Genetic characterization of Portuguese brown trout (Salmo trutta L.) and comparison with other European populations

Abstract– Allozyme and other protein loci were examined to study the genetic structure of Portuguese brown trout ( Salmo trutta ) populations. A total of 247 individuals from three tributaries of the Lima hydrological basin and a hatchery, all located in northern Portugal, were analyzed. Four of 22 protein coding loci were found to be polymorphic: CK-A1^*, GPI-A2^*, MPI-2^* and TF^*. A new allelc at the latter locus was found in Atlantic populations. The data obtained for Portuguese brown trout were compared with published data for 14 European populations and three hatchery stocks. Six polymorphic loci (CK-A1^*, GPI-A2^*, GPI-B2^*, LDH-C^*, ME^* and MPI-2^*) were used in a cluster analysis. This showed the similarity of Portuguese natural populations and northern Iberian populations and that Portuguese hatchery fish have an autochthonous origin, distinct from that of other Atlantic hatchery stocks.     

Effect of glyphosate on the tripartite symbiosis formed by Glomus intraradices,Bradyrhizobium japonicum,and genetically modified soybean

Most soybeans grown in North America are genetically modified (GM) to tolerate applications of the broad-spectrum herbicide glyphosate; as a result, glyphosate is now extensively used in soybean cropping systems. Soybean roots form both arbuscular mycorrhizal (AM) and rhizobial symbioses. In addition to individually improving host plant fitness, these symbioses also interact to influence the functioning of each symbiosis, thereby establishing a tripartite symbiosis. The objectives of this study were to (1) estimate the effects of glyphosate on the establishment and functioning of AM and rhizobial symbioses with GM soybean, and (2) to estimate the interdependence of the symbioses in determining the response of each symbiosis to glyphosate. These objectives were addressed in two experiments; the first investigated the importance of the timing of glyphosate application in determining the responses of the symbionts and the second varied the rate of glyphosate application. Glyphosate applied at recommended field rates had no effect on Glomus intraradices or Bradyrhizobium japonicum colonization of soybean roots, or on soybean foliar tissue [P]. N₂-fixation was greater for glyphosate-treated soybean plants than for untreated-plants in both experiments, but only when glyphosate was applied at the first trifoliate soybean growth stage. These data deviate from previous studies estimating the effect of glyphosate on the rhizobial symbiosis, some of which observed negative effects on rhizobial colonization and/or N₂-fixation. We did observe evidence of the response of one symbiont (stimulation of N₂-fixation following glyphosate) being dependent on co-inoculation with the other; however, this interactive response appeared to be contextually dependent as it was not consistent between experiments. Future research needs to consider the role of environmental factors and other biota when evaluating rhizobial responses to herbicide applications.     

Litter production and nutrient deposition from native woody species in the Brazilian semi-arid region

Agroforestry Systems - The use of native trees in agroecosystems is a promising way to increase litter deposition and nutrient cycling and foster the recovery of degraded areas, especially in...     

Higher feeding diets effects on age and liveweight gain at puberty in crossbred Nelore × Hereford heifers

This study was conducted to determine the age and liveweight at puberty of 120 crossbred beef heifers submitted to four diets to achieve predetermined weight gains (kg/day): 0.5 (G500; n?=?32), 0.75 (G750; n?=?32), 1.0 (G1000; n?=?29), and 1.25 (G1250; n?=?27). Animals were classified depending on their level of crossing between Nelore (N) and Hereford (H): 25%N-75%H, 37.5%N-63.5%H, 43.7%N-56.7%H, 50%N-50%H, and 75%N-25%H. Reproductive evaluation was performed at the beginning and at the end of the experimental period by ultrasonography and hormone analysis. The average age and liveweight at puberty were 388.0?±?1.9 days and 331.4?±?1.3 kg, respectively. Animals from the 25%N-75%H group reached puberty earlier than heifers from other genetic groups supplemented with G1250 diet (P?<?0.05). Heifers with higher degree of Nelore (75%N-25%H) fed with G1000 diet showed estrus 42 days prior to mating, but only 57 % reached puberty at mating (P?<?0.05). Heifers with follicles of higher diameter reached puberty (P?<?0.05) earlier. Higher average daily weight gain showed a positive effect on follicular diameter and IGF-I level at puberty (P?<?0.01). Concentrations of GH were lower in heifers fed G1250 compared to G1000 diet (P?<?0.05). There was a significant interaction between nutritional level and insulin levels at puberty (P?<?0.01). We demonstrated the relationship between IGF-I and average daily gain on the onset of puberty in heifers. In conclusion, heifers submitted to the higher feeding level showed a higher follicular diameter and were younger at puberty.     

Feeding ecology of fishes associated with Egeria spp. patches in a tropical reservoir, Brazil

Abstract –  This research characterised feeding ecology of fishes associated with patches of Egeria najas and Egeria densa , two submerged macrophytes, in Rosana Reservoir, Upper Paraná River basin, Brazil. Fishes were sampled using a 1 m² throw trap in patches of different macrophyte biomass and in three diel periods during a wet season. Fish diet (10 species) was primarily composed of autochthonous items (zooplankton, algae and aquatic insect larvae). Almost all intra-specific diet patterns had moderate to low levels of diet similarity, indicating a high variability in diet. Some species showed no diel patterns in feeding activity, whereas others were primarily diurnal or nocturnal. No differences in feeding activity were observed among habitats of medium and high macrophyte biomasses, and species tended to feed on the same items among these habitats. The most abundant fish species demonstrated low inter-specific diet overlap and appear not to compete for food resources. We suggest that Egeria patches are feeding grounds and shelter for small-sized fish species.     

Flavonol glycosides from leaves of Costus spiralis.

A new flavonol diglycoside, 3,5-dihydroxy-7,4'-dimethoxyflavone 3-O-neohesperidoside (1), together with four known flavonol 3-O-neohesperidosides were isolated from the leaves of Costus spiralis and their structures were elucidated by a combination of spectroscopic methods and chemical reactions.     

The Tripartite Symbiosis Formed by Indigenous Arbuscular Mycorrhizal Fungi, Bradyrhizobium japonicum and Soya Bean Under Field Conditions

The tripartite symbiosis formed by indigenous arbuscular mycorrhizal fungi (AMF), Bradyrhizobium japonicum (Kirchner) Jordan and soya bean (Glycine max L. Merr. cv. Evans) was investigated under field conditions to test the hypotheses that: (i) the tripartite symbiosis enhances nodulation and nodule activity; and (ii) its establishment does not rely on improved phosphorus (P) uptake through the fungal partner. Soil tillage was used to produce treatments with contrasting AMF colonization potentials while the amount of B. japonicum inoculum was kept constant. Nodulation, AMF colonization and the P and nitrogen (N) nutrition of plants were evaluated at 10 and 51 (full‐bloom) days after emergence. N₂ fixation was estimated by the difference method and by the isotopic dilution method. At the early stage of plant growth, AMF hyphal colonization and nodulation were, respectively, 16 % and 33 % greater in plants from untilled than from rototilled soil. The establishment of the tripartite symbiosis was observed under field conditions, and factors other than P nutrition were critical to its formation. However, the tripartite symbiosis did not promote N₂ fixation under the high soil P conditions of this study.     

Can the arbuscular mycorrhizal fungus Glomus intraradices actively mobilize P from rock phosphates?

Plants colonized by arbuscular mycorrhizal (AM) fungi have been shown to respond positively to the application of insoluble forms of inorganic phosphorus (P) such as rock phosphates (RPs). The mechanism(s) underlying such responses remain(s) unknown and although it has been hypothesized, there is no experimental support for the production of chelating agents by AM fungal hyphae. Here we investigate whether AM fungi can solubilize P from RPs and transfer it to plant roots. Using root-organ cultures of Daucus carrota L. inoculated or not with Glomus intraradices Schenk & Smith and containing P from different RP sources, we predicted that: (1) roots inoculated with G. intraradices would take up more P than those uninoculated; that (2) the amount of P taken up by roots through G. intraradices would be positively correlated with the RP reactivity; and that (3) G. intraradices would have access to RP through localized alterations of pH and/or by the production of organic acid anions that may act as chelating agents. The RP reactivity was positively correlated with P uptake. However, mycorrhizal roots grew initially slower and did not respond differently to any P treatment than those uninoculated. There was no evidence of localized changes in pH in proximity of G. intraradices hyphae, indicating that responses to RP by mycorrhizal plants observed in previous studies do not appear to result from the release of H⁺ ions alone or in combination with organic acid anions.     

Accumulation of specific flavonoids in soybean (Glycine max (L.) Merr.) as a function of the early tripartite symbiosis with arbuscular mycorrhizal fungi and Bradyrhizobium japonicum (Kirchner) Jordan

This study is the first report assessing the effect of soil inoculation on the signalling interaction of Bradyrhizobium japonicum, arbuscular mycorrhizal fungi (AMF) and soybean plants throughout the early stages of colonisation that lead to the tripartite symbiosis. In a study using soil disturbance to produce contrasting indigenous AMF treatments, the flavonoids daidzein, genistein and coumestrol were identified as possible signals for regulating the establishment of the tripartite symbiosis. However, it was unclear whether soil disturbance induced changes in flavonoid root accumulation other than through changing the potential for AMF colonization. In this study, soil treatments comprising all possible combinations of AMF and B. japonicum were established to test whether (1) modifications in root flavonoid accumulation depend on the potential for AMF colonization, and (2) synthesis and accumulation of flavonoids in the roots change over time as a function of the early plant-microbial interactions that lead to the tripartite symbiosis. The study was comprised of two phases. First, maize was grown over 3-week periods to promote the development of the AM fungus Glomus clarum. Second, the interaction between soybean, G. clarum and B. japonicum was evaluated at 6, 10, 14 and 40 days after plant emergence. Root colonization by G. clarum had a positive effect on nodulation 14 days after emergence, producing, 30% more nodules which were 40% heavier than those on roots solely inoculated with B. japonicum. The tripartite symbiosis resulted in 23% more N₂ being fixed than did the simpler symbiosis between soybean and B. japonicum. The presence of both symbionts changed accumulation of flavonoids in roots. Daidzein and coumestrol increased with plant growth. However, development of the tripartite symbiosis caused a decrease in coumestrol; accumulation of daidzein, the most abundant flavonoid, was reduced in the presence of AMF.     

In vitro inhibition of the activation of Pro-matrix Metalloproteinase 1 (Pro-MMP-1) and Pro-matrix metalloproteinase 9 (Pro-MMP-9) by rice and soybean Bowman-Birk inhibitors

The in vitro inhibitory activity of the rice Bowman-Birk inhibitor (rBBI) or soybean Bowman-Birk inhibitor (sBBI) against trypsin-catalyzed activation of pro-matrix metalloproteinase 1 or 9 (pro-MMP-1 or pro-MMP-9), respectively, was investigated using electrophoresis with silver staining, heparin-enhanced zymography, biotinylated gelatin, Biotrak assay, and fluorescence quenched substrate hydrolysis. rBBI at concentrations of 0.08-0.352 mg/mL dose-dependently inhibited the in vitro activation of 45 microg/mL pro-MMP-1 by trypsin. Heparin-enhanced zymography analysis of pro-MMP-1, trypsin-activated MMP-1, and a mixture of pro-MMP-1-trypsin-rBBI showed clear zones associated with trypsin-activated MMP-1 and the absence of clear zones in lanes containing pro-MMP-1 or a mixture of pro-MMP-1, trypsin, and rBBI. The results of the Biotrak assay also indicated that rBBI dose-dependently suppressed the activation of pro-MMP-1 by trypsin. sBBI dose-dependently inhibited the activation of 100 microg/mL of pro-MMP-9 by trypsin. Biotinylated gelatin assays demonstrated that pro-MMP-9 or pro-MMP-9 in the presence of trypsin and BBI did not hydrolyze gelatin, whereas p-aminophenylmercury acetate (APMA)-activated MMP-9 and trypsin-activated MMP-9 caused significant hydrolysis of gelatin. Quenched fluorescence substrate hydrolysis for total MMP activity showed that pro-MMP-1 or pro-MMP-9 did not hydrolyze the substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2; active MMP-1 or MMP-9 hydrolyzed the substrate, but lower substrate hydrolysis was obtained when pro-MMP-1 or pro-MMP-9 was incubated with trypsin in the presence of increasing concentrations of rBBI. The results are discussed in light of the role of MMP-1 and MMP-9 in the process of angiogenesis and the potential of rBBI or sBBI as a functional food ingredient.