Robert M. Boddey

The contributions of nitrogen-fixing crop legumes to the productivity of agricultural systems

Data collated from around the world indicate that, for every tonne of shoot dry matter produced by crop legumes, the symbiotic relationship with rhizobia is responsible for fixing, on average on a whole plant basis (shoots and nodulated roots), the equivalent of 30–40 kg of nitrogen (N). Consequently, factors that directly influence legume growth (e.g. water and nutrient availability, disease incidence and pests) tend to be the main determinants of the amounts of N2 fixed. However, practices that either limit the presence of effective rhizobia in the soil (no inoculation, poor inoculant quality), increase soil concentrations of nitrate (excessive tillage, extended fallows, fertilizer N), or enhance competition for soil mineral N (intercropping legumes with cereals) can also be critical. Much of the N2 fixed by the legume is usually removed at harvest in high-protein seed so that the net residual contributions of fixed N to agricultural soils after the harvest of legumegrain may be relatively small.Nonetheless, the inclusion of legumes in a cropping sequence generally improves the productivity of following crops. Whilesome of these rotational effects may be associated with improvements in availability of N in soils, factors unrelated to N also play an important role. Recent results suggest that one such non-N benefit may be due to the impact on soil biology of hydrogenemitted from nodules as a by-product of N2, fixation.

Biological nitrogen fixation associated with sugar cane

A recent15N dilution/N balance study confirmed that certain sugar cane varieties are capable of obtaining large contributions of nitrogen from plant-associated N2 fixation. It was estimated that up to 60 to 80% of plant N could be derived from this source, and under good conditions of water and mineral nutrient supply, it may be possible to dispense with N fertilization of these varieties altogether. The recently discovered bacterium,Acetobacter diazotrophicus, apparently responsible for this N2 fixation associated with the plants, has unique physiological properties for a diazotroph, such as tolerance to low pH, and high sugar and salt concentrations, lack of nitrate reductase, and nitrogenase activity which tolerates short-term exposure to ammonium. Furthermore, it also behaves as an endophyte, in that it is unable to infect sugar cane plants unless through damaged tissue or by means of VA mycorrhizae and is propagated via the planting material (stem pieces).

The success of BNF in soybean in Brazil

Approximately forty years after commercial cropping of soybean in Brazil began, the total area under this crop has reached over 13 M ha with a mean productivity of 2400 kg ha−1. Soybean varieties introduced from the USA and varieties rescued from early introductions in Brazilian territory were part of the Brazilian soybean-breeding programme which spread the crop from high to low latitudes. Disease-resistance, pest-resistance, tolerance to low fertility soils, as well as production of plants with pods sufficiently high above the ground for efficient mechanical harvesting, were all aims of the programme. Although BNF was not explicitly considered as a trait for selection in the breeding/selection programme, maximisation of biological nitrogen fixation (BNF) was favoured by conducting selection and breeding trials on soils low in N, in which the seeds were inoculated with efficient Bradyrhizobium inoculants but without N fertiliser application. Several efficient imported Bradyrhizobium strains were found to be unable to compete with native soil micro-flora and other previously-introduced Bradyrhizobium strains. Surprisingly, after being in the soil for many years one or two of these strains had become more competitive while maintaining their high BNF capacity. Today, these strains are included amongst the recommended Brazilian inoculants and have promoted significant improvements in grain yields. The breeding of soybeans in conditions that made grain yield highly dependent on BNF, and the continuous attention paid to the selection of Bradyrhizobium strains appropriate for the newly released varieties, have been the main contributors to todays high yields and their great benefit to the Brazilian economy. There seems to be no reason why this ongoing research programme should not serve as an appropriate model to improve BNF inputs to grain legumes in other countries of the world.

The Potential of C4 Perennial Grasses for Developing a Global BIOHEAT Industry

Unprecedented opportunities for biofuel development are occurring as a result of rising fossil fuel prices, the need to reduce greenhouse gases, and growing energy security concerns. An estimated 250 million hectares (ha) of farmland could be utilized globally to develop a bioenergy industry if efficient and economical perennial biomass crops and bioenergy conversion systems are employed. In temperate zones, C4 or warm-season grass research and development efforts have found switchgrass (Panicum virgatum) and Miscanthus capable of producing biomass yields of 10 to 20 oven dried tonnes (ODT)/ha/yr, while in tropical areas Erianthus and napier grass (Pennisetum purpureum) are producing 25 to 35 ODT/ha/yr. The potential to annually produce 100 barrels of oil energy equivalent/ha with a 25:1 energy output to input ratio appears achievable with high-yielding, N-fixing warm-season grasses grown on marginal lands in the tropics. Commercialization of densified herbaceous plant species has been slow because of the relatively high alkali and chlorine contents of the feedstocks, which leads to clinker formation and the fouling of boilers. This challenge can be overcome by improving biomass quality through advances in plant breeding and cultural management to reduce the chlorine, alkali, and silica content and through the use of new combustion technologies. Warm-season grasses can be readily densified provided suitable grinding and densification equipment and pressure are utilized. The major advantages of producing densified warm-season grasses for BIOHEAT include: it is the most efficient strategy to use marginal farmlands in most temperate and tropical climates to collect solar radiation; it has an excellent energy balance; the feedstocks can be used conveniently in a variety of energy applications; and it is relatively environmentally friendly. Densified warm-season grass biofuels are poised to become a major global fuel source because they can meet some heating requirements at less cost than all other alternatives available today.

Quantification of biological nitrogen fixation associated with sugar cane using a 15N aided nitrogen balance

A nitrogen balance study was performed on four commercial sugar cane cultivars grown in pots containing 64 kg of soil from a sugar cane growing area, with the objective of quantifying possible contributions of biological nitrogen fixation (BNF) to the plants. Distillery (stillage) waste from an alcohol distillery was added to half of the pots, and all treatments were replicated 10 times. The pots were maintained in the field and amended with the equivalent of 80 kg N ha−1 of 15N-labelled urea fertilizer. After 12 months of growth the plants accumulated the equivalent of between 10 and 24% of the total N in the soil and added fertilizer. In the next 9 months of growth, with no more N additions, the plants further accumulated the equivalent of between 8.5 and 19% of the original soil + fertilizer N. The cultivar CB 47–89 accumulated significantly more N than the other cultivars. This difference was greatest in the absence of distillery waste, and in this treatment the 15N enrichment of this cultivar was almost half that of the others, which suggests that there was a contribution of unlabelled N from the air to this cultivar via BNF. After 21 months of growth the N content of the soil and roots was determined on 5 replicates. These analyses revealed that the planted plots lost between 7.5 and 12.5 g of N from soil + fertilizer, but all accumulated more than this in plant tissue. In view of the fact that unplanted plots lost between 10 and 14 g of N during this period, the data suggest that all of the cultivars received some N from associated BNF. In the case of the cultivar CB 47–89, in the absence of distillary waste the plants removed approx. 10 g of N from the soil and fertilizer but accumulated almost 35 g of N. These data constitute the first direct evidence of very significant contributions of plant-associated BNF to a sugar cane cultivar although further experiments must be performed to determine whether such large contributions would be encountered under field conditions.

Effect of inoculation ofAzospirillum spp. on nitrogen accumulation by field-grown wheat

SummaryTwo experiments were performed to examine the effects of inoculation of field grown wheat with various Azospirillum strains. In the first experiment the soil was sterilized with methyl bromide to reduce the Azospirillum population and15N labelled fertilizer was added to all treatments. Two strains ofAzospirillum brasilense isolated from surface sterilized wheat roots and theA. brasilense type strain Sp7 all produced similar increases in grain yield and N content. From the15N and acetylene reduction data it was apparent that these increases were not due to N2 fixation.In the second experiment performed in the same (unsterilized) soil, twoA. brasilense strains (Sp245, Sp246) and oneA. amazonense strain (Am YTr), all isolated from wheat roots, produced responses of dry matter and N content while the response to the strain Sp7 was much smaller. These data confirm earlier results which indicate that if natural Azospirillum populations in the soil are high (the normal situation under Brazilian conditions), strains which are isolated from wheat roots are better able to produce inoculation responses than strains isolated from other sources.The inoculation of a nitrate reductase negative mutant of the strain Sp245 produced only a very small inoculation response in wheat. This suggests that the much greater inoculation response of the original strain was not due to N2 fixation but to an increased nitrate assimilation due to the nitrate reductase activity of the bacteria in the roots.

Nitrogen fixation associated with grasses and cereals: Recent progress and perspectives for the future

Over the last 20 years many new species of N2-fixing bacteria have been discovered in association with grasses, cereals and other non-nodulating crops. Virtually all of these bacteria are microaerophylic, fixing N2 only in the presence of low partial pressures of oxygen. Until a few years ago much attention was focussed on members the genus Azospirillum and it was assumed that N2 fixation was restricted to the rhizosphere or rhizoplane of the host plants. Through the use of N balance and 15N techniques it has been shown that in the case of lowland rice, several tropical pasture grasses and especially sugar cane, the contributions of biological N2 fixation (BNF) are of agronomic significance.

Nitrogen fixation associated with grasses and cereals: recent results and perspectives for future research

Over the last few years research in the area of biological nitrogen fixation (BNF) associated with cereals and grasses has become divided into two areas. On the one hand there have been a large number of reports of responses of field-grown plants to inoculation with N2-fixing bacteria, principallyAzospirillum spp. On the other hand there have been several reports of significant contributions of associated BNF to the nutrition of several crops, including wetland rice, sugar cane and some forage grasses. However, where BNF contributions have definitely been established no certain information is available as to the diazotrophic organisms responsible. Furthermore, certain recent reports indicate that, at least in some cases, responses of plants to inoculation withAzospirillum spp. have been shown not to be due to BNF contributions.In this paper we review some recent progress in this field, particularly at our institute in Rio de Janeiro, concerning specificity of selected Azospirillum strains in the infection of cereal roots and the promotion of responses in the host plants. The possible mechanisms of plant response are discussed including the possibility that plant growth substances or bacterial nitrate reductase are involved. The application of15N and N balance techniques to the quantification of plant associated BNF are considered and the possible strategies that may be adopted to further the understanding of true N2-fixing plant/diazotroph associations.The recent discovery of many more plant-associated N2-fixing bacteria suggests that further research in this area may eventually lead to the development of such associations with applications for agricultural productivity.

The contribution of biological nitrogen fixation for sustainable agricultural systems in the tropics

Abstract The pressing need to increase food production in the tropics to feed the burgeoning population of the Third World requires that crop yields ha−1 must be increased without prejudicing the resource base for future generations. Biological nitrogen fixation (BNF), especially that associated with legumes, has great potential to contribute to productive and sustainable agricultural systems for the tropics, but more research is required to investigate how biologically fixed N, and the increased BNF contributions resulting from research innovations, can be incorporated into viable agricultural systems to increase crop or pasture yields and to substitute N fertiliser inputs. A majority of the soils of the humid and semi-humid tropics have mineral fractions composed of 1:1 lattice clays or sesquioxides of relatively low capacity to retain nutrients (CEC) and water (WHC). It is the soil organic matter (SOM) which has high CEC (after liming) and WHC, and soils under undisturbed climax vegetation are usually high in organic matter which is responsible for their fertility. The key to the long term fertility of such soils is to maintain their soil organic matter by the preservation of crop residues and the selection of suitable crop rotations or fallows. In this review we examine several types of agricultural systems utilised in the tropics ranging from pastures, ley cropping, zero-till rotations as well as green manuring and discuss the management options that can be adopted to preserve their agricultural productivity through the strategic use of legumes in these systems, and their effects on pasture and crop yields. The introduction of forage legumes into tropical pastures can increase and sustain their productivity, with only modest inputs of lime and P and K fertilisers. Similarly, crop and pasture rotations (ley cropping) maintain SOM and soil fertility and crop yields can benefit greatly from the introduction of pasture legumes into the ley. Continuous cropping under minimum or zero tillage can maintain soil cover, and stimulate the retention of SOM, such that nutrient losses are often minimal, and legume derived N can be efficiently transferred to subsequent crops. The options for the resource-poor small-holder to efficiently utilise biologically fixed N as a N supply for cereal grains are more limited and need more attention from researchers as well as less neglect from government organisations. The addition of lime and P fertiliser in modest quantities in many under-developed regions could make large contributions to increased crop yields. If such modest fertiliser inputs were to be combined with suitable crop rotations including green manure or grain legume crops, larger increases in crops yields could be achieved on a sustainable basis, but in many regions agricultural extension services are non-existent and poor farmers have little access to even these basic chemical inputs.

Nodulation and nitrogen fixation by Mimosa spp. in the Cerrado and Caatinga biomes of Brazil

*An extensive survey of nodulation in the legume genus Mimosa was undertaken in two major biomes in Brazil, the Cerrado and the Caatinga, in both of which there are high degrees of endemicity of the genus. *Nodules were collected from 67 of the 70 Mimosa spp. found. Thirteen of the species were newly reported as nodulating. Nodules were examined by light and electron microscopy, and all except for M. gatesiae had a structure typical of effective Mimosa nodules. The endosymbiotic bacteria in nodules from all of the Mimosa spp. were identified as Burkholderia via immunolabelling with an antibody against Burkholderia phymatum STM815. *Twenty of the 23 Mimosa nodules tested were shown to contain nitrogenase by immunolabelling with an antibody to the nitrogenase Fe- (nifH) protein, and using the delta(15)N ((15)N natural abundance) technique, contributions by biological N(2) fixation of up to 60% of total plant N were calculated for Caatinga Mimosa spp. *It is concluded that nodulation in Mimosa is a generic character, and that the preferred symbionts of Brazilian species are Burkholderia. This is the first study to demonstrate N(2) fixation by beta-rhizobial symbioses in the field.