Involvement of Bradyrhizobium japonicum denitrification in symbiotic nitrogen fixation by soybean plants subjected to flooding

Denitrification by Bradyrhizobium japonicum bacteroids contributes to nitric oxide (NO) production within soybean nodules in response to flooding conditions. However, the physiological relevance of NO production by denitrification in B. japonicum-Glycine max symbiosis is still unclear. In this work, soybean plants were inoculated with B. japonicum strains lacking the nirK or norC genes which encode the copper-containing nitrite reductase and the c-type nitric oxide reductase enzymes, respectively. 14 days flooding increased nodule number of plants inoculated with the WT and norC strains, but not of plants inoculated with the nirK mutant. However, nodule dry weight was not affected by 14 days flooding regardless of the strain used for inoculation. Supporting this observation, individual nodule growth was significantly higher in plants inoculated with nirK than those inoculated with WT or norC after 14 days flooding. Nodule functioning was strongly inhibited by flooding since leghemoglobin content of the nodules induced by any of the strains was significantly decreased after 7 or 14 days flooding compared to control plants. However, this effect was more relevant in nodules of plants inoculated with the WT or norC mutant than in those inoculated with the nirK mutant. Nitrogen fixation was also estimated by analyzing nitrogen content derived from biological nitrogen fixation in shoots, using the ¹⁵N isotope dilution technique. By using this approach, we observed that the negative effect of 14 days flooding on nitrogen fixation was more pronounced in plants inoculated with the norC mutant. However, nitrogen fixation of plants inoculated with nirK showed the highest tolerance to 14 days flooding. These findings allowed us to demonstrate the previously proposed hypothesis which suggests that NO formed by copper-containing nitrite reductase in soybean nodules, in response to flooding, has a negative effect on nitrogenase activity. We propose that inoculation of soybeans with a B. japonicum nirK mutant, which does not produce NO from nitrate, increases the tolerance of symbiotic nitrogen fixation to flooding.     

Effects of Rapid Cooling Prior to Freezing on the Quality of Canine Cryopreserved Spermatozoa

The aim of this study was to evaluate the effects of rapid cooling prior to freezing on frozen-thawed canine sperm quality. In experiment 1, centrifuged ejaculates from 6 dogs were pooled, split into 4 aliquots and cryopreserved by the Uppsala procedure using different cooling rates (control, cooling speed 18 C/90 min and average cooling rate 0.2 C/min; rapid, cooling speed 18 C/8 min and average cooling rate 2.25 C/min) in combination with 2 glycerol addition protocols (fractionated or unfractionated). In experiment 2, centrifuged ejaculates from 4 dogs were processed individually using the same cooling rates described in experiment 1 in combination with an unfractionated glycerol addition protocol. Each of the experiments was replicated 5 times. Sperm quality was evaluated after 30 and 150 min of post-thawing incubation at 38 C. Total motility (TM), progressive motility (PM) and quality of movement parameters were assessed using a computerized system, and sperm viability (spermatozoa with intact plasma and acrosome membranes) was assessed using flow cytometry (H-42/PI/FITC-PNA). Values for TM, PM, viable spermatozoa and the quality of movement parameters after thawing were not significantly affected by the cooling rate. The interaction between the cooling rate and the added glycerol protocol was not significant. There were significant differences among the males (P<0.01) in the sperm quality parameters evaluated after thawing. The interaction between the males and the cooling rate was not significant. In conclusion, canine spermatozoa can be cryopreserved using the Uppsala method at an average cooling rate of 2.25 C/min prior to freezing together with addition of fractionated or unfractionated glycerol.     

Effects of selection for color intensity on antioxidant capacity in maize (Zea mays L.)

Maize kernels with diverse colors have pigments and other substances with antioxidant capacity that can be highly beneficial for human health. Our objectives were to analyze antioxidant capacity of the different maize kernel colors, to investigate if selection for color intensity increases antioxidant capacity and to check the effect of bakery processing on antioxidant capacity. Pigment content was directly related to antioxidant capacity in maize kernels and also to the hydrophilic fraction, while the lipophilic fraction was not related to antioxidant capacity. Visual selection for kernel color increased anthocyanin content from 0.55 to 1.13 μmol/g of fresh weight in one unique cycle of selection. The traditional method for making maize bread out of whole maize flour has not significant effects on pigment content or antioxidant capacity. Therefore, maize bread made out of black or purple maize provides significant amounts of antioxidant substances potentially beneficial for human health.     

Boron increases pollen quality,pollination, and fertility of different genetic lines of pepper

Using different genetic lines of pepper, this study analyses the effect of boron (B) on pollen quality and its capacity to pollinate and fertilize. Three different B concentrations (100, 150, and 200 mg L^?1) were applied to six genetic lines, evaluating the in vitro pollen quality and pollen formation. In a second experiment, the highest and the lowest B concentrations were applied to two low-pollen genetic lines, evaluating the growth of the pollen tubes in flowers pollinated with treated pollen. The higher B concentrations significantly improved pollen quality and formation in vitro, while improved pollination and fecundity was observed after monitoring the pollen tubes. For all variables, the specific genetic line also showed some effect. This technique represents a tool for improving pollen in manual hybrid seed programs.     

Sampling soil and sediment depth profiles at a fine resolution with a new device for determining physical,chemical and biological properties: the Fine Increment Soil Collector (FISC)

Purpose

Many environmental investigations (empirical and modelling) and theories are based on reliable information on the depth distribution of physical, chemical and biological properties in soils and sediments. However, such depth profiles are not easy to determine using current approaches, and, consequently, new devices are needed that are able to sample soils and sediments at fine resolutions.

Materials and methods

We have designed an economic, portable, hand-operated surface soil/sediment sampler—the Fine Increment Soil Collector (FISC)—which allows for the close control of incremental soil/sediment sampling and for easy recovery of the material collected by a simple screw-thread extraction system. This innovative sampling system was developed originally for the beryllium-7 (⁷Be) approach in soil and sediment redistribution research. To ensure reliable estimates of soil erosion and sediment deposition from ⁷Be measurements, the depth distribution of this short-lived fallout radionuclide in soil/sediment at the resolution of millimetres is a crucial requirement. This major challenge of the ⁷Be approach can be met by using the FISC.

Results and discussion

We demonstrate the usefulness of the FISC by characterising the depth distribution of ⁷Be at increments of 2.5 mm for a soil reference site in Austria. The activity concentration of ⁷Be at the uppermost increment (0–2.5 mm) was ca. 14 Bq kg^?1 and displayed decreasing activity with depth. Using most conventional sampling devices (i.e. the scraper-plate system), the most accurate depth increment would have been 10 mm, and the activity concentration at the surface would have been considerably lower. Consequently, coarser sampling would have influenced estimates of ⁷Be-derived soil erosion and deposition. The potential application for other soil/sediment properties, such as nutrients (e.g. phosphorus), contaminants and carbon are also discussed.

Conclusions

By enabling soil and sediment profiles to be sampled at a depth resolution of millimetres, the FISC has the potential to provide key information when addressing several environmental and geoscientific issues, such as the precise depth distributions of soil/sediment nutrients, contaminants and biological properties.     

Afforestation method affects the isotopic composition of planted Pinus halepensis in a semiarid region of Spain

We used an isotopic approach to evaluate the effects of three afforestation methods on the ecophysiology of an Aleppo pine plantation in semiarid Spain. The site preparation methods tested were excavation of planting holes (H), subsoiling (S), and subsoiling with addition of urban solid refuse to soil (S + USR). Five years after plantation establishment, trees in the S + USR treatment were over three times larger than those in the S treatment, and nearly five-fold larger than those planted in holes. Differences in tree biomass per hectare were even greater due to disparities in initial planting density and pine tree mortality among treatments. Pine trees in the S + USR treatment showed higher foliar P concentration, δ¹³C and δ¹⁵N than those in the S or H treatments. Foliar δ¹⁵N data proved that trees in the S + USR treatment utilized USR as a source of nitrogen. Foliar δ¹³C and δ¹⁸O data suggest that improved nutrient status differentially stimulated photosynthesis over stomatal conductance in the pine trees of the S + USR treatment, thus enhancing water use efficiency and growth. In the spring of 2002, trees in the S + USR treatment exhibited the most negative predawn water potentials of all the treatments, indicating that the rapid early growth induced by USR accelerated the onset of intense intra-specific competition for water. The results of this study have implications for the establishment and management of Aleppo pine plantations on semiarid soils. Planting seedlings at low density and/or early thinning of pine stands are strongly recommended if fast tree growth is to be maintained beyond the first few years after USR addition to soil. Foliar C, O and N isotope measurements can provide much insight into how resource acquisition by trees is affected by afforestation techniques in pine plantations under dry climatic conditions.     

Anticholinesterase and pediculicidal activities of monoterpenoids

The repetitive and inadequate application of pediculicidal products frequently results in the development of resistance to these compounds. Essential oils are a promising alternative to synthetic insecticides, although their mode of action remains to be explored. It has been proposed that one possible target of the essential oils is the inhibition of acetylcholinesterase (AChE). The role of monoterpenoids as possible AChE inhibitors and their relationship with the toxicity was investigated both in vitro and in vivo. Inhibition of electric eel AChE activity showed that the most effective inhibitor was 1,8-cineole with IC(50) 6 x 10(-3) M. The inhibition of AChE activity of head louse homogenate by 1,8-cineole showed IC(50) 7.7 x 10(-2) M. The intoxication symptoms of head lice exposed to vapors of 1,8-cineole was recorded before the in vivo head louse AChE inhibition assay. No correlation was found between neurotoxic symptoms and inhibition of AChE activity.