Culture of bovine bone marrow progenitor cells in vitro.

In vitro methylcellulose cultures of bovine bone marrow progenitor cells were developed. An existing technique described for bovine species was compared to a method for human tissue and further adapted during subsequent experiments. Bovine bone marrow samples were collected at the slaughterhouse, and mononuclear cells were separated by gradient centrifugation (1.077 g/ml specific density and 400 g). The use of 3% bovine leucocyte-conditioned medium, produced by stimulation of blood lymphocytes with 4 microg/ml concanavalin A and harvested on day 4 of culture, gave better results than the use of supernatant of the human bladder carcinoma 5637, which is widely used in human bone marrow cultures. However, bovine leucocyte-conditioned medium was not added to erythroid cultures because inhibitory effects were observed. Erythroid colonies were stimulated with erythropoietin, and hemin was added to enable microscopic identification. Reduced oxygen tension was necessary to induce growth of erythroid colonies. This was not necessary for myeloid cultures. In conclusion, the results of this study show that the growth of myeloid and erythroid colonies in methylcellulose-based medium requires different culture conditions, which are different from the culture conditions for human cells.     

Effect of mentor pollen on pistil-pollen incongruities among species of Cucumis L.

Summary The merits of mentor pollen in certain interspecific hybridizations in the genus Cucumis L. were evaluated. African wild cucumber species Cucumis metuliferus Naud. and C. africanus L. and Asian species C. sativus var. hardwickii Alef and var. sikkimensis Hook, were reciprocally crossed. Pollen tube growth was arrested halfway down the style in all combinations except in C. sativus × C. africanus. Mentor pollen irradiated with 100 and 200 krad grew through the style and into the ovules in all species. Fruitset was obtained in all cases when mentor pollen was used either alone or in mixtures. Fruits set after self pollination with mentor pollen yielded only seeds without embryos.In several crosses, aided by mentor pollen, a number of relatively large seeds (with enlarged embryosac) were obtained. A sample of these embryosacs contained a globular structure like an embryo. Embryosacs with embryo-like structures were explanted on several different media but no development was obtained.     

Effects of gamma irradiation on in vitro pollen germination of different Cucumis species

Summary Pollen grains of 14 Cucumis accessions were irradiated with 0, 1, 2, 3 or 4 kGy acute gamma rays and germinated in vitro directly afterwards. Pollen germination was significantly reduced by increasing irradiation dose for all species, except C. melo var. agrestis. Pollen tube growth was generally reduced likewise. Pollen of two C. anguria subspecies was most sensitive to irradiation. Sensitivity of the pollen with respect to pollen tube growth in relation to irradiation dose was inversely related to total DNA amount per nucleus. In vitro germination was not related to DNA amount per nucleus. Results show that the examined Cucumis species, especially C. melo var. agrestis, are sufficiently resistant to irradiation to be used as donor species for in vivo egg cell transformation of the cucumber.     

In vitro pollen germination and tube growth of tomato (Lycopersicon esculentum Mill.) and its relation with plant growth

Summary In vitro pollen germination at 10°C, 14°C and 22°C of four groups of two pure line tomato varieties was compared with their plant growth at 19°C D/10°C N under controlled environmental conditions. Generally, pollen germination was slow at 10°C but after 6 h the percentage of germination was similar to that at 22°C. Maximum germination was obtained at 14°C already after 4 h. The longest pollen tubes occurred at 22°C. The two varieties within each group differed significantly from each other for percentage of pollen germination. For one group, this difference was greater at 10°C than at 22°C. The varieties in two groups also differed significantly for pollen tube growth. These differences in pollen tube growth rate were greater at 22°C than at 10°C. There was no clear relationship between pollen germination, pollen tube growth and plant growth in any of the four pairs of varieties. The results are discussed with regard to the possibility of pollen selection at low temperature in order to improve the efficiency of breeding for growth at low temperature.     

Relationships between African species of the genus Cucumis L. Estimated by the production,vigour and fertility of F1 hybrids

Summary The production, vigour, and fertility of F₁ hybrids between nine African species of the genus Cucumis L. were studied as a measure of the relationships between the species. Hybrid plants were obtained from 29 out of the 72 possible cross combinations. Two F₁ hybrids died as seedlings, and 27 hybrids were raised to maturity. Pollen production and stainability varied greatly amongst these hybrids, as did fruit and seed set following self pollination and backcrossing with either parental species. The fruit shape of the hybrids was always intermediate between that of both parental species.Two species appeared to be closely related: C. prophetarum L. and C. anguria var. longipes A. Meeuse. Most other species produced highly to moderately fertile F₁ hybrids with at least one other species. C. metuliferus Naud. produced only sterile hybrids with C. zeyheri Sond. 2x. The results sustain the recent taxonomic classification of the genus (Jeffrey, 1980).     

Report on the 43rd Annual Meeting of the DPG-Working Group Nematology

Journal of Plant Diseases and Protection -     

Importing plants into the Netherlands: an assessment of the risk of plant parasitic nematodes and a survey on their entry with adhering soil

Few plant parasitic nematodes are currently included in the European Union (EU) regulated pest lists. However, many plant parasitic nematodes not present in the EU are known to be damaging and present a risk to Europe. Therefore a study was performed to assess the risk of importing plant parasitic nematodes with the soil attached to plants for planting, with emphasis on plants in pots. Prior to the survey, a list of harmful plant parasitic nematodes not present within Europe was prepared per continent based on a literature review. This resulted in a Risk List of 26 species. Additionally, over a 3‐year period a survey was carried out on plants for planting imported to the Netherlands with adhering soil, focusing on all plant parasitic nematodes and with special attention to the nematodes on the Risk List. A total of 258 soil samples were studied, originating from 54 different plant species and 20 different countries. Despite the small number of samples, several quarantine nematodes and species from the Risk List were detected, including new nematode–plant combinations. This survey illustrates that plants with adhering soil can be a pathway for the introduction of listed nematodes, including those from the Risk List, to the EU/EPPO region. This information might encourage a more risk‐based approach to performing import inspections.     

Flow cytometric analysis of the DNA content of bovine and human bone marrow cells

The defence against infection in high-yielding dairy cows is correlated with the number and function of circulating neutrophils and depends on their production in bone marrow. Therefore, the DNA content of isolated bone marrow cell suspensions from 7 calves, 7 cows and 14 humans was assayed by flow cytometry. Bovine sternal bone marrow samples were collected within 30 min of death, and human marrow samples were collected by sternal puncture and aspiration. Mononucleated cells were isolated by gradient centrifugation. In the bone marrow samples from calves and cows, 35 +/- 2.6% and 31.8 +/- 1.5% of the isolated bone marrow cells respectively were in the S/G2/M-phase. The difference between calves and cows was not significant. In the human samples, only 12 +/- 0.8% of the cells were in the S/G2/M-phase. A significant (P < 0.001) difference was observed between the two species. These results indicated that the proliferative, in activity of haematopoietic cells is significantly higher in cattle than in humans.     

Analysis of differences in growth of cucumber genotypes under low light conditions in relation to night temperature

Summary Growth of four breeding lines and two cultivars of slicing cucumber with different growth at low temperature and light intensity was analysed in relation to night temperature. Plants were grown from 17 days after sowing at 20°C day temperature and 20, 15 or 10°C night temperature (NT), a light intensity of 24 W/m² visible radiation and a daylength of 8 hours. Data from six successive harvests, ending 47 days after sowing, were analysed according to a MANOVA procedure.RGR, NAR, LAR, SLA and LWR generally decreased with time. Ln W and RGR were largely similar at 20 and 15°C NT, but reduced at 10°C NT. LAR and SLA decreased at each lower NT, whereas LWR increased. NAR was only marginally affected by NT. Significant genotypic differences occurred for 1n W, LAR, SLA, LWR, stem length and percentage of dry matter in leaves and stems at all NT's and for RGR and NAR only at 20°C. Genotypic differences for 1n W could not be explained by differences for RGR, but by differences for initial plant weight.Significant genotype × night temperature interactions were present for LWR. Breeding lines increased LWR in response to lower NT, while that of cultivars remained fairly constant. Implications for breeding are discussed.Abbreviations and definitions L plant length - LA leaf area - W dry weight of the shoot - RGR relative growth rate (the increase in W per unit of time and unit of W) - NAR net assimilation rate (the increase in W per unit of time and unit of LA) - LAR leaf area ratio (the leaf area at time t per unit of W) - SLA specific leaf area (the leaf area at time t per unit of leaf weight) - LWR leaf weight ratio (the leaf weight at time t per unit of W) - RGR NAR × LAR - LAR SLA × LWR - MANOVA multivariate analysis of variance