Contribution of biological nitrogen fixation to Elephant grass (Pennisetum purpureum Schum.)

Aims

Because of its high dry matter (DM) productivity, elephant grass (Pennisetum purpureum) is an ideal candidate for biomass production for biofuel production if low N fertilizer rates are used to avoid high fossil fuel inputs. The objective of this study was to investigate the potential of different elephant grass genotypes to obtain contributions of plant-associated biological N₂ fixation (BNF).

Methods

Three field experiments with 4 or 5 different genotypes were conducted on low-fertility Acrisols, two in Rio de Janeiro State and one in Espirito Santo for the evaluation of DM and N accumulation and ¹⁵N abundance.

Results

DM and N accumulation rates of four genotypes in the two experiments in Rio State stabilized at high levels after 2?years of growth. In all experiments the spontaneously-occurring weeds in the plots were significantly higher in ¹⁵N abundance than the elephant grass genotypes. The lower ¹⁵N abundance of the elephant grass was shown not to be due to lower δ¹⁵N abundance at depth in the soil.

Conclusions

Four of the grass genotypes obtained between 18 and 70% of their N from BNF amounting to inputs of between 36 and 132?kg N ha^?1?yr^?1.     

Methods for the study of nitrogen assimilation and transport in grain legumes

Summary The study of nitrogen assimilation and transport in grain legumes is a necessary preliminary to selecting plant cultivars for breeding programs aimed at increasing the contribution of nitrogen fixation to plant production and seed yield. Methods used in such studies are spread widely throughout the literature and have not always been sufficiently tested. This paper presents a comprehensive set of reliable techniques particularly adapted for use in laboratories in developing countries which lack sophisticated and expensive instruments. A continuous flow system for the measurement of N₂ fixation (acetylene reduction) using simple and inexpensive components is described, as are techniques for the estimation of nodule respiration, hydrogen evolution and plant nitrate reductase activity. A number of colorimetric techniques have been adapted for the analysis of nitrate-, amino-, amide-, ureide- and total-N in small volumes of xylem (bleeding) sap.

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Resumen El estudio de la asimilación y del transporte de nitrógeno en leguminosas de grano es un preámbulo necesario para la selección de cultivares obtenidos en programas de mejora que tengan como objetivo el incremento de la contribución de la fijación de nitrógeno a la producción de plantas y de semillas. Los métodos utilizados en dichos estudios estan dispersos en la literatura y no siempre se han comprobado lo suficiente. Este trabajo presenta un amplio conjunto de técnicas seguras, que se adaptan particularmente bien para su uso en laboratorios de países en desarrollo que carecen de instrumental caro y sofisticado. Se describe un sistema de flujo continuo para la medida de la fijación de N₂ (reducción de acetileno) que utiliza componentes sencillos y baratos, tembién se describen técnicas para estimar la respiración de los nódulos, la evolución del hidrógeno y la actividad nitratò-reductasa de la planta. Se han adaptado varios métodos colorimétricos para el análisis del Nitrógeno total y del Nitrógeno combinado (en forma de aminas, amidas y derivados de la urea) contenidos en pequeños volumenes de savia bruta (xilema).

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Résumé L'étude de l'assimilation et du transport de l'azote dans les légumineuses à graines est une nécessité préliminaire pour la sélection des cultivars destinés aux croisements visant à accroître la fixation de l'azote pour la croissance des plantes et le rendement en graines. Les méthodes employées pour ces études sont éparpillées dans la littérature et n'ont pas été suffisamment testées. Dans cet article, il est présenté une étude d'ensemble de méthodes fiables, particulièrement adaptées aux laboratoires des pays en voie de développement qui ne di sposent pas d'instrumentation sophistiquée et coûteuse. Il est notamment décrit un système simple et peu onéreux de flux continu pour mesurer la fixation de l'azote par réduction de l'acétylène, ainsi que des techniques pour déterminer la respiration des nodules, le dégagement d'hydrogéne et l'activité nitrate-réductase des plantes. Plusieurs techniques colorimétriques ont été adaptées à l'analyse de l'azote nitrique, aminé, uréidique et total, dans de faibles volumes de suc de xylème prélevé par saignée.

     

Estimation of biological nitrogen fixation associated withBrachiaria andPaspalum grasses using15N labelled organic matter and fertilizer

Summary Six pasture grasses,Paspalum notatum cv batatais,P. notatum cv pensacola,Brachiaria radicans, B. ruziziensis, B. decumbens andB. humidicola, were grown in concrete cylinders (60 cm diameter) in the field for 31 months. The soil was amended with either a single addition of¹⁵N labelled organic matter or frequent small (2 kg N. ha^–1) additions of¹⁵N enriched (NH₄)₂SO₄. In the labelled fertilizer treatment soil analysis revealed that there was a very drastic change in¹⁵N enrichment in plant-available nitrogen (NO ₃ ^– +NH ₄ ⁺ ) with depth. The different grass cultivars recovered different quantities of applied labelled N, and evidence was obtained to suggest that the roots exploited the soil to different depths thus obtaining different¹⁵N enrichments in soil derived N. This invalidated the application of the isotope dilution technique to estimate the contribution of nitrogen fixation to the grass cultivars in this treatment. In the labelled organic matter treatment the¹⁵N label in the plant-available N declined at a decreasing rate during the experiment until in the last 12 months the decrease was only from 0.274 to 0.222 atom % excess. There was little change in¹⁵N enrichment of available N with depth, hence it was concluded that although the grasses recovered different quantities of labelled N, they all obtained virtually the same¹⁵N enrichment in soil derived N. Data from the final harvests of this treatment indicated thatB. humidicola andB. decumbens obtained 30 and 40% respectively of their nitrogen from N₂ fixation amounting to an input of 30 and 45 kg N.ha^–1 year^–1 respectively.     

A molecular and evolutionary study of the beta-globin gene family of the Australian marsupial Sminthopsis crassicaudata

Beta-globin gene families in eutherians (placental mammals) consist of a set of four or more developmentally regulated genes which are closely linked and, in general, arranged in the order 5'-embryonic/fetal genes- adult genes-3'. This cluster of genes is proposed to have arisen by tandem duplication of ancestral beta-globin genes, with the first duplication occurring 200 to 155 MYBP just prior to a period in mammalian evolution when eutherians and marsupials diverged from a common ancestor. In this paper we trace the evolutionary history of the beta-globin gene family back to the origins of these mammals by molecular characterization of the beta-globin gene family of the Australian marsupial Sminthopsis crassicaudata. Using Southern and restriction analysis of total genomic DNA and bacteriophage clones of beta-like globin genes, we provide evidence that just two functional beta-like globin genes exist in this marsupial, including one embryonic- expressed gene (S.c-epsilon) and one adult-expressed gene (S.c-beta), linked in the order 5'-epsilon-beta-3'. The entire DNA sequence of the adult beta-globin gene is reported and shown to be orthologous to the adult beta-globin genes of the North American marsupial Didelphis virginiana and eutherian mammals. These results, together with results from a phylogenetic analysis of mammalian beta-like globin genes, confirm the hypothesis that a two-gene cluster, containing an embryonic- and an adult-expressed beta-like globin gene, existed in the most recent common ancester of marsupials and eutherians. Northern analysis of total RNA isolated from embryos and neonatals indicates that a switch from embryonic to adult gene expression occurs at the time of birth, coinciding with the transfer of the marsupial from a uterus to a pouch environment.      

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).     

Acetylene reduction in the rhizosphere of rice: Methods of assay

Summary A method is described for thein situ assay of acetylene reduction activity in the rhizosphere of rice involving the use of inexpensive, lightweight equipment. The assay is quick to set up and thus many replicates can be run simultaneously. Control of any acetylene-reducing blue-green algae in the assay chamber is achieved with a photosynthetically active herbicide which has no measurable effect on rhizosphere activity. Results using this method are compared with those from an excised root assay of the same plants. Only a very weak correlation between the two methods was found.     

Alleviation of water and osmotic stress-induced changes in nitrogen metabolizing enzymes in <Emphasis Type="Italic">Triticum aestivum</Emphasis> L. cultivars by potassium

Present communication reports laboratory and pot experiments conducted to study the influence of water and osmotic stress on nitrogen uptake and metabolism in two wheat (Triticum aestivum L) cultivars with and without potassium supplementation. Polyethylene glycol 6000-induced osmotic stress/restricted irrigation caused a considerable decline in the activity of nitrate reductase, glutamate synthase, alanine and aspartate aminotransferases, and glutamate dehydrogenase. Potassium considerably improved nitrogen metabolism under normal water supply conditions and also resulted in amelioration of the negative impact of water and osmotic stresses indicating that potassium supplementation can be used as a potential tool for enhancing the nitrogen use efficiency in wheat for exploiting its genetic potential.     

The effect of the presence of a forage legume on nitrogen and carbon levels in soils under Brachiaria pastures in the Atlantic forest region of the South of Bahia,Brazil

The impact of forest clearance, and its replacement by Brachiaria pastures, on soil carbon reserves has been studied at many sites in the Brazilian Amazonia, but to date there appear to be no reports of similar studies undertaken in the Atlantic forest region of Brazil. In this study performed in the extreme south of Bahia, the changes in C and N content of the soil were evaluated from the time of establishment of grass-only B. humidicola and mixed B. humidicola/Desmodium ovalifolium pastures through 9 years of grazing in comparison with the C and N contents of the adjacent secondary forest. The decline in the content of soil C derived from the forest (C3) vegetation and the accumulation of that derived from the Brachiaria (C4) were followed by determining the ¹³C natural abundance of the soil organic matter (SOM). The pastures were established in 1987, 10 years after deforestation, and it was estimated that until 1994 there was a loss in forest-derived C in the top 30 cm of soil of approximately 20% (9.1 Mg C ha^–1). After the establishment of the pastures, C derived from Brachiaria accumulated steadily such that at the final sampling (1997) it was estimated 13.9 Mg ha^–1 was derived from this source under the grass-only pasture (0–30 cm). Samples taken from all pastures and the forest in 1997 to a depth of 100 cm showed that below 40 cm depth there was no significant contribution of the Brachiaria-derived C and that total C reserves under the grass/legume and the grass-only pastures were slightly higher than under the forest (not significant at P=0.05). The more detailed sampling under the pastures showed that to a depth of 30 cm there was significantly (P<0.05) more C under the mixed pasture than the grass-only pasture. It was estimated that from the time of establishment the apparent rate of C accumulation (0–100 cm depth) under the grass/legume pastures (1.17 Mg ha^–1 yr^–1) was almost double that under the grass-only pastures (0.66 Mg ha^–1 yr^–1). The data indicated that newly incorporated SOM derived from the Brachiaria had a considerably higher C:N ratio than that present under the forest.     

Integrated effects of abiotic stresses on inoculant performance, legume growth and symbiotic dependence estimated by 15N dilution

Temperature, water, salinity, sodicity, acidity and nutrient disorders are major abiotic stresses that can affect legume growth or the establishment and function of the legume-Rhizobium symbiosis. We have examined the literature where the application of the ¹⁵N isotope dilution methodology permits the effect of individual abiotic stresses to be independently and quantitatively separated into plant growth-mediated and BNF (biological N₂ fixation)-mediated components. The response of the symbiosis to a particular stress depends on a host of factors, including legume genotype, cultivar, Rhizobium inoculant, climatic conditions, and the duration, timing and severity of the stress. Published data are analysed in terms of the above variables and their interactions. As a general rule, severe stress inhibits both legume dry matter (DM) and the proportional dependence of the legume on BNF as a source of N. The symbiosis is resilient to low to moderate stress, but there may still be a penalty on legume DM. Gaps in knowledge are identified, and general guidelines on the identification and amelioration of abiotic stresses are provided.