Transgenic fertile soybean plants derived from somatic embryos transformed via the combined DNA-free particle bombardment and <Emphasis Type="Italic">Agrobacterium</Emphasis> system

An Agrobacterium-mediated transformation procedure for soybean [Glycine max L. Merrill] proliferating somatic embryos is here described. The Agrobacterium tumefaciens LBA4404 strain harboring pTOK233, pCAMBIA1390-olp or pH7WG2Dwrky plasmids was used to mediate gene transfer into the plant genome. Prior to Agrobacterium inoculation, proliferative soybean embryogenic clusters were microwounded by DNA-free tungsten particle bombardment. Three independent transformation experiments were performed. In Experiment I, 26 transgenic plants were obtained from a unique clone of cv Bragg, while 580 plants were recovered from 105 clones of cv IAS5. In Experiment II, a single hygromycin-resistant clone of cv BRSMG68 Vencedora was recovered and gave rise to five plants. In Experiment III, 19 plants of cv Bragg and 48 plants of IAS5 were recovered, representing five and 14 independent transformation events, respectively. PCR and Southern analyses confirmed the transgenes’ integration into plant genomes. Transgenic plants were fertile. They flowered, set pods and seeds. Transgene segregation in two T₁ progenies fits the Mendelian pattern (3:1 transgenic:non-transgenic plants). This is the first report of transgenic fertile soybean plants obtained from somatic embryogenic tissues transformed by the system that combines DNA-free particle bombardment and Agrobacterium.     

Phenological and physiological evaluation of first and second cropping periods of sorghum and maize crops

Environmental conditions influence phenology and physiological processes of plants. It is common for maize and sorghum to be sown at two different periods: the first cropping (spring/summer) and the second cropping (autumn/winter). The phenological cycle of these crops varies greatly according to the planting season, and it is necessary to characterize the growth and development to facilitate the selection of the species best adapted to the environment. The aim of this study was to characterize phenological phases and physiological parameters in sorghum and maize plants as a function of environmental conditions from the first cropping and second cropping periods. Two parallel experiments were conducted with both crops. The phenological characterization was based on growth analyses (plant height, leaf area and photoassimilate partitioning) and gas exchange evaluations (net assimilation rate, stomatal conductance, transpiration and water-use efficiency). It was found that the vegetative stage (VS) for sorghum and maize plants was 7 and 21 days, respectively, longer when cultivated during the second cropping. In the first cropping, the plants were taller than in the second cropping, regardless of the crop. The stomatal conductance of sorghum plants fluctuated in the second cropping during the development period, while maize plants showed decreasing linear behaviour. Water-use efficiency in sorghum plants was higher during the second cropping compared with the first cropping. In maize plants, in the second cropping, the water-use efficiency showed a slight variation in relation to the first cropping. It was concluded that the environmental conditions as degree-days, temperature, photoperiod and pluvial precipitation influence the phenology and physiology of both crops during the first and the second cropping periods, specifically cycle duration, plant height, leaf area, net assimilation rate, stomatal conductance and water-use efficiency, indicating that both crops respond differentially to environmental changes during the growing season.     

Selection proposal of cultivars of spring barley in the years from 2001 to 2004, using Joint Regression Analysis

Joint Regression Analysis is a widely used technique in the analysis of series of experiments for cultivar comparison. We show how this technique may be used in the selection of recommended cultivars. We apply this technique to a series of experiments on spring barley cultivars carried out from 2001 to 2004. The data in these designs were kindly made available by the Central Institute for Supervising and Testing in Agriculture in Brno, Czech Republic.     

A field evaluation of anthelmintics for control of cyathostomes of horses in Brazil

The anthelmintic efficacy of ivermectin (IVM), piperazine citrate (PPZ) and fenbendazole (FBZ) alone, and combinations of FBZ with varying dosages of PPZ, was evaluated for the control of strongyles of Mangalarga equids. In all cases of positive strongly egg counts per gram of faeces (EPG) before and after treatment, pure populations of cyathostome larvae with eight gut cells were consistently observed in faecal cultures. The faecal egg count reductions (FECR) were evaluated between Day 7 and Day 56 post-treatment. Significant differences (P less than or equal to 0.05) were found between Day 7 and Day 49 when comparing the mean EPG values of IVM with those of the other anthelmintics. PPZ alone and the combination of FBZ (7.5 mg kg-1) and PPZ (55 mg base kg-1) did not show significant differences during the trial; both groups exhibiting an FECR of greater than 90% between Day 7 and Day 30. Horses treated with the combinations of FBZ (7.5 mg kg-1) and PPZ at either 40 or 25 mg base kg-1, and with FBZ alone at either 7.5 or 15 mg kg-1, showed an FECR of less than 90% 2 weeks after treatment. The emergence of a degree of resistance of strongyles to FBZ and PPZ in a segment of the cyathostomes in this population is suggested.     

Spatio-temporal impacts of roads on the persistence of populations: analytic and numerical approaches

Roads can have drastic impacts on wildlife populations. Although there is wide recognition of the negative impacts caused by roads and a wealth of practical studies, there is a lack of theoretical work that can be used to predict the impact of road networks or to implement mitigation measures. Here, using Skellam’s diffusion model, we develop analytic and numerical approaches to analyze the impact of road networks on the survival of populations. Our models show that the viability of a population is determined not only by road density but also by the size and shape of patches. Accordingly, we studied the minimum size of a patch to sustain a population with given diffusion and growth parameters. We provide simple formulas to estimate the minimum patch size, and illustrate the importance of shape with square and rectangular patches. Our models also allow the estimation of time to extinction after road construction for a population in a patch smaller than that of the minimum size. Finally, using numerical computations we illustrate how the spatial arrangement of fences strongly affects both the equilibrium density and the spatial distribution of populations, and that not all fence layouts are equally effective. We anticipate that our methods provide a tool to assess the impact of geometrical features of road networks on wildlife and that they can be used to design mitigation measures to prevent the decline and extinction of populations in an anthropogenically disturbed landscape.     

Comparison of a rebound and an applanation tonometer for measuring intraocular pressure in normal rabbits

Objective To compare intraocular pressure (IOP) measurements made on healthy adult rabbits without the effect of tranquilizers using the new applanation tonometer, Tono‐Pen Avia^®, and the rebound tonometer Tonovet^®. Methods Intraocular pressure was measured throughout the day (6:00, 9:00, 12:00, 15:00, and 18:00 h) in 38 adult New Zealand White rabbits (76 eyes). The animals were 20 males and 18 females, with a mean weight of 3.5 kg and an average age of 6 months. A complete ocular exam (including Schirmer tear test, fluorescein staining, slit‐lamp biomicroscopy, and direct ophthalmoscopy) was performed on all animals at the beginning of the trial. Rebound tonometry was performed, and after 10 min, anesthetic drops were instilled and applanation tonometry was carried out. IOP values obtained using the two techniques were analyzed statistically. Results The mean IOP was 9.51 ± 2.62 mmHg with Tonovet^®, and 15.44 ± 2.16 mmHg with the Tono‐Pen Avia^®. Significant differences between measurements with the two tonometers were observed (P < 0.001). The linear regression equation describing the relationship between the two tonometers was y = 0.4923x + 10.754 (y = Tonovet^® and x = Tono‐Pen Avia^®). High IOPs were recorded in the early measurements (6:00), but the average IOPs from both devices were statistically similar throughout the day (P = 0.086). The correlation coefficient was r² = 0.357. No significant difference in IOP regarding gender was observed. Conclusion The Tono‐Pen Avia^® recorded higher levels of IOP compared with the Tonovet^®. Early in the day, the IOP of rabbits was higher than later in the day, regardless of the tonometer used.