Molecular marker diversity and bacterial wilt resistance in wild <Emphasis Type="Italic">Solanum commersonii</Emphasis> accessions from Uruguay

Solanum commersonii is a wild tuber-bearing species native to Uruguay with high potential for use in potato breeding programs. Little is known about the genetic diversity within this wild species and the relationship with the resistance to the bacterial pathogen Ralstonia solanacearum. We studied 30 S. commersonii clonal accessions, 20 of which were collected from geographically different areas across the country, while the other ten were grown from seeds from a single plant. Resistance against R. solanacearum was tested and different levels of resistance were found, ranging from delayed wilting to asymptomatic reactions. The genetic variation and the relationships among individuals in this germplasm collection were studied by different molecular markers: Random Amplified Polymorphic DNA (RAPD), Amplified Fragment Length Polymorphism (AFLP) and Microsatellites or Simple Sequence Repeats (SSR). AFLP markers generated the largest number of total and polymorphic fragments per assay unit while SSR revealed the highest frequency of polymorphic bands (100%), followed by AFLP (96.2%) and RAPD (89.4%). In contrast, when comparing the number of different genetic profiles generated, the SSR markers exhibited the lowest discriminatory power. The clustering pattern obtained with the three marker systems showed a similar distribution of the S. commersonii germplasm revealing a high correlation between the three methods employed. All three dendrograms grouped most of the accessions into two main clusters, containing the same accessions regardless of the marker type. Bacterial wilt resistant accessions were present in both clusters. Accessions originated from different seeds of the same plant were grouped within one of the major clusters, and differed in the response to R. solanacearum revealing segregation of resistance. Furthermore, the distribution in two main clusters showed high correspondence with the geographical origin of the accessions, from the north and south of the country, and with the subspecies malmeanum and commersonii morphologically identified.     

A recombinant form of chagasin from Trypanosoma cruzi: inhibitory activity on insect cysteine proteinases

BACKGROUND: The activity of the major digestive cysteine proteinase detected in the intestinal tract of larvae of the bean weevil, Acanthoscelides obtectus (Say), was efficiently inhibited by the well-characterized cysteine proteinase synthetic inhibitor E-64 and also by a recombinant form of chagasin (r-chagasin), a tight-binding cysteine proteinase inhibitor protein from Trypanosoma cruzi. RESULTS: Incorporation of r-chagasin into an artificial diet system at 0.1 g kg(-1) retarded growth rate, decreased larval survival and led to complete mortality of A. obtectus at the end of the trial. The observed differences in growth rates occurred particularly in the first and second development stages. Artificial seeds containing high levels of r-chagasin (0.5-30 g kg(-1)) completely inhibited larval penetration. CONCLUSION: Together, the results reported in this paper support the hypothesis that the inhibitory activity of r-chagasin towards the major insect gut cysteine proteinase in vitro and in vivo is an accurate prediction of its insecticidal effects. The selectivity of this inhibitor against insect digestive proteinases supports the key role in parasite virulence by affecting the endogenous proteinase activity in its natural host.     

Mini-cuttings technique: a new ex vitro method for clonal propagation of sweetgum

An efficient production system, which can quickly produce high quality plants, is shown in this study of Liquidambar styraciflua. The mini-cuttings technique was used for ex vitro clonal propagation of three sweetgum clones. The semi-hydroponics system for production of propagules was also tested, and the concentration of indole-3-butyric acid (IBA) for rooting was evaluated. Three adult clones, produced according to conventional cutting processes, were used as mini-stumps and maintained in semi-hydroponics systems of medium size sand. Ten collections of sprouts were prepared to evaluate the production of semi-hydroponics systems (mini-clonal hedges). Sprouts from collection number six were used at different concentrations of IBA solution (0; 500; 1,000; 3,000 and 6,000 mg l⁻¹) to test the mini-cuttings technique. The high survival rates of mini-stumps after ten collections (96.3%), and high annual production per square meter (~2,953 mini-cuttings) showed that the mini-clonal hedges are viable to conduct sweetgum mini-stumps. The formation of 62.5% of healthy plants after hardening under outdoor conditions showed that sweetgum could be successfully propagated by the mini-cuttings technique.     

Above-ground biomass dynamics after reduced-impact logging in the Eastern Amazon

Changes in above-ground biomass (AGB) of 17 1 ha logged plots of terra firme rain forest in the eastern Amazon (Brazil, Paragominas) were monitored for four years (2004–2008) after reduced-impact logging. Over the same time period, we also monitored two 0.5 ha plots in adjacent unlogged forest. While AGB in the control plots changed little over the observation period (increased on average 1.4 Mg ha⁻¹), logging resulted in immediate reductions in ABG that averaged 94.5 Mg ha⁻¹ (±42.0), which represented 23% of the 410 Mg ha⁻¹ (±64.9) present just prior to harvesting. Felled trees (dbh > 55 cm) accounted for 73% (±15) of these immediate losses but only 18.9 Mg ha⁻¹ (±8.1) of biomass was removed in the extracted logs. During the first year after logging, the annual AGB balance (annual AGB gain by recruitment and growth − annual AGB loss by mortality) remained negative (−31.1 Mg ha⁻¹ year⁻¹; ±16.7), mainly due to continued high mortality rates of damaged trees. During the following three years (2005–2008), average net AGB accumulation in the logged plots was 2.6 Mg ha⁻¹ year⁻¹ (±4.6). Post-logging biomass recovery was mostly through growth (4.3 ± 1.5 Mg ha⁻¹ year⁻¹ for 2004–2005 and 6.8 ± 0.9 Mg ha⁻¹ year⁻¹ for 2005–2008), particularly of large trees. In contrast, tree recruitment contributed little to the observed increases in AGB (1.1 ± 0.6 Mg ha⁻¹ year⁻¹ for 2004–2005 and 3.1 ± 1.3 Mg ha⁻¹ year⁻¹ for 2005–2008). Plots with the lowest residual basal area after logging generally continued to lose more large trees (dbh ≥70 cm), and consequently showed the greatest AGB losses and the slowest overall AGB gains. If 100% AGB recovery is desired and the 30-year minimum cutting cycle defined by Brazilian law is adhered to, current logging intensities (6 trees ha⁻¹) need to be reduced by 40–50%. Such a reduction in logging intensity will reduce financial incomes to loggers, but might be compensated for by the payment of environmental services through the proposed REDD (reduced emissions from deforestation and forest degradation) mechanism of the United Nations Framework Convention on Climate Change.     

Subsidy or stress? Tree structure and growth in wetland forests along a hydrological gradient in Southern Europe

In forested wetlands, hydrology exerts complex and sometimes compensatory influences on tree growth. This is particularly true in semi-arid ecosystems, where water can be both a limiting resource and a stressor. To better understand these relationships, we studied hydrologic and edaphic controls on the density, growth, tree architecture and overall productivity of forested wetlands dominated by the tree species Alnus glutinosa and Salix atrocinerea in Southern Europe. We sampled 49 plots set within 21 stands in the Atlantic coastal zone of the Iberian Peninsula, and quantified woody composition, size structure (diameter and height), and radial growth using dendrochronology. Plots were grouped into three saturation classes to compare tree growth characteristics (tree density, degree of sprouting, live basal area and productivity) across levels of saturation. We used Principal Component Analysis (PCA) to create integrated explanatory factors of hydrology, soil nutrient status and soil texture for use in linear mixed models to predict stand characteristics. Increased site saturation favoured a shift in species dominance from Alnus to Salix and resulted in a higher degree of multi-stemmed tree architecture (‘shrubbiness’), particularly for Alnus. Radial growth was negatively correlated with long-term soil saturation; however, annual productivity on a per-tree basis varied by species. Alnus growth and tree density were negatively correlated with waterlogging and fine-textured soils, possibly due to anaerobiosis in the rooting zone. In contrast, Salix growth was more influenced by nutrient limitation. Overall site productivity as measured by annual basal area increment decreased with prolonged saturation. In summary, soil saturation appears to act as a chronic stressor for tree species in this ecosystem. However, these species persist and maintain a dominant canopy position in the most permanently flooded patches through increased sprouting, albeit at a reduced rate of overall biomass accumulation relative to well-drained sites. The diversity in functional responses among wetland forest species has important implications for the conservation and management of these ecosystems. The sustainable management of these ecosystems is directly tied to their vulnerability to changing hydrological conditions. Non-equilibrium modifications to the hydrologic regime from land use and climate change, which are particularly severe in semi-arid regions, may further decrease productivity, integrity and resilience in these stress-adapted communities.     

Influence of Superovulatory Protocols on In Vitro Production of Nellore (Bos indicus) Embryos

There are indications in the literature that delaying the period between ovarian superestimulation and ovum pick up (OPU) would induce follicles to a condition of initial atresia, which could be beneficial to oocyte development. In this work, we compared three protocols for OPU and in vitro production (IVP) of embryos, in Nellore cattle. Nellore cows (n = 18) were randomly allocated in three groups: Group 1 (OPU), Group 2 [Follicle stimulating hormone (FSH) and OPU] and Group 3 (FSH deprivation and OPU). Three OPUs were performed, and the animals were switched to a different group each time (crossover), in such a way that at the end of the experiment all cows received the 3 protocols. At random stage of the oestrous cycle (D‐2), all follicles ≥ 6 mm were aspirated to induce a new follicular wave 2 days afterwards (D0). In Group 1, OPU was performed on D2 and oocytes were processed to IVP. In Group 2, starting on D0, cows were superstimulated (FSH, Folltropin^®, 30 mg administered daily, i.m., during three consecutive days, total dose = 180 mg), and 6 h after the last FSH dose, they received exogenous luteinizing hormone (LH) (12.5 mg, i.m., Lutropin^®, D3). The OPU was performed 6 h after LH administration, i.e. 12 h after the last dose of FSH. Animals in Group 3 received the same treatment as those in Group 2, except that LH was administered 42 h after the last dose of FSH, and OPU occurred 6 h later. Therefore, in this group, follicles were deprived of FSH at 48 h. Both cleavage and blastocyst rates were similar (p > 0.05, anova ) among oocytes from Groups 1, 2 and 3, respectively: 77.4% (144/185) and 42.70% (79/185); 75.54% (105/139) and 31.65% (44/139); 63.52% (101/159) and 33.33% (53/159). However, hatched blastocyst rate was higher (p < 0.01) in Group 1 (30.27%, 56/185) when compared with Group 2 (11.51%, 16/139) or 3 (15.72%, 25/159). It is concluded that, contrary to previous work on European breeds (Bos taurus), ovarian superstimulation associated with deprivation of FSH and OPU (Group 3) did not increase IVP of Nellore embryos (Bos indicus). On the contrary, the highest hatched blastocyst rates were observed in oocytes from non‐superstimulated cows.     

Modulation of cutaneous inflammation induced by ticks in contrasting phenotypes of infestation in bovines

Tick saliva contains molecules that are inoculated at the site of attachment on their hosts in order to modulate local immune responses and facilitate a successful blood meal. Bovines express heritable, contrasting phenotypes of infestations with the cattle tick, Rhipicephalus (Boophilus) microplus: breeds of Bos taurus indicus are significantly more resistant than those of Bos taurus taurus. Tick saliva may contain molecules that interfere with adhesion of leukocytes to endothelium and resistant hosts may mount an inflammatory profile that is more efficient to hamper the tick's blood meal. We show in vitro that adhesion of peripheral blood mononuclear cells to monolayers of cytokine-activated bovine umbilical endothelial cells was significantly inhibited by tick saliva. The inflammatory response to bites of adults of R. microplus mounted by genetically resistant and susceptible bovine hosts managed in the same pasture was investigated in vivo. The inflammatory infiltrates and levels of message coding for adhesion molecules were measured in biopsies of tick-bitten and control skin taken when animals of both breeds were exposed to low and high tick infestations. Histological studies reveal that cutaneous reactions of resistant hosts to bites of adult ticks contained significantly more basophils and eosinophils compared with reactions of the susceptible breed. Expression of the adhesion molecules – intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and P-selectin – was higher in adult-infested skin of susceptible hosts undergoing low infestations compared to resistant hosts; when host was exposed to high infestations expression of these adhesion molecules was down-regulated in both phenotypes of infestations. Expression of leukocyte adhesion glycoprotein-1 (LFA-1) was higher in skin from susceptible hosts undergoing low or high infestations compared to resistant hosts. Conversely, higher levels of E-selectin, which promotes adhesion of memory T cells, were expressed in skin of resistant animals. This finding may explain the resistant host's ability to mount more rapid and efficient secondary responses that limit hematophagy and infestations. The expression profiles observed for adhesion molecules indicate that there are differences in the kinetics of the inflammatory reactions mounted by resistant and susceptible hosts and the balance between tick and host is affected by the number of tick bites a host receives. We show that the contrasting phenotypes of infestations seen in bovines infested with R. microplus are correlated with differences in the cellular and molecular composition of inflammatory infiltrates elicited by bites with adult ticks.     

Mixed germ cell tumour of the spinal cord in a young dog

A spinal cord mixed germ cell tumour was identified in a two-year-old, female Airedale terrier with a history of progressive paraplegia. At necropsy, a discrete pale pink spinal cord mass was identified at the L5 vertebra, which extensively invaded the associated spinal cord segments L6 and L7. Histologically, the tumour was located within the subarachnoid space and invaded the spinal cord. It was composed of three different types of cells: small round cells, forming clusters or islands, similar to germ cells identified in testicular germinomas; large cells with eosinophilic or amphophilic cytoplasm, forming trabeculae; and a third, rarer type of well differentiated epithelial cells, forming acinar or tubular structures. Immunohistochemical examination revealed reactivity to antibodies against alpha-fetoprotein and broad spectrum cytokeratins. The composition, cytological features and immunohistochemical staining of this neoplasm are similar to germ cell tumours previously reported in the suprasellar region and in the eye of dogs.