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.     

A developmentally regulated neuraminidase activity in Trypanosoma cruzi

The human pathogen Trypanosoma cruzi (Y strain) contains a neuraminidase activity that varies widely in the different developmental stages of the parasite. The specific neuraminidase activity of infective trypomastigotes obtained from tissue culture and from the bloodstream of infected mice is 7 to 15 times higher than that of the acellular culture forms. Amastigotes were devoid of enzyme activity. The enzyme has a pH optimum of 6.0 to 6.5. Live trypanosomes released sialic acid from human erythrocytes and plasma glycoproteins. Several sialyl compounds were hydrolyzed by the parasite, but the best substrate was the protein orosomucoid. Erythrocytes from infected mice with T. cruzi parasitemia were agglutinated by peanut lectin and the hemagglutination titer was correlated with the degree of parasitemia.     

Synthetic clonal reproduction through seeds

Cloning through seeds has potential revolutionary applications in agriculture, because it would allow vigorous hybrids to be propagated indefinitely. However, asexual seed formation or apomixis, avoiding meiosis and fertilization, is not found in the major food crops. To develop de novo synthesis of apomixis, we crossed Arabidopsis MiMe and dyad mutants that produce diploid clonal gametes to a strain whose chromosomes are engineered to be eliminated after fertilization. Up to 34% of the progeny were clones of their parent, demonstrating the conversion of clonal female or male gametes into seeds. We also show that first-generation cloned plants can be cloned again. Clonal reproduction through seeds can therefore be achieved in a sexual plant by manipulating two to four conserved genes.     

Pericarp total protein profiles as molecular markers of tomato fruit quality traits in two segregating populations

The tomato fruit quality results of biochemical and physiological changes that occur during the ripening process. Although, the pericarp total protein profiles are less polymorphic than DNA-based markers the polymorphism in those could be directly associated with fruit quality traits. The aim of this work was to identify associations between polymorphic polypeptides from fruit pericarp at two ripening stages and fruit quality traits evaluated in two segregating populations of tomato. A cross between a normal ripening cultivar of Solanum lycopersicum (C, Caimanta) and a genotype carrying the nor (non ripening) gene (N) as well as a cross between Caimanta and a cherry type tomato of S. lycopersicum var. cerasiforme (Ce) showed genetic variance for several fruit quality traits such as fruit weight, shape, solids soluble content, acidity, color and fruit shelf life. The quantitative variations observed at phenotypic level had correspondence with the polymorphism detected in the protein profiles. Indeed, the polymophic polypeptides associated with quality fruit traits and fruit shelf life would be useful to assist tomato breeding programs as protein molecular markers.     

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.     

Stem modeling and simulation of conversion of cork oak stems for quality wood products

Recent research is underway to study cork oak (Quercus suber L.) wood potential for production of wood components. A total of 35 stems of young and mature cork oaks were sampled, live sawn into flitches, scanned using VTT’s WoodCIM^®, and the measured data computed by VTT’s PuuPilot software, allowing stem 3D reconstruction. Sawing simulations were run for 0.5 m and 1 m logs and the whole stem. Sawn products were (1) planks, (2) parquet, (3) lamparquet, and (4) external component of multilayer planks. Cork oak stems showed a moderate to small taper (mean 24 mm/m). Curviness varied between straight to significantly crooked stems (mean value 40 mm) Batch yields for the tested products ranged 25–43% and 37–50% for 0.5 m logs of young and mature trees, respectively; for 1 m logs, batch yields ranged 19–41% and 25–54%. When using the whole stem, batch yields were lower, ranging 11–38% and 15–50%. Higher yields were obtained for all log lengths and samples for production of lamparquet, parquet, and multilayer component.