Improved formation of regenerable callus in isolated microspore culture of maize: impact of carbohydrates, plating density and time of transfer.

Pure fractions of maize (Zea mays L.) microspores at various densities were exposed to defined media containing different concentrations of maltose and sucrose. In general, lower carbohydrate concentrations (60, 90 g/l) yielded higher frequencies of embryo-like structures than a high concentration (120 g/l). Optimum cell density seemed to depend on the genotype, but densities above 80,000 microspores/ml led to reduced embryogenesis in all genotypes tested. Direct comparison of maltose and sucrose as carbohydrate source in the induction medium clearly demonstrated the superiority of maltose with regard to the regeneration frequency. For two out of three genotypes tested, maltose also enhanced the formation of embryo-like structures. The time of embryo transfer to callus induction media had a significant effect on regeneration frequency.

Improved production of doubled haploids by colchicine application to wheat (Triticum aestivum L.) anther culture.

The aim of this study was to optimize the in vitro chromosome-doubling procedure in wheat anther culture. Colchicine, at concentrations of 100-5000 mg/l, was added to the induction medium for 1-5 days. Beneficial effects were obtained with concentrations of 100 and 1000 mg/l colchicine. With time, significant reductions in embryo-like structures as well as higher doubling indices were found. Similar results were obtained with the high- and low-responding genotypes. Colchicine (100 mg/l), added 5 and 20 days after inoculation for 1 and 3 days increased the induction response, but this value was reduced when colchicine was added 10 or 15 days after inoculation. The doubling effect was similar to the control, except for a significant increase with the 3-day application 20 days after inoculation. The highest success index was reached when colchicine was added to the culture medium after 20 days.

[Efficient use of sleep pillows in patients suffering from non-organic sleep disorders--a pilot study]. / Effizienter Einsatz von Schlafkissen bei Personen mit nicht-organischen Schlafstörungen--eine Pilotuntersuchung.

OBJECTIVE: Sleep disorders may critically affect working performance and quality of life. Sleep pillows have been traditionally used to overcome such disorders. Scientifically based clinical trials to demonstrate the efficacy are missing. METHODS: 28 patients with problems falling asleep and/or staying asleep not related to psychiatric or organic diseases were investigated in an accredited sleep laboratory. The diagnosis was confirmed by polysomnography. After 2 and 4 weeks of treatment the polysomnography was repeated to document any influences by the sleep pillows. RESULTS: The polysomnographic records showed a monotonic trend to regain an age-related distribution of the non-REM sleep stages. The REM sleep phase increased nearly twofold; however, the norm values were not reached within the 4-week period of treatment. Sleep pillows of intensity 2 were superior to those of intensity 1; a further increase to intensity 3 did not create any additional effect. CONCLUSION: The results demonstrate an effective treatment of non-complicated sleep disorders with sleep pillows, which has been shown with objective measurements in a sleep laboratory.

Effects of L-proline and post-plating temperature treatment on Maize (Zea mays L.) anther culture.

Comparison of different post-plating temperature regimes with a control treatment (27° C) revealed that a short-term cold (8/14°C:2/2 days or 14°C:4 days) as well as a heat treatment (30°C:14 days) increased the production of embryro-like-structures (ELS) from cultured maize anthers. The beneficial effects of short-term cold treatments were magnified 2-3 times when L-proline (PROL) was added to the induction medium (125-500 mg/L). In the best treatment (14°C:4 days, 125 mg/L L-proline) one genotype produced 143.5 ELS/100 anthers. Anthers subjected to high temperature (30°C:4 days, 30°C:7 days, 30°C:14 days) generally showed a lower response than did cold treated anthers, although genotypic differences were observed. Regeneration frequency did not appear to be affected by the presence of L-proline in the induction medium.

Colchicine-mediated chromosome doubling during anther culture of maize (Zea mays L.).

Efficient methods of chromosome doubling are critical for the production of microspore-derived, doubled-haploid (=DH) plants, especially if, as in maize anther culture, spontaneous chromosome doubling occurs infrequently. In the present study, colchicine (5-1000 mg/l) was added to the induction medium and maize anthers were incubated in the colchicine-containing medium for different durations (1-7 days). In order to improve overall anther culture response, the culture temperature was adjusted to 14°C during the first 7 days. Colchicine applied at low concentration, i.e. 5 mg/l (7 days), or for short duration, i.e. 1-3 days (250 mg/l), showed beneficial effects on the formation of embryolike structures (=ES) and thus led to increased plant production, but was comparatively ineffective regarding chromosome doubling. Optimal doubling effects were observed when anthers had been exposed to culture medium containing 250 and 1000 mg/l of colchicine (7 days); in these treatments the doubling index (=DI), defined as the quotient of the number of DH plants and the number of totally regenerated plants in a specific treatment, rose to 0.56 and 0.53, respectively, compared to 0.20 in the untreated control. However, colchicine administered at concentrations higher than 250 mg/l seemed to be detrimental to general plant production; thus, in spite of a high DI, the overall DH plant production was even lower than in the control treatment. Maximum DH plant production for three different genotypes was accomplished with culture medium containing 250 mg/l of colchicine (7 days). With the best-responding genotype (ETH-M 36) a DH plant production of 9.9 DH plants/100 anthers was accomplished, i.e. a 7-fold increase compared to the non-treated anthers. This is the first report on efficient chromosome doubling in anther culture by subjecting anthers to colchicinecontaining induction medium during a post-plating cold treatment. Chromosome doubling as described here becomes an integral part of the maize anther culture protocol and thus represents a rapid and economical way to produce DH plants.

Significance of genetic and environmental aspects in the field cultivation of Hypericum perforatum.

Agronomical and biochemical parameters of seven Hypericum perforatum (St. John's wort) accessions grown at three experimental sites in Switzerland were followed over a two year period (1995-1996). Significant effects of environmental (= site) and genetic factors (= accession) on flowering dates, plant length, and plant dry matter production (= plant yield) were observed in both years; rankings of sites and accessions with regard to plant yield were similar in both years despite the fact that the first year crop contributed only a minor part to the overall yield of both years together. Maximum dry matter production per year reached 159 dt/ha for the total plant and 54 dt/ha for the flowering segment (i.e. the pharmaceutically relevant, upper segment of the plants comprising the majority of flowers). HPLC analysis of the constituents covered eight secondary metabolites (amentoflavone, biapigenin, hyperforin, hypericin, hyperosid, pseudohypericin, quercetin, rutin). Generally, secondary metabolite contents were significantly lower in the first year of cultivation ranging from 12% (hyperosid) to 83% (hyperforin) of the contents measured in the 1996-crop. Significant genetic effects on the production of all tested secondary metabolites (except biapigenin) were observed in 1996 whereas environmental effects appeared to be less distinct (except for amentoflavone and pseudohypericin). In conclusion, genetic factors strongly affected plant yield as well as secondary metabolite content in H. perforatum cultivation; the availability of genetically superior plant material next to improved agrotechnological methods therefore is supposed to become a key factor for successful future field production.

Autoclaved and filter sterilized liquid media in maize anther culture: significance of activated charcoal.

Medium sterilization techniques (autoclaving, filter sterilization and separate sterilization of medium components), combined with preculture exposure to activated charcoal (AC) were evaluated for effects on maize anther culture response. The addition of AC to filter sterilized medium had no effect on the number of embryo-like-structures (ES) produced. For autoclaved medium, pre-culture AC treatment resulted in a 3-fold increase in ES yield over medium lacking AC. When AC was included, autoclaved medium was more productive than filter sterilized medium. Autoclaved media without AC gave lower response than filter sterilized medium. Separate sterilization of sucrose or FeEDTA was beneficial for media autoclaved in the absence of AC. However, when all components were autoclaved together in the presence of AC, there was no advantage to separate sterilization. The maximum ES frequency (224.6 ES/100 anthers) was obtained with the genotype ETH-M 52 cultured in autoclaved medium which had been exposed to AC (5 g/L) for 96 h prior to culture initiation. It is supposed that the higher ES frequencies observed with AC-treated, autoclaved media were due to the availability of glucose and fructose following heat-induced hydrolysis of sucrose and the AC-mediated adsorption of inhibitory compounds produced during autoclaving.