Evaluation of the API 50CH and API ZYM systems for rapid characterization of Clavibacter michiganensis subsp. sepedonicus, causal agent of potato ring rot

API 50CH and API ZYM systems were used to characterize fifty-three strains of Clavibacter michiganensis subsp. sepedonicus from different geographic locations and several reference strains of the same and different species, including other potato pathogens. Clavibacter michiganensis subsp. sepedonicus strains displayed a high level of homogeneity, both in carbohydrate utilization and in enzymatic activity. Using API 50CH and API ZYM it was possible to differentiate C. michiganensis subsp. sepedonicus strains from the remaining taxa analysed in this study, which included representative strains of the other subspecies of C. michiganensis as well as other bacterial pathogens affecting potatoes. Therefore, these systems could be used as an effective method to characterize C. michiganensis subsp. sepedonicus. Such a procedure would constitute an alternative system to the conventional nutritional and physiological identification tests currently included in the official methods employed in the European Union to detect and identify this bacterium. The results obtained with the API systems agreed with the current taxonomic classification of C. michiganensis, clearly separating sepedonicus from the other subspecies belonging to this species.     

Plant growth,yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on four rootstocks

Vegetative growth, yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on ‘Rangpur’ lime, ‘Swingle’ citrumelo, ‘Orlando’ tangelo, and ‘Cleopatra’ mandarin were evaluated under subtropical climate of Northern São Paulo State, Brazil, from 2000 through 2006 harvest seasons. ‘Fallglo’ mandarin trees had the highest cumulative yield on ‘Rangpur’ lime, and the smallest on ‘Swingle’ citrumelo and ‘Orlando’ tangelo. Plants of this cultivar had the highest yield efficiency on ‘Rangpur’ lime, and the lowest on ‘Orlando’ tangelo. ‘Sunburst’ mandarin trees began to bear fruits later than ‘Fallglo’ mandarin trees, with no differences in yield induced by the rootstocks. ‘Cleopatra’ mandarin induced the most vigorous growth in ‘Fallglo’ mandarin as compared to plants on ‘Swingle’ citrumelo. On the other hand, the largest trees of ‘Sunburst’ mandarin were registered on ‘Orlando’ tangelo, and the smallest on ‘Rangpur’ lime. ‘Sunburst’ mandarin had higher alternate bearing than ‘Fallglo’ regardless the rootstock. Fruit weight and juice content were not affected by the rootstock. These two mandarin scion cultivars may be considered adequate alternatives to produce good fruit quality for the fresh fruit market. ‘Cleopatra’ mandarin and ‘Rangpur lime are suitable rootstocks for ‘Fallglo’ mandarin, whereas all rootstocks evaluated are adequate for ‘Sunburst’ mandarin.     

Assessment of real-time PCR as a method for determining the presence of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> in different Solanaceae cultivars

Real-time PCR was used to detect and quantify Verticillium dahliae and to assess the susceptibility of four Capsicum annuum cultivars (Luesia, Padrón, SCM331 and PI201234) and the Capsicum chinense cv. C118 to this pathogen. The symptoms which developed after infection included stunting and yellowing, and were more acute in the cv. SCM331, which also suffered defoliation in later stages of the disease and in C118, which suffered severe stunting. Quantification of the pathogen DNA in roots 23 and 34 days post-inoculation (dpi) revealed that there were significantly higher amounts of Verticillium dahliae DNA in C118 than in the other cultivars, followed by SCM331, Padrón and PI201234. The lowest amounts of fungal DNA in roots were found in Luesia. In hypocotyls, the highest amounts of fungal DNA were found in SCM331, while Luesia, Padrón and PI201234 had much lower amounts, and C118 had intermediate levels. When a compatible versus an incompatible system was studied, using the near-isogenic tomato lines LA3030 (susceptible) and LA3038 (resistant to V. dahliae), we were able to detect fungal DNA in both lines. As expected, the fungus/plant DNA ratio was lower in LA3038 than in LA3030 and it decreased with time in LA3038. The amount of Verticillium dahliae DNA in the roots of LA3030 remained constant between days 23 and 34 post-inoculation, but increased 10-fold in collars. Finally, when real-time PCR was applied as a diagnostic method to samples from pepper plants, soil and water collected from farms in northwest Spain, we were able to detect V. dahliae DNA in these samples even when symptoms of the disease were not evident.     

<Emphasis Type="Italic">Amaranthus leaf mottle virus</Emphasis>: 3′-end RNA sequence proves classification as distinct virus and reveals affinities within the genus <Emphasis Type="Italic">Potyvirus</Emphasis>

Amaranthus leaf mottle virus (AmLMV) was classified as a member of the genus Potyvirus on the basis of its particle morphology, serology, and biological properties (Casetta et al., 1986). Based on these properties, an Amaranthus viridis-infecting virus isolated in Spain, causing mottle and leaf blistering as well as reduced growth has been identified as AmLMV. The 3′ terminal genomic region of this and a reference isolate from Italy has been sequenced and reveals a 95% nucleotide identity between the two isolates. The sequenced part comprises the coat protein with 281 amino acids and 315 nucleotides of the 3′ untranslated region (UTR) preceding a polyadenylated tail. Pairwise comparisons and phylogenetic analysis of the nucleotide and deduced amino acid sequences of the CP and 3′ UTR of the cloned cDNAs with those of other potyviruses shows that AmLMV is a distinct potyvirus closely related to Potato virus Y.     

Evaluation of Feed Attractants in Juvenile Senegalese Sole,Solea senegalensis

The Senegalese sole, Solea senegalensis, is a flatfish with increasing interest in the European aquaculture. However, one of the main limitations found in this species is related to the scarce attraction to commercial aquafeeds, thus determining low feed intake in fish farms. The aim of this study was to evaluate four ingredients of marine origin (fish meal hydrolysate [FMH], polychaete meal [POM], mussel meal [MUM], and squid meal [SQM]) as potential attractants for juvenile Senegalese sole. Four experimental feeds containing 10% of the potential attractants, plus a control diet without these ingredients, were evaluated. A total of 150 fish were distributed into ten 40‐L tanks (duplicate tanks per feed). A Latin square design was carried out, and hence all the experimental diets were offered to each tank in successive periods of 7 d. Fish were fed twice a day (morning and afternoon), and the feeding behavior of the animals was assessed by direct observations. Behavioral pattern assessed was classified as distant orientation, near orientation, or continuation of intake. The acceptability index for each behavioral phase and experimental feeds used was established for each observation. No behavioral differences could be noticed between the morning and the afternoon feedings. FMH showed the greatest attractant potential than among the ingredients assessed, and consequently this ingredient yielded the higher feed intake rates. On the other hand, POM was not the best attractant for juvenile Senegalese sole.     

Pulmonary recruitment of neutrophils and bacterial clearance in mice inoculated with aerosols of Pasteurella haemolytica or Staphylococcus aureus.

Pulmonary alveolar macrophages are considered to be the main phagocytic cell of the pulmonary defense mechanism. However recent studies indicate that neutrophils may also participate in the defense against inhaled bacteria. The aim of this investigation was to study in mice the correlation between numbers of phagocytic cells in the bronchoalveolar space and the pulmonary clearance of bacteria. White mice were exposed to aerosols of Pasteurella haemolytica (n = 129) or Staphylococcus aureus (n = 129) in three different experimental replicates. Another group of mice (n = 22) was sham exposed to an aerosol of sterile phosphate buffered solution in a single replicate. Animals were sacrificed at various times postaerosolization. The numbers of neutrophils and alveolar macrophages in lung lavages and the pulmonary bacterial clearance rates were determined and statistically analysed. No significant differences (p greater than 0.05) were observed in the rates of pulmonary clearance between the two genera of bacteria, but P. haemolytica had a significant (p less than 0.05) replicate effect. The number of alveolar macrophages was not significantly affected by either bacteria or phosphate buffered solution. Exposure to P. haemolytica resulted in dramatic, significant (p less than 0.01) but transient increases in neutrophils in the bronchoalveolar space as well as a significant (p less than 0.01) increase in the weights of lung. The correlation between neutrophils and clearance was positive for P. haemolytica but negative for S. aureus. These results indicate that both species of bacteria are rapidly eliminated from the lung despite a rather different cellular response.     

Changes in hydrophilic and lipophilic antioxidant activity and related bioactive compounds during postharvest storage of yellow and purple plum cultivars

Eight plum cultivars (four dark-purple and four yellow) were harvested at the commercial ripening stage, and changes of fruit quality properties were evaluated during cold storage and subsequent shelf-life, with special emphasis on bioactive compounds (phenolics, anthocyanins and carotenoids) and antioxidant activity (TAA). From the eight plum cultivars, four showed the typical climacteric ripening pattern (‘Blackamber’, ‘Larry Ann’, ‘Golden Globe’ and ‘Songold’) while four behaved as suppressed-climacteric types (‘Golden Japan’ ‘Angeleno’, Black Diamond’ and ‘TC Sun’), the latter being described for the first time. At harvest, large variations in phytochemicals and antioxidant activity were found among cultivars in peel and pulp tissues, although phytochemical concentration and antioxidant activity were higher in the peel than in the flesh (2–40-fold depending on the bioactive compound). During storage, increases in total phenolics for all cultivars (peel and pulp), in total anthocyanin content in the peel of the dark-purple plums, and total carotenoids in the peel and pulp of the yellow cultivars were observed. This behaviour of the bioactive compounds was reflected in TAA changes, since hydrophilic-TAA (H-TAA) was correlated with both phenolics and anthocyanins, while lipophilic-TAA (L-TAA) was correlated with carotenoids. L-TAA comprised about 30–50% of the TAA in plum tissues. Carotenoids and phenolics (and among them the anthocyanins) could be the main lipophilic and hydrophilic compounds contributing to L-TAA and H-TAA, respectively. No significant loss of bioactive compounds and TAA occurred during prolonged plum storage. Moreover, for a better evaluation of the antioxidant potential of plums, the contribution to carotenoids should not be overlooked.     

Fruit set dependence on carbohydrate availability in citrus trees

We generated source-sink imbalances by defoliation and sucrose supplementation by stem injection, to investigate effects of carbohydrate availability on fruitlet growth and abscission in cv. Okitsu of Satsuma mandarins (Citrus unshiu (Mak.) Marc.). Partial defoliation promoted fruitlet abscission, whereas sucrose supplementation increased citrus fruit set by more than 10%. Moreover, when applied together, sucrose supplementation counteracted the effect of partial defoliation on fruit set. When sucrose was supplied continuously from flowering until harvest, it increased the concentrations of soluble and insoluble sugars in fruits. We conclude that fruit set in citrus is highly dependent on carbohydrate availability.     

Changes in understory species diversity during theNothofagus pumilio forest management cycle

Human commercial activities must be compatible with sustainable management to achieve species conservation in the natural environment. After harvesting, diversity and abundance of understory species differ substantially from the pre-harvest situation. Have knowledge of the changes that logging produces in all components of the system allows us to develop management alternatives with reduced impacts. The objective of this work was to evaluate changes in density, biomass and diversity of understory species along a forest shelterwood cut system management cycle in aNothofagus pumilio forest. Six forest management situations (postharvesting stands and different phases of forest development) were sampled in a pureN. pumilio forest of Tierra del Fuego (Argentina) of middle-high site class (fromclass I toIII). Species number, density and biomass were modified substantially after a shelterwood cut. These were being greatest after cutting and falling substantially when the forest canopy closes during theinitial growth phase, returning to values similar to this virgin forest when themature phase was reached. The impact of the forest management on understory species diversity is analyzed and discussed. The silvicultural practices applied toN. pumilio forest in Tierra del Fuego permit the introduction of alien flora to the system, and produce great changes during the forest management cycle. To Celina Matteri, Itatí Ferraro, and Rosa Kofalt for their aid in taxonomical identification. To Juan Manuel Cellini, Ricardo Vukasovic and Gregorio Martínez Pastur for their participation in the field work; and to the sawmillLos Castores, Universidad Nacional de la Patagonia Austral, Centro Austral de Investigaciones Científicas and Instituto Nacional de Tecnología Agropecuaria for the institutional and financial support.     

Biomass dynamics of Erythrina lanceolata as influenced by shoot-pruning intensity in Costa Rica

Pruning of agroforestry trees, while reducing shade of the crops, usually reduces both biomass production and nitrogen fixation. Short pruning cycles are often not sustainable on the long run, because tree production declines over subsequent pruning cycles. We compared biomass and labile carbohydrate dynamics of Erythrina lanceolata Standley (Papilionaceae) shade trees under total and partial pruning regimes in a vanilla (Vanilla planifolia L.) plantation in South-western Costa Rica. The highest biomass production was measured in the unpruned control, followed by trees with 50% of the leaf pruned every three months, while total pruning every six months resulted in the lowest biomass pruduction. In the more productive treatments, a higher proportion of the production was in branches. Because, the N content of woody branches was high, they were important for nitrogen cycling. In the partial pruning treatment more nitrogen was returned to the soil from litter and woody branches than from pruned leaf. Sugar concentrations were not different between treatments and the dynamics of non-structural carbohydrates (sugar and starch) seems to depend more on plant phenology than pruning treatment. However, the starch concentrations in the total pruning were lower than in the other treatments.This revised version was published online in November 2005 with corrections to the Cover Date.