Exogenous application of thiamin promotes growth and antioxidative defense system at initial phases of development in salt-stressed plants of two maize cultivars differing in salinity tolerance

The effects of thiamin (Thi) applied as seed soaking or foliar spray on some key physiological parameters were investigated in two differentially salt-responsive maize (Zea mays L.) cultivars, DK 5783 and Apex 836 F1, exposed to saline stress in two different experiments. An initial experiment (germination experiment) was designed to identify appropriate doses of Thi which could lessen the deleterious effects of salt on plants and screen all available maize cultivars for their differential tolerance to salt stress (100 mM NaCl). The seeds of nine maize cultivars were soaked for 24 h in solutions containing six levels of Thi (25, 50, 75, 100, 125 and 150 mg l^?1). Based on the results obtained from the germination experiment, maize cultivar DK 5783 was found to be the most salt tolerant and Apex 836 as the most sensitive cultivar. Also, of six Thi levels used, two levels (100 and 125 mg l^?1) were chosen for subsequent studies. In the second experiment (glasshouse experiment), two maize cultivars, DK 5783 (salt tolerant) and Apex 836 (salt sensitive) were subjected to saline regime (100 mM NaCl) and two levels of Thi (100 and 125 mg l^?1) applied as foliar spray. Salt stress markedly suppressed shoot and root dry mass, total chlorophylls (“a” + “b”), leaf water potential and maximum fluorescence yield (Fv/Fm) in the plants of both maize cultivars, but it increased proline accumulation, leaf osmotic pressure, malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) concentrations, electrolyte leakage (EL) as well as activities of some key antioxidant enzymes, superoxide dismutase (SOD; EC. 1.15.1.1), peroxidase (POD; EC. 1.11.1.7) and catalase (CAT; EC. 1.11.1.6). Salt-induced reduction in plant growth parameters was higher in the salt-sensitive cultivar, Apex 836, which was found to be associated with relatively increased EL, and MDA and H₂O₂ levels, and decreased activities of the key antioxidant enzymes. Application of Thi as seed soaking or foliar spray partly mitigated the deleterious effects of salinity on plants of both maize cultivars. The most promising effect of Thi on alleviation of adverse effects of salt stress on maize plants was found when it was applied as foliar spray at 100 mg l^?1. Thiamin application considerably reduced tissue Na⁺ concentration, but improved those of N, P, Ca²⁺ and K⁺ in the salt-stressed maize plants. Exogenously applied thiamin-induced growth improvement in maize plants was found to be associated with reduced membrane permeability, MDA and H₂O₂ levels, and altered activities of some key antioxidant enzymes such as CAT, SOD and POD as well as increased photosynthetic pigment concentration under saline regime.     

Transformation of Gaeumannomyces graminis with β‐Glucuronidase Reporter and Hygromycin Resistance Genes

Take‐all disease is caused by Gaeumannomyces graminis, (Sacc.) Arx & D. Olivier, a soil‐borne fungus, which colonizes the root and crown tissue of many members of the Poaceae plant family. This fungus is able to grow along the surface of roots as darkly pigmented runner hyphae, which has the ability to penetrate the root. Here, we describe a genetic transformation of G. graminis var. graminis by using polyethylene glycol (PEG)‐based protoplast transformation. Fungus cells were transformed with a plasmid, pHPG, containing the gusA reporter gene that codes for β‐glucuronidase (GUS) and the hph gene for hygromycin resistance as the selectable marker. A de novo transformant selection assay was developed to identify the putative transformants that were expressing the hph gene. In addition, the transformed cells maintained the ability to infect the plant tissues. The GUS‐expressing fungus can be used to study fungal infection processes including fungal penetration, colonization and the role(s) of melanin during pathogenesis. Thus, this study is the first report of G. graminis var. graminis transformed with a visibly detectable reporter gene that provides a useful tool to a better understanding of host–Gaeumannomyces interactions.     

Production of pullulanase by free and immobilized cells of <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> NRC22 in batch and continuous cultures

The production of extracellular pullulanase by Bacillus licheniformis NRC22 was investigated using different fermentation modes. In batch culture maximal enzyme activity of 18 U/ml was obtained after 24 h of growth. In continuous fermentation by the free cells, maximal reactor productivity (4.15 KU/l/h) with enzyme concentration of 14.8 U/ml and specific productivity of 334.9 U/g wet cells/h was attained at a dilution rate of 0.28/h, over a period of 25 days. B. licheniformis NRC22 cells were immobilized on Ca-alginate. The immobilization conditions with respect to matrix concentration and cell load was optimized for maximal enzyme production. In repeated batch operation, the activity of the immobilized cells was stable during the 10 cycles and the activity remained between 9.8 and 7.7 U/ml. Continuous production of pullulanase by the immobilized cells was investigated in a packed–bed reactor. Maximal reactor productivity (7.0 KU/h) with enzyme concentration of 16.8 U/ml and specific productivity of 131.64 U/g wet cells/h was attained at dilution rate of 0.42/h. The enzyme activity in the effluent started to decline gradually to the level of 8.7 U/ml after 25 days of the operation.     

The RecQ helicase AtRECQ4A is required to remove inter-chromosomal telomeric connections that arise during meiotic recombination in Arabidopsis

RecQ helicases are a conserved group of proteins with a role in the maintenance of genome integrity. In Saccharomyces cerevisiae (budding yeast), meiotic recombination is increased in the absence of the RecQ helicase Sgs1. Here we investigated the potential meiotic role of the Sgs1 homologue AtRECQ4A and the closely related AtRECQ4B. Both proteins have been shown to function during recombination in somatic cells, but so far their meiotic role has not been investigated. Both AtRECQ4A and AtRECQ4B were expressed in reproductive tissues. Although immunolocalization studies showed that AtRECQ4A associates with recombination intermediates, we found no evidence that its loss or that of AtRECQ4B had a significant effect on meiotic cross-overs, suggesting functional redundancy with other RECQ family members. Nevertheless, pollen viability decreased in Atrecq4A, resulting in a reduction in fertility, although this was not the case in Atrecq4B. Cytological analysis revealed chromatin bridges between the telomeres of non-homologous chromosomes in Atrecq4A at metaphase I, in some instances accompanied by chromosome fragmentation at anaphase I. The bridges required telomeric repeats and were dependent on meiotic recombination. Immunolocalization confirmed the association of AtRECQ4A with the telomeres during prophase I, which we propose enables dissolution of recombination-dependent telomeric associations. Thus, this study has identified a hitherto unknown role for a member of the RECQ helicase family during meiosis that contributes to the maintenance of chromosome integrity. As telomere structure is generally conserved, it seems likely that these associations may arise during meiosis in other species, where they must also be removed.     

Migraine headaches among university students using id migraine test as a screening tool

Background  

Migraine is a significant health problem, especially for the young people, due to its frequency and accompanying morbidity, causing disability and loss of performance. In this study, our aim was to determine the prevalence of migraine headaches among university students in Edirne, a Turkish city.     

Gap junctions mediate STAT5-independent β-casein expression in CID-9 mammary epithelial cells

Crosstalk between gap junction intracellular communication (GJIC), STAT5 and OCT-1 in gap junction (GJ)-dependent β-casein expression was investigated. CID-9 mammary cells plated with prolactin on non-adherent substratum (poly-HEMA) expressed β-casein independent of STAT5 only in the presence of the GJIC inducer, cAMP. Nuclear STAT5 levels were not detectable. By contrast, cells on EHS-drip expressed β-casein in a STAT5-dependent manner and nuclear STAT5 levels were up-regulated. A 75 kDa OCT-1 isoform was detected in conditions that induced β-casein expression regardless of substratum. Interestingly, 40 and 28 kDa OCT-1 isoforms were induced in cells on polyHEMA with cAMP. Electrophoretic mobility shift assays (EMSA) for OCT-1 revealed two band shifts in cells on polyHEMA with cAMP and on EHS-drip, which were repressed by the GJIC inhibitor, 18α-GA. These studies demonstrated that mammary cells on polyHEMA expressed β-casein in response to prolactin in a pathway that involves GJIC and OCT-1 and is independent of STAT5 nuclear translocation.