Cytochemical and histochemical characterization of cotyledonary bodies from Pharbitis nil seedlings

Cytological and histochemical characterization of the structures from which an obscure substance is secreted via open stomata to the abaxial surface of Japanese morning glory (Pharbitis nil Choisy cv. Violet) cotyledons has been carried out. Observation of intact cotyledons using the light microscope revealed randomly distributed semi-transparent structures. These structures, which were shown to be the same as those previously described as giant oil cells are referred to here as cotyledonary bodies. These bodies can be eas- ily isolated and purified after enzymatic digestion of the cotyledons. Using different staining procedures we have confirmed that each cotyledonary body originates from an individual mesophyll cell dur- ing embryo development. Purified bodies consist of (i) a thick shell- like envelope; (ii) a transparent, hydrophilic zone; (iii) a hydropho- bic core. Hydrophobic contents of the bodies were readily extracted with methanol and shown to contain fatty acids and phenolic com- pounds using the gas chromatography/mass spectrometry (GC/MS) technique. Methanolic extracts of cotyledonary bodies showed high fluorescence with two excitation and emission maxima. Using a flu- orescence microscope we have shown that the bodies isolated from seedlings grown in continuous light, conditions non-inductive for flowering, and those grown under conditions inductive for flowering (a single 16 h, long dark period) have different fluorescence emis- sion spectra. Different levels of free Ca 2+ inside cotyledonary bodies isolated from light-grown and single dark-period treated P. nil seed- lings were found using the fluorescent calcium indicator dye Fluo-3 under a confocal scanning laser microscope. On the basis of these observations we speculate that cotyledonary bodies could be in- volved in floral induction

Photomorphogenic mutants of tomato

Photomorphogenesis of tomato is being studied with the aid of mutants which are either modified in their photore- ceptor composition or in their signal transduction chain(s) . Several mutants affecting the phytochrome family of photoreceptors, some of which appear deficient for specific genes encoding phytochrome apoproteins have been isolated . In addition, other mutants, including transgenic lines overexpressing phytochrome A, exhibit exaggerated photomorphogenesis during de-etiolation . Anthocyanin biosynthesis and plastid development are being used as model systems for the dissection of the complex interactions among photomorphogenic photoreceptors and to elucidate the nature of their transduction chains

Effect of the 5-HTTLPR polymorphism on affective temperament, depression and body mass index in obesity

Background and aim: Many studies show high prevalence of affective disorders in obese patients. Affective temperament is a subclinical manifestation of such conditions. The 5-HTT gene encoding the serotonin transporter may be involved in both mood and eating dysregulation. The aim of this study was to investigate the influence of a polymorphism in the 5-HTT gene on affective temperament types, depressive symptoms and Body Mass Index (BMI) in obese patients. Methods: This study involved 390 patients (237 females, and 153 males) with obesity. The TEMPS-A questionnaire, Beck Depression Inventory (BDI) and Hamilton Depression Rating Scale (HDRS) were used to evaluate affective temperaments and prevalence of depression. DNA was obtained for serotonin transporter gene-linked polymorphism (5-HTTLPR) genotyping. Results: In obese patients S/S genotype was associated with depressive and L/L with cyclothymic temperament. Subjects with L/L genotype presented significantly higher BMI and greater intensity of depressive symptoms in BDI and HDRS. Females scored higher in anxious and depressive, while males in hyperthymic, cyclothymic and irritable temperaments. Females scored higher in BDI (subjective depression) while males in HDRS (objective depression). Limitations: TEMPS-A, BDI and HDRS are frequently used in studies on affective disorders. However, these methods do not examine all dimensions of mood and personality. Conclusions: In obese patients S allele of 5-HTTLPR was associated with development of depressive temperament while L allele corresponded with greater obesity and prevalence of depression. Different mechanisms may be involved in manifestation of depression in males and females with obesity

Impact of IL-28B polymorphisms on pegylated interferon plus ribavirin treatment response in children and adolescents infected with HCV genotypes 1 and 4

IL-28B polymorphisms are predictors of response to therapy in adults infected with hepatitis C. We do not know whether they are markers of response to therapy in children and adolescents. The aim of this study was to determine whether single-nucleotide polymorphisms (SNPs) in the IL-28B gene could influence the probability of response to therapy compared with other known baseline prognostic factors and correlate with clinical findings in pediatric patients infected with hepatitis C virus (HCV) genotypes 1 or 4. We determined three SNPs of IL-28B (rs12979860, rs12980275, and rs8099917) in 82 patients with chronic HCV infection treated with pegylated interferon alpha and ribavirin (peg-IFNα/RBV). Treatment response and clinical data were analyzed. Overall, sustained virological response (SVR) was achieved by 45 % of patients infected with difficult-to-treat HCV genotypes 1 and 4. Except for IL-28B polymorphisms, there was no association of SVR with any other clinical data. IL-28B rs12979860 CC [odds ratio (OR), 6.81; p = 0.001] and rs8099917 TT (OR, 3.14; p = 0.013) genotypes were associated with higher SVR rates. IL-28B rs12980275 was not significantly associated with SVR ( p = 0.058). Only the distribution between CC and CT-TT genotypes of rs12979860 significantly differentiated patients achieving early virological response (EVR) (OR, 10.0; p = 0.011). Children with the rs12979860 CC genotype had significantly higher baseline viral load compared with CT-TT patients ( p = 0.010). In children and adolescents chronically infected with HCV genotypes 1 and 4, IL-28B rs12979860 and rs8099917 polymorphisms were the only predictors of response to peg-IFN/RBV

Analiza zdolnosci do kwitnienia regenerantow Carlina acaulis subsp. simplex

Regenerated plants of Carlina acaulis subsp. simplex induced on shoot tips and fragments of hypocotyls, cotyledons and roots were used as an experimental material. Explants were isolated from 10-day-old, sterile seedlings and were put on growth media supplemented with BA(3 mg × dm⁻³), andNAA(0,l mg × dm⁻³). Plantlets were acclimatized to ex vitro conditions and planted to the field. Analysis of flowering ability, inflorescence stem morphology, and survival level was the objective of the study. The plants regenerated from shoot tips and cotyledons were able to flower in the first year after acclimatization, however no vital seeds were found, while in the case of hypocotyl- and root-regenerated plants flowering appeared in the second year after acclimatization. Number of flowering-able plants grew in time, reaching 100% level. Few percent of inflorescence stems displayed branches ending with additional capitula. The number of this type of plants decreased in successive years, while the average length of inflorescence stem increased. In the case of intensively flowering plants, the survival rate decreased in 3 consecutive years.Materiał eksperymentalny stanowiły regeneranty Carlina acaulis subsp. simplex, indukowane na wierzchołkach wzrostu oraz fragmentach hypokotyli, liścieni i korzeni. Eksplanty były izolowane z 10-dniowych sterylnych siewek i wykładane na pożywki uzupełnione w BA (3 mg × dm⁻³) i NAA (0,1 mg × dm⁻³). Regeneranty były aklimatyzowane do warunków ex vitro i wysadzane do gruntu. Celem badań była analiza: zdolności do kwitnienia, morfologii pędu kwiatostanowego oraz poziomu przeżywalności w 3 kolejnych sezonach wegetacyjnych. Rośliny zregenerowane z wierzchołków wzrostu i liścieni były zdolne do kwitnienia już w pierwszym roku po aklimatyzacji, ale nie stwierdzono żywotnych nasion. Natomiast rośliny zregenerowane z fragmentów hypokotyli i korzeni były zdolne do kwitnienia w drugim roku po aklimatyzacji. Liczba roślin zdolnych do kwitnienia rosła w kolejnych latach osiągając poziom 100%. Niewielki odsetek roślin wykazywał rozgałęzione pędy kwiatostanowe zakończone dodatkowymi koszyczkami. Liczba tego typu roślin zmniejszała się w kolejnych latach, rosła natomiast średnia długość pędów kwiatostanowych. W przypadku roślin intensywnie kwitnących po 3 latach uprawy spadał wskaźnik przeżywalności

Shoot multiplication and in vitro rooting of Carlina onopordifolia Basser

This paper is the first published report describing micropropagation of Carlina onopordifolia, using shoot tip and hypocotyl explants. The explants were excised from 10-day-old seedlings and transferred to proliferation medium supplemented with 6-benzylaminopurine (BA; 1.0 or 3.0 mg l-1), kinetin (Kn; 1.0 or 3.0 mg l-1) or zeatin (ZEA; 1.0 or 3.0 mg l-1) in combination with naphthaleneacetic acid (NAA; 0.1 mg l-1). The shoot tips were significantly better than hypocotyls as initial material for shoot regeneration. For shoot multiplication, MS medium supplemented with BA proved superior to the other cytokinins tested. Medium supplemented with 1.0 mg l-1 BA gave the highest shoot propagation frequency (66.9%) and number of shoots per explant (2.5). Single shoots were separated from each other and rooted on MS supplemented with IBA for the whole period of culture, with longor short-pulse IBA application. The highest rooting frequency (84.8%) and root number (18.8) were for shortpulse (1 min) 1000 mg l-1 IBA solution. The higher IBA concentration stimulated callus formation and the development of short roots. The shoots were transferred to MS medium without growth regulators. Survival was highest (54.4%) for the plants from the short-pulse 100 mg l-1 IBA treatment. After 8 weeks of acclimatization the plantlets were removed to field conditions and grew normally