Physiological and biochemical differences among Ulmus minor genotypes showing a gradient of resistance to Dutch elm disease

Dutch elm disease (DED) spread across Europe and North America in the 20th century killing most natural elm populations. Today, breeding programmes aim at identifying, propagating and studying elm clones resistant to DED. Here, we have compared the physiology and biochemistry of six genotypes of Ulmus minor of variable DED resistance. Leaf gas exchange, water potential, stem hydraulic conductivity and biochemical status were studied in 5‐year‐old trees of AB‐AM2.4, M‐DV2.3, M‐DV2 × M‐CC1.5 and M‐DV1 and 6‐year‐old trees of VA‐AP38 and BU‐FL7 before and after inoculation with Ophiostoma novo‐ulmi. Leaf water potential and net photosynthesis rates declined, while the percentage loss of hydraulic conductivity (PLC) increased after the inoculation in susceptible trees. By the 21st day, leaf predawn and midday water potential, stomatal conductance to water vapour and net photosynthesis rates were lower, and PLC was higher in trees of susceptible (S) genotypes inoculated with the pathogen than in control trees inoculated with water, whereas no significant treatment effect was observed on these variables in the resistant (R) genotypes. Fourier transform infrared spectroscopy analyses revealed a different biochemical profile for branches of R and S clones. R clones showed higher absorption peaks that could be assigned to phenolic compounds, saturated hydrocarbons, cellulose and hemicellulose than S clones. The differences were more marked at the end of the experiment than at the beginning, suggesting that R and S clones responded differently to the inevitable wounding from inoculation and repeated sampling over the experimental course. We hypothesize that a weak activation of the defence system in response to experimental wounding can contribute to the susceptibility of some genotypes to O. novo‐ulmi. In turn, the decline in shoot hydraulic conductivity and leaf carbon uptake caused by the infection further exacerbates tree susceptibility to the fungus.     

Factors involved in Agrobacterium tumefaciens-mediated gene transfer into Pinus nigra Arn. ssp. salzmannii (Dunal) Franco

Cotyledons from dissected sterile embryos of salgareño pine (Pinus nigra Arn. ssp. salzmannii (Dunal) Franco) were inoculated with different disarmed Agrobacterium tumefaciens strains harbouring the binary vector p35SGUSint. The transient expression of a β-glucuronidase gene (uidA) was studied, using a histochemical staining procedure. Nineteen days after inoculation, the activity of β-glucuronidase was detected in epidermal and subepidermal layers of cotyledonary explants. The EHA105 strain harbouring a disarmed agropine-type Ti-plasmid (pTiBO542) was the most effective for gene transfer of the uidA gene. The effects of exudates and extracts from 0-day-old embryos on induction of vir gene expression in A. tumefaciens were also examined. The results of this study showed that salgarño pine embryo exudates contain a substance(s) that induce vir gene expression, in similar way to that observed with 100 μM acetosyringone (AS).All these findings suggest that T-DNA processing and transfer might take place when Agrobacterium infects suitable tissues of salgareño pine.     

Sclerotinia resistance in sunflower: I. Genotypic variations of hybrids in three environments of Argentina

Sclerotinia sclerotiorum infections on sunflower capitula produce white rot, one of the most dangerous diseases sunflower bears in all humid areas. Therefore, disease quantification on developed genetic materials is a very important action in sunflower breeding. Given the horizontal type of resistance these evaluations should be made in different environments. Several hybrids obtained after crossing seven female × seven male sunflower inbred lines in a factorial mating design were evaluated for resistance to white rot in two locations in the southeast of the province of Buenos Aires, Argentina. Disease incidence and the symptom appearance index indicated significant genotypic effects and genotype × environment (G × E) interactions. Genotypic effects were a greater source of variance than the G × E interaction effects. The G × E interactions only represented changes in magnitude. These results were used to identify the best genotypes for both resistance factors. Four hybrids showed low disease incidence values under both artificial and natural infections, of which only two had high values of the symptom appearance index at both locations. These hybrids are considered to have adequate resistance attributes for the ascosporic penetration and the mycelium extension in the capitulum tissue and could therefore be sown in the province of Buenos Aires without increasing risks of S. sclerotiorum attacks.     

Molecular Analysis of Phytophthora infestans (Mont.) de Bary from Chapingo,Mexico. Phylogeographic Referential

An Andean origin of Phytophthora infestans was originally based on written records, and recently inferred from mitochondrial and nuclear gene genealogies, which emphasize a common South American origin of the European isolates, and absence of the I-b haplotype in Mexico (recently identified in Tlaxcala, Mexico). In order to compare such results, we sequenced the nuclear genes β-tubulin, Ras, and Intron Ras, and rDNA ITS regions in 24 P. infestans genotypes isolated, identified, and selected, during three crop growing cycles in Chapingo, Mexico. The pathogen showed homology and 100 % sequence similarities to the ones reported for the Andes and Europe β-tubulin, Ras, and Intron Ras genes. Results for the ITS region were inconclusive. The molecular oomycete similarities from the three geographical regions (South America, Europe, and Mexico), and the Mexican genotypes characteristics reported in the species supports the hypothesis of the location of P. infestans diversity and a center of dispersal in the central highlands of Mexico.     

In vitro release of angiotensin-converting enzyme inhibitors, peroxyl-radical scavengers and antibacterial compounds by enzymatic hydrolysis of glycated gluten

The combined effect of gluten glycation and proteolysis on the release of compounds exhibiting in vitro angiotensin-converting enzyme (ACE) inhibitory, antioxidant and antibacterial activities was investigated. Model systems consisting of wheat gluten and glucose were heated at 120 °C for 45 min, 150 °C for 30 min and 220 °C for 30 min to produce various Maillard reaction products mimicking reactions occurring in bread crusts. Progress of the Maillard reaction was estimated through indirect measurement of Amadori compounds as 2-furoylmethyl-amino acids. Glycation was followed by digestion with Pronase E and ultrafiltration. The anti-hypertensive activity was measured as the ability to inhibit the activity of angiotensin I-converting enzyme involved in hypertension regulation. The Oxygen Radical Absorbance assay was used to measure the peroxyl radical scavenging capacity of the products and their effect on microbial growth of Escherichia coli ATCC 25922 and Staphlococcus aureus ATCC 25923 was also studied. Advanced products of the reaction enhanced the antioxidant and antibacterial properties of gluten hydrolysates and decreased the overall ACE inhibitory activity. Ultrafiltration provided a useful method for separating compounds (< 3000 Da) with ACE inhibitory activity and advanced Maillard reaction products (>3000 Da) which scavenged peroxyl radicals and inhibited the microbial growth.     

Pigmentation, carotenoids, lipid peroxides and lipid composition of skin of red porgy (Pagrus pagrus) fed diets supplemented with different astaxanthin sources

A feeding experiment was carried out to determine the efficiency of different commercial sources, chemical forms and levels, of dietary astaxanthin, to appropriately pigment the red porgy (Pagrus pagrus) skin. According to this, total carotenoid content, profiles and chemical forms present in the skin were determined. In order to establish the potential for antioxidant protecting role of astaxanthin supplemented diets, peroxide levels and lipid composition of skin were also determined.

Red porgy alevins were fed six dietary treatments in triplicate; a basal diet (B) without carotenoids; two diets (N25 and N50) formulated to supply either 25 or 50 mg kg^− 1 of an esterified source of astaxanthin (Haematococcus pluvialis, NatuRose™); two diets (CP25 and CP50) with either 25 or 50 mg kg^− 1 of unesterified astaxanthin (Carophyll® Pink); and a positive control diet (B + S) proved as a successful pigmenting-diet in previous experiences (B + S, 88% basal diet:12% frozen shrimp) [Cejas, J., Almansa, E., Tejera, N., Jerez, S., Bolaños, A., Lorenzo, A., 2003. Effect of dietary supplementation with shrimp on skin pigmentation and lipid composition of red porgy (P. pagrus) alevins. Aquaculture 218, 457–469].

All fish fed carotenoid supplemented diets displayed a pink-coloured skin after 4 months of feeding in contrast to the greyish appearance displayed by fish fed the basal diet not supplemented with carotenoids (B). Furthermore, astaxanthin diesters were the major carotenoid in the skin of pink fish. A second carotenoid, tentatively identified as tunaxanthin diester, was also detected. The best results in terms of skin natural reddish hue, total carotenoid and astaxanthin contents were found by using the esterified forms of dietary astaxanthin (N25, N50 and B + S). Interestingly, the lowest levels of lipid peroxides were found in the fish fed these three treatments. However, no effect of treatment on lipid composition was found. In conclusion, red porgy alevins are able to efficiently utilise dietary natural or synthetic astaxanthin, and deposit this pigment in its esterified form to acquire an acceptable pink-coloured skin compared to that of the wild fish.     

Hydraulic Retention Time Influence on Improving Flocculation in the Activated Sludge Processes Through Polyelectrolytes

Hydraulic retention time (HRT) influence improving sludge flocculation with adding the polyelectrolytes (non-ionic, anionic, and cationic) was studied on an activated sludge (AS) system fed with synthetic domestic wastewater (SDW), dairy industry wastewater (DIW), and caramel industry wastewater (CIW). The sludge volumetric index, food/microorganism ratio (F/M), and mixed liquor volatile suspended solids at different HRTs (6, 8 and 10 h) were monitored on an experimental model. Results showed that both SDW and IW had the best sludge flocculation conditions at 8 h and 100 mL of non-ionic polyelectrolyte (0.2 mg L^?1). In addition, this phenomenon reached the organic matter removal efficiencies of 95.9, 95.7, and 94.2% for SDW, DIW, and CIW, respectively. Therefore, optimum HRT increased the organic matter removal efficiencies by 10%, sludge concentration by 37% (22–55%), and F/M ratio by 70%. Moreover, the polyelectrolytes used in AS improved the sludge flocculation by 2.9 times.     

Effect of hypo- and hyper-saline conditions on osmolarity and fatty acid composition of juvenile shrimp Litopenaeus vannamei (Boone, 1931) fed low- and high-HUFA diets

Litopenaeus vannamei (Boone) grown in ponds are exposed to salinities of less than 5 g L^?1 during inland shrimp culture or to more than 40 g L^?1 from evaporation and reduced water exchange in dry, hot climates. However, dietary requirements for shrimp grown in low or high salinities are not well defined, particularly for fatty acids. Feeding shrimp postlarvae with highly unsaturated fatty acids (HUFA) enhances tolerance to acute exposure to low salinity, as a result of better nutritional status, or/and specific effects of HUFA on membrane function and osmoregulation mechanisms. This study analysed the effect of HUFA supplementation (3% vs. 34%) on L. vannamei juveniles reared for 21 days at low (5 g L^?1), medium (30 g L^?1) and high salinities (50 g L^?1). Juveniles grown at 5 g L^?1 had lower survival compared with controls (30 g L^?1) or shrimp grown at 50 g L^?1, but no significant effect on survival was observed as a result of HUFA enrichment. In contrast, growth was significantly lower for shrimp grown at 50 g L^?1, but this effect was compensated by the HUFA‐enriched diet. Osmotic pressure in haemolymph was affected by salinity, but not by HUFA enrichment. Shrimp fed HUFA‐enriched diets had significantly higher levels of eicosapentaenoic acid and docosahexaenoic acid in hepatopancreas and gills. These results demonstrate that growth at high salinities is enhanced with diets containing high HUFA levels, but that HUFA‐enriched diets have no effect on shrimp reared at low salinities.     

Intimal thickening of coronary arteries in the rabbitfish,Chimaera monstrosa L. (Chondrichthyes: Holocephali)

This study is the first to report the occurrence of intimal thickening of coronary arteries in a holocephalan, namely the rabbitfish, Chimaera monstrosa. The sample studied consisted of five hearts from rabbitfish with body weights ranging between 12 g and 1116 g. The specimens were fixed in formalin, in methanol:acetone:water (MAW), or in paraformaldehyde and were examined by brightfield and polarization light microscopy. Coronary arteries from three larger animals displayed focal intimal thickenings, which were located in the left main coronary artery trunk, at the level of the cono‐ventricular junction, and in several intramyocardial ventricular arteries. The vascular changes were characterized by myointimal proliferation, breakage or absence of the inner elastic lamina, and, in one case, by increased collagen within the myointimal proliferation. Taking into account the severity of the coronary arterial changes, their location, and the body weight of the affected animals, we conclude that (1) these changes are age‐related in the rabbitfish and (2) local intense mechanical tension seems to be an important factor in their formation in this species.