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.     

Perspectives on the use of a seaweed extract to moderate the negative effects of alternate bearing in apple trees

Summary

Alternate bearing is a serious problem in many apple varieties. In fact, each year, apple trees have to be thinned in order to ensure a correct and constant balance between the reproductive and vegetative tendencies of the plant. In organic fruit growing, there is no alternative to the expensive and time-consuming process of hand-thinning. Under these conditions, the use of natural bio-stimulants which enhance the efficiency of nutrient uptake, plant fitness to different pedoclimatic conditions, and plant tolerance to stress, offers new opportunities to reduce the negative effects of alternate bearing. The aim of this work was to evaluate the effects of Actiwave^®, a metabolic enhancer derived from the alga, Ascophillum nodosum, on ‘Fuji’ apple trees affected by experimentally-imposed alternate bearing. The experiments were carried in open field conditions over four consecutive years. Although the effect of Actiwave^® was erratic from year-to-year, the compound generally reduced those problems linked to alternate bearing. Actiwave^® had its most significant effect on alternate bearing plants that were also affected by nutrient deprivation due to the absence of fertilisation. In these trees,Actiwave^® decreased the oscillations in yield between “on” and “off” years and increased the average fruit weight on plants affected by too-high a crop load. Treated trees also showed higher leaf chlorophyll contents (increased by 12%), with a consequent increase in the rates of photosynthesis and respiration. On the other hand, under standard conditions, in well-fertilised balanced plants, Actiwave^® did not have any significant effect. This evidence corroborates the hypothesis that Actiwave^® may be a useful tool to reduce alternate bearing in organic and low-input farming.     

Hunting or habitat degradation? Decline of primate populations in Udzungwa Mountains,Tanzania: An analysis of threats

Hunting and habitat degradation are universal threats to primates across the tropics, thus deciphering the relative impact of threats on population relative abundance is critical to predicting extinction risk and providing conservation recommendations. We studied diurnal primates over a period of nearly 6 years in the Udzungwa Mountains of Tanzania, a site of global importance for primate conservation. We assessed how population relative abundance of five species (of which two are endemic and IUCN-Endangered) differed between two forest blocks that are similar in size and habitat types but contrast strongly in protection level, and how abundance changed during 2004–2009. We also measured habitat and disturbance parameters and, in the unprotected forest, evaluated hunting practices. We found significant differences in primates’ abundance between protected and unprotected forests, with the greater contrast being the lower abundance of colobine monkeys (Udzungwa red colobus and Angolan colobus) in the unprotected forest. At this site moreover, colobines declined to near-extinction over the study period. In contrast, two cercopithecines (Sanje mangabey and Sykes’ monkey) showed slightly higher abundance in the unprotected forest and did not decline significantly. We argue that escalating hunting in the unprotected forest has specifically impacted the canopy-dwelling colobus monkeys, although habitat degradation may also have reduced their abundance. In contrast, cercopithecines did not seem affected by the current hunting, and their greater ecological adaptability may explain the relatively higher abundance in the unprotected forest. We provide recommendations towards the long-term protection of the area.     

Daily variations of serum lipids in Ovis aries under different lighting and feeding conditions

The aim of this study was to discriminate the impact of lighting and feeding conditions on the regulation of lipid metabolism in Ovis aries. Six clinically healthy female Comisana ewes, not pregnant and not lactating were kept under different environmental conditions: 12:12 light-dark (LD) cycle, constant darkness and fasting. Blood samples were collected at the end of each period of treatment every 4 h for a 24-h period. Blood concentration of triglycerides, total cholesterol and non-esterified fatty acid showed robust daily rhythmicity in ewes maintained under 12:12 LD cycle and fed ad libitum. Conversely, either constant darkness or fasting had a deep impact on all lipid parameters investigated. Our results suggest that lighting and feeding conditions have an impact on daily variations in lipid metabolism in ewes.     

The influence of gamete co-incubation length on the in vitro fertility and sex ratio of bovine bulls with different penetration speed

The objectives of this work were to evaluate whether the sperm penetration speed is correlated to the in vitro fertility and whether adapting the gamete co-incubation length to the kinetics of the bull improves in vitro fertility and affects the sex ratio. In vitro matured oocytes were co-incubated with spermatozoa from four different bulls (A-D). At various post-insemination (p.i.) times (4, 8, 12, 16 and 20 h), samples of oocytes were fixed and stained with DAPI for nuclei examination, while the remaining ones were transferred into culture to evaluate embryo development. The blastocysts produced were sexed by PCR. Two bulls (A and B) had faster kinetics than the others (C and D), as shown by the higher penetration rates recorded at 4 h p.i. (43%, 30%, 11% and 6%, respectively for bulls A, B, C and D; p<0.01). The differences in the kinetics among bulls did not reflect their in vitro fertility. The incidence of polyspermy was higher for faster penetrating bulls (36%, 24%, 16% and 4%, respectively for bulls A, B, C and D; p<0.01) and at longer co-incubation times (0%, 16%, 19%, 30% and 34%, respectively at 4, 8, 12, 16 and 20 h p.i.; p<0.01). The fertilizing ability of individual bulls may be improved by adapting the co-incubation length to their penetration speed. A sperm-oocyte co-incubation length of 8 h ensured the greatest blastocyst yields for the two faster penetrating bulls. On the contrary, 16 h co-incubation was required to increase (p<0.01) cleavage rate of the two slower bulls. Bulls with a faster kinetics did not alter the embryo sex ratio towards males. The female/male (F/M) ratios recorded were 2.1, 1.4, 1.2, 1.3 and 1.6, respectively at 4, 8, 12, 16 and 20 h p.i.     

Comparative evaluation of PRRS virus infection in vaccinated and naïve pigs

The purpose of this study was to evaluate the time-course of the immune response to a field Porcine Respiratory and Reproductive Syndrome virus (PRRSV) strain in PRRS-naïve, untreated pigs, as well as in four groups of age and breed-matched pigs injected with a live attenuated PRRS vaccine, its adjuvant, an inactivated PRRS vaccine and an irrelevant, inactivated Porcine Circovirus type 2 (PCV2) vaccine, respectively. PRRSV infection was confirmed in all groups by PCR and antibody assays. The antibody response measured by ELISA took place earlier in pigs injected with the live attenuated vaccine, which also developed a much stronger serum-neutralizing antibody response to the vaccine strain. Yet, no clear protection was evidenced in terms of viremia against the field virus strain, which showed 11.1% nucleotide divergence in ORF7 from the vaccine strain. In vitro, the interferon (IFN)-γ response to PRRSV was almost absent on PVD 60 in all groups under study, whereas the prevalence of interleukin (IL)-10 responses to PRRSV was fairly high in PCV2-vaccinated animals, only. Results indicate that distinct patterns of immune response to a field PRRSV strain can be recognized in PRRS-vaccinated and naïve pigs, which probably underlies fundamental differences in the development and differentiation of PRRSV-specific immune effector cells.     

Intra-varietal genetic diversity of the grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) cultivar ‘Nero d’Avola’ as revealed by microsatellite markers

The Sicilian grape cultivar ‘Nero d’Avola’ is among the oldest and most cultivated in the island, taking part in the production of several red wines exported worldwide, including DOC wines (Etna Rosso and Cerasuolo di Vittoria). Due to the ancient origin and repeated clonally propagation of the cultivar, phenotypic variability has been observed. Clone identification in this important cultivar has so far relied on phenotypic and chemical traits analyses, often affected by environmental conditions. Genetic markers, such as microsatellites, are particularly useful for cultivar identification, parentage testing, pedigree reconstruction and population structure studies. In the present paper, microsatellites were used to analyze the intra-varietal genetic diversity among 118 plants of ‘Nero d’Avola’, collected in 30 vineyards displaced in different areas of Sicily. Out of 22 microsatellites, 11 showed polymorphism among samples and 15 different phylogenetic groups were identified. Results show that ‘Nero d’Avola’ actually comprises different genetic profiles, although most of clones share a common origin.     

Leaf and ecosystem response to soil water availability in mountain grasslands

Climate change is expected to affect the Alps by increasing the frequency and intensity of summer drought events with negative impacts on ecosystem water resources. The response of CO₂ and H₂O exchange of a mountain grassland to natural fluctuations of soil water content was evaluated during 2001-2009. In addition, the physiological performance of individual mountain forb and graminoid plant species under progressive soil water shortage was explored in a laboratory drought experiment. During the 9-year study period the natural occurrence of moderately to extremely dry periods did not lead to substantial reductions in net ecosystem CO₂ exchange and evapotranspiration. Laboratory drought experiments confirmed that all the surveyed grassland plant species were insensitive to progressive soil drying until very low soil water contents (<0.01 m³ m⁻³) were reached after several days of drought. In field conditions, such a low threshold was never reached. Re-watering after a short-term drought event (5 ± 1 days) resulted in a fast and complete recovery of the leaf CO₂ and H₂O gas exchange of the investigated plant species. We conclude that the present-day frequency and intensity of dry periods does not substantially affect the functioning of the investigated grassland ecosystem. During dry periods the observed “water spending” strategy employed by the investigated mountain grassland species is expected to provide a cooling feedback on climate warming, but may have negative consequences for down-stream water users.