Development of micropropagated plants from different clones of Senecio x hybridus in relation to BAP concentration and temperature in vitro

Summary

Selected mature seedling plants were used as stock plants and about half of them were successfully micropropagated. There was no correlation between the ability of petiole expiants to form adventitious shoots and the capacity of these shoots to multiply. Growth and development varied between the micropropagated clones, originating from one seedling each. The colour of the flowers was always constant within a clone. Developmental time, foliage height and the numbers of shoots, leaves and flowers were influenced by the BAP concentration used in vitro. The temperature during the in vitro phase affected the development time and the height of foliage and inflorescences. Increased BAP concentration (from 0 or 0.1 mg l^?1 to 1 mg H) resulted in plants with a longer development time to anthesis and more shoots, leaves and flowers. Plants raised on 5 mg l^?1 BAP developed slowly, resulting in stunted growth, low foliage height, few leaves and flowers. Consequently, it is possible to use the BAP concentration in vitro to regulate the growth of the progeny crop. After transfer to soil in a growth chamber, plants grown at 10°C in vitro flowered earlier, and had lower foliage and higher inflorescences than plants grown at 21°C. This observation indicates that flowering may be promoted by low temperatures in vitro.     

Modelling low molecular weight organic acid dynamics in forest soils

Low molecular weight organic acids such as citrate and oxalate have been hypothesized to play a key role in rhizosphere ecology and pedogenesis. A mathematical site-specific model, DYNLOW, was constructed to describe the temporal and spatial dynamics of these organic acids in coniferous forest soils using the modelling software STELLA^®. Experimentally derived values for biodegradation, adsorption, and daily values of soil temperature, moisture and hydrological flow were used to parameterize the model. The model describes the dynamics and downward movement of oxalate and citrate through the horizons (O, AE, E, Bhs, Bs) of three podzolic soil profiles in Sweden. After calibration, the model predicted average soil solution organic acid concentrations ranging from <1 to 90 μM, which was in agreement with experimental measurements (<1 to 116 μM). The model results indicated that microbial degradation of organic acids was in quantitative terms the biggest process regulating soil solution concentrations. Primary production rates of organic acid in the soil were predicted to be high (<1 to 1250 nmol g⁻¹ soil d⁻¹) in comparison to the amount present at steady state in the soil solution pool (<0.1 to 240 nmol g⁻¹ soil). The downward transfer of organic acids between soil horizons due to mass flow was predicted to be a small flux (<0.1 to 3% of the total loss) compared to that lost by microbial biodegradation. The model predicted that the amount of basal soil respiration that could be attributable to the microbial turnover of organic acids was on average 19±22% of the basal CO₂ production across all sites and horizons for citrate and 7±7% for oxalate. The model results are discussed in the context of pedogenesis, forest soil respiration and organic matter production.     

Endosperm structure affects the malting quality of barley (Hordeum vulgare L.)

Twenty-seven barley (Hordeum vulgare L.) samples collected from growing sites in Scandinavia in 2001 and 2002 were examined to study the effect of endosperm structure on malting behavior. Samples were micromalted, and several malt characteristics were measured. Samples were classified as having a mealier or steelier endosperm on the basis of light transflectance (LTm). Because endosperm structure is greatly dependent on protein content, three barley sample pairs with similar protein contents were chosen for further analysis. During malting, the steelier barley samples produced less root mass, but showed higher respiration losses and higher activities of starch-hydrolyzing enzymes. Malts made from steelier barley had a less friable structure, with more urea-soluble D hordein and more free amino nitrogen and soluble protein. The reason for these differences may lie in the structure or localization of the hordeins as well as the possible effects of endosperm packing on water uptake and movement of enzymes.     

Flavonoids in baby spinach (Spinacia oleracea L.): changes during plant growth and storage

The variation in flavonoid concentration and composition was investigated in baby spinach (Spinacia oleracea L.) cv. Emilia sown on three occasions, each harvested at three growth stages at 6-day intervals. After harvest, leaves were stored in polypropylene bags at 2 or 10 degrees C. Flavonoids were analyzed by reversed phase HPLC. Twelve flavonoid peaks were detected. The main flavonoid, making up on average 43% of the total flavonoid concentration, was identified as 5,3',4'-trihydroxy-3-methoxy-6:7-methylenedioxyflavone-4'-glucuronide. Four other flavonoids each contributed 7-12% of the total flavonoid content. Total flavonoid content was relatively stable during normal retail storage conditions, although some of the individual flavonoid compounds showed considerable variation. The youngest plants had the highest flavonoid concentration, indicating that by harvesting the baby spinach a few days earlier than the current commercial stage of harvest, the flavonoid concentration in the product may be increased and the content of potentially health-promoting compounds enhanced.     

Accurate determination of oosporein in fungal culture broth by differential pulse polarography

A simple and accurate differential pulse polarographic method has been developed for the determination of oosporein in the culture broth of the fungus Beauveria brongniartii. This hydroxybenzoquinone derivative is the only major secondary metabolite secreted by this entomopathogenic fungus, which is used as biological pest control agent (BCA) against Melolontha melolontha larvae. It can be found in the host organism as well as in the formulated product. The polarographic behavior of oosporein was examined in various buffer systems over the pH range 3-10. In Britton-Robinson buffer/methanol solution (3:7 v/v, pH 5.5) the differential pulse polarograms exhibited reproducible peaks at E(p) = -0.18 V vs silver/silver chloride/potassium chloride (3 M). Under these conditions, a plot of peak height vs concentration of oosporein was found to be linear over the range 5.9 x 10(-)(7) to 2.5 x 10(-)(5) M (0.18-7.74 microg mL(-)(1); r = 0.9998). The detection limit was calculated to be 54 ng mL(-)(1). To evaluate the concentration of oosporein, the standard addition method was applied. The analysis of oosporein in the culture broth led to a mean value of 524.9 microg mL(-)(1) broth with a relative standard deviation (S(rel)) of +/-2.6%. The proposed polarographic method is accurate, not time-consuming, and it is of low cost because no separation steps are necessary.     

Seven distinct dietary patterns identified among pregnant Finnish women--associations with nutrient intake and sociodemographic factors

OBJECTIVES: To identify and describe dietary patterns in a cohort of pregnant women and investigate whether the dietary patterns are associated with dietary intake and sociodemographic factors. DESIGN: Mothers entering the Finnish Type 1 Diabetes Prediction and Prevention (DIPP) Nutrition Study in 1997-2002 were retrospectively asked to complete a food-frequency questionnaire concerning their diet during pregnancy. Principal components analysis was used to identify dietary patterns. SETTING: Finland. SUBJECTS: Subjects were 3730 women with a newborn infant carrying increased genetic susceptibility to type 1 diabetes mellitus. RESULTS: Seven factors were identified and named. Energy intake correlated positively with 'Healthy', 'Fast food', 'Traditional bread', 'Traditional meat' and 'Coffee' patterns and inversely with the 'Alcohol and butter' pattern. Intake of dietary fibre correlated positively with 'Healthy', 'Traditional bread' and 'Low-fat foods' patterns and inversely with the 'Alcohol and butter' pattern. The seven dietary patterns seemed to account for relatively large proportions of the variance in energy and nutrient intakes except for the intake of vitamin D, vitamin C, carotenoids and calcium. Maternal age and higher level of education were associated with higher scores on 'Healthy', 'Low-fat foods' and 'Alcohol and butter' patterns. CONCLUSION: Principal components analysis produced seven dietary patterns which may be useful for further research concerning maternal diet and health outcomes among both mothers and their offspring.     

Diatoms in peat - Dominant producers in a changing environment?

Changes in hydrology and temperature can induce rapid changes in boreal wetland ecosystems. Factors such as hydrosere, permafrost, climate and human interference may disturb the prevailing mire vegetation, whereby a new dominant assemblage can develop. At the transition from one vegetation type to another, the old vegetation may be suppressed, die out or start to decay, and some time may pass until a new mire vegetation is fully established. Here, we demonstrate that diatoms may thrive during such transitions, creating isolated and shallow peat layers with significantly elevated biogenic silica content. Biogenic silica and other nutrients that would otherwise be lost during mineralization in runoff are in this way retained in the ecosystem. Our results imply that silica storage originating from diatoms can be expected to increase in today's rapidly changing boreal wetlands. The impacts on transport of Si through boreal watersheds are currently unknown.     

Primary infection protects pigs against re-infection with Lawsonia intracellularis in experimental challenge studies

In two separate trials pigs were experimentally infected with Lawsonia intracellularis at 5-6 weeks of age followed by antibiotic treatment and resolution of the primary infection and then re-inoculated at 12-13 weeks of age. A treatment-control group of pigs received the primary infection and antibiotic treatment only, and served as control for the antibiotic treatment of the primary infection. A challenge-control group of pigs received the second inoculation dose only at 12-13 weeks of age to control infectivity of the challenge-dose and susceptibility of pigs to L. intracellularis at this age. Pigs were monitored for shedding of L. intracellularis in faeces by PCR, and for the development of antibodies and responses of acute phase proteins in serum. The presence of L. intracellularis antigen in the intestinal mucosa was examined in post mortem samples by immunohistochemistry. In both trials primary infected pigs were protected from infection after challenge inoculation as evidenced by absence of faecal shedding of L. intracellularis, lack of changes in acute phase protein concentrations after challenge and with low levels of bacterial antigen in the intestinal mucosa of re-inoculated pigs comparable to that of the treatment-control pigs. In contrast, challenge-control pigs shed L. intracellularis in faeces, had L. intracellularis antigen extensively present within all layers of the intestinal mucosa and developed a significant acute phase protein response in serum after the experimental infection. The acute phase protein response to L. intracellularis infection was detected as an increased rise in the serum concentrations of C-reactive protein and haptoglobin from day-6 post infection, and increased serum concentrations of haptoglobin were generally seen 2-3 weeks after inoculation both at 5-6 and 12-13 weeks of age. In conclusion substantial protection against L. intracellularis infection was found in the re-inoculated pigs in contrast to the development of infection in age-matched control pigs. The acute phase protein responses reflected both the observed protection against L. intracellularis infection upon secondary challenge and that increased resistance to the infection develops with age.     

Co-incubation with IL-18 potentiates antigen-specific IFN-γ response in a whole-blood stimulation assay for measurement of cell-mediated immune responses in pigs experimentally infected with Lawsonia intracellularis

The whole-blood interferon-gamma (IFN-γ) assay is a quantitative in vitro assay for a direct read-out of Ag-specific cell-mediated immune (CMI) responses to infectious diseases. The IFN-γ assay is robust in severe intracellular infections like Brucella or mycobacteria, but more difficult to evaluate for less severe or immunocompromising infections. Here we investigated the performance of the assay when recombinant co-stimulatory cytokines IL-12 and/or IL-18 were added along with Ag or PBS to cultures of whole-blood from pigs infected with Lawsonia intracellularis. In pigs recovering from a natural infection, addition of rIL-12 or rIL-18 alone increased the Ag-specific IFN-γ release while addition of both cytokines resulted in increased IFN-γ release also in PBS cultures. In analyses after experimental infections with L. intracellularis, significant increased levels of Ag-specific IFN-γ production were measured in Ag+rIL-18 cultures from infected pigs compared to the background response in PBS+rIL-18 control samples (p<0.01) or to Ag+rIL-18 cultures from non-inoculated control pigs (p<0.05). Flow cytometry identified two lymphocyte subsets as the Ag-specific IFN-γ producers. The highest IFN-γ production was by CD4(+)CD8(+) cells while a more numerous population of CD4(-)CD8(+) cells produced lower amounts of IFN-γ in response to rIL-18 and L. intracellularis Ag.