of N through BNF. Reducing soil N supply to the legume by suitable soil and crop management can increase BNF. Much of the N in legume biomass might be removed from the land in an economic crop produce. As biomass is removed, the likelihood of obtaining a positive soil N balance diminishes. Nonetheless, use of legumes rather than non-legumes is likely to contribute higher quantities of N to a subsequent rice crop. A whole-system approach to N management will be necessary to capture and effectively use soil and atmospheric sources of N in the lowland rice ecosystem.IRRI-NifTAL-IFDC joint contribution.     

Biological nitrogen fixation in mixed legume/grass pastures

Biological nitrogen fixation (BNF) in mixed legume/grass pastures is reviewed along with the importance of transfer of fixed nitrogen (N) to associated grasses. Estimates of BNF depend on the method of measurement and some of the advantages and limitations of the main methods are outlined. The amounts of N fixed from atmospheric N₂ in legume/grass pastures throughout the world is summarised and range from 13 to 682 kg N ha^-1 yr^-1. the corresponding range for grazed pastures, which have been assessed for white clover pastures only, is 55 to 296 kg N ha^-1 yr^-1.Biological nitrogen fixation by legumes in mixed pastures is influenced by three primary factors; legume persistence and production, soil N status, and competition with the associated grass(es). These factors and the interactions between them are discussed. Legume persistence, production and BNF is also influenced by many factors and this review centres on the important effects of soil moisture status, soil acidity, nutrition, and pests and disease.Soil N status interacts directly with BNF in the short and long term. In the short-term, increases in soil inorganic N occurs during dry conditions and where N fertiliser is used, and these will reduce BNF. In the long-term, BNF leads to accumulation of soil N, grass dominance, and reduced BNF. However, cyclical patterns of legume and grass dominance can occur due, at least in part, to temporal changes in plant-available N levels in soil. Thus, there is a dynamic relationship between legumes and grasses whereby uptake of soil N by grass reduces the inhibitory effect of soil N on BNF and competition by grasses reduces legume production and BNF. Factors affecting the competition between legumes and grasses are considered including grass species, grazing animals, and grazing or cutting management.Some fixed N is transferred from legumes to associated grasses. The amount of N transferred below-ground, predominantly through decomposition of legume roots and nodules, has been estimated at 3 to 102 kg N ha^-1 yr^-1 or 2 to 26% of BNF. In grazed pasture, N is also transferred above-ground via return in animal excreta and this can be of a similar magnitude to below-ground transfer.Increased BNF in mixed legume/grass pastures is being obtained through selection or breeding of legumes for increased productivity and/or to minimise effects of nutrient limitations, low soil moisture, soil acidity, and pests and disease. Ultimately, this will reduce the need to modify the pasture environment and increase the role of legumes in low-input, sustainable agriculture.     

Biological N2 Fixation in wetland rice fields: Estimation and contribution to nitrogen balance

This paper 1) reviews improvements and new approaches in methodologies for estimating biological N₂ fixation (BNF) in wetland soils, 2) summarizes earlier quantitative estimates and recent data, and 3) discusses the contribution of BNF to N balance in wetland-rice culture.Measuring acetylene reducing activity (ARA) is still the most popular method for assessing BNF in rice fields. Recent studies confirm that ARA measurements present a number of problems that may render quantitative extrapolations questionable. On the other hand, few comparative measures show ARA's potential as a quantitative estimate. Methods for measuring photodependent and associative ARA in field studies have been standardized, and major progress has been made in sampling procedures. Standardized ARA measurements have shown significant differences in associative N₂ fixation among rice varieties.The ¹⁵N dilution method is suitable for measuring the percentage of N derived from the atmosphere (% Ndfa) in legumes and rice. In particular, the ¹⁵N dilution technique, using available soil N as control, appears to be a promising method for screening rice varieties for ability to utilize biologically fixed N. Attempts to adapt the ¹⁵N dilution method to aquatic N₂ fixers (Azolla and blue-green algae [BGA]) encountered difficulties due to the rapid change in ¹⁵N enrichment of the water.Differences in natural ¹⁵N abundance have been used to show differences among plant organs and species or varieties in rice and Azolla, and to estimate Ndfa by Azolla, but the method appears to be semi-quantitative.Recent pot experiments using stabilized ¹⁵N-labelled soil or balances in pots covered with black cloth indicate a contribution of 10–30 kg N ha^-1 crop^-1 by heterotrophic BNF in flooded planted soil with no or little N fertilizer used.Associative BNF extrapolated from ARA and ¹⁵N incorporation range from 1 to 7 kg N ha^-1 crop^-1. Straw application increases heterotrophic and photodependent BNF. Pot experiments show N gains of 2–4 mg N g^-1 straw added at 10 tons ha^-1.N₂ fixation by BGA has been almost exclusively estimated by ARA and biomass measurements. Estimates by ARA range from a few to 80 kg N ha^-1 crop^-1 (average 27 kg). Recent extensive measurements show extrapolated values of about 20 kg N ha^-1 crop^-1 in no-N plots, 8 kg in plots with broadcast urea, and 12 kg in plots with deep-placed urea.Most information on N₂ fixed by Azolla and legume green manure comes from N accumulation measurements and determination of % Ndfa. Recent trials in an international network show standing crops of Azolla averaging 30–40 kg N ha^-1 and the accumulation of 50–90 kg N ha^-1 for two crops of Azolla grown before and after transplanting rice. Estimates of % Ndfa in Azolla by ¹⁵N dilution and delta ¹⁵N methods range from 51 to 99%. Assuming 50–80% Ndfa in legume green manures, one crop can provide 50–100 kg N ha^-1 in 50 days. Few balance studies in microplots or pots report extrapolated N gains of 150–250 kg N ha^-1 crop^-1.N balances in long-term fertility experiments range from 19 to 98 kg N ha^-1 crop^-1 (average 50 kg N) in fields with no N fertilizer applied. The problems encountered with ARA and ¹⁵N methods have revived interest in N balance studies in pots. Balances are usually highest in flooded planted pots exposed to light and receiving no N fertilizer; extrapolated values range from 16 to 70 kg N ha^-1 crop^-1 (average 38 kg N). A compilation of balance experiments with rice soil shows an average balance of about 30 kg N ha^-1 crop^-1 in soils where no inorganic fertilizer N was applied.Biological N₂ fixation by individual systems can be estimated more or less accurately, but total BNF in a rice field has not yet been estimated by measuring simultaneously the activities of the various components in situ. As a result, it is not clear if the activities of the different N₂-fixing systems are independent or related. A method to estimate in situ the contribution of N₂ fixed to rice nutrition is still not available. Dynamics of BNF during the crop cycle is known for indigenous agents but the pattern of fixed N availability to rice is known only for a few green manure crops.     

Field response of rice paddy crop to Azospirillum inoculation: physiology of rhizosphere bacterial communities and the genetic diversity of endophytic bacteria in different parts of the plants

The response of rice plants to the application of inoculant containing two Azospirillum brasilense strains was studied under field conditions. The experiment was performed as three treatments with four replicates in randomized complete blocks arranged as plots of 60 m² in an area on a Vertic Argiudol soil type in the province of Entre Ríos, Argentina. The bacterial rhizosphere community and also the diazotrophic isolates obtained from control and inoculated rice plants were analyzed in relation to their physiology and biological nitrogen fixation (BNF). The MPN of diazotrophs in the rhizosphere varied during the ontogenic cycle. The patterns of distribution of the microbial physiological activities obtained by principal component analysis of community-level physiological profiles (CLPP) showed differences in the utilization of carbon sources by the rhizosphere communities among treatments. Although the analyses of DGGE 16S and nifH profiles have not indicated that the inoculation influenced the genetic diversity of bacterial communities among treatments, they revealed that the banding profiles were altered in different parts of the rice plant by each Azospirillum inoculation treatment. These observations suggest that physiological responses of plant tissues to the inoculation may have occurred. According to agronomic parameters of each treatment, the Azospirillum inoculation increased aerial biomass at the tillering and grain-filling stages. Although the N content accumulated in rice plants increased by 16 and 50 kg ha^?1, the BNF contribution could not be estimated under our experimental conditions by the ¹⁵N balance technique. Based on this field inoculation experiment to rice plants, it is noteworthy that our data suggest that due to Azospirillum inoculation the increase of total N accumulated in rice plants could be a tool to help farmers to improve production and maintain high input of plant residues, providing more organic matter to the soil and guaranteeing sustainability of the system.     

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.     

Application of 15N natural abundance technique for evaluating biological nitrogen fixation in oil palm ecotypes at nursery stage in pot experiments and at mature plantation sites

A range of different species of diazotrophic bacteria has been found in tissues and the rhizosphere of oil palm plants, suggesting a potential to benefit from biological nitrogen fixation (BNF). A few studies have confirmed that plantlets at nursery stage can benefit significantly from BNF after inoculation with Azospirillum spp. but no data are available regarding the benefit from naturally-occurring diazotrophic bacteria in oil palm. The results described here were derived from two pot trials laid out under controlled conditions with plantlets from two important regions for palm oil production in Brazil, as well as from different field sites of mature oil palm plantations. The ¹⁵N natural abundance technique was employed to estimate plant dependence on BNF (%Ndfa) by the different ecotypes grown in soil and previously characterized as hosting diazotrophic bacteria. From both pot trials it was possible to identify some ecotypes of high potential for N₂-fixation that reached in some cases approximately 50%Ndfa. However, the accuracy of measurement still needs to be improved using more suitable reference plants for pot experiments. Values of δ ¹⁵N signals from oil palm and reference plants in the field were inconclusive concerning any benefit from BNF to oil palm, owing to apparently high temporal and spatial variability of δ ¹⁵N of the plant-available N in the heterogeneous soil matrix for the different palm and reference plant tested.     

Nitrogen fixation in legumes and actinorhizal plants in natural ecosystems: values obtained using 15N natural abundance

Background: Nitrogen fixation has been quantified for a range of crop legumes and actinorhizal plants under different agricultural/agroforestry conditions, but much less is known of legume and actinorhizal plant N₂ fixation in natural ecosystems.

Aims: To assess the proportion of total plant N derived from the atmosphere via the process of N₂ fixation (%Ndfa) by actinorhizal and legume plants in natural ecosystems and their N input into these ecosystems as indicated by their ¹⁵N natural abundance.

Methods: A comprehensive collation of published values of %Ndfa for legumes and actinorhizal plants in natural ecosystems and their N input into these ecosystems as estimated by their ¹⁵N natural abundance was carried out by searching the ISI Web of Science database using relevant key words.

Results: The %Ndfa was consistently large for actinorhizal plants but very variable for legumes in natural ecosystems, and the average value for %Ndfa was substantially greater for actinorhizal plants. High soil N, in particular, but also low soil P and water content were correlated with low legume N₂ fixation. N input into ecosystems from N₂ fixation was very variable for actinorhizal and legume plants and greatly dependent on their biomass within the system.

Conclusions: Measurement of ¹⁵N natural abundance has given greater understanding of where legume and actinorhizal plant N₂ fixation is important in natural ecosystems. Across studies, the average value for %Ndfa was substantially greater for actinorhizal plants than for legumes, and the relative abilities of the two groups of plants to utilise mineral N requires further study.     

氮肥和种植密度对达乌里胡枝子的生长与生物固氮的影响

氮供给和种植密度是影响植物生长的两个重要因素。豆科植物因其生物固氮能力而在受到氮限制的生态系统中具有重要作用, 氮含量增加促进植物生长的同时也会抑制豆科植物的生物固氮能力, 种植密度会通过种内竞争影响豆科植物的生长和生物固氮能力, 然而少有研究关注氮肥添加和种植密度对豆科植物生长和生物固氮能力的影响。该研究以达乌里胡枝子(Lespedeza davurica)为研究对象, 通过温室盆栽实验, 探究氮肥和种植密度对其生长和生物固氮的影响。实验设置4个氮添加水平(0、5、10、20 g·m^-2·a^-1)和3种种植密度(1、3、6 Ind.·pot^-1, 约32、96、192 Ind.·m^-2)。结果发现: 1)施肥和密度增加均影响了达乌里胡枝子的生长。叶片碳(C)、氮(N)含量、净光合速率随施氮量增加而增加, 氮添加也促进了植物的生长, 当施氮量为10 g·m^-2·a^-1时植物产量达到最大。叶片C、N含量、净光合速率随种植密度增加而下降, 密度增加可以促进每盆的总生物量, 但对单个植株的生长有负效应。2)氮肥对根瘤形成有抑制作用, 但种植密度增加会缓解氮肥对生物固氮能力带来的“氮阻遏”。该实验条件下, 当施氮量为10 g·m^-2·a^-1, 种植密度为3 Ind.·pot^-1, 或施氮量为5 g·m^-2·a^-1, 种植密度为6 Ind.·pot^-1时, 能最大程度发挥“施氮增产”和种植密度缓解“氮阻遏”的作用。氮添加降低了达乌里胡枝子的根瘤生物量和对根瘤形成的投资(根瘤生物量占总生物量的比例), 从而抑制达乌里胡枝子的生物固氮。种植密度增加导致达乌里胡枝子因种内竞争增加而使资源获取受限, 从而增加对根瘤的投资和根瘤生物量来获得更多来自大气中的氮。3)结构方程结果显示, 氮肥和种植密度通过直接或间接作用, 解释了64%的达乌里胡枝子生物量变化和42%的根瘤生物量变化。上述结果表明合理优化豆科植物的施肥量和种植密度可能对人工草地种植以及退化草地恢复管理具有重要意义。     

Evidence from field nitrogen balance and 15N natural abundance data for the contribution of biological N2 fixation to Brazilian sugarcane varieties

Aims

In Brazil N fertilization of sugarcane (Saccharum spp.) is low compared to most other countries. ¹⁵N-aided studies and the occurrence of many N₂-fixing bacteria associated with cane plants suggest significant contributions from biological N₂ fixation (BNF). The objective of this study was to evaluate BNF contributions to nine cane varieties under field conditions using N balance and ¹⁵N natural abundance techniques.

Methods

The field experiment was planted near Rio de Janeiro in 1989, replanted in 1999 and harvested 13 times until 2004. Soil total N was evaluated at planting and again in 2004. Samples of cane leaves and weeds for the evaluation of ¹⁵N natural abundance were taken in 2000, 2003 and 2004.

Results

N accumulation of the commercial cane varieties and a variety of Saccharum spontaneum were persistently high and N balances (60 to 107?kg?N ha^?1?yr^?1) significantly (p?<?0.05) positive. The δ¹⁵N of leaf samples were lower than any of the weed reference plants and data obtained from a greenhouse study indicated that this was not due to the cane plants tapping into soil of lower ¹⁵N abundance at greater depth.

Conclusion

The results indicate that the Brazilian varieties of sugarcane were able to obtain at least 40?kg?N ha^?1?yr^?1 from BNF.     

Investigating patterns of symbiotic nitrogen fixation during vegetation change from grassland to woodland using fine scale δ15N measurements

Biological nitrogen fixation (BNF) in woody plants is often investigated using foliar measurements of δ¹⁵N and is of particular interest in ecosystems experiencing increases in BNF due to woody plant encroachment. We sampled δ¹⁵N along the entire N uptake pathway including soil solution, xylem sap and foliage to (1) test assumptions inherent to the use of foliar δ¹⁵N as a proxy for BNF; (2) determine whether seasonal divergences occur between δ¹⁵N_{xylem sap} and δ¹⁵N_{soil inorganic N} that could be used to infer variation in BNF; and (3) assess patterns of δ¹⁵N with tree age as indicators of shifting BNF or N cycling. Measurements of woody N‐fixing Prosopis glandulosa and paired reference non‐fixing Zanthoxylum fagara at three seasonal time points showed that δ¹⁵N_{soil inorganic N} varied temporally and spatially between species. Fractionation between xylem and foliar δ¹⁵N was consistently opposite in direction between species and varied on average by 2.4‰. Accounting for these sources of variation caused percent nitrogen derived from fixation values for Prosopis to vary by up to ～70%. Soil–xylem δ¹⁵N separation varied temporally and increased with Prosopis age, suggesting seasonal variation in N cycling and BNF and potential long‐term increases in BNF not apparent through foliar sampling alone.     

矿质养分输入对森林生物固氮的影响

生物固氮是森林生态系统重要的氮素来源,并且在全球氮循环中占有重要的地位。近代以来,因人类活动加剧而导致氮沉降的增加以及其它矿质养分元素(如磷、钼、铁等)输入的改变已成为影响森林生态系统生物固氮的重要因素之一,并引起了学术界的普遍关注。综述了国内外关于森林生物固氮对矿质养分输入的响应及机理。主要内容包括:(1)森林生物固氮的概念及主要的测定方法;(2)矿质养分输入对森林生物固氮的影响。整体上讲,氮素输入抑制了森林生物固氮,磷和其他营养元素输入则表现为促进作用。氮和磷、磷和微量元素同时添加均提高了森林的固氮量;(3)矿质养分改变森林生物固氮的机理。包括生物作用机制(如改变地表层固氮菌的数量或群落丰度、改变结瘤植物的根瘤生物量和附生植物的丰度或盖度)和环境作用机制(如引起土壤酸化、改变碳源物质的含量);(4)探讨了矿质养分输入对森林生物固氮影响研究中所存在的问题,并对未来该领域的研究提出建议。     

Azospirillum amazonense inoculation: effects on growth, yield and N2 fixation of rice (Oryza sativa L.)

Bacteria of the genus Azospirillum are considered to be plant-growth promoting bacteria (PGPR) and stimulate plant growth directly either by synthesising phyto-hormones or by promoting nutrition by the process of biological nitrogen fixation (BNF). Although this genus extensively studied, the effects of inoculation and the possible BNF contribution of the Azospirillum amazonense specie are very scarce. The aim of this study was to isolate, characterise and evaluate auxin production and nitrogenase activity of this species and to select, by inoculation of rice plants, promising isolates based on their ability to improve plant growth, yield and the BNF contribution. One hundred and ten isolates obtained from rice were characterised and grouped according to colony features. Forty-two isolates, confirmed as A. amazonense by the fluorescent in situ hybridization (FISH) technique, were tested for auxin production and nitrogenase activity in vitro. Subsequently plant growth promotion related to plant nutrition effect was evaluated, in vitro and in greenhouse experiments. The BNF contribution to plant growth was evaluated using the ¹⁵N isotope dilution technique. All A. amazonense strains tested produced indoles, but only 10% of them showed high production, above 1.33 μM mg protein⁻¹. The nitrogenase activity also was variable and only 9% of isolates showed high nitrogenase activity and the majority (54%) exhibited a low potential. The inoculation of selected strains in rice under gnotobiotic conditions reduced the growth of root and aerial part when compared to the control, showing the negative effects of excess of phytohormone accumulation in the medium. However, in the greenhouse experiment, inoculation of strains of A. amazonense increased grain dry matter accumulation (7 to 11.6%), the number of panicles (3 to 18.6%) and nitrogen accumulation at grain maturation (3.5 to 18.5%). BNF contributions up to 27% were observed for A. amazonense Y2 (wild type strain). The data presented here is the first report that the PGPR effect of A. amazonense for rice plants grown under greenhouse conditions is mainly due the BNF contribution as measured by ¹⁵N isotope dilution technique, in contrast to the hormonal effect observed with other Azospirillum species studied.     

Biological nitrogen fixation in resource-poor agriculture in South Africa

Farm lands of resource-poor communities in South Africa are depleted of nutrients due to continuous mono-cropping, limited use of fertilisers, and sometimes leaching caused by high rainfall. Despite the well-known advantages of biological nitrogen fixation (BNF) in cropping systems, less than 10% of the grain crops planted annually in these areas are legumes. Using a participatory research and development approach, resource-poor farmers were introduced to conservation agriculture (CA) practices, including BNF, that promoted zero (or reduced) tillage, increased retention of soil cover, as well as crop diversification. Because crop rotation and intercropping of legumes with cereals are known to contribute to soil fertility while enhancing food security, resource-poor farmers from various Provinces in South Africa were trained on the benefits of legume culture for eight years. As a result, these resource-poor farmers did not only get training in inoculation techniques, but were also supplied with inoculants for use on their farms. Data collected from Farmers Demonstration Trials at Belvedere, Dumbarton and Lusikisiki, showed that the grain and fodder yield of maize planted after legumes, and maize intercropped with legumes, were comparable to those of maize receiving high N fertilizer dose (i.e. 54 kg N at planting and 54 kg N as top-dressing). The same data further showed thatRhizobium inoculation, when combined with application of low levels of P and K, significantly increased crop yields within farmers’ trial plots. BNF therefore offers a great opportunity for resource-poor farmers in South Africa to increase their crop yields and thus improve the quality of their livelihoods through the adoption of affordable and sustainable biological technologies that enhance soil fertility.     

Estimation of residual N effect of faba bean and pea on two succeeding cereals using15N methodology

A field experiment was conducted using¹⁵N methodology to study the effect of cultivation of faba bean (Vicia faba L.), pea (Pisum sativum L.) and barley (Hordeum vulgare L.) on the N status of soil and their residual N effect on two succeeding cereals (sorghum (Sorghum vulgare) followed by barley). Faba bean, pea and barley took up 29.6, 34.5 and 53.0 kg N ha^–1 from the soil, but returned to soil through roots only 11.3, 10.8 and 5.7 kg N ha^–1, respectively. Hence, removal of faba bean, pea and barley straw resulted in a N-balance of about –18, –24, and –47 kg ha^–1 respectively. A soil nitrogen conserving effect was observed following the cultivation of faba bean and pea compared to barley which was of the order of 23 and 18 kg N ha^–1, respectively. Cultivation of legumes resulted in a significantly higher A_N value of the soil compared to barley. However, the A_N of the soil following fallow was significantly higher than following legumes, implying that the cultivation of the legumes had depleted the soil less than barley but had not added to the soil N compared to the fallow. The beneficial effect of legume cropping also was reflected in the N yield and dry matter production of the succeeding crops. Cultivation of legumes led to a greater exploitation of soil N by the succeeding crops. Hence, appreciable yield increases observed in the succeeding crops following legumes compared to cereal were due to a N-conserving effect, carry-over of N from the legume residue and to greater uptake of soil N by the succeeding crops when previously cropped to legumes.     

Functional groups' performances as influenced by nitrogen,phosphorus and nodule inhibition of legumes

Aims Nitrogen (N) and phosphorus (P) constitute essential elements for plant growth and their availability influence species diversity in herbaceous plant communities. Legumes exhibit relatively high abundance in N-limited soils. Moreover, the legumes' N:P ratios are much higher than those of the other plant species grown in the same site, probably because they are able to fix atmospheric N 2. The objective of this study was to determine how the relative proportion in N and P availability and the restriction of legumes to fix atmospheric N 2 affect: (i) the primary productivity of plant species, (ii) species composition and (iii) N and P concentrations of species.Methods In an outdoor experiment, mixtures containing grasses, legumes and non-legume forbs were established in 32 containers under four soil treatments (control, N addition, P addition and disinfected soil), in a completely randomized design with eight replicates. Plant growth was examined when N and P were limited in the control soil:sand mixture, in a pot experiment sown with Plantago lanceolata .Important findings The pot experiment indicated that both N and P were limiting for the growth of P. lanceolata. Soil treatments affected primary productivity and species composition. Legumes had a relatively high abundance in the control and their growth was favoured, especially that of Medicago sativa, by P addition. Grasses' growth was increased by the addition of N. Inhibition of rhizobia resulted in poor growth of legumes and concomitant higher growth of grasses, in comparison to the control. The N:P ratios of non-legume species differed between treatments and were always higher in the legume species, even in the disinfected soil. The latter provides evidence that the high N concentrations found in legumes are a physiological characteristic of this specific group of plants.     

Screening legume green manures as nitrogen sources to succeeding non-legume crops

A buried bag incubation technique was proposed to monitor N release from soil and decomposing green manure. The technique would facilitate not only the screening of legumes as sources of N but also measurement of the N supplying capacity of soils. Several tropical legumes were incorporated into field plots followed either by maize (Zea mays L.) or by bare fallow. Soil samples from the plow layer containing the incorporated green manure were placed in low density polyethylene bags and buried within the plow layer under the maize crop for in situ incubation. Periodic withdrawal of the bags was accompanied by fallow soil profile sampling. Above ground N accumulation by maize was equally well correlated to N release measured by either method although the bag technique required much less labor. Supplemental experiments suggested that N accumulation in the bags was reduced due to inadequate O₂ diffusion but only when O₂ demand was high and soil water potential was high. The results show that in situ bag incubation alone or together with fallow soil sampling can be used to estimate the N supplying potential of soil and leguminous residues.