The biochemical transformation of oak (Quercus robur) leaf litter consumed by the pill millipede (Glomeris marginata)

Soil macrofauna play an essential role in the initial comminution and degradation of organic matter entering the soil environment and yet the chemical effects of digestion on leaf litter are poorly understood at the molecular level. This study was undertaken to assess the selective chemical transformations that saprophagous soil invertebrates mediate in consumed leaf litter. A number of pill millipedes (Glomeris marginata) were fed oak leaves (Quercus robur) after which the biomolecular compositions (lipids and macromolecular components) of the leaves and millipede faeces were compared using a series of wet chemical techniques and subsequent analysis by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). It was found that the concentrations of short chain (<C₂₀) n-alkanoic acids, sterols and triacylglycerols reduced dramatically in the millipede faeces relative to the leaf litter. Hydrolysable carbohydrates and proteins both decreased in concentration in the faeces, whereas similar yields of phenolic components were observed for the cupric oxidation products of lignin, although the oxygenated functionalities were affected by passage through the millipede gut, yielding a more highly condensed state for lignin. This shows that the chemical composition of fresh organic matter entering the soil is directly controlled by invertebrates feeding upon the leaf litter and as such that they are key contributors to the early stages of diagenesis in terrestrial soils.     

Extreme neutral genetic and morphological divergence supports classification of Adriatic three-spined stickleback (Gasterosteus aculeatus) populations as distinct conservation units

The fact that the intraspecific genetic differentiation in neutral genetic markers and genes coding for adaptive traits are not typically correlated has caused a great deal of conceptual and practical trouble in delimitation of conservation units. Although the importance of combining information on adaptive genetic divergence with information on historical and recent gene flow in the delimitation of conservation units has been recognized, integrated empirical studies to this end are still rare. We explored the evidence for the specific conservation status of two freshwater three-spined stickleback (Gasterosteus aculeatus) populations on the Adriatic side of the Balkan Peninsula by comparing their phenotypic and genetic characteristics to those of other representative European populations. Apart from focusing on the neutral genetic divergence in mitochondrial DNA sequences and microsatellite markers, we also compared the patterns of morphological differentiation (i.e. bony armour development) resulting from adaptation to freshwater environments. The Balkanic populations formed two distinct groups with regard to neutral genetic variation and had the least developed bony armour of all the examined populations. All morphometric analyses identified the two Balkanic populations as phenotypically – and hence most likely also ecologically – clearly distinct from other European three-spined stickleback populations. These results suggest that the two Balkanic populations (River Neretva and River Zeta) fulfil the most stringent criteria (i.e. lack of genetic and ecological exchangeability) to be classified as conservation units distinct from other European three-spined stickleback populations.     

Homing ability widens the sphere of influence of the earthworm Lumbricus terrestris L.

We report that the deep-burrowing earthworm Lumbricus terrestris L. possesses short-range homing capability. For 2 months we followed the soil surface activity of 18 field-collected L. terrestris in an experimental laboratory population. We recorded 26 cases where an individual after completely leaving its burrow returned to the burrow by backing along its own outward trail. Maximally, homing occurred from a distance of 0.70 m and after more than 3 h on the soil surface. The evolution of homing ability in L. terrestris is most likely related to the importance of the burrow for survival and reproduction. This adaptation also guarantees individuals an expanded foraging area, which we believe can partly explain the remarkable efficiency of L. terrestris in the burial of surface litter.     

Purification and characterization of a methylene urea-hydrolyzing enzyme from Rhizobium radiobacter (Agrobacterium tumefaciens)

Slow-release fertilizers are gaining acceptance to increase fertilizer use efficiency and reduce environmental impact. The release of nitrogen from methylene urea, a common slow release N fertilizer, is controlled by microbial decomposition. An enzyme hydrolyzing slow-release nitrogen fertilizer, methylene urea, was purified from Rhizobium radiobacter (Agrobacterium tumefaciens) to homogeneity using a four-step purification procedure with an overall yield of 3%. The active enzyme has a molecular mass of approximately 180 kDa determined by size exclusion chromatography, and the SDS page of the purified protein indicated three subunits of different sizes (62, 34 and 32 kDa). The N-terminal amino acid sequence of the 62 kDa fragment indicates identity with urease subunits from Mycobacterium tuberculosis (73%) and Helicobacter pylori (71%). However, for the internal amino acid sequences of the 62 kDa fragment no matches with known proteins were found. Some internal peptides in the smaller subunits (32 and 34 kDa) are homologous to urease subunits and unknown proteins in Agrobacterium tumefaciens. Based on the kinetic properties, substrate selectivity, and inhibition characteristics, the novel enzyme (MUase) is an intracellular enzyme complex with urease activity. The enzymatic mechanism of methylene urea breakdown was studied using a novel LC-MS method for MU analysis, which indicates that all cold-water soluble nitrogen forms of methylene urea are subjected to hydrolysis, and the hydrolysis proceeds via methylurea, urea and other yet unidentified hydrolysis-products, suggesting that the isolated enzyme complex performs a multistep hydrolysis. The microbiological and molecular data is useful in determining the soil factors affecting the efficacy of methylene urea as a slow release fertilizer in agricultural production systems.     

Conserving marginal populations of the food plant (Impatiens noli-tangere) of an endangered moth (Eustroma reticulatum) in a changing climate

Impatiens noli-tangere is scarce in the UK and probably only native to the Lake District and Wales. It is the sole food plant for the endangered moth Eustroma reticulatum. Significant annual fluctuations in the size of I. noli-tangere populations endanger the continued presence of E. reticulatum in the UK. In this study, variation in population size was monitored across native populations of I. noli-tangere in the English Lake District and Wales. In 1998, there was a crash in the population size of all metapopulations in the Lake District but not of those found in Wales. A molecular survey of the genetic affinities of samples in 1999 from both regions and a reference population from Switzerland was performed using AFLP and ISSR analyses. The consensus UPGMA dendrogram and a PCO scatter plot revealed clear differentiation between the populations of I. noli-tangere in Wales and those in the Lake District. Most of the genetic variation in the UK (H_T=0.064) was partitioned between (G_ST=0.455) rather than within (H_S=0.034) regions, inferring little gene flow occurs between regions. There was similar bias towards differentiation between metapopulations in Wales, again consistent with low levels of interpopulation gene flow. This contrasts with far lower levels of differentiation in the Lake District which suggests modest rates of gene flow may occur between populations. It is concluded that in the event of local extinction of sites or populations, reintroductions should be restricted to samples collected from the same region. We then surveyed climatic variables to identify those most likely to cause local extinctions. Climatic correlates of population size were sought from two Lake District metapopulations situated close to a meteorological station. A combination of three climatic variables common to both sites explained 81-84% of the variation in plant number between 1990 and 2001. Projected trends for these climatic variables were used in a Monte Carlo simulation which suggested an increased risk of I. noli-tangere population crashes by 2050 at Coniston Water, but not at Derwentwater. Implications of these findings for practical conservation strategies are explored.     

Litter decomposition and nitrogen mineralization in newly formed gaps in a Danish beech (Fagus sylvatica) forest

Gap formation is suggested as an alternative forest management approach to avoid extreme changes in the N cycle of forest ecosystems caused by traditional management practises. The present study aimed to investigate the effect of gap formation on N availability in beech litter and mineral soil on sites, which experienced only little soil disturbance during tree harvest. N pools, litter decomposition, and N mineralization rates in mineral soil were studied in two gaps (17 and 30 m in diameter) in a 75-year-old managed European beech (Fagus sylvatica L.) forest in Denmark and related to soil temperature (5 cm depth) and soil moisture (15 cm depth). Investigations were carried out during the first 2 years after gap formation in measurement plots located along the north-south transect running through the centre of each gap and into the surrounding forest.An effect of gap size was found only for soil temperatures and litter mass loss: soil temperatures were significantly increased in the northern part of the large gap during the first year after gap formation, and litter mass loss was significantly higher in the smaller gap. All other parameters investigated revealed no effect of gap size. Nitrification, net mineralization, and soil N concentrations tended to be increased in the gaps. Cumulative rates of net mineralization were two fold higher in the gaps during the growing season (June-October), but a statistically significant increase was found only for soil NH₄-N concentrations during this period. Forest floor parameters (C:N ratios, mass loss, N release) were not significantly modified during the first year after gap formation, neither were the total C content nor the C:N ratio in mineral soil at 0-10 cm depth.     

Size, symbiotic effectiveness and genetic diversity of field pea rhizobia (Rhizobium leguminosarum bv. viciae) populations in South Australian soils

Field pea (Pisum sativum L.) is widely grown in South Australia (SA), often without inoculation with commercial rhizobia. To establish if symbiotic factors are limiting the growth of field pea we examined the size, symbiotic effectiveness and diversity of populations of field pea rhizobia (Rhizobium leguminosarum bv. viciae) that have become naturalised in South Australian soils and nodulate many pea crops. Most probable number plant infection tests on 33 soils showed that R. l. bv. viciae populations ranged from undetectable (six soils) to 32×10³ rhizobia g⁻¹ of dry soil. Twenty-four of the 33 soils contained more than 100 rhizobia g⁻¹ soil. Three of the six soils in which no R. l. bv. viciae were detected had not grown a host legume (field pea, faba bean, vetch or lentil). For soils that had grown a host legume, there was no correlation between the size of R. l. bv. viciae populations and either the time since a host legume had been grown or any measured soil factor (pH, inorganic N and organic C). In glasshouse experiments, inoculation of the field pea cultivar Parafield with the commercial Rhizobium strain SU303 resulted in a highly effective symbiosis. The SU303 treatment produced as much shoot dry weight as the mineral N treatment and more than 2.9 times the shoot dry weight of the uninoculated treatment. Twenty-two of the 33 naturalised populations of rhizobia (applied to pea plants as soil suspensions) produced prompt and abundant nodulation. These symbioses were generally effective at N₂ fixation, with shoot dry weight ranging from 98% (soil 21) down to 61% (soil 30) of the SU303 treatment, the least effective population of rhizobia still producing nearly double the growth of the uninoculated treatment. Low shoot dry weights resulting from most of the remaining soil treatments were associated with delayed or erratic nodulation caused by low numbers of rhizobia. Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) fingerprinting of 70 rhizobial isolates recovered from five of the 33 soils (14 isolates from each soil) showed that naturalised populations were composed of multiple (5-9) strain types. There was little evidence of strain dominance, with a single strain type occupying more than 30% of trap host nodules in only two of the five populations. Cluster analysis of RAPD PCR banding patterns showed that strain types in naturalised populations were not closely related to the current commercial inoculant strain for field pea (SU303, ≥75% dissimilarity), six previous field pea inoculant strains (≥55% dissimilarity) or a former commercial inoculant strain for faba bean (WSM1274, ≥66% dissimilarity). Two of the most closely related strain types (≤15% dissimilarity) were found at widely separate locations in SA and may have potential as commercial inoculant strains. Given the size and diversity of the naturalised pea rhizobia populations in SA soils and their relative effectiveness, it is unlikely that inoculation with a commercial strain of rhizobia will improve N₂ fixation in field pea crops, unless the number of rhizobia in the soil is very low or absent (e.g. where a legume host has not been previously grown and for three soils from western Eyre Peninsula). The general effectiveness of the pea rhizobia populations also indicates that reduced N₂ fixation is unlikely to be the major cause of the declining field pea yields observed in recent times.     

Hybridization mechanisms between the endangered marble trout (Salmo marmoratus) and the brown trout (Salmo trutta) as revealed by in-stream experiments

Populations of the marble trout (Salmo marmoratus) are critically declining due to introgression by brown trout (S. trutta) strains. Hybrids between the two forms are fertile and presently predominant in most rivers of the species’ range.The involved hybridization mechanisms have been studied through two large scale in-stream experiments (Driselpoh and Stopnikarca) as 50% of each species have been stocked at the age of one year in fishless streams, each fish being individually marked. Diagnostic molecular markers were applied to test a partial reproductive isolation between the two species. Emphasis was put on survival and growth patterns of stocked fish (parental generation) as well as on fish hatched within the stream (F₁ generation).No evidence of a partial reproductive barrier between the two species was observed. Survival of the parental generation depended on the year in both streams, as well as on the species in Stopnikarca, but was identical for both species in Driselpoh. In both streams survival of instream hatched individuals measured from 0+ to 2+ was lower for brown trout. In Driselpoh, F₁ 0+, 1+ and 2+ hybrids were larger than pure individuals. Larger hybrids were only observed in Stopnikarca when analyses focused on individuals in inter-specific competition suggesting that heterosis and stress effects may explain the observed size differences.Our results point out important ecological differences between marble and brown trout and have shown that hybridization can easily take place. The findings indicate that high F₁ hybrid presence, survival and heterosis effects may impede marble trout rehabilitation in the area.     

Quantification of the contribution of biological nitrogen fixation to Cratylia mollis using the N natural abundance technique in the semi-arid Caatinga region of Brazil

This study was performed to investigate the capacity of the woody perennial Cratylia mollis, a legume endemic to the semi-arid region of the North-East of Brazil, to nodulate, and obtain N from BNF using the ¹⁵N natural abundance technique. To estimate the ¹⁵N abundance of the N-derived from soil, the leaves of several (4-6) non-legume and non-nodulating legume species growing in close proximity to the Cratylia were analysed for δ¹⁵N. The δ¹⁵N values of these reference plants were high (from +9 to +16‰) and showed relatively small differences between species at each site/sampling time. At the irrigated site at both samplings, and at the non-irrigated site sampled in the rainy season, the ¹⁵N abundance of the Cratylia mollis leaves was far lower (+1 to +5‰) strongly suggesting that the legume obtained large proportional contributions from BNF. As was to be expected, no nodules were found on the Cratylia plants at the non-irrigated site in the dry season, and the ¹⁵N abundance of the Cratylia mollis plants were very similar to that of the reference plants, consistent and nodulation and BNF being limited by water deficit at this time.     

Nutrient release from decomposing leaf mulches of karité (Vitellaria paradoxa) and néré (Parkia biglobosa) under semi-arid conditions in Burkina Faso, West Africa

Information on decomposition and nutrient release from leaf litter of trees in agroforestry parkland systems in Sub-Saharan Africa is scarce despite the significant role of these trees on soil fertility improvement and maintenance. Decomposition and nutrient release patterns from pruned leaves of the two most common species of parklands of the semi-arid zone of West Africa: Vitellaria paradoxa C.F. Gaertn (known locally as karité) and Parkia biglobosa (Jacq.) Benth. (known locally as néré), were investigated by a litter-tube study in Burkina Faso. Litter quality, methods of leaf exposure to the soil and combination with fertilizers were the factors studied. Leaves of néré had a higher nutrient content (C, N, P, Ca) and contained more ash and lignin than leaves of karité. Karité leaves had a greater content of K, cellulose and polyphenols. The pruned leaves of karité and néré showed two distinct decomposition patterns. Néré leaves decomposed more rapidly, with less than 32% of the initial weight remaining after the rainy season (4 months) while karité leaves decomposed more slowly with 43% of the leaf litter remaining after the rainy season. Addition of urea and compost did not significantly affect the rate of decomposition. Significant interaction was observed between species and method of leaf exposure for the first sampling date. Leaf litter of néré buried in soil gave the highest weight loss (34% of the initial mass in 1 month) compared with exposed leaf litter of néré and karité, and buried leaf litter of karité. Except for N, nutrient release patterns were similar for both species but the nutrient release rates were higher for néré leaves compared with karité leaves. N was immobilised in karité leaves most likely due to low N and high phenolic content. The rate of nutrient release from karité leaves followed the general trend K>P>N.     

Field survival of the phytostimulator Azospirillum lipoferum CRT1 and functional impact on maize crop, biodegradation of crop residues, and soil faunal indicators in a context of decreasing nitrogen fertilisation

Heavy nitrogen fertilisation is often implemented in maize cropping systems, but it can have negative environmental effects. Nitrogen-fixing, phytohormone-producing Azospirillum plant growth-promoting rhizobacteria (PGPR) have been proposed as crop inoculants to maintain high yield when decreasing nitrogen fertilisation. In this context, agronomic and ecological effects of the inoculation of maize seeds with the PGPR Azospirillum lipoferum CRT1 were studied in two consecutive years. The inoculant was recovered from maize at 10⁵ CFU g⁻¹ root or higher. Inoculation enhanced root growth and development based on results of root biomass, rooting depth and/or parameters describing root system architecture, and a transient positive effect on shoot height was observed in the first year. Inoculation did not increase yield, but reducing mineral nitrogen fertilisation had only a minor effect on yield. This suggests that the lack of positive effect of the PGPR on yield was due to the fact that the whole field was heavily fertilised in years prior to the start of the experiment. Soil nitrogen levels decreased during the 2 years of the study, and the inoculant had no effect on residual soil nitrogen levels at harvest. Inoculation had no impact on Fusarium symptoms and concentration of the mycotoxin deoxynivalenol in maize kernels, but both were influenced by the interaction between inoculation and nitrogen fertilisation level. Inoculation did not influence meso/macrofaunal soil populations, but had a small but significant effect (smaller than the effect of added nitrogen) on decomposition, nitrogen mineralisation and mesofaunal colonisation of maize leaves (in litter bags). Overall, the ecological impact of seed inoculation with the PGPR A. lipoferum CRT1 was small, and its magnitude was smaller than that of chemical nitrogen fertilisation.     

First indications for the involvement of strigolactones on nodule formation in alfalfa (Medicago sativa)

Strigolactones have recently been suggested to be phytohormones that are present in all plants. Strigolactones are released by roots into the rhizosphere, stimulating the seed germination of parasitic plants such as Striga spp. and Orobanche spp. and play a crucial role in the interaction between plants and symbiotic arbuscular mycorrhizal fungi.By applying different concentrations of the synthetic strigolactone analogue GR24 to alfalfa (Medicago sativa) inoculated with Sinorhizobium meliloti we could show that in alfalfa nodulation is positively affected by the presence of the strigolactone analogue GR24. Moreover, we could show that this increased nodulation cannot be linked with a stimulatory effect of GR24 on the growth or the expression of nod genes of S. meliloti.Putative mechanisms operating in the plant in response to the addition of GR24 and leading to increased nodule formation by rhizobia are discussed.     

Changes in soil aggregation and glomalin-related soil protein content as affected by the arbuscular mycorrhizal fungal species Glomus mosseae and Glomus intraradices

Arbuscular mycorrhizal (AM) fungi are key organisms of the soil/plant system, influencing soil fertility and plant nutrition, and contributing to soil aggregation and soil structure stability by the combined action of extraradical hyphae and of an insoluble, hydrophobic proteinaceous substance named glomalin-related soil protein (GRSP). Since the GRSP extraction procedures have recently revealed problems related to co-extracting substances, the relationship between GRSP and AM fungi still remains to be verified. In this work the hypothesis that GRSP concentration is positively correlated with the occurrence of AM fungi was tested by using Medicago sativa plants inoculated with different isolates of Glomus mosseae and Glomus intraradices in a microcosm experiment. Our results show that (i) mycorrhizal establishment produced an increase in GRSP concentration - compared to initial values - in contrast with non-mycorrhizal plants, which did not produce any change; (ii) aggregate stability, evaluated as mean weight diameter (MWD) of macroaggregates of 1-2 mm diameter, was significantly higher in mycorrhizal soils compared to non-mycorrhizal soil; (iii) GRSP concentration and soil aggregate stability were positively correlated with mycorrhizal root volume and weakly correlated with total root volume; (iv) MWD values of soil aggregates were positively correlated with values of total hyphal length and hyphal density of the AM fungi utilized.The different ability of AM fungal isolates to affect GRSP concentration and to form extensive and dense mycelial networks, which may directly affect soil aggregates stability by hyphal enmeshment of soil particles, suggests the possibility of selecting the most efficient isolates to be utilized for soil quality improvement and land restoration programs.     

Impacts of earthworms and arbuscular mycorrhizal fungi (Glomus intraradices) on plant performance are not interrelated

Earthworms and arbuscular mycorrhizal fungi (AMF) might interactively impact plant productivity; however, previous studies reported inconsistent results. We set up a three-factorial greenhouse experiment to study the effects of earthworms (Aporrectodea caliginosa Savigny and Lumbricus terrestris L.) and AMF (Glomus intraradices N.C. Schenck & G.S. Sm.) on the performance (productivity and shoot nutrient content) of plant species (Lolium perenne L., Trifolium pratense L. and Plantago lanceolata L.) belonging to the three functional groups grasses, legumes and herbs, respectively. Further, we investigated earthworm performance and plant root mycorrhization as affected by the treatments. Our results accentuate the importance of root derived resources for earthworm performance since earthworm weight (A. caliginosa and L. terrestris) and survival (L. terrestris) were significantly lower in microcosms containing P. lanceolata than in those containing T. pratense. However, earthworm performance was not affected by AMF, and plant root mycorrhization was not modified by earthworms. Although AMF effectively competed with T. pratense for soil N (as indicated by δ¹⁵N analysis), AMF enhanced the productivity of T. pratense considerably by improving P availability. Remarkably, we found no evidence for interactive effects of earthworms and AMF on the performance of the plant species studied. This suggests that interactions between earthworms and AMF likely are of minor importance.     

Soil texture and irrigation influence the transport and the development of Pasteuria penetrans, a bacterial parasite of root-knot nematodes

The transport of the spores of Pasteuria penetrans was studied in three contrasted textured soils (a sandy, a sandy-clay and a clay soils), cultivated with tomato, inoculated with juveniles of Meloidogyne javanica and watered with 25 or 150 mm day⁻¹. One month after inoculation of the nematodes, 53% of the spores inoculated were leached by water flow in the sandy soil but only 14% in the sandy-clay soil and 0.1% in the clay soil. No nematodes survived in the clay soil, while the population was multiplied both in the sandy and in the sandy-clay soils. But juveniles of M. javanica were more infected by P. penetrans in the sandy-clay soil than in the sandy soil. Comparing different combinations of bare soils containing 1.1-57% of clay showed that the best spore percolation and retention balance occurred in soils amended with 10-30% clay. However, the spore recoveries decreased when the soil was enriched with more than 30% clay. The role of clay particles on the extractability of spores and on their availability to attach to the nematode cuticle in the soil is discussed.     

Inferring species distributions in the absence of occurrence records: An example considering wolverine (Gulo gulo) and Canada lynx (Lynx canadensis) in New Mexico

Information about geographic distributions is required for species conservation and management. Ultimately, this information is derived from records of occurrence. However, the reliability and availability of occurrence records are variable. A conceptual framework for evaluating the reliability of occurrence records is provided. Only records associated with physical evidence, especially a museum voucher specimen, are considered verified. However, errors in species identification or location are possible even for verified records. In addition, biases exist in occurrence records because they generally are collected haphazardly. Other sources of bias include sampling error associated with small areas or range limits and aspects of the species’ biology that make it unlikely to be documented. A practical method is provided for interpreting a species’ distribution in a particular area given a paucity of reliable occurrence records. Factors that must be considered for including such areas of interest within the range of a species include: (1) plausible reason for the paucity of records; (2) continuous suitable habitat between the area of interest and localities of reliable occurrence; and (3) absence of biogeographic breaks in the distribution of other organisms with similar evolutionary histories. The possible distribution of wolverine (Gulo gulo) and Canada lynx (Lynx canadensis) in New Mexico provides a case study of this approach. It is concluded that the mountains of north-central New Mexico should be considered within the natural range of wolverine and Canada lynx.     

Attraction to prey and prey-associated odours by the predatory mite Hypoaspis aculeifer in a soil experimental system

In terms of food searching and trophic interactions among soil organisms volatile signalling plays a crucial role. Still, volatile-based foraging decisions are poorly understood, especially as the dispersal and perception of cues is more restricted within the soil pore space than in simple laboratory approaches where volatiles disperse freely in the air. Focusing on the tritrophic interaction between the soil fungus Trichoderma viride, the fungivorous collembolan Folsomia candida and the predatory mite Hypoaspis aculeifer, we tested the effect of prey and prey-associated volatile signals on the food choice decision of the predatory mite in a laboratory experiment under soil conditions as well as the possible occurrence and relevance of grazing-induced indirect defence signals. The mites showed a clear preference for ungrazed and grazed fungal mycelium. However, we found no evidence of induced indirect defence as ungrazed mycelium was preferred to grazed fungus.     

Reconciling the dichotomy between single species and ecosystem conservation: black kites (Milvus migrans) and eutrophication in pre-Alpine lakes

In recent times there has been a growing dichotomy between preservation of single species and broader, ecosystem-based approaches to conservation. Freshwater habitats are among the most highly human-impacted ecosystems. We used a long-term data set on black kites (Milvus migrans), a threatened raptor dependent on aquatic habitats, to explore ways to reconcile single species and biodiversity-driven approaches to conservation. In the pre-Alpine lakes of northern Italy, black kite populations showed medium-low density, extremely low breeding success and widespread declines. Spatio-temporal variations showed density and breeding success to be positively related to ecosystem productivity (as estimated by phosphorus concentrations in lakes), availability of aquatic habitats and grassland, and to be negatively related to extent of farmland and fish harvest by professional fishermen. Fish species richness, used as a surrogate of biodiversity, was highest in one oligotrophic lake, but on average increased with increasing ecosystem productivity (i.e. lake eutrophication). Given expected future declines in ecosystem productivity, kite conservation will be helped by enhancing populations of alternative prey in terrestrial habitats (e.g. through incentives for grassland), and higher regulation of fish harvest. On the other hand, ecosystem management may profit from the use of black kites or other aerial piscivores as indicators of biodiversity and of diffuse ecosystem stress, such as sustainability of fishing practices.     

New insights into the origins and evolution of rhizobia that nodulate common bean (Phaseolus vulgaris) in Brazil

It is generally accepted that there are two major centers of genetic diversification of common beans (Phaseolus vulgaris L.): the Mesoamerican (Mexico, Colombia, Ecuador and north of Peru, probably the primary center), and the Andean (southern Peru to north of Argentina) centers. Wild common bean is not found in Brazil, but it has been grown in the country throughout recorded history. Common bean establishes symbiotic associations with a wide range of rhizobial strains and Rhizobium etli is the dominant microsymbiont at both centers of genetic diversification. In contrast, R. tropici, originally recovered from common bean in Colombia, has been found to be the dominant species nodulating field-grown common-bean plants in Brazil. However, a recent study using soil dilutions as inocula has shown surprisingly high counts of R. etli in two Brazilian ecosystems. In the present study, RFLP-PCR analyses of nodABC and nifH genes of 43 of those Brazilian R. etli strains revealed unexpected homogeneity in their banding patterns. The Brazilian R. etli strains were closely similar in 16S rRNA sequences and in nodABC and nifH RFLP-PCR profiles to the Mexican strain CFN 42^T, and were quite distinct from R. etli and R. leguminosarum strains of European origin, supporting the hypothesis that Brazilian common bean and their rhizobia are of Mesoamerican origin, and could have arrived in Brazil in pre-colonial times. R. tropici may have been introduced to Brazilian soils later, or it may be a symbiont of other indigenous legume species and, due to its tolerance to acidic soils and high temperature conditions became the predominant microsymbiont of common bean.