Comparative lime requirements of tropical and temperate legumes

Summary In a field trial using a continuous function design, calcium carbonate was applied to a nitrogen deficient Hawaiian Oxisol at rates which increased along the plant row from 0 to 22 tons/ha, producing a gradient in soil pH from 4.7 to 7.1. Inoculated legumes representing 18 species were grown.Lime response curves showed no distinct general difference between tropical and temperate legumes. Within each group, individual species varied. Responsiveness rankings of species varied depending on criterion of response.The species ranked as follows according to the amount of lime needed for 90% of the maximum attained yield: Coronilla varia (16 tons/ha) > Leucaena leucocephala (11) > Phaseolus vulgaris, Medicago sativa (9–10) > Glycine max var. Kanrich (7) > Glycine wightii var. Cooper, Lotus corniculatus (6) > Glycine wightii var. Tinaroo, Trifolium repens, Trifolium subterraneum (5) > Desmodium canum, Dolichos axillaris, Glycine max var. Kahala (4) > Arachis hypogea, Desmodium intortum, Vigna sinensis (1–2) > Stylosanthes fruticosa, Stylosanthes guyanensis (0.1).The species ranked as follows according to the magnitude of yield increase due to lime: C. varia (20-fold increase) > T. repens (9-fold) > L. leucocephala, D. axillaris, M. sativa (6-fold) > G. wightii var. Tinaroo (5-fold) > P. vulgaris (4-fold) > G. wightii var. Cooper, L. corniculatus (3-fold) > D. canum, D. intortum, T. subterraneum (2-fold) > A. hypogea, G. max, S. fruticosa, S. guyanensis, V. sinensis (30–50%).Improved calcium availability could account for plant responses to lime rates over 3 to 4 tons/ha. Only 4 tons CaCO₃ sufficed to raise soil pH to 6 and depress solution aluminum and manganese to low concentrations, but 6 tons were needed to raise soluble calcium to 1 meq/l and 20 tons to raise it to 3 meq/l.Journal series No. 1953 of the Hawaii Agricultural Experiment Station.Department of Soils and Plant Nutrition, University of California, Davis.Department of Agronomy and Soil Science, University of Hawaii, Honolulu.     

Nodulation and nitrogen fixation by fast- and slow-growing rhizobia strains of soybean on several temperate and tropical legumes

Three slow-growingBradyrhizobium japonicum (G₃, USDA-110 and KUL-150) of diverse origins and two fast-growing strains ofRhizobium fredii (USDA-192 and USDA-193) were tested with a cropped soybean (Glycine max L. Merrill) cultivar, two cowpeas (Vigna unguiculata), one mung-bean (Phaseolus radiata), one winged-bean (Psophocarpus tetragonolobus) and one field bean (Phaseolus vulgaris) varieties.TheR. fredii strains nodulated and fixed Nitrogen as effectively as the strains ofB. japonicum in a modern european soybean cultivar, namely Fiskeby V. The other western bred soybeans tested were not nodulated by theseR. fredii strains. All of the soybean rhizobia produced nodules in both cowpeas and in mung-bean; theR. fredii strains showed effective N₂-fixation in the cowpeas, particularly USDA-193, yielding shoot dry weights greater than those from theB. japonicum. The symbiotic performance of theR. fredii strains with soybean and other legumes indicated that they should be placed in an intermediate group between the slow-growingB. japonicum and cowpearhizobium sp.The hydrogen uptake activites suggested a possible host effect on the expression of such genes in one out of theB. japonicum strains tested. Furthermore, the slow-growing rhizobia showed significantly higher nitrate-reduction than theR. fredii in the nodules.     

Field experiments on nitrogen fixation by nodulated legumes

Summary Phosphate increased nitrogen uptake by lucerne appreciably on a saline soil. Nitrogenous fertiliser or inoculation with an effective strain ofRhizobium meliloti did not increase the yield significantly. In soils where indigenousRhizobium japonicum was absent inoculation increased soybean yields and the additional fixed nitrogen removed by soybeans amounted to 40 to 120 kg ha⁻¹. Gram and groundnut also responded to Rhizobium inoculation in field trials.     

An appraisal of an IBP experiment on nitrogen fixation by nodulated legumes

Summary Two field experiments on symbiotic nitrogen fixation by nodulated legumes are described, one relatively complex in design and the other very simple. An appraisal has been made of the feasibility of each experiment as a model for world-wide trials intended to provide data about the contribution to nitrogen fixation by useful legumes that can be made by indigenous strains of Rhizobium. It is concluded that more extensive use will be made of the trial and the necessary information secured if the model is of simple design.     

Algal nitrogen fixation in temperate regions

Summary A review of the few earlier investigations of nitrogen fixation by blue-green algae in temperate soils is presented. The extent of fixation by free-living algae in soil at three locations near Uppsala is discussed and compared with fixation by symbiotic algae living as phycobionts or cephalodia in lichens.     

Effect of salt stress on nodulation and nitrogen fixation in legumes

It is now well established that almost all phases of root nodule development in legumes are adversely affected by saline conditions in the rooting medium. There is also a general agreement that the rhizobia are more tolerant to salt stress than the host plant, but they show considerable strain variability in growth and survival under saline conditions. Inhibitory effect of salinity on nodulation has been attributed to decrease in rhizobial colonisation and shrinkage and lack of root hair formation. Salt stress also induces premature senescence of already formed nodules. Both N2-fixation activity and nodule respiration are inhibited sharply on exposure of plants to saline conditions. The decrease in N2-fixation has been ascribed to direct effect on nitrogenase activity or an indirect effect through decrease in leghemoglobin content, respiratory rate, malate concentrations in nodules and photosynthate availability. Salinity increases oxygen diffusion resistance in the nodules and alters their ultrastructure. Decrease in N2-fixation in nodules under salinity is also accompanied by parallel decrease in the activity of H2O2-scavenging enzymes like catalase, ascorbate peroxidase and the level of antioxidants like ascorbic acid. Nodules appear to undergo osmoregulation under saline conditions by accumulating physiologically compatible solutes like proline, sugars (pinnitol) and lactic acid. The intensity of the adverse effects of salinity on nodule functioning depends on plant species, rhizobial strain, duration of exposure to saline conditions, nature, concentration and mode of salt application.     

Effect of light and atmospheric carbon dioxide concentration on nitrogen fixation by herbage legumes

Summary Lucerne, red clover and white clover were grown at two atmospheric concentrations of CO₂ (300 and 1000 μl l⁻¹) and the effects on N₂ fixation, nodule mass/number and root/shoot dry matter production determined. Pea plants were similarly evaluated as a comparison with grain legumes. CO₂ enrichment increased N₂ fixation activity in all cases but activity/unit nodule mass was significantly increased only in the pea. The enhancement of N₂ fixation in herbage legumes by CO₂ enrichment reflected an increase in nodule mass which in turn was attributed to increased nodule number, and results show that under the experimental conditions obtaining here photosynthate supply did not limit nodule N₂ fixation in these plants though it was limiting in the case of peas. White clover growing in a 6 and 14 hour photoperiod was studied for response of the N₂ fixing system to light. Long photoperiod (14 hour) plants assayed at constant temperature (20°C) did not show a significant response to light at the end of the dark period either in terms of fixation per plant or per unit nodule mass, in contrast with short photoperiod (6 hour) plants which showed significant responses. Short photoperiod plants compensated for reduced photosynthates by maintaining only half the root nodule mass and fixation activity of 14 hour photoperiod plants though plants in both systems supported similar rates of N₂ fixation per unit mass of nodule during the photoperiod. Comparison of N₂ fixation activities in whole and decapitated plant systems indicates the importance of shoot reserves for sustaining nitrogenase activity in white clover during short-term interruption of photosynthesis. These results support the conclusion of the CO₂ enrichment studies, that herbage legumes have the potential for supplying their nodule photosynthate requirements for sustaining optimum rates of N₂ fixation and excess carbon supply is used solely to promote further nodulation. Nodules of short photoperiod white clover plants were less efficient in N₂ fixation in that they evolved more H₂ relative to N₂ (C₂H₂) reduced than did long photoperiod plants.     

Effect ofAzospirillum inoculation on nitrogen fixation and growth of several winter legumes

Summary Inoculation of naturally nodulatedPisum sativum L. (garden pea) withAzospirillum in the greenhouse caused a significant increase in nodule numbers above controls. Field inoculation of garden peas in the winter 1981–1982 andCicer arietinum L. (chick pea), in winter 1982–1983, withAzospirillum one week after plant emergence, produced a significant increase in seed yield, but did not affect plant dry matter yield. ForVicia sativa L. (vetch) grown in soil in the greenhouse and in the field for forage, winter 1980–1981, inoculation significantly increased dry matter yield, %N, N-content, and acetylene reduction (nitrogen fixation) activity. InHedysarum coronarium L. (sulla clover), winter 1981–1982, inoculated with both its specificRhizobium (by the slurry method) andAzospirillum, 7 days after emergence, there was an increase in acetylene reduction above controls inoculated withRhizobium alone. These results suggest that it is possible, under conditions tested in this work, to increase nodulation, nitrogen fixation, and crop yields of winter legumes by inoculation withAzospirillum.     

Methods for measuring biological nitrogen fixation in grain legumes

To assure proper management and fully realize the benefits of the legume-Rhizobium symbiosis it is necessary to be able to quantify the amount of nitrogen fixed. Having measured the effectiveness of atmospheric N₂ fixation the macro- or micro-symbionts as well as agronomic factors can be manipulated with the objective to maximize biological nitrogen fixation. A suitable method to quantify nitrogen fixation is therefore necessary in any programme aiming at increasing N₂ fixation, like the one being reported in this volume. There are several methods available to quantify nitrogen fixation and most of the commonly used ones are described in the present paper listing their advantages and disadvantages.     

Factors in the physical and biological environment affecting nodulation and nitrogen fixation by legumes

Summary The effects of root temperature on the four main stages of nodule formation and function are reviewed. Compared with results obtained under optimal conditions, lower root temperatures retard root hair infection more than they affect nodule initiation, nodule development (including bacteroid tissue development and degeneration), or nitrogen assimilation. Higher root temperatures upset the formation of bacteroid tissue and hasten its degeneration. Tropical and subtropical legumes have higher minimum temperatures for nodule formation than temperate species. Low and high shoot temperatures affect nodulation and nitrogen fixation, but the effect is less severe than that of similar root temperatures. Various approaches to minimise adverse environmental effects are considered. These include the selection of appropriate biological material (both host plants and bacterial strains) for the prevailing conditions, and the adoption of management practices designed to utilise the environment or to minimise its adverse effects. The importance of increase in bacteroid volume in relation to increase in rate of nitrogen fixation is considered, and the concept of compensation in nodule production and activity is examined. The limited information on defoliation effects on the nodulation of both temperate and tropical legumes is reviewed and aspects requiring examination are discussed.     

15N2 measurement of nitrogen fixation by legumes and actinorhizals: theory and practice

A gas-tight chamber has been constructed to calibrate the ¹⁵N isotope dilution method against direct ¹⁵N₂ measurements. The theoretical basis for such estimates is given, and the practical problems associated with the experiments are discussed.     

Effects of desiccation and rehydration on nitrogen fixation by epiphylls in a tropical rainforest

Nitrogen fixation rates by epiphyllous microorganisms are affected by desiccation. Rates from leaf samples which had been dried for 12 h were 0.66 ng N/10 cm²/h. In contrast, rates from leaves which had been kept continuously wet were 18.69 ng N/10 cm²/h. Leaf samples which had been rehydrated for 2 and 4 h showed intermediate fixation rates (4.26 and 9.76 ng N/10 cm²/h, respectively). Epiphyllous bryophytes maintain moist conditions on the leaf surface and thus create a microenvironment suitable for prolonged fixation by the microorganisms.     

Germination, field establishment patterns and nitrogen fixation of indigenous legumes on nutrient-depleted soils

Integrating N₂-fixing indigenous legumes in smallholder farming systems has potential to alleviate some of the major soil fertility constraints associated with lack of nitrogen (N) inputs in many parts of Sub-SaharanAfrica. Studies were conducted under low (450–650 mm yr^?1) and high (>800 mm yr^?1) rainfall areas in Zimbabwe to investigate the establishment and nitrogen fixation patterns of fifteen indigenous legume species. The legume seeds were broadcast in mixtures at 120 seeds m^?2 species^?1 during 2004/05 and 2005/06 rainfall seasons.Eriosema ellipticum, Crotalaria ochroleuca andC. pallida had emergence rates above 15% compared with <10% forTephrosia radicans andIndigofera astragalina. Seed hardness accounted for >50% germination failure, while low viability explained 10–30%.Crotalaria ochroleuca andC. pallida attained a maximum biomass of 5–9 t ha^?1 (dry weight) over six months, while species that reached peak biomass over three months (e.g.C. cylindrostachys andC. glauca) gave lowest yields of ≈0.5 t ha^?1. Biennials,Neonotonia wightii, E. ellipticum and Tephrosia radicans, exhibited slow growth rates and only attained their maximum biomass of ≈2 t ha^?1 in the second season. The legumes derived 60–99% of their N from the atmosphere, fixing 5–120 kg N ha^?1 under low rainfall and 78–267 kg N ha^?1 under high rainfall. These findings suggest that the legumes could contribute in restoring productivity of soils continuously cultivated with little or no nutrient inputs in most of Zimbabwe and similar agro-ecologies in SubSaharan Africa.     

Structure of root nodules and nitrogen fixation in Egyptian wild herb legumes

Five wild herb legumes (Trifolium resupinatum, Melilotus indica, Medicago intertexta, Trigonella hamosa, and Alhagi murarum) were collected from cultivated lands of the Nile Valley, and compared with clover (Trifolium alexandrinum), a cultivated forage legume. The wild herb legumes exhibited great variation in nodulation percentage, nodule number, nodule mass and acetylene reduction activity with regard to locality. Nodulation of T. resupinatum and M. indica ranged between 50 - 100% and 33 - 100%, respectively, compared to 50 - 100% for T. alexandrinum. The number of nodules formed on T. resupinatum was 9 - 128 and that of M. indica 6 - 39, compared to 13 - 122 nodule per plant for T. alexandrinum. Nodule mass was correlated with nodule number. In M. indica, a small number of nodules was compensated with high specific nitrogenase activity. The herb legumes formed nodules of small size, varying shape (globose, cylindrical, branched, etc.), and of different types (crotalaroid and astragaloid). Microscopic examination of root-nodules from T. resupinatum, M. indica and M. intertexta, showed that these legumes formed indeterminate and effective nodules, containing apical meristems, central symbiotic tissue with characteristic zonation and peripheral vascular bundles. The nodules harboured bacteroids with pleiomorphic morphology. This revised version was published online in July 2006 with corrections to the Cover Date.     

Biological nitrogen fixation and phosphate solubilization by bacteria isolated from tropical soils

Introduction

In addition to fixing atmospheric nitrogen, some bacterial isolates can also solubilize insoluble phosphates, further contributing to plant growth.

Aims

The objectives of this study were the following: isolate, select, and identify nodulating bacteria in the cowpea that are efficient not only in biological nitrogen fixation (BNF) but also in the solubilization of insoluble inorganic phosphates; identify and quantify the organic acids produced; and establish the relationship between those acids and the solubilizing capacity.

Methods

The bacteria were captured from two soils containing high concentrations of insoluble phosphorus from the cities of Lavras and Patos de Minas, using the cowpea [Vigna unguiculata (L.) Walp.] as bait. We obtained 78 strains, which were characterized according to their cultural attributes in culture medium 79 with the strains UFLA 03-84, INPA 03-11B, and BR3267 (approved by the Ministry of Livestock and Supply Agriculture—MAPA, as inoculants for the cowpea) and Burkholderia cepacia (LMG1222^T), which was used as a positive control for phosphate solubilization. Strains that were selected for their efficiency in both processes were identified by 16S rDNA sequence analysis. We evaluated the symbiotic efficiency (BNF) in a greenhouse and the solubilization efficiency of CaHPO₄, Al(H₂PO₄)₃, and FePO₄.2H₂O in solid and liquid GELP media. Strains that excelled at the solubilization of these phosphate sources were also evaluated for the production of the following organic acids: oxalic, citric, gluconic, lactic, succinic, and propionic.

Results

The presence of Acinetobacter, Bacillus, Firmicutes, Microbacterium, Paenibacillus, and Rhizobium was detected by 16S rDNA sequencing and analysis. Bacterial strains obtained from cowpea nodules varied greatly in the efficiency of their BNF and phosphate solubilization processes, especially in the strains UFLA 03-09, UFLA 03-10, UFLA 03-12, and UFLA 03-13, which were more efficient in both processes. More strains were able to solubilize insoluble inorganic calcium and iron phosphates in liquid medium than in solid medium. The production of organic acids was related to the solubilization of CaHPO₄ and FePO₄.2H₂O for some strains, and the type and concentration of the acid influenced this process.

Conclusions

These are the first results obtained with bacterial isolates from tropical soils in which the production of organic acids was detected and quantified to examine the solubilization of insoluble inorganic phosphates.     

Distinct Bradyrhizobium [corrected] communities nodulate legumes native to temperate and tropical monsoon Australia

Geographic isolation and growing climate aridity played major roles in the evolution of Australian legumes. It is likely that these two factors also impacted on the evolution of their root-nodule bacteria. To investigate this issue, we applied a multilocus sequence analysis (MLSA) approach to examine Bradyrhizobium isolates originating from temperate areas of Western Australia (WA) and the tropical-monsoon area of the Northern Territory (NT). The isolates were mostly collected from the nodules of legumes belonging to tribes, genera and species endemic or native to Australia. Phylogenetic analyses of sequences for the housekeeping atpD, dnaK, glnII, gyrB, recA and 16S rRNA genes and nodulation nodA gene revealed that most isolates belonged to groups that are distinct from non-Australian Bradyrhizobium isolates, which is in line with earlier studies based on 16S rRNA gene sequence analyses. Phylogenetic analysis of the nodA data allowed identification of five major Clades among the WA and NT isolates. All WA isolates grouped in a subgroup I.1 of Clade I with strains originating from temperate eastern Australia. In contrast, the NT isolates formed part of Clades I (subgroup I.2), III (subgroup III.3), IV, V and X. Of these nodA clades, Clade I, Clade IV, Clade X presumably have an Australian origin. Overall, these data demonstrate that the impact of geographic isolation of the Australian landmass is manifested by the presence of numerous unique clusters in housekeeping and nodulation gene trees. In addition, the intrinsic climate characteristics of temperate WA and tropical-monsoon NT were responsible for the formation of distinct legume communities selecting for unrelated Bradyrhizobium groups.