Genetic Variability and Structure of Canadian Populations of the Sapstain Fungus Ceratocystis resinifera

ABSTRACT Genomic DNA was extracted from 129 isolates of Ceratocystis resinifera, a species belonging to the C. coerulescens complex, and 19 polymorphic random amplified polymorphic DNA markers were used to study the population genetic structure of this fungus. The analysis suggested a moderate value for genetic diversity (H(S) = 0.209). However, when monomorphic markers and rare alleles, representing 89 markers, also were included in the calculation, the genetic diversity of Canadian populations of C. resinifera appeared to be much lower (H(S) = 0.045). This could be explained by two hypotheses: (i) recent introduction of this species into North America and (ii) clonal reproduction (by selfing). No specialization by C. resinifera for coniferous tree species was observed based on genetic differentiation index between isolates sampled from Pinus and Picea spp. and on phylogenetic analysis using Dice coefficient of association. In spite of a low genetic diversity, a very high genetic differentiation was observed among the nine geographical populations studied (F(ST) = 20.8%). The genetic differences were especially striking when populations from Eastern Canada were compared with populations from Western Canada (phiST = 0.27%; P < 0.001), suggesting that a geographic reproductive barrier occurs in Central Canada. This barrier may be the consequence of a weak migration of insect vectors of C. resinifera due to reduced presence of hosts in the Canadian Great Plains, where extensive agriculture occurs. However, results from pairwise F(ST) matrix and phylogeny of haplotypes suggest that the barrier is not totally impenetrable because some gene flow occurred from the west and from the east in the Big River (Saskatchewan) population located in the middle of the Great Plains.     

Immune response and antigen recognition in non-pregnant ewes experimentally infected with Neospora caninum tachyzoites

The cellular and humoral responses as well as the antigen recognition during the acute stage of a Neospora caninum (NC) infection were investigated in non-pregnant ewes. The experimentally infected ewes developed specific lymphoproliferative and humoral responses within 2 weeks post-infection (PI). The magnitude of the cellular response showed large variations between animals. A significant decrease in the proliferative response to Con A mitogen and N. caninum, Toxoplasma gondii (TG) antigens was recorded on day 21 post-infection (PI). The humoral response and the pattern of antigen recognition were similar among infected ewes. Proteins of 44, 42, 40, 39 and 28 kDa were intensively recognized by the infected animals during the experiment. The 42 and 28 kDa antigens should be considered as useful for the diagnostic of N. caninum infection, as the intensity of recognition infection of the other antigens had decreased markedly 8 weeks post-infection. For some antigens a sequential recognition was recorded. The 59, 54 and 38-37 kDa proteins were frequently recognized by infected sera during the first weeks of the infection, but recognition of these antigens was absent or rare at the end of the experiment. These antigens could be related to the acute stage of the infection.     

Seasonal patterns of cadmium accumulation in Arrhenatherum elatius (Poaceae): Influence of mycorrhizal and endophytic fungal colonisation

We investigated the influence of arbuscular mycorrhizae (AM) and dark septate fungi (DSF) colonisation on cadmium (Cd) accumulation in Arrhenatherum elatius from heavy metal-contaminated sites. AM colonisation disappeared when Cd concentrations in soil increased, while DSF infection was weak but constant throughout the experiment indicating that soil heavy metals are toxic to AM but not to DSF. AM colonisation was greatest when plant Cd concentrations were highest providing evidence that AM colonisation may influence Cd accumulation. In addition, the disappearance of AM and the concomitant reduction of Cd in shoots during seed maturation result in our suggestion that seasonal variation in AM may play a role in protecting developing seeds from soil pollution.     

Transtracheal aspiration in the diagnosis of pulmonary blastomycosis (17 cases: 2000-2005)

Blastomyces dermatitidis is a common etiologic agent of fungal pneumonia in dogs. Definitive diagnosis is based on cytologic demonstration of the organism in affected tissues. Fluid obtained through transtracheal aspiration has previously been reported to have a low diagnostic yield for B. dermatitidis organisms. This retrospective study identified B. dermatitidis organisms in 76% of samples when transtracheal aspiration was performed in 17 nonsedated dogs with pulmonary blastomycosis. Transtracheal aspiration is a noninvasive and simple procedure that should be considered as an early diagnostic test whenever blastomycosis is a differential diagnosis in dogs with pulmonary disease.     

Intake and digestibility in sheep and chemical composition during different seasons of some West African browse species

Foliage of Afzelia africana, Pterocarpus erinaceus and Khaya senegalensis, from 10 trees per species, was collected every two weeks during the late dry, rainy and cool season to determine the seasonal effects on chemical composition. Fifteen rams of the Djallonké breed, weighing on average 20.0 kg, were used to evaluate the voluntary intake and digestibility of hay of A. gayanus, foliage of A. africana (as a sole feed), and A. africana, P. erinaceus and K. senegalensis offered with 30% of the diet as A. gayanus hay. The crude protein (CP) content of A. africana, and P. erinaceus decreased significantly from the late dry season to the cool season when that of K. senegalensis tended to increase. The mean CP of A. africana, P. erinaceus and K. senegalensis differed significantly (173 g, 139 g and 114 g/kg DM, respectively). The DM intake of A. africana offered with hay (571 g/d) or as a sole feed (598 g/d) were not significantly different, but was higher than that of P. erinaceus (428 g/d) and K. senegalensis (298 g/d). The digestibility calculated by difference of DM and CP of A. africana (582 g/kg DM and 795 g/kg CP, respectively) did not differ significantly from A. africana as a sole feed, but were higher than for the other species. The nutritive value of A. africana seems to justify the high preference of herders for this species.     

RADIX: rhizoslide platform allowing high throughput digital image analysis of root system expansion

Background

Phenotyping of genotype-by-environment interactions in the root-zone is of major importance for crop improvement as the spatial distribution of a plant’s root system is crucial for a plant to access water and nutrient resources of the soil. However, so far it is unclear to what extent genetic variations in root system responses to spatially varying soil resources can be utilized for breeding applications. Among others, one limiting factor is the absence of phenotyping platforms allowing the analysis of such interactions.

Results

We developed a system that is able to (a) monitor root and shoot growth synchronously, (b) investigate their dynamic responses and (c) analyse the effect of heterogeneous N distribution to parts of the root system in a split-nutrient setup with a throughput (200 individual maize plants at once) sufficient for mapping of quantitative trait loci or for screens of multiple environmental factors. In a test trial, 24 maize genotypes were grown under split nitrogen conditions and the response of shoot and root growth was investigated. An almost double elongation rate of crown and lateral roots was observed under high N for all genotypes. The intensity of genotype-specific responses varied strongly. For example, elongation of crown roots differed almost two times between the fastest and slowest growing genotype. A stronger selective root placement in the high-N compartment was related to an increased shoot development indicating that early vigour might be related to a more intense foraging behaviour.

Conclusion

To our knowledge, RADIX is the only system currently existing which allows studying the differential response of crown roots to split-nutrient application to quantify foraging behaviour in genome mapping or selection experiments. In doing so, changes in root and shoot development and the connection to plant performance can be investigated.

     

A microplate assay to measure soil microbial biomass phosphorus

Quantification of phosphorus (P) concentrations in microbial biomass is required to better understand how P immobilization and turnover in soils are controlled by environmental and anthropogenic factors. Soil microbial biomass P (MBP) is generally extracted using the chloroform fumigation-direct extraction procedure and then analysed for P using the ammonium molybdate-ascorbic acid method on a flow injection analysis (FIA) system. Our objective was to determine whether a microscale malachite green method on a microplate system would provide as accurate MBP analysis as the ascorbic acid method on an FIA system. Twelve soils were collected from agricultural fields in southwestern Quebec, fumigated with chloroform and extracted with 0.5 M NaHCO₃ (pH 8.5). The dissolved inorganic phosphorus (DIP) concentration in fumigated soils was not affected by the method of analysis, and results from the two systems of analysis were significantly correlated (r =0.998, P <0.05). The MBP concentrations in these agricultural soils were between 0.36 and 60.05 g P g^–1, consistent with other published values. Our results indicate that MBP can be assessed equally well with the malachite green method using a microplate system as with the ascorbic acid method on an FIA system. The microplate system is rapid and requires smaller volumes of samples and reagents than the FIA system, thus reducing the quantity of waste produced. We conclude that the microscale malachite green method could be applied to measure the MBP concentration in a wide range of soils with good sensitivity, reproducibility and accuracy.     

The effect of energy reserves and cryoprotectants on overwintering mortality in Mercenaria mercenaria notata (Say 1822) at two tidal levels

The objective of this study is to identify possible causes of the high winter mortality noted in juvenile Mercenaria mercenaria notata in eastern Canada. The percentage mortality, shell growth, concentration of energy reserves, and production of cryoprotectant molecules were compared between notata and native quahogs kept at intertidal and subtidal levels. Overwintering mortality of the notata strain reached 47.2% and was 3–9 times higher than that of the native strain. Shell increment was higher in the native than in the notata strain and also at the intertidal than at the subtidal level. The quahogs from the subtidal zone had a higher concentration of protein than those from the intertidal zone in August and April but a lower concentration in February. The notata strain had a lower concentration of lipids and glucose (34.9 and 0.21 mg g⁻¹ dry weight) than the native strain (42.2 and 0.28 mg g⁻¹ dry weight). Thermal hysteresis was detected in none of the quahog groups. High winter mortality in the notata strain seems to be caused, in part, by a lower capacity to stock lipid compared with the native strain. The higher concentration of glucose in the native strain may favour survival in cold water by helping to reduce the freezing point of the animals’ fluids.     

Fucoidan Structure and Its Impact on Glucose Metabolism: Implications for Diabetes and Cancer Therapy

Fucoidans are complex polysaccharides derived from brown seaweeds which consist of considerable proportions of L-fucose and other monosaccharides, and sulphated ester residues. The search for novel and natural bioproduct drugs (due to toxicity issues associated with chemotherapeutics) has led to the extensive study of fucoidan due to reports of it having several bioactive characteristics. Among other fucoidan bioactivities, antidiabetic and anticancer properties have received the most research attention in the past decade. However, the elucidation of the fucoidan structure and its biological activity is still vague. In addition, research has suggested that there is a link between diabetes and cancer; however, limited data exist where dual chemotherapeutic efforts are elucidated. This review provides an overview of glucose metabolism, which is the central process involved in the progression of both diseases. We also highlight potential therapeutic targets and show the relevance of fucoidan and its derivatives as a candidate for both cancer and diabetes therapy.     

Potential of Exopolysaccharide from Porphyridium marinum to Contend with Bacterial Proliferation,Biofilm Formation,and Breast Cancer

Exopolysaccharide (EPS) from marine microalgae are promising sources of a new generation of drugs. However, lot of them remain to be discovered and tested. In this study, EPS produced by Porphyridium marinum and its oligomers prepared by High Pressure Homogenizer have been tested for different biological activities, i.e., antibacterial, anti-fungal and antibiofilm activities on Candida albicans, as well as for their effects on the viability of murine breast cancer cells. Results have shown that all EPS samples present some biological activity. For antibacterial and antibiofilm activities, the native EPS exhibited a better efficiency with Minimum Inhibitory Concentration (MIC) from 62.5 µg/mL to 1000 µg/mL depending on the bacterial strain. For Candida albicans, the biofilm formation was reduced by about 90% by using only a 31.3 µg/mL concentration. Concerning breast cancer cells, lower molar masses fractions appeared to be more efficient, with a reduction of viability of up to 55%. Finally, analyses of polymers composition and viscosity measurements were conducted on all samples, in order to propose hypotheses involving the activities caused by the intrinsic properties of polymers.