Localization of Associative Cyanobacteria on the Roots of Epiphytic Orchids

This work is the first study of the localization of phototrophic microorganisms in the rhizoplane and velamen of epiphytic orchids, namely, on the aerial and substrate roots of Acampe papillosa and Dendrobium moschatum and on the aerial roots of Phalaenopsis amabilis and Dendrobium phalaenopsis. The composition of the bacterial community on the plant roots depended on the conditions of plant growth. Under conditions simulating the climate of moist tropical forests, the aerial roots proved to be populated with phototrophic microorganisms, among which cyanobacteria predominated. Interlaced fungal hyphae and filamentous cyanobacteria formed a sheath on the surface of the aerial roots. The nitrogen-fixing capacity of the sheath of the aerial roots was studied on the example of P. amabilis.     

Diversity of Rhizobium-Phaseolus vulgaris symbiosis: overview and perspectives

Common bean (Phaseolus vulgaris) has become a cosmopolitan crop, but was originally domesticated in the Americas and has been grown in Latin America for several thousand years. Consequently an enormous diversity of bean nodulating bacteria have developed and in the centers of origin the predominant species in bean nodules is R. etli. In some areas of Latin America, inoculation, which normally promotes nodulation and nitrogen fixation is hampered by the prevalence of native strains. Many other species in addition to R. etli have been found in bean nodules in regions where bean has been introduced. Some of these species such as R. leguminosarum bv. phaseoli, R. gallicum bv. phaseoli and R. giardinii bv. phaseoli might have arisen by acquiring the phaseoli plasmid from R. etli. Others, like R. tropici, are well adapted to acid soils and high temperatures and are good inoculants for bean under these conditions. The large number of rhizobia species capable of nodulating bean supports that bean is a promiscuous host and a diversity of bean-rhizobia interactions exists. Large ranges of dinitrogen fixing capabilities have been documented among bean cultivars and commercial beans have the lowest values among legume crops. Knowledge on bean symbiosis is still incipient but could help to improve bean biological nitrogen fixation.     

Mineral nutrition and growth of tropical maize as affected by soil acidity

Soil constraints linked to low pH reduce grain yield in about 10% of the maize growing area in tropical developing countries. The aim of this research was to elucidate the reasons for this maize yield reduction on an oxisol of Guadeloupe. The field experiment had two treatments: the native non-limed soil (NLI, pH 4.5, 2.1 cmol Al kg^–1, corresponding to 20% Al saturation), and the same soil limed 6 years prior to the experiment (LI, pH 5.3, 0 cmol Al kg^–1). The soils were fertilized with P and N. The above-ground biomass, root biomass at flowering, grain yield and yield components, leaf area index (LAI), light interception, radiation-use-efficiency (RUE), P and N uptake, soil water storage, and soil mineral N were measured during the maize cycle. The allometric relationships between shoot N concentration, LAI and above-ground biomass in LI were similar to those reported for maize cropped in temperate regions, indicating that these relationships are also useful to describe maize growth on tropical soils without Al toxicity. In NLI, soil acidity severely affected leaf appearance, leaf size and consequently the LAI, which was reduced by 60% at flowering, although the RUE was not affected. Therefore, the reduction in the above-ground biomass (30% at flowering) and grain yield (47%) were due to the lower LAI and light interception. At flowering, the root/shoot ratio was 0.25 in NLI and 0.17 in LI, and the root biomass in NLI was reduced by 64% compared to LI. Nitrogen uptake was also reduced in NLI in spite of high soil N availability. Nevertheless, shoot N concentration vs aboveground biomass showed a typical decline in both treatments. In NLI, the shoot P concentration vs above-ground biomass relationship showed an increase in the early stages, indicating that P uptake and root-shoot competition for the absorbed P in the early plant stages controlled the establishment and the development of the leaf area.     

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 today's 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 effects of fire on soil nitrogen associated with patches of the actinorhizal shrub Ceanothus cordulatus

Nitrogen is a limiting resource in many temperate forests and nitrogen-fixing plants are usually limited to the early stages of post-disturbance succession. In fire-dependent Sierra Nevada forests, however, Ceanothus cordulatus is relatively abundant even in old-growth forest conditions which are at least partly maintained by fire. We conducted a field experiment to determine if soil beneath Ceanothus patches represent `resource islands' of available N which persist after fire. Nine plots containing discrete patches of Ceanothus, Arctostaphylos patula (manzanita; chosen as a non N-fixing reference species), and bare forest floor were subjected to either a low-intensity (n=3) or high-intensity (n=3) bum treatment, or remained unburned as controls (n=3). Soil temperatures during the bum were monitored by a network of thermocouples placed at the surface of the mineral soil and at ca. 10 cm depths. Soil samples were collected from the organic horizon, 0-10 cm and 15-25 cm depths within each patch type immediately before burning and 2 days, and 6, and 11 months after. Soil moisture, total C and N, and ammonium and nitrate concentrations were determined in the laboratory. Before the burn, Ceanothus patches were significantly enriched in total and inorganic N in the organic horizon relative to the other patch types. A sharp increase in inorganic N was observed in all patch types and depths immediately following burning, but by 6 months after the burn, Ceanothus patches were significantly enriched relative to the surrounding patch types and remained so at months. Resprouting Ceanothus patches will continue to be an important source of a limiting nutrient in this fire-prone ecosystem.     

Ammonia-oxidizing bacteria on root biofilms and their possible contribution to N use efficiency of different rice cultivars

Ammonia-oxidizing bacteria (AOB) populations were studied on the root surface of different rice cultivars by PCR coupled with denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH). PCR-DGGE of the ammonium monooxygenase gene (amoA) showed a generally greater diversity on root samples compared to rhizosphere and unplanted soil. Sequences affiliated with Nitrosomonas spp. tended to be associated with modern rice hybrid lines. Root-associated AOB observed by FISH were found within a discrete biofilm coating the root surface. Although the total abundance of AOB on root biofilms of different rice cultivars did not differ significantly, there were marked contrasts in their population structure, indicating selection of Nitrosomonas spp. on roots of a hybrid cultivar. Observations by FISH on the total bacterial community also suggested that different rice cultivars support different bacterial populations even under identical environmental conditions. The presence of active AOB in the root environment predicts that a significant proportion of the N taken up by certain rice cultivars is in the form of NO₃ ^–-N produced by the AOB. Measurement of plant growth of hydroponically grown plants showed a stronger response of hybrid cultivars to the co-provision of NH₄ ⁺ and NO₃ ^–. In soil-grown plants, N use efficiency in the hybrid was improved during ammonium fertilization compared to nitrate fertilization. Since ammonium-fertilized plants actually receive a mixture of NH₄ ⁺ and NO₃ ^– with ratios depending on root-associated nitrification activity, these results support the advantage of co-provision of ammonium and nitrate for the hybrid cultivar.     

Soil type affects nitrogen conservation in foliage of small Pinus sylvestris L. trees

Nutrient conservation in plants and soil fertility may be intricately linked. We studied nitrogen conservation in small Scots pine (Pinus sylvestrisL.) trees growing in stands on organogenic Dystric Histosols and on mineral Podzols. Nitrogen-resorption efficiency (NRE) and proficiency (NRP) of senescent needles, and mean residence time of nitrogen (MRT) were studied in relation to needle surface area, needle longevity, and leaf mass per area (LMA). Trees on Podzols had higher nitrogen concentration in green needles than the trees on Dystric Histosols, but the nitrogen concentration of yellowing needles was similar for trees on both soil types. NRE averaged 65±3.5% (mean±SD) and 56±7.2% for the trees on Podzols and Dystric Histosols, respectively. Neither NRP (0.44±0.05% and 0.35±0.07%, respectively) nor MRT (8.4±2.3 and 6.1±1.2 years) differed significantly between the stands on the two soil types. Mean needle surface area was significantly smaller in trees on Dystric Histosols (76±29 mm²) than on Podzols (131±38 mm²), whereas needle longevity varied between 2 and 4 years independently of the soil type. Trees invested, on average, the same amount of dry matter per unit of needle area on both soil types. Growth of trees, measured as increment of shoot length, was more restricted on Dystric Histosols (55±18 mm yr^–1) than on Podzols (184±44 mm yr^–1). The results of the correlation analysis applied to pooled data were inconsistent with the relations between traits of stress resistance syndrome observed in inter-specific comparisons. The study indicated that Scots pine trees relocated nitrogen from senescent foliage more efficiently on mineral Podzols than on organogenic Dystric Histosols, but the minimum nitrogen concentration of needles appeared to be similar on both soil types.     

The influence of dissolved nutrients and particulate organic matter quality on microbial respiration and biomass in a forest stream

1. Although dissolved nutrients and the quality of particulate organic matter (POM) influence microbial processes in aquatic systems, these factors have rarely been considered simultaneously. We manipulated dissolved nutrient concentrations and POM type in three contiguous reaches (reference, nitrogen, nitrogen + phosphorus) of a low nutrient, third‐order stream at Hubbard Brook Experimental Forest (U.S.A). In each reach we placed species of leaves (mean C : N of 68 and C : P of 2284) and wood (mean C : N of 721 and C : P of 60 654) that differed in elemental composition. We measured the respiration and biomass of microbes associated with this POM before and after nutrient addition. 2. Before nutrient addition, microbial respiration rates and biomass were higher for leaves than for wood. Respiration rates of microbes associated with wood showed a larger response to increased dissolved nutrient concentrations than respiration rates of microbes associated with leaves, suggesting that the response of microbes to increased dissolved nutrients was influenced by the quality of their substrate. 3. Overall, dissolved nutrients had strong positive effects on microbial respiration and fungal, but not bacterial, biomass, indicating that microbial respiration and fungi were nutrient limited. The concentration of nitrate in the enriched reaches was within the range of natural variation in forest streams, suggesting that natural variation in nitrate among forest streams influences carbon mineralisation and fungal biomass.     

Effects of harvesting on nutrient leaching in a Norway spruce (Picea abies Karst.) ecosystem on a Lithic Leptosol in the Northern Limestone Alps

On a heavily karstified site in the Northern Limestone Alps (Austria), nutrient budgets and leaching in Norway spruce stands were investigated along a chronosequence (clearcut, 10-year-old plantation (25% cover of planted and naturally regenerated spruce and larch, 75% weed cover) and mature stand). The soils were Lithic Leptosols on very pure limestone. Nutrient fluxes were studied during three growth periods (4–5 months each). Despite of inorganic nitrogen inputs from precipitation between 5 and 10 kg ha^–1, inorganic nitrogen output with seepage water from the mature stand and the regeneration plot was only 0.5–1.2 kg ha^–1 during these periods. In the first and second growth periods after clearcut, inorganic N fluxes with seepage increased to 20 and 30 kg ha^–1, respectively, declining in the third growth period to 8 kg ha^–1. DON output during the growth period was between 3 and 6 kg ha^–1 in the mature stand and 7 and 11 kg ha^–1 in the clearcut as well as in the regeneration plot. K output rates achieved 30 kg ha^–1 in the first, 20 kg ha^–1 in the second and 9 kg ha^–1 in the third growth period after clear-cutting while output rates during the growth periods were less than 2 kg ha^–1 in the mature stand and in the regeneration plot. K pools in the humus layer were only 150–210 kg ha^–1, total K pools including above and below ground biomass in the mature stand were 360 kg ha^–1. Thus, post-harvest hydrological losses comprise a substantial depletion of K for this specific ecosystem. Since precipitation is high in this area (1400 mm a^–1), forest growth is limited by nutrient rather than by water supply. Needle analyses already indicate a deficient potassium supply. Harvesting and post-harvesting losses of K in combination with elevated nitrogen deposition may have negative influences on the stability of forest stands on the studied sites.     

Nitric oxide production in tobacco leaf cells: a generalized stress response?

The function of nitric oxide (NO), a gaseous free radical emitted by many plants, is incompletely understood. In the present study the hypothesis that NO generation, like that of the reactive oxygen species, occurs as a general response to different environmental cues was tested. Leaf peels and mesophyll cell suspensions of Nicotiana tabacum cv. Xanthi were loaded with the NO‐specific fluorophore, diaminofluorescein, and subjected to an abiotic stressor. Light stress and mechanical injury had no apparent effect on NO production. In contrast, high temperatures, hyperosmotic stress, salinity and epi‐illumination in a microscope all led to rapid surges in NO‐induced fluorescence. The fluorescence originated from cells of the palisade mesophyll and across all epidermal cell types, including guard cells, subsidiary cells, and long and short trichomes. Fluorescence was evident first in the plastids, then in the nucleus and finally throughout the cytosol. Nicotiana plumbaginifolia cell suspensions expressing the calcium reporter aequorin provided evidence that, under hyperosmotic stress, NO participates in the elevation of free Ca²⁺ in the cytoplasm. The physiological significance of NO production in response to abiotic stressors is discussed.