Occurrence and frequency of putatively Y chromosome linked DNA markers in Cannabis sativa L.

DNA from female and male hemp (Cannabis sativa L.) plants belonging to nine different varieties were screened with180 RAPD primers in a search for sex-associated DNA markers. About 1500bands were produced in total, nine primers were found yielding one or two DNA bands amplified in all nine male DNA bulks and absent in all female DNA bulks. These putatively male-associated markers were then scored in three different F1progenies, deriving from a cross between a common male parent and three different female plants. The sex of the progeny was accurately scored on the basis of the floral phenotype, and the presence of the nine male-associated markers was verified by RAPD analysis. In all three progenies examined, all the male plants showed the DNA markers previously identified by bulk segregant analysis (BSA) on the hemp varieties, while all the female plants lacked them. The fact that the association between these markers and the staminate phenotype is found when examining male plants of distantly related varieties, and that such linkage is never broken when different progenies are examined, strongly supports the hypothesis that the markers found are physically located on the Y chromosome, in a region excluded from recombination during meiosis. Another marker was shown to be present in the male parent, in all the male plants of each progeny, and in 50% of the female progenies, while it was absent in the female parent; the possible occurrence of markers deriving from multiple amplification sites of the genome is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effect of Azospirillum inoculants on arbuscular mycorrhiza establishment in wheat and maize plants

Plant growth-promoting rhizobacteria and arbuscular mycorrhizal (AM) fungi represent two main groups of beneficial microorganisms of the rhizosphere. The role of different strains of Azospirillum on AM fungi development was evaluated by measuring the percentage of AM colonisation of the root system in durum wheat and maize plants, grown under both greenhouse and field conditions. The effect of wild-type Azospirillum brasilense strain Sp245 and genetically modified (GM) derivatives overproducing indole-3-acetic acid was assessed at greenhouse level in (1) three different cultivars of durum wheat, in the presence of indigenous AM fungi and (2) maize plants artificially inoculated with Glomus mosseae and Glomus macrocarpum. In addition, the establishment of natural AM fungal symbiosis was evaluated using Azospirillum lipoferum CRT1 in maize plants at field level. Despite the stimulatory effect of the different Azospirillum inocula on root growth, no significant differences in AM colonisation were found, independently of the AM fungus involved, either in wheat or in maize plants. Similarly, GM A. brasilense, which strongly stimulates root development, did not affect AM formation. Although these results were obtained in conditions in which the mycorrhization rate was moderate (15–30%), overall considered they indicate that the use of wild-type or GM Azospirillum phytostimulators does not alter mycorrhization.     

Semiautomated determination of pesticides in water using solid phase extraction disks and gas chromatography-mass spectrometry

A method based on semiautomated solid phase extraction using octadecyl-bonded silica disks and gas chromatography-mass spectrometry, operated in selected ion monitoring mode, allows detection and quantification of approximately 100 pesticides and transformation products in drinking water. Samples (500 mL) were passed through the disk, and the retained pesticides were eluted with acetone and ethyl acetate. Typical recoveries for pesticides at 0.1 microg L(-1) in water were in the range of 72-120% with relative standard deviations less than 20%. Calibration curves were linear over the range of 0.025-0.5 microg mL(-1) (equivalent to a concentration range in drinking water of 0.05-1.0 microg L(-1)).     

A Preliminary Study of the Biological Control of Strongyles Affecting Equids in a Zoological Park

The main goal in this research was to determine the beneficial effect of incorporating biological procedures in parasite control programs for equids in zoological parks. Two trials were developed for Equus quagga, E asinus , and E africanus asinus. The first trial (September 2010 to August 2011) consisted of chemotherapy only (ivermectin plus praziquantel), and the second trial (September 2011 to September 2012) consisted of administration of chemotherapy and chlamydospores of the nematophagous fungus Arthrobotrys (Duddingtonia) flagrans. The effect of these measures was evaluated by the estimation of the reduction in the fecal egg counts (FECR). In the first trial, 100% FECR values were achieved 15 days after treatment in all the animals. The egg reappearance period (ERP) was 2-3 months for the equids, and all of them were passing strongyle eggs in the feces at 2-4 months after their deworming. In the second experiment, the FECR values were 100% in the three species. ERPs of 3 months in the European donkeys, 4 months in the Africans, and 6 months in the zebras were recorded. All the equids had positive results for the coprological flotation test 4-8 months after anthelmintic administration. This preliminary study demonstrates the incorporation of chlamydospores of nematophagous fungus, as A flagrans appears highly promising for reduction of the infective stages of the strongyles affecting captive animals, but the experimental design precludes true determination of whether the treatment is fully efficacious.     

Genetic diversity of a <Emphasis Type="Italic">Coffea</Emphasis> Germplasm Collection assessed by RAPD markers

Genetic diversity and relationships within and among nine species of Coffea, one species of Psilanthus and the Piatã hybrid from the Coffee Germplasm Collection of Instituto Agronômico de Campinas (IAC), Brazil were assessed using RAPD markers. Genetic diversity and relationships were evaluated by proportion of polymorphic loci (P), Shannon’s genetic index (H′ and G′_ST) and clustering analysis. The overall RAPD variation among all accessions was mostly partitioned between rather than within species. However, C. canephora and C. liberica showed a high genetic diversity within the species (\({\underline{\hbox{H}'}} \) _sp = 0.414 and \({\underline{\hbox{H}'}} \) _sp = 0.380, respectively) and this was highly structured (high \({\underline{\hbox{G}'}} \) _ST). Genetic diversity from C. congensis and C. arabica was also structured, but with lower levels of genetic diversity (\({\underline{\hbox{H}'}} \) _sp = 0.218 and \({\underline{\hbox{H}'}} \) _sp = 0.126, respectively). The results were consistent with agronomic and molecular studies and demonstrated that the IAC Coffea Collection is representative of the phylogenetic structure observed in the genera. This study devises sampling strategies for coffee germplasm collections and provides genetic diversity parameters for future comparisons among them.     

Effects of different cooking methods on nutritional and physicochemical characteristics of selected vegetables

The objective of the present study was to evaluate the effect of three common cooking practices (i.e., boiling, steaming, and frying) on phytochemical contents (i.e., polyphenols, carotenoids, glucosinolates, and ascorbic acid), total antioxidant capacities (TAC), as measured by three different analytical assays [Trolox equivalent antioxidant capacity (TEAC), total radical-trapping antioxidant parameter (TRAP), ferric reducing antioxidant power (FRAP)] and physicochemical parameters of three vegetables (carrots, courgettes, and broccoli). Water-cooking treatments better preserved the antioxidant compounds, particularly carotenoids, in all vegetables analyzed and ascorbic acid in carrots and courgettes. Steamed vegetables maintained a better texture quality than boiled ones, whereas boiled vegetables showed limited discoloration. Fried vegetables showed the lowest degree of softening, even though antioxidant compounds were less retained. An overall increase of TEAC, FRAP, and TRAP values was observed in all cooked vegetables, probably because of matrix softening and increased extractability of compounds, which could be partially converted into more antioxidant chemical species. Our findings defy the notion that processed vegetables offer lower nutritional quality and also suggest that for each vegetable a cooking method would be preferred to preserve the nutritional and physicochemical qualities.     

Preliminary Analysis of the Results of Selective Therapy Against Strongyles in Pasturing Horses

Control of horse parasites often omits application of measures to eradicate the free-living stages in pastures and frequently relies on chemotherapy only. Selective therapy was used for Spanish Sport horses grazing either in the same pasture (continuous) or in rotated meadows. In each group, equines exceeding a cutoff value of 300 strongyle eggs per gram of feces received ivermectin or moxidectin. Efficacy of the treatment was assessed by estimating reduction of fecal egg counts and the number of horses shedding parasite eggs (PHR). Coprocultures revealed presence of the cyathostomins Cyathostomum and Gyalocephalus spp. In all treated groups, a 100% value for both reduction of fecal egg counts and PHR against cyathostomins was obtained, and PHR values ranged from 100% to 12%. The longest strongyle egg reappearance period was observed in horses undergoing rotation grazing and receiving ivermectin (9 weeks), compared with a 6-week period recorded for the other treated equines. Our results seem to point that the efficacy of selective therapy in equine herds could be reduced if the horses with fecal egg counts below the threshold value (thus not receiving chemotherapy) remain grazing in the same pastures with the treated ones. It is strongly suggested that interested parties consider performing periodic fecal analyses to monitor fecal egg counts, together with the percentage of horses passing eggs in feces, to improve the effect of this procedure.     

Anti-Phytophthora cinnamomi activity of Phlomis purpurea plant and root extracts

Phlomis purpurea (Lamiaceae), found in Quercus suber and Quercus ilex ssp. rotundifolia forest habitats in southern Portugal, is a non-host for the oomycete Phytophthora cinnamomi, the main biotic factor involved in cork oak and holm oak decline in the Iberian Peninsula. The effect of P. purpurea crude ethanol root extract was evaluated in vitro on P. cinnamomi mycelial growth, sporangial production, zoospore release and germination as well as on chlamydospore production and viability. The protection of cork oak against infection by the pathogen was also evaluated in planta. At 10 mg ml^?1, in vitro inhibition of the pathogen structures was 85–100 %. In addition, P. purpurea plants were shown to protect Q. suber and Q. ilex from P. cinnamomi infection and to reduce the inoculum potential in glasshouse trials, indicating the ability to reduce root infection by the pathogen. The results suggest that P. purpurea has the potential to reduce disease spread and that their root extracts could provide candidate substances for control of the important pathogen, P. cinnamomi.