Protein Composition of Seminal Plasma in Fractionated Stallion Ejaculates

Seminal plasma (SP) contains several types of compounds derived from the epididymides and accessory glands. The aim of this study was to examine the protein composition of different ejaculate fractions. Trial I: fractionated ejaculates were collected from two normal and two subfertile stallions. Samples containing pre‐sperm fluid and the first sperm‐rich jets (HIGH‐1), the main sperm‐rich portion (HIGH‐2), the jets with low sperm concentrations (LOW), and a combined whole‐ejaculate (WE) sample was centrifuged, and the SP was filtered and frozen. A part of each SP sample was stored (5°C, 24 h) with spermatozoa from HIGH‐2 and skim milk extender. Sperm motility was evaluated after storage in extender mixed with the stallion’s own SP or SP from one of the other stallions (sperm from a normal stallion stored in SP from a subfertile stallion and vice versa). Protein composition was analysed using reverse‐phase liquid chromatography (RP‐HPLC), N‐terminal sequencing and mass spectrometry. The area‐under‐the‐curve (AUC) was used for quantitative comparison of proteins within fractions. Trial II: semen samples were collected from seven stallions. Fractions with the highest (HIGH) and lowest (LOW) sperm concentrations and WE samples were examined using SDS‐PAGE and densitometry. No significant differences emerged between fractions in the AUC‐values of the Horse Seminal Protein‐1 (HSP‐1) and HSP‐2 peaks, or the peak containing HSP‐3 and HSP‐4 (HSP‐3/4). Levels of HSP‐1, HSP‐2 and HSP‐3/4 were not significantly correlated with total sperm motility, progressive sperm motility or average path velocity after storage. Significant differences between ejaculate fractions in the amount of different protein groups present in SP were not found in Trial I; but in Trial II, the proteins in the 60–70 kDa range were more abundant in LOW than in HIGH and WE, indicating that this band contained proteins derived mainly from the seminal vesicles, which produce most of the SP in LOW.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Artificial reproduction of wild and cultured barbel (Barbus barbus, Cyprinidae) under controlled conditions

The aim of this work was to compare the effects of controlled reproduction of cultured and wild common barbel, Barbus barbus (L.). Preparations containing different GnRH analogues and dopamine receptor antagonists (Ovopel, Ovaprim) as well as human chorionic gonadotropin (hCG) (in the case of cultured fish) were applied and their influence on ovulation, spermiation and quality of gametes obtained was determined. No differences in the qualitative or quantitative parameters of semen were found between fish stimulated with different hormonal preparations and those not receiving hormonal stimulation. The high suitability of Ovaprim for ovulation induction in (cultured and wild) barbel was confirmed. The highest synchronisation of ovulation was obtained after the application of Ovopel (18 ± 3 h), but the best results of controlled reproduction (expressed as the percentage of ovulations and survival of embryos) were obtained by applying Ovaprim (83.2 ± 4.1). A significantly higher percentage of ovulation was obtained in cultured fish (80-90%) than in wild fish (< 25%).     

Effect of Catch Cropping on Phosphorus Bioavailability in Comparison to Organic and Inorganic Fertilization

ABSTRACT

Phosphorus (P) is a limited resource and its efficient use is a main task in sustainable agriculture. In a 3-year field experiment the effects of catch cropping [oil radish (Raphanus sativus), buckwheat (Fagopyrum esculentum), serradella (Ornithopus sativus), ryegrass (Lolium westerwoldicum), and phacelia (Phacelia tanacetifolia)] of organic fertilization (cattle manure and biowaste compost) and of inorganic fertilization (Triple-Superphosphate) on plant and soil parameters were investigated on a P-poor loamy sand in Northeast Germany. The catch crops were sown in September and remained on the plots until next spring. Then the main crops oilseed rape (Brassica napus), spring barley (Hordeum vulgare), or spring wheat (Triticum aestivum) were cultivated. The yield and P uptake of the main crops were determined. Furthermore, in the soil the organic matter content, pH, phosphorus (P) in soil solution (Psol), double-lactate and oxalate P content, P sorption capacity, and degree of P saturation were measured. All applied forms of fertilizer affected the P contents in soil and the yields and P uptakes of main crops. For green fertilization especially phacelia was found to contribute to the P supply of the main crops, since it increased the P uptake as well as the P contents in soil significantly. The cultivation of ryegrass led to a reduction of the P availability in soil. For example, in average of the three years the Psol content was 0.35 mg L^{? 1}when phacelia was cultivated and 0.22 mg L^{? 1} when ryegrass was cultivated. The cultivation of phacelia had a comparable effect on soil and plant parameters as the organic and mineral fertilization. An improved P availability and P utilization by catch cropping can reduce the need for external P input which may help to save the limited P resources worldwide.     

Genetic variation in Polish strains of Gremmeniella abietina

Thirty-nine strains of Gremmeniella abietina were isolated between 1980 and 2005 from diseased or symptomless Pinus sylvestris and Pinus nigra, or sporadically Pinus jeffreyi and Pinus armandii from four regions of Poland with varying climatic conditions. Thirty-five of the strains were genetically similar to the G. abietina A type found in Scandinavia, whereas four were similar to the B type. On the basis of random amplified microsatellite markers, very high intrapopulation and interpopulation genetic variation was detected among the strains. The average value of the Jaccard genetic similarity coefficients among strains was 0.56 for A type and 0.47 for B type, and the monomorphic loci partition was small, at 14% of all analysed loci for the A type. The degree of genetic variation of the fungal strains within each of the four regions was similar, showing no significant differences. The Jaccard coefficient for the strains isolated from P. sylvestris and P. nigra differed significantly, at 0.57 and 0.54 respectively. The genetic distance among A type strains from the investigated regions, expressed using Nei’s coefficient, was not correlated with geographic distance; however, it was highly dependent on elevation (correlation coefficient r = 0.92).     

Geographical variation of solanidane aglycone glycoalkaloids in the wild potato species Solanum chacoense Bitter

The Colorado Potato Beetle is a serious pest of the cultivated potato. Natural resistance has been found in a few wild species, including Solanum chacoense Bitter, in which the resistance is attributed to the presence of leptine glycoalkaloids. Production and accumulation of these compounds within S. chacoense varies widely and appears to be inherited in a quantitative fashion, but high leptine-producing clones occur rarely. In the present study, 15 different accessions from various locations and altitudes of origin within central to northern Argentina and Paraguay were analyzed for foliar glycoalkaloid (leptine, leptinine, solanine, chaconine) content. The objective was to infer the frequency of leptine production in ecogeographically distinct S. chacoense accessions, and to ascertain any possible association between leptine levels/concentrations and ecogeographical location. Leptines were detected in 8 of the 15 accessions, and the amounts within each accession varied widely. Most of the leptine-containing accessions originated from western Argentina except two in province Córdoba in central Argentina. There was no relationship between elevational level and leptine, but there was a negative trend with total glycoalkaloids (TGA) and elevation, due to solanine and chaconine levels which decreased with increasing elevation. In addition, nine unidentified, putative glycoalkaloids were detected, in very high proportions in some individuals and accessions. This study raises interesting questions about glycoalkaloid distribution, helps provide direction for new avenues of leptine and glycoalkaloid research, and proposes a systematic, ecogeographically based method for bioprospecting genes controlling rare plant secondary compounds.     

Performance of Colorado potato beetle larvae,Leptinotarsa decemlineata (Say), reared on synthetic diets supplemented withSolanum glycoalkaloids

Glycoalkaloids are a class of secondary compounds (nitrogenous, steroidal glycosides), ubiquitously distributed throughout the Solanaceae. Numerous studies (in planta) have shown that certain glycoalkaloids, e.g., α-tomatine, solanocardenine, and leptine, have a negative impact on performance of the Colorado potato beetle,Leptinotarsa decemlineata (Say). However, the presence of other secondary compounds, as well as the nutritional status of the plants used, are additional variables in such experiments. By rearing Colorado potato beetle, from egg to prepupal stage, on a synthetic diet supplemented with the glycoalkaloids α-tomatine, α-chaconine, α-solanine, leptine I and the steroidal aglycone solanidine, we have been able to further establish the effects (ex planta) of these compounds on the Colorado potato beetle. Leptine I displayed a dose-dependent negative activity against the Colorado potato beetle (as measured by larval weight gain and tune to molt), when assayed at 0.31, 0.62 and 1.23 mM concentrations; however, by the fourth stadium no effect was found. When Colorado potato beetle were fed a higher concentration of leptine I (2.4 mM), there was a sustained effect in all stadia. At 2.4 mM, leptine I displayed a greater negative impact on Colorado potato beetle growth and development than did α-tomatine. α-Chaconine at 2.4 mM did not impair Colorado potato beetle performance relative to Colorado potato beetle reared on control diets. α-Chaconine plus α-solanine, at concentrations commonly found inSolanum tuberosum L. foliage (0.6 and 0.3 mM respectively), did not impair Colorado potato beetle performance. The steroidal aglycone solanidine (2.4 mM) had a significant negative impact on Colorado potato beetle performance. Results are discussed in the context of host-plant resistance and insect-plant interactions.     

The effect of a seaweed concentrate on the in vitro growth and acclimatization of potato plantlets

Summary The seaweed concentrate ‘Kelpak’ is used commercially in the greenhouse and field to improve plant quality. ‘Kelpak’ was added to the in vitro culture medium of potato cv. ‘BP1’ and also applied as a leaf/soil drench immediately after transplanting. The addition of 0.25% seaweed concentrate to the medium improved plantlet quality and led to better establishment in the greenhouse. No beneficial effect of seaweed concentrate in the tissue culture medium was observed if a second cutting was part of the micropropagation process. Additional application of 0.5% seaweed concentrate in the greenhouse to plantlet cuttings derived from tissue culture was not beneficial.     

The use of image analysis to study developmental variation in micropropagated potato (Solanum tuberosum L.) plants

Summary Potato leaf morphology changes during plant development with the phase shift from vegetative growth to flowering. Image analysis can detect differences in leaf morphology and has been used here to distinguish differences in leaf morphology between potato crops derived from seed tubers and minitubers and between crops derived from different micropropagation protocols. Further, leaf shape parameters can be used to determine the relative maturity of crops. This finding is of economic importance since differences in plant development, for example delayed flowering, are associated with yield parameters. It is hypothesised that image analysis of established microplants can be used as an early evaluation of micropropagation protocols for potato.