Selection,characterization and genetic analysis of laboratory mutants of <Emphasis Type="Italic">Botryotinia fuckeliana</Emphasis> (<Emphasis Type="Italic">Botrytis cinerea</Emphasis>) resistant to the fungicide boscalid

Resistance to the fungicide boscalid in laboratory mutants of Botryotinia fuckeliana (Botrytis cinerea) was investigated. The baseline sensitivity to boscalid was evaluated in terms of colony growth (EC₅₀ = 0.3–3 μg ml⁻¹; MIC = 10–30 μg ml⁻¹) and conidial germination (EC₅₀ = 0.03–0.1 μg ml⁻¹; MIC = 1–3 μg ml⁻¹) tests. Mutants were selected in vitro from wild-type strains of the fungus on a fungicide-amended medium containing acetate as a carbon source. Mutants showed two different levels of resistance to boscalid, distinguishable through the conidial germination tests: low (EC₅₀ ∼ 0.3 μg ml⁻¹, ranging from 0.03 to 1 μg ml⁻¹; MIC > 100 μg ml⁻¹) and high (EC₅₀ > 100 μg ml⁻¹) resistance. Analysis of meiotic progeny from crosses between resistant mutants and sensitive reference strains showed that resistant phenotypes were due to mutations in single major gene(s) inherited in a Mendelian fashion, and linked with both the Daf1 and Mbc1 genes, responsible for resistance to dicarboximide and benzimidazole fungicides, respectively. Gene sequence analysis of the four sub-units of the boscalid-target protein, the succinate dehydrogenase enzyme, revealed that single or double point mutations in the highly conserved regions of the iron-sulphur protein (Ip) gene were associated with resistance. Mutations resulted in proline to leucine or phenylalanine replacements at position 225 (P225L or P225F) in high resistant mutants, and in a histidine to tyrosine replacement at position 272 (H272Y) in low resistant mutants. Sequences of the flavoprotein and the two transmembrane sub-units of succinate dehydrogenase were never affected.     

Isolation and characterization of Avenin-like protein type-B from durum wheat

Proteomic techniques were applied for the investigation of “avenin-like protein type-B” in tetraploid wheat. Protein was extracted from endosperm, according to a classical chromatographic procedure and its identity confirmed by liquid chromatography-tandem mass spectrometry analysis. The characterization of avenin-like protein type-B was in good agreement with the gene-derived sequence, with the exception of glutamine as N-terminus. The presence in the glutenin extract and the high number of cysteine residues suggested that avenin-like protein type-B is integrated into gluten polymers via inter-chain disulphide bonds. This raises the question of whether the protein could play a role in determining the functional properties of gluten.     

Effects of two fractions of inspired oxygen on lung aeration and gas exchange in cats under inhalant anaesthesia

ObjectiveTo compare the effects of two fractions of inspired oxygen (FiO₂) (0.4 and 1) on lung aeration and gas exchange during general anaesthesia in cats.Study designRandomized, blinded, controlled study.AnimalsThirty healthy, mixed breed, client owned female cats.Materials and methodsCats were premedicated intramuscularly with acepromazine (0.03 mg kg^?1) and medetomidine (0.015 mg kg^?1). Anaesthesia was induced with propofol (5 mg kg^?1) and, after orotracheal intubation, maintained with isoflurane carried by either 100% oxygen (G100, n = 15) or an oxygen-air mixture with 40% oxygen (G40, n = 15). All cats were placed in dorsal recumbency and breathed spontaneously throughout the entire procedure. Following surgery (ovariectomy), a spiral computed tomography (CT) of the thorax was performed, arterial oxygen (PaO₂) and carbon dioxide (PaCO₂) tensions were measured and alveolar-arterial gradient of oxygen [P(A-a)O₂] calculated. The CT images were analysed for lung aeration by the analysis of radiograph attenuations (Hounsfield units, HU), according to the following classification: hyperinflated area (-1000 to -900 HU), normally aerated area (-900 to -500 HU), poorly aerated area (-500 to -100 HU) and non-aerated area (-100 to +100 HU). The groups were compared using one-way anova.ResultsCompared to G100, the normally-aerated lung area was significantly greater and the poorly-aerated and non-aerated areas were significantly smaller in G40. PaCO₂ was similar in both groups. PaO₂ and P(A-a)O₂ were significantly higher in G100. In both groups, pulmonary atelectasis developed preferentially in the caudal lung fields.ConclusionIn cats anaesthetised with isoflurane, the administration of an FiO₂ of >0.9 significantly impaired lung aeration and gas exchange as compared to an FiO₂ of 0.4.Clinical relevanceAn FiO₂ of 0.4 may better preserve lung aeration and gas exchange in anaesthetised spontaneously breathing cats but monitoring is essential to ensure oxygenation is adequate.     

Closed cycle subirrigation with low concentration nutrient solution can be used for soilless tomato production in saline conditions

Closed cycle soilless techniques can be adopted to minimize water and fertilizer losses in greenhouse cultivation. There is a general lack of information regarding the soilless cultivation of vegetables with closed cycle subirrigation techniques, specifically when using saline water. In this study, a trough bench subirrigation system (SUB), with two fertilizer concentrations (“100%”, containing 9.8 mol m⁻³ N-NO₃, 1.6 mol m⁻³ P-H₂PO₄, 8.7 mol m⁻³ K⁺, 2.8 mol m⁻³ Ca⁺, 1.8 mol m⁻³ Mg⁺, 4 mol m⁻³ S-SO₄, and “70%”, containing 70% of the macronutrient concentration) in the nutrient solution (NS), was compared with open cycle drip-irrigation (DRIP with “100%” NS). For all the three treatments, NS was prepared using rain water (0.05 dS m⁻¹) and adding NaCl (1 g L⁻¹), in order to simulate moderate saline irrigation water. The effect of the treatments on tomato (Solanum lycopersicum L.) plant growth, yield, fruit quality, water use efficiency (WUE) and fertilizer consumption was evaluated. Substrate and recirculating NS composition were also studied. Subirrigation, regardless of NS concentration, reduced plant height (by 30 cm), leaf area (by 1411 cm²), total fresh and dry weight (by 429 and 48.5 g plant⁻¹, respectively) but not dry matter percentage of the whole plant, with respect to DRIP. Yield was reduced when plants were subirrigated with the higher concentrated NS, but no differences with open cycle DRIP were recorded when the lower NS concentration was used in SUB. Fruit quality was not affected by irrigation system or NS concentration. The higher WUE was obtained with subirrigation. NaCl accumulated similarly over the crop cycle in recirculating NS of both SUB treatments and in growing substrates of all the three treatments. Higher salt concentration was found in subirrigated substrates, in particular in the upper part of the substrate profile. Fertilizers accumulated in the subirrigated substrates when the higher NS concentration was used, but not when the NS concentration was reduced by 30%. The results of this study indicate that tomato can be grown successfully in a closed cycle subirrigation system, using saline water, by reducing the fertilizer NS concentration normally used with traditional open cycle systems.     

Dietary Supplementation of Oregano and Sage Dried Leaves on Performances and Meat Quality of Rabbits

The aim of this research was to evaluate the dietary supplementation 1% （w/w） of oregano and sage dried leaves on performances and meat quality of broiler rabbits. A feeding trial, which lasted 48 d, was carried out on 105 male Bianca Italiana rabbits randomly divided in seven groups and fed ad libitum. At the end of the trial ten animals per group were slaughtered and samples of dorsal muscle were taken in order to perform laboratory analysis. Mortality rate did not statistically differ between groups. Growth performances of animals fed diets supplemented with aromatic plants were higher （P〈0.05） than those of animals of control group, whereas carcass parameters were not affected by treatments excepting for the slaughter weight that showed the same trend as growth performances. Meat quality traits, oxidative lipid stability and fatty acid proifle were not inlfuenced by aromatic plant supplementation. In conclusion, oregano and sage in form of dried leaves can be used in rabbit without adverse effects on performance, carcass characteristics and meat quality traits.     

A new set of microsatellite markers for Fragaria species and their application in linkage analysis

Summary

This study reports the development of 68 new microsatellite markers. Of these, 45 were obtained, together with 20 others already published, from an AC-enriched genomic library of the wild strawberry Fragaria vesca. The 68 markers were tested for transportability to the cultivated strawberry F. ananassa ‘Miss’ and 83% gave positive amplifications. Twenty pairs of primers were selected and tested for their transportability to 16 Fragaria taxa and eight species of Rosaceae (peach, almond, apricot, European and Sino-Japanese plums, sweet and sour cherry, apple). The average proportion of primers amplifying loci in Fragaria was 69%, while the transportability to Rosaceae was very low and resulted in null amplification for 80% of the primer pairs. In addition, 23 microsatellite markers were developed from F. ananassa ‘expressed sequence tags’ databases. A total of 141 primer pairs from these and published primers, were tested for polymorphism in the two parents (91.333.2 and ‘Snovit’; both belonging to F. vesca) of a full sib population of 46 individuals. Fifty-eight percent of the primers were discarded because they were monomorphic, or were difficult to interpret, or their allelic conformation was not useful for mapping. The segregation of 73 primers was tested in the progeny and a partial map of the female parent was constructed, based on the segregation of 66 useful markers that were ordered into eight linkage groups of which four had from seven to 14 markers.