Genome-wide analysis of plastome sequence variation and development of plastidial CAPS markers in common potato and related <Emphasis Type="Italic">Solanum</Emphasis> species

The plastome sequence of the European cultivated potato, Solanum tuberosum subsp. tuberosum (tbr, GenBank accession no. DQ386163), was compared with that of S. bulbocastanum, a wild potato relative (blb, GenBank accession no. DQ347958), in order to characterize the degree and type of variability in different genomic regions, and develop molecular markers relevant to genetics, breeding and biotechnology of potato. One hundred forty-two and 251 PICs (Potentially Informative Characters) were found in coding and non-coding sequences (NCSs), respectively. Further, while variation in coding regions was almost exclusively due to nucleotide substitutions, 25% of PICs in NCSs of tbr and blb were due to indels, most of them mononucleotide or longer tandem repeats (micro and minisatellites). Four intergenic regions were selected for further analyses in other 16 tuber-bearing Solanum species. The rps16-trnQ ^UUG gene spacer was found to be the most variable, forty-six PICs in this region distinguishing 18 haplotypes. Analysis of haplotype relationships, based on variability in the four intergenic regions, confirmed that the most primitive species from Central America were the most distant to S. tuberosum. Finally, polymorphic sites in the same regions were used to develop a set of CAPS (Cleaved Amplified Polymorphic Sequences) markers for species/cytoplasm identification in Solanum spp.     

Chemical composition and pasting properties of einkorn (Triticum monococcum L. subsp. monococcum) whole meal flour

Einkorn (Triticum monococcum L. subsp. monococcum) is an ancient wheat appreciated for its high proteins, carotenoids and tocols content. To better understand its potential for human consumption and food preparation, composition and pasting properties of 65 einkorn accessions, of different geographical origin but cultivated at S. Angelo Lodigiano (Italy), were evaluated. Eight Triticum turgidum and seven Triticum aestivum cultivars, belonging to different subspecies, were assessed as controls.On average, the einkorn samples had light seeds (25.0 mg/kernel), extra-soft texture (238.3 g), high protein content (18.2%), high ash content (2.35%), low SDS sedimentation volume (25.6 ml), high carotenoids (8.36 μg/g) and high yellow pigments (8.46 μg/g). The pasting parameters studied were peak viscosity (average: 2426 cP), breakdown (765 cP), final viscosity (2788 cP) and setback (1126 cP). Total amylose (25.7%) and total starch (65.5%) were also measured. Significant differences from the controls and a broad variation for all the traits analysed were observed. Einkorns from diverse geographical areas showed different mean values for all characters, apart from ash content.The good pasting properties, coupled with high proteins and carotenoids content, suggest that einkorn is particularly suited for the production of baby and specialty foods.     

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.     

Serologic and molecular characterization of tickborne pathogens in lions (Panthera leo) from the Fasano Safari Park, Italy.

Lions (Panthera leo) are an endangered species threatened by illegal hunting, habitat loss, and infectious diseases. Little is known about the tick-borne pathogens that infect lions and could contribute to population declines. The objective of this study was to characterize Rickettsia spp., Anaplasma phagocytophilum, and Coxiella burnetii infections in 10 lions from the Fasano Safari Park in Italy by serology, polymerase chain reaction, and sequence analysis. Although animals did not show clinical signs of tick-borne diseases, evidence of infection with C. burnetii, spotted fever group Rickettsia sp., and A. phagocytophilum were found in 50%, 20%, and 10% of the lions, respectively. One of the lions tested positive for all three pathogens. This study is the first report of molecular evidence of infection with C. burnetii, Rickettsia sp., and A. phagocytophilum in lions and provides evidence that these felids become infected and serve as hosts for tick-transmitted bacteria.     

Segregation distortion in doubled haploid lines of barley (Hordeum vulgare L.) detected by simple sequence repeat (SSR) markers

A total of 147 simple sequence repeat (SSR) markers (including86 barley and 61 wheat microsatellite markers) were tested for their segregation in a doubled haploid (DH) and an F₂ population of barley. The DH population consisted of 71 doubled haploid lines, developed from F₁ plants of a cross between Tadmor and WI2291using isolated microspore culture technique. A genetic linkage map consisting of 43 microsatellite markers was constructed using the DH population. Particularly on chromosome 4H microsatellite markers showed distorted segregation ratios. Segregation of DH lines based on molecular markers were compared with segregation of 92 F₂ lines from the same cross. The proportion of loci deviating from the expected monogenic segregation ratios in the DH population was significantly higher (19/43loci, 44%) than in the F₂ population (7/43 loci, 16%). The deviation was biased towards the WI2291 parent alleles. In line with this observation, WI2291 was found to perform better than Tadmor in regenerating green plantlets with the isolated microspore-culture technique. This revised version was published online in July 2006 with corrections to the Cover Date.     

Enhanced iron reduction by iron supplement: A strategy to reduce methane emission from paddies

As an option for mitigating methane emissions from rice paddies the effects of ferrihydrite application to an experimental field plot at the beginning of the growth season was studied. Methane emissions during the vegetation period of rice were significantly lower (50%) in the fertilized plot compared to the non-supplemented control plot. Although toxic effects of iron are known to occur with wetland plants including rice, our field experiment showed no deterioration of agrophysiological data. Grain yield, harvest index, and iron content of grains were not different in the two plots. Therefore, we propose that iron application is a suitable strategy to reduce methane emission from rice paddies.     

Colour Flow Mapping examination: An useful screening test for the early diagnosis of ductus venosus patency in canine newborns

Ductus venosus (DV) closure plays a key role in hepatic circulation adaptation to postnatal metabolic function, and DV patency might develop a congenital portosystemic shunt (CPSS). The noninvasive Color Flow Mapping (CFM) examination, a validated method to diagnose CPSS in adult dogs, is routinely performed to assess DV closure after birth in humans. This study aimed to describe the feasibility of the ultrasonographic evaluation of the DV after birth and to determine its closure time in healthy Great Dane neonates. Patency of DV in serial Color Flow Mapping (CFM) examinations and bodyweight (BW) were recorded on Days 0‐3‐6‐9 in 24 neonates that were classified as having patent (PDV) or closed ductusvenosus (CDV) basing on CFM signal presence/absence. Since the 3rd day, DV diameter was recorded. Data were analysed by ANOVA (p < 0.05). All dogs resulted healthy 1 year later. The number of PDV and CDV puppies at birth was not different on Day 3 (24 and 0 vs. 22 and 2, PDV and CDV, respectively), whereas it resulted different on Days 6 (24 and 0 vs. 14 and 10) and 9 (24 and 0 vs. 0 and 24); on Day 3, it was different compared to Days 6 and 9; on Day 6, it was different from Day 9. Reduction of DV diameter resulted positively related to neonatal BW growth. The CFM evaluation of DV closure after birth in Great Dane puppies represents a feasible technique. Present results suggest the time of functional closure in normal neonates within 9 days after birth. Thus, CFM examination, as an early screening test for DV patency evaluation, performed 10 days after birth, may identify suspicious dogs at risk that would require further investigations. Further studies are needed to deepen the role of a delayed closure in low bodyweight and preterm puppies.     

Effects of glandular trichomes on the development of Phytophthora infestans infection in potato (S. tuberosum)

S. berthaulltii Hawkes, a wild potato species, possesses two types of glandular trichomes on its leaf surface: types A and B wich confer resistance to insects. Preliminary results indicated that the presence of glandular trichomes also confers resistance to Phytophthora infestans, the causal agent of late blight. In this paper we report the evaluation of trichome characteristics (density and exudate activities) performed on 8 lines of potato, including the control varieties Desirée and Tropicana, 5 hybrid lines derived from intercrosses and backcrosses between S. tuberosum and S. berthaultii and one accession of S. berthaultii. Whole plants of the 8 genotypes were inoculated with a spore suspension of Phytophthora infestans in order to evaluate their resistance to late blight. The resultant disease incidence correlated negatively with type A trichome density and with polyphenol-oxidase (PPO) activity of type A trichome glands. The removal of type B trichomes exudate by leaf dipping in ethanol did not influence the disease development after artificial inoculation of detached leaves. Type B trichome exudate was extracted and the fraction corresponding to Fatty Acids Sucrose Esters (FASE) was recovered after Thin Layer Chromatography and tested on the fungal mycelium in vitro. The FASE exhibited a slight inhibitory effect on mycelium growth. The results of the experiments are discussed, and several hypotheses regarding the possible role of types A and B trichomes are formulated. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.