Application of biotechnological tools to Quercus improvement

The genus Quercus, which belongs to the family Fagaceae, is native to the northern hemisphere and includes deciduous and evergreen species. The trees of the different species are very important from both economic and ecological perspectives. Application of new technological approaches (which span the fields of plant developmental biology, genetic transformation, conservation of elite germplasm and discovery of genes associated with complex multigenic traits) to these long-rotation hardwoods may be of interest for accelerating tree improvement programs. This review provides a summary of the advances made in the application of biotechnological tools to specific oak species. Significant progress has been made in the area of clonal propagation via organogenesis and somatic embryogenesis (SE). Standardized procedures have been developed for micropropagating the most important European (Q. robur, Q. petarea, Q. suber) and American (Q. alba, Q. bicolor, Q. rubra) oaks by axillary shoot growth. Although regenerated plantlets are grown in experimental trials, large-scale propagation of oak species has not been carried out. The induction of SE in oaks from juvenile explants is generally not problematic, although the use of explants other than zygotic embryos is much less efficient. During the last decade, enormous advances have been made in inducing SE from selected adult trees, mainly specimens of pedunculate oak (Q. robur) and cork oak (Q. suber). Advances in the understanding of the maturation and germination steps are required for better use of embryogenic process in clonal forestry. Quercus species are late-maturing and late-flowering, exhibit irregular seed set, and produce seeds that are recalcitrant to storage by conventional procedures. Vitrification-based cryopreservation techniques were used successfully in somatic embryos of pedunculate oak and cork oak, and an applied genbank of cork oak selected genotypes is now under development. The feasibility of genetic transformation of pedunculate oak and cork oak somatic embryos by means of co-culture techniques with several strains of Agrobacterium tumefaciens has also been demonstrated. To date, most research on the genomics of Quercus species has concerned population genetics. Approaches using functional genomics to examine the molecular and cellular mechanisms that control organogenesis and or somatic embryogenesis are still scarce, and efforts on the isolation and characterization of genes related to other specific traits should be intensified in the near future, as this would help improve the practical application of clonal forestry in recalcitrant species such as oaks.     

Economic Impacts of Zebra Chip in Idaho,Oregon, and Washington

Zebra Chip disease vectored by the potato psyllid Bactericera cockerelli (?ulc) was first reported in Idaho and the Columbia Basin of Oregon and Washington in 2011. Since then growers have incurred significant costs for managing the disease. Thus, we conducted an expert opinion survey to estimate expenditure on insecticides dedicated to controlling potato psyllids in the largest potato producing regions of Idaho, Oregon, and Washington. Results highlight a total of about 9 million US dollars spent on active ingredients targeted at psyllid control. When application costs are added to the cost of insecticides, expenditures total about 11 million US dollars.     

Importance of chip selection and elaboration process on the aromatic composition of finished wines

The evolution of volatile compounds extracted from wood while being macerated for 1 month with four different commercial chips (different geographical origins and toasting degrees) was studied. Furthermore, the effect of the microoxygenation process between alcoholic and malolactic fermentation also was studied. The wood volatile compounds in wines macerated with the four types of chips evolved in the same way. However, the amounts of compounds extracted depended on the type of chip used. There were differences in the levels of vanillin, cis-whiskey lactone, furfural, trans-isoeugenol, and cis-isoeugenol in wines in accordance with the type of wood chips (French or American), and the last two compounds along with 5-methyl furfural presented differences that were directly related to the toast level. However, no effects of microoxygenation treatment on the extraction of volatile compounds extracted from chips were observed. Therefore, the results obtained in this study highlight the importance of chip selection on the aromatic characteristics of finished wines.     

Acute phase protein response in experimental canine leishmaniasis

Acute phase proteins (APPs) have been proposed as useful markers for the diagnosis and monitoring of treatment of dogs infected by Leishmania infantum. However, the kinetics and behavior of these proteins in canine leishmaniasis is still unknown. The aim of this study was to monitor the kinetics of APPs in dogs experimentally infected with L. infantum, before, during and after therapy against canine leishmaniasis. Levels of serum haptoglobin, serum amyloid A and C-reactive protein from 6 infected beagles, positive by both PCR and parasite culture, were monitored for 7 months post-infection. The dogs were then treated for 3 months with allopurinol (20 mg mg/kg/day PO), and their response to therapy was followed for 11 additional months. Levels of Immunoglobulins G and M were recorded during these 21 months and compared. Experimental infection with L. infantum amastigotes induced an increase in all APPs studied which was statistically significant 2 months after infection for all proteins. Clinical recovery was accompanied by a significant decrease of all APPs 1 month after the beginning of treatment. However, differences were found between the APPs in both magnitude and duration of serum level elevations. The increase in total IgG and IgM was delayed in comparison to APPs and contrarily to the APPs, these immunoglobulins did not significantly decrease with treatment. In conclusion, the results of this study suggest that APPs could be used as early markers for disease as well as for monitoring the response to treatment in canine leishmaniasis.     

Pistil abortion is related to ovary mass in olive (Olea europaea L.)

Pistil abortion is considered to be an evolutionary adaptation to save resources in andromonoecious species, balancing the number of pistils with the resources available. It follows that large-fruited species/cultivars should have greater levels of pistil abortion. Working in olive, we found that pistil abortion, expressed as percentage of staminate flowers, was positively correlated to the average ovary mass at bloom, across cultivars with different ovary/fruit mass. Since ovary mass correlated with fruit mass, pistil abortion also correlated with fruit mass. The absolute number of perfect flowers per inflorescence was negatively correlated with both ovary mass and pistil abortion, while the number of staminate flowers per inflorescence increased with both parameters. The total number of flowers (i.e. staminate + perfect) was not correlated to ovary mass or pistil abortion, suggesting that male function was not affected. The results support the hypothesis that pistil abortion is related to competition for resources among ovaries, and suggest that genetic differences in pistil abortion among olive cultivars may be explained by their different pistil mass and sink strength.     

AVES POLYOMAVIRUS 1 IN ARA CHLOROPTERA AND ECLECTUS RORATUS WITH DISCLOSURE OF FULL GENOMIC SEQUENCES

Aves polyomavirus 1 (APyV) is the causative agent of acute disease in birds and causes high mortality rates in nestlings. Infections have been reported worldwide in a significant number of caged bird species, such as parrots, caiques, budgerigars, lovebirds, and macaws. However, the number of complete viral sequences available in public databases is scarce, especially those with associated clinical histories. In this study, the clinical, pathological, epidemiological, and molecular characteristics of 2 APyV strains detected in Portugal are described. In the autumn of 2015, a 2-month-old female green-winged macaw (Ara chloroptera) from a small breeder died with signs of dehydration, weight loss, and depression, raising the suspicion of polyomavirus infection. Histopathological analysis revealed lesions compatible with APyV infection, and the presence of polyomavirus in several organs was confirmed by polymerase chain reaction. From the cohabitants tested (n = 14), 1 eclectus parrot (Eclectus roratus), which was more than 1-year-old, was also APyV DNA-positive. The full genomic sequences of the 2 strains were obtained and found identical, suggesting a single introduction in the premises and the occurrence of subsequent infections by the same virus. When compared with sequences of other APyVs available in public databases, high nucleotide similarity percentages were obtained, confirming the close genetic relationship among polyomaviruses worldwide. Interestingly, strain APV7, detected in a white-bellied parrot in 2008 in Japan, was the closest strain to those isolated in this report. Attempts to isolate the virus in eggs and cell lines failed. Phylogenetic analysis was performed by the Bayesian method to determine the phylogenetic classification of the macaw and parrot strains. Both clustered into group V, together with other strains from different bird species with no host or spatial-temporal relationships being observed.     

Two monoclonal antibodies for the recognition of Mytilus spp. larvae: studies on cultured larvae and tests on plankton samples

Monoclonal antibodies (mAbs) were generated against 2-day-old mussel larvae in an attempt to develop a rapid and rigorous method for the identification of mussel larvae in field plankton samples. Previously, we have shown that two of these mAbs recognised Galician Mytilus galloprovincialis obtained from monospecific cultures, but did not recognise the larvae of other bivalve species present in that area. To assess the possibility of using these mAbs in routine assays for measuring the abundance of mussel larvae in plankton, studies on cultured mussel larvae, at different stages of development, and tests on bivalve larvae from plankton samples were carried out. Initially, to see whether the two mAbs also recognise other mussel larval stages, they were tested against mussel larvae of different ages obtained from monospecific cultures. The results indicate that both antibodies stain all the stages tested, even 1-month-old postlarvae. In addition, we also demonstrate that these mAbs also recognise other forms of Mytilus. Both antibodies bind to M. galloprovincialis larvae from the Mediterranean Sea and M. edulis larvae. Finally, and more significantly, studies on field plankton samples were performed to confirm if both mAbs are really mussel-specific, and do not cross-react with larvae of any other bivalve species existing in the plankton. The results presented here clearly indicate that our two monoclonal antibodies specifically recognise the mussel larvae in field plankton samples from different geographical regions, but not the larvae of any other bivalve species. Thus, these monoclonal antibodies could be used for routine monitoring of mussel larvae in plankton samples from different sources.