Tagging the juvenile locus in soybean [Glycine max (L.) Merr.] with molecular markers

Soybean cultivars carrying the `long juvenile trait' show a delayed flowering response under short day conditions. The incorporation of this character into genotypes of agronomic interest may allow a broader range of sowing dates and latitudes for a single cultivar adaptation. The objective of this work was to identify molecular markers linked to the juvenile locus in soybean. Experiments were carried out using two pairs of near isogenic lines(NILs) differing in the presence of the long juvenile trait, and RAPD markers. Four hundred primers were first screened to find polymorphism associated with the trait. Additional differences between NILs were sought by digesting the genomic DNA with five restriction enzymes. Polymorphic fragments detected between NILs were tested for linkage to the juvenile locus in the corresponding F₂ segregating populations. Marker bc357-HaeIII was linked (χ²L = 46.316) to the juvenile locus with an estimated recombination frequency of 0.13 ± 0.03in one of the genetic backgrounds studied. The fragment was cloned, sequenced and converted into a SCAR marker. Moreover,bc357-HaeIII was used as RFLP probe. Both, SCAR and RFLP generated markers linked to the juvenile locus in the two genetic backgrounds analysed. Results presented in this work can be utilised for both, the localisation of the gene associated with the character and for tagging the juvenile trait in soybean breeding programs. This revised version was published online in August 2006 with corrections to the Cover Date.     

Carbon sequestration in a limestone quarry mine soil amended with sewage sludge

To reclaim a limestone quarry, 200 and 400 Mg/ha of municipal sewage sludge were mixed with an infertile calcareous substrate and spread as mine soil in 1992. Soil samples were taken 1 week later and again after 17 yr of mine soil rehabilitation so as to assess changes in the amount and persistence of soil organic carbon (SOC). Sludge application increased SOC as a function of the sludge rate at both sampling times. Seventeen years after the sludge amendments, the nonhydrolysable carbon was increased in the 400 Mg/ha of sludge treatment. The recalcitrance of SOC was less in sludge‐amended soils than in the control treatment at the initial sampling, but 17 yr later this trend had reversed, showing qualitative changes in soil organic carbon. The CO₂‐C production had not differed between treatments, yet the percentage of mineralized SOC was less in the high sludge dose. When the size of active (C_active) and slow (C_slow) potentially mineralizable C pools was calculated by curve fitting of a double‐exponential equation, the proportion of C_active was observed to be smaller in the 400 Mg/ha sludge treatment. Soil aggregate stability, represented by the mean weight diameter of water‐stable soil aggregates, was significantly greater in mine soil treated with the high dose of sludge (18.5%) and SOC tended to be concentrated in macro‐aggregates (5–2 mm). Results suggest that SOC content in sludge‐amended plots was preserved due by (i) replacement of the labile organic carbon of sludge by more stable compounds and (ii) protection of SOC in aggregates.     

Metabolisable energy value and digestibility of fat and fatty acids in linseed determined with growing broiler chickens

1. In the first experiment, the AMEN of linseed was evaluated at rates of 40, 80, 160 and 240 g/kg in a basal diet with broiler chickens aged 28 d using the total collection method. 2. Increasing the concentration of linseed decreased the AMEN of diet, from 11.78 to 8.75 MJ/kg DM, and AMEN of linseed, from 16.64 to -2.96 MJ/kg DM. The negative values clearly indicated that linseed interacted with the other dietary ingredients and impaired their energy utilisation. 3. In the second experiment, a semipurified diet with graded addition of linseed (80, 120, 160 and 240 g/kg DM) was evaluated for crude fat and fatty acid apparent digestibilities. The coefficients decreased from 0.602 to 0.359 for crude fat and from 0.661 to 0.352 for individual fatty acids as the level of linseed increased. 4. In both experiments, a quadratic regression model explained the relationship between data better than the linear model. 5. The presence of mucilage and other antinutritional factors in linseed might explain the results obtained in this study.     

Serologic response of red-legged partridges (Alectoris rufa) after oral inoculation with Toxoplasma gondii oocysts

Thirty 5-month-old red-legged partridges (Alectoris rufa) reared in battery were divided into five groups: 4 birds in group A, 14 birds in group B, 4 birds in group C, 4 birds in group D and 4 birds in group E, and were inoculated orally with 10, 50, 10(2), 10(3) and 10(4) oocysts of the OV-51/95 strain of Toxoplasma gondii, respectively. During the experiment, blood samples from all birds were drawn every 3-7 days and at necropsy. Serologic response was measured by the modified agglutination test (MAT) and the latex agglutination test (LAT). One bird from each group was killed at 44, 58, 65 and 72 days after inoculation (DAI). From 72 DAI to the end of the experiment, surviving partridges from group B were killed at weekly intervals. The last partridges were sacrified 100 DAI. MAT was the most sensitive and specific test for detecting T. gondii antibodies in the birds. First positive titers were detected by MAT in all sera on 7 DAI, but titers by LAT did not appear until 13 DAI. Antibody titers detected by MAT on 7 DAI were higher in the partridges with the largest inocula (10(3) or 10(4) oocysts) than those inoculated with 10, 50 or 10(2) oocysts. All surviving birds developed a serologic response to T. gondii, with maximum titers of 512-32,768 in the MAT on 13-17 DAI, and positive titers persisted at least until 100 DAI. To the contrary, LAT reveals only very low antibody titers even in partridges inoculated with the highest dose of T. gondii.     

Relationship between DNA fragmentation of equine granulosa cells and oocyte meiotic competence after in vitro maturation

The acquisition of equine oocyte developmental capacity is ensured by the follicular environment, such as granulosa cells, which could reflect the meiotic development potential of immature oocytes. This study evaluated the relationship between DNA fragmentation of granulosa cells, using the chromatin dispersion test, and equine oocyte meiotic development after in vitro maturation. Granulosa cells and cumulus–oocytes complexes (n = 50) were recovered from slaughterhouse‐derived ovaries. Oocytes were in vitro matured, stained and evaluated under fluorescence microscopy. Maturation rates were classified into outstanding, medium and poor levels of maturation using 25th and 75th percentiles as thresholds. For DNA assessment, each sample was processed with the Ovoselect^® kit (Halotech DNA). High, low and total DNA fragmentation percentages were compared among levels of maturation rates by ANOVA, followed by Duncan test. Results were expressed as mean ± SE. Total and high DNA fragmentation rates of granulosa cells were significantly higher (p < 0.05) in follicles whose oocytes had reached outstanding maturation level than those originating from follicles whose oocytes had reached poor maturation level. In conclusion, the DNA fragmentation analysis of equine granulosa cells can be a valuable test to identify equine oocytes showing the best meiotic competence after in vitro maturation.     

Redox cycling induces spermptosis and necrosis in stallion spermatozoa while the hydroxyl radical (OH•) only induces spermptosis

Oxidative stress is a major factor explaining sperm dysfunction of spermatozoa surviving freezing and thawing and is also considered a major inducer of a special form of apoptosis, visible after thawing, in cryopreserved spermatozoa. To obtain further insights into the link between oxidative stress and the induction of apoptotic changes, stallion spermatozoa were induced to oxidative stress through redox cycling after exposure to 2‐methyl‐1,4‐naphthoquinone (menadione), or hydroxyl radical formation after FeSO₄ exposure. Either exposure induced significant increases (p < 0.05) in two markers of lipid peroxidation: 8‐iso‐PGF_2α and 4‐hydroxynonenal (4‐HNE). While both treatments induced changes indicative of spermptosis (caspase‐3 activation and decreased mitochondrial membrane potential) (p < 0.01), menadione induced sperm necrosis and a dramatic reduction in motility and thiol content in stallion spermatozoa. Thus, we provided evidence that oxidative stress underlies spermptosis, and thiol content is a key factor for stallion sperm function.     

Morphological and genetic diversity of Pura Raza Español horse with regard to the coat colour

Gene mutations influencing melanocytes also impact on physiological and behavioural functions. In this study, we investigated their association with four different coat colours in the Pura Raza Español (PRE) horse using morphological traits and molecular datasets. Four different subpopulations were identified according to individual coat colour: grey, bay, chestnut and black. Coat colour significantly associated with morphological measurements. Observed and expected heterozygosity values were low in grey compared with the other three subpopulations, suggesting the presence of unique ancestral alleles probably arisen by genetic drift and selection mechanism effects. Nei's distance demonstrated a clear division among subpopulations, the grey being the most divergent group. Gene flow estimates were similar, showing the lowest values in grey. Divergence times among subpopulations assessed with the average square distance suggested that grey was the original PRE population which diverged from bay, chestnut and black. Our results also demonstrated a clear morphological differentiation according to coat colour. The close genetic structure of bay and chestnut PRE subpopulations and the clear differences in most morphological traits of grey and chestnut PRE mares would suggest the pleiotropic effect of genomic regions determining coat colour in horses. However, further analysis including genomic information would be necessary to elucidate the mechanisms involved.     

Organizational learning through participatory research: CIP and CARE in Peru

Participatory research (PR) has been analyzed and documented from different points of view, with emphasis on the benefits generated for farmers. The effect of PR on organizational learning has, however, received little attention. This paper analyzes the interaction between a research and a development institution, the International Potato Center (CIP) and CARE in Peru, respectively, and makes the case that PR can contribute to creating a collaborative learning environment among organizations. The paper describes the evolution of the inter-institutional collaborative environment between the two institutions for more than a decade, including an information-transfer period (1993–1996), an action-learning period (1997–2002), and a social-learning period (2003–2007). Several lessons learned from each period are described, as are changes in institutional contexts and stakeholders’ perceptions. The case shows that research and development-oriented organizations can interact fruitfully using PR as a mechanism to promote learning, flexibility in interactions, and innovation. Interactions foster the diffusion of information and the sharing of tacit knowledge within and between organizations, which in turn influences behavior. However, the paper also argues that long-term inter-organizational interactions are needed to facilitate learning, which can be used to influence the way organizations implement their interventions in a constantly changing environment.     

Partial resistance to a Fusarium root disease in Egyptian white lupin landraces

A Fusarium sp. root pathogen of lupin is the causal agent of the most important disease that affects the cultivation of white lupin (Lupinus albus L.) in Egypt. The aim of our research was to investigate whether host resistance to Fusariumroot disease was available in Egyptian landraces ofLupinus albus. Five Fusarium isolates collected from white lupin samples in Egypt were tested with an Egyptian landrace and a French cultivar. The most aggressive isolate was used to screen an additional 15 Egyptian landraces, two cultivars released in Egypt after selection among landraces, one Polish cultivar, and two French cultivars. The assessment of host response to Fusarium was performed in a field, and under controlled conditions in a greenhouse pot experiment. Most landraces and the two Egyptian cultivars showed better resistance with good accordance between field and pot experiment. This experiment showed that Egyptian genetic resources of white lupin possess partial resistance to Fusarium root rot. Egyptian germplasm may be an alternative genetic source for incorporating partial resistance to Fusarium root rot into the breeding pools. This revised version was published online in July 2006 with corrections to the Cover Date.