<Emphasis Type="Italic">n</Emphasis>-Alkanes variability in the diazotrophic cyanobacterium <Emphasis Type="Italic">Anabaena cylindrica</Emphasis> in response to NaCl stress

n-Alkanes pattern in response to NaCl stress has been studied in the cyanobacterium Anabaena cylindrica. Saturated hydrocarbons were separated and identified by gas chromatography-mass spectrometry (GC-MS) using serially coupled capillary column. Light chain n-alkanes in the range of C₉–C₁₇ (43%) and heavy chain n-alkanes in range of C₁₇–C₂₃ (34%) and C₂₃–C₃₁ (23%) were identified as the major components of total hydrocarbons in the NaCl adapted cells of A. cylindrica. In contrast, NaCl-untreated cells of A. cylindrica had dominance of only long chain n-alkanes in the range of C₂₃–C₃₁ comprising about 94% of its total n-alkanes. The persistence of high level (43%) of short chain n-alkanes (C₉–C₁₇) in NaCl adapted cells of A. cylindrica as compared to its negligible level (0.2%) in NaCl untreated counterpart clearly indicates that NaCl stress causes the A. cylindrica to shift towards the synthesis of short chain n-alkanes.     

<Emphasis Type="Italic">Acanthamoeba castellanii</Emphasis> an environmental host for <Emphasis Type="Italic">Shigella dysenteriae</Emphasis> and <Emphasis Type="Italic">Shigella sonnei</Emphasis>

The interaction between Shigella dysenteriae or Shigella sonnei and Acanthamoeba castellanii was studied by viable counts, gentamicin assay and electron microscopy. The result showed that Shigella dysenteriae or Shigella sonnei grew and survived in the presence of amoebae for more than 3 weeks. Gentamicin assay showed that the Shigella were viable inside the Acanthamoeba castellanii which was confirmed by electron microscopy that showed the Shigella localized in the cytoplasm of the Acanthamoeba castellanii. In conclusion, the relationship between Shigella dysenteriae and Shigella sonnei with Acanthamoeba castellanii is symbiotic, and accordingly free-living amoebae may serve as a transmission reservoir for Shigella in water.     

Fluctuating asymmetry of <Emphasis Type="Italic">Amphibalanus</Emphasis> (<Emphasis Type="Italic">Balanus</Emphasis>) <Emphasis Type="Italic">amphitrite</Emphasis> (Cirripedia: Thoracica) in association with shore height and metal pollution

The level of fluctuating asymmetry (FA), which is defined as random deviations from perfect bilateral symmetry in the morphological traits of an organism, increases with increasing developmental instability, and it may be used as an indicator of environmental and/or genetic stresses. This study attempted to relate FA levels in the opercular plates of the barnacle Amphibalanus (Balanus) amphitrite with shore height and body trace metal concentrations. Barnacles were collected from both low and mid shores at six coastal locations with various degrees of marine pollution in Hong Kong. Four opercular traits, namely scutum length, tergum length, scutum width, tergum width were measured in the specimen while concentrations of five common trace metals (Cd, Cu, Cr, Mn and Zn) were determined in their body tissues using inductively coupled plasma-atomic emission spectrophotometry. Among the four traits, only tergum length and scutum width fulfilled the assumption of FA while their measurement errors were low. Across all sites, mid-shore A. amphitrite consistently exhibited a significantly higher FA level in scutum width than that in low-shore barnacles, but FA levels in tergum length were similar between the two shore heights. These results suggested that FA was trait-dependent, and mid-shore barnacles were possibly under high physical stress, such as desiccation and high temperature, leading to high developmental instability. Although no positive association was observed between FA and metal contamination in low-shore A. amphitrite, FA level in tergum length significantly increased with decreasing body concentration of manganese (Mn) in these barnacles. Such a negative relationship may be explained by the mechanism of bioaccumulation and physiological role of Mn in A. amphitrite with respect to the formation of barnacle shell plates. Handling editor: T. P. Crowe     

<Emphasis Type="Italic">Susto</Emphasis> and<Emphasis Type="Italic"> Nervios</Emphasis>: Expressions for Stress and Depression

Folk illnesses that are cultural constructions of psychological distress offer a vehicle for the cross-cultural study of stress and stress-related morbidity. This study explores the relationship between the Latin American folk illnesses susto and nervios and mental health. We hypothesize that these folk illnesses are distinct and that there is a stronger association between current levels of stress and depressive symptoms with past experience of nervios than with susto, because the cultural constructions of these folk illnesses reflect chronic and acute concepts of distress, respectively. Interviews were conducted in Guadalajara, Mexico, where participants responded to questions about their socio-demographic characteristics, stress, depressive symptoms, and whether they had experienced susto or nervios. Susto and nervios are very prevalent and occur across sociodemographic subgroups, with the exception that nervios occurred more often in women (p < 0.05). Susto was significantly associated with stress and depressive symptoms (p < 0.05), but nervios had a much stronger association (p < 0.0001), even after controlling for gender. Susto and nervios were expressions of psychological distress; most of those with depression reported susto and/or nervios. This study validates the link between these folk illnesses and stress and depression and may, ultimately, facilitate cross-cultural research on stress.     

<Emphasis Type="Italic">Ty</Emphasis>1<Emphasis Type="Italic">/copia</Emphasis>- and <Emphasis Type="Italic">Ty</Emphasis>3<Emphasis Type="Italic">/gypsy</Emphasis>-like DNA sequences in <Emphasis Type="Italic">Helianthus </Emphasis>species

Two repeated DNA sequences isolated from a partial genomic DNA library of Helianthus annuus, p HaS13 and p HaS211, were shown to represent portions of the int gene of a Ty3 /gypsy retroelement and of the RNase-Hgene of a Ty1 /copia retroelement, respectively. Southern blotting patterns obtained by hybridizing the two probes to BglII- or DraI-digested genomic DNA from different Helianthus species showed p HaS13 and p HaS211 were parts of dispersed repeats at least 8 and 7 kb in length, respectively, that were conserved in all species studied. Comparable hybridization patterns were obtained in all species with p HaS13. By contrast, the patterns obtained by hybridizing p HaS211 clearly differentiated annual species from perennials. The frequencies of p HaS13- and p HaS211-related sequences in different species were 4.3x10(4)-1.3x10(5) copies and 9.9x10(2)-8.1x10(3) copies per picogram of DNA, respectively. The frequency of p HaS13-related sequences varied widely within annual species, while no significant difference was observed among perennial species. Conversely, the frequency variation of p HaS211-related sequences was as large within annual species as within perennials. Sequences of both families were found to be dispersed along the length of all chromosomes in all species studied. However, Ty3 /gypsy-like sequences were localized preferentially at the centromeric regions, whereas Ty1/ copia-like sequences were less represented or absent around the centromeres and plentiful at the chromosome ends. These findings suggest that the two sequence families played a role in Helianthusgenome evolution and species divergence, evolved independently in the same genomic backgrounds and in annual or perennial species, and acquired different possible functions in the host genomes.     

Diversification of the barley and wheat <Emphasis Type="Italic">blt101</Emphasis>/<Emphasis Type="Italic">wpi6</Emphasis> promoters by the <Emphasis Type="Italic">Xumet</Emphasis> element without affecting stress responsiveness

BLT101-family plasma membrane proteins are found in a wide range of organisms from bacteria to nematodes and are involved in the regulation of cellular cation concentration under stress conditions. A comparison of the promoter regions of barley blt101 and its wheat ortholog, wpi6, revealed highly conserved nucleotide sequences between both genes and a unique insertion of a Xumet element in the blt101 promoter. The Xumet insertion occurred between a putative abscisic acid-responsive element (ABRE) and the dehydration-responsive element/c-repeat (DRE/CRT) within the blt101 promoter. However, blt101 and wpi6 were induced similarly in response to ABA, drought and low temperature, suggesting that the insertion does not affect promoter functions. The Xumet insertion in the blt101/wpi6 promoter region was detected in five barley cultivars, but absent in two wheat cultivars tested, suggesting that the insertion is barley-specific. Genomic Southern blot analysis revealed a large number of Xumet sequences interspersed in the barley genome, whereas only one or very few copies are present in the wheat genome. The data suggested that an expansion in copy number of Xumet elements occurred in the barley genome through evolution.     

Polymorphism of the <Emphasis Type="Italic">CONSTANS</Emphasis> gene in <Emphasis Type="Italic">Brassica</Emphasis> plants

Using a direct amplification of genomic DNA from two Brassica rapa forms, we obtained two homologs of the CONSTANS gene, which controls the photoperiodic induction of flowering in Arabidopsis plants. The cloned fragments of B. rapa genome were identified as members of the CONSTANS-LIKE1 class. By aligning the nucleotide sequences of the CONSTANS gene and its homologs, three classes, CONSTANS, CONSTANS-LIKE1, and CONSTANS-LIKE2, were distinctly discerned by their primary structure. The pattern of restriction fragment length polymorphisms (RFLP) of the CONSTANS homologs in B. carinata, B. juncea, B. napus, B. nigra, B. oleracea, and B. rapa were genome-specific; in addition, the CONSTANS homologs were classified by plant geographic origin, and we assume that such classification is related to plant photoperiodic response.Translated from Fiziologiya Rastenii, Vol. 52, No. 2, 2005, pp. 274–281.Original Russian Text Copyright © 2005 by Martynov, Khavkin.This revised version was published online in April 2005 with a corrected cover date.     

Trehalose-Producing Enzymes MTSase and MTHase in <Emphasis Type="Italic">Anabaena</Emphasis> 7120 Under NaCl Stress

Salt tolerance, a multigenic trait, necessitates knowledge about biosynthesis and function of candidate gene(s) at the cellular level. Among the osmolytes, trehalose biosynthesis in cyanobacteria facing NaCl stress is little understood. Anabaena 7120 filaments exposed to 150 mm NaCl fragmented and recovered on transfer to –NaCl medium with the increased heterocysts frequency (7%) over the control (4%). Cells failed to retain Na⁺ beyond a threshold [2.19 mm/cm³ (PCV)]. Whereas NaCl-stressed cells exhibited a marginal rise in K⁺ (1.1-fold) only at 30 h, for Na⁺ it was 130-fold at 48 h over cells in control. A time-course study (0–54 h) revealed reduction in intracellular Na⁺ beyond 48 h [0.80 mm/cm³ (PCV)] suggestive of ion efflux. The NaCl-stressed cells showed differential expression of maltooligosyltrehalose synthase (MTSase; EC 5.4.99.15) and maltooligosyltrehalose trehalohydrolase (MTHase; EC 3.2.1.141) depending on the time and the extent of intracellular Na⁺ buildup.     

<Emphasis Type="Italic">Agrobacterium</Emphasis><Emphasis Type="Italic">tumefaciens</Emphasis>-mediated transformation of callus cells of <Emphasis Type="Italic">Crataegus</Emphasis><Emphasis Type="Italic">aronia</Emphasis>

A genetic transformation system has been developed for callus cells of Crataegus aronia using Agrobacterium tumefaciens. Callus culture was established from internodal stem segments incubated on Murashige and Skoog (MS) medium supplemented with 5 mg l⁻¹ Indole-3-butyric acid (IBA) and 0.5 mg l⁻¹ 6-benzyladenine (BA). In order to optimize the callus culture system with respect to callus growth and coloration, different types and concentrations of plant growth regulators were tested. Results indicated that the best average fresh weight of red colored callus was obtained on MS medium supplemented with 2 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg l⁻¹ kinetin (Kin) (callus maintenance medium). Callus cells were co-cultivated with Agrobacterium harboring the binary plasmid pCAMBIA1302 carrying the mgfp5 and hygromycin phosphotransferase (hptII) genes conferring green fluorescent protein (GFP) activity and hygromycin resistance, respectively. Putative transgenic calli were obtained 4 weeks after incubation of the co-cultivated explants onto maintenance medium supplemented with 50 mg l⁻¹ hygromycin. Molecular analysis confirmed the integration of the transgenes in transformed callus. To our knowledge, this is the first time to report an Agrobacterium-mediated transformation system in Crataegus aronia.     

The <Emphasis Type="Italic">Caenorhabditis</Emphasis><Emphasis Type="Italic">briggsae</Emphasis> genome contains active <Emphasis Type="Italic">CbmaT1</Emphasis> and <Emphasis Type="Italic">Tcb1</Emphasis> transposons

The maT clade of transposons is a group of transposable elements intermediate in sequence and predicted protein structure to mariner and Tc transposons, with a distribution thus far limited to a few invertebrate species. We present evidence, based on searches of publicly available databases, that the nematode Caenorhabditis briggsae has several maT-like transposons, which we have designated as CbmaT elements, dispersed throughout its genome. We also describe two additional transposon sequences that probably share their evolutionary history with the CbmaT transposons. One resembles a fold back variant of a CbmaT element, with long (380-bp) inverted terminal repeats (ITRs) that show a high degree (71%) of identity to CbmaT1. The other, which shares only the 26-bp ITR sequences with one of the CbmaT variants, is present in eight nearly identical copies, but does not have a transposase gene and may therefore be cross mobilised by a CbmaT transposase. Using PCR-based mobility assays, we show that CbmaT1 transposons are capable of excising from the C. briggsae genome. CbmaT1 excised approximately 500 times less frequently than Tcb1 in the reference strain AF16, but both CbmaT1 and Tcb1 excised at extremely high frequencies in the HK105 strain. The HK105 strain also exhibited a high frequency of spontaneous induction of unc-22 mutants, suggesting that it may be a mutator strain of C. briggsae.     

GA signaling and <Emphasis Type="Italic">CO/FT</Emphasis> regulatory module mediate salt-induced late flowering in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Flowering timing is very important for the reproductive success of higher plants. However, effects of salt on plant flowering and the underlying molecular mechanisms are largely unknown. Here, we show that salt stress delays flowering in Arabidopsis in a dose-dependent manner. Mild salt stress (≤50 mM NaCl) promoted and prolonged the vegetative growth, whereas high salinity (≥100 mM NaCl) largely delayed or inhibited the transition from vegetative growth to reproductive development. The gibberellin (GA)-pathway plays an important role in this phenotype, and application of exogenous GA could restore late flowering induced by salt. In addition, the CONSTANS (CO)/FLOWERING LOCUS T (FT) module may also play a critical role in mediating the effects of salt on flowering. The mRNA abundance of CO was significantly reduced by salt stress in a dose-dependent manner. The constans (co-2) mutants did not respond to moderate salt stress, whereas over-expressing CO manifested no delay in flowering time in response to salinity. Expression of FT, SOC1 and LFY in the downstream of the pathways was also reduced by salt according to dose. Moreover, salt-sensitive mutant salt overly sensitive3 (sos3) exhibited greater sensitivity in flowering, further suggesting that ion disequilibrium mediates salt-induced late flowering. Kexue Li and Youning Wang contributed equally to this report.     

<Emphasis Type="Italic">Galleria</Emphasis><Emphasis Type="Italic">mellonella</Emphasis> are Resistant to <Emphasis Type="Italic">Pneumocystis</Emphasis><Emphasis Type="Italic">murina</Emphasis> Infection

Studying Pneumocystis has proven to be a challenge from the perspective of propagating a significant amount of the pathogen in a facile manner. The study of several fungal pathogens has been aided by the use of invertebrate model hosts. Our efforts to infect the invertebrate larvae Galleria mellonella with Pneumocystis proved futile since P. murina neither caused disease nor was able to proliferate within G. mellonella. It did, however, show that the pathogen could be rapidly cleared from the host.     

Revision of the <Emphasis Type="Italic">Serrulati</Emphasis> species of <Emphasis Type="Italic">Penstemon</Emphasis> section <Emphasis Type="Italic">Saccanthera</Emphasis> subsection <Emphasis Type="Italic">Serrulati</Emphasis>

A revision of Penstemon sect. Saccanthera subsect. Serrulati includes a new species (P. salmonensis), a new variety (P. triphyllus var. infernalis), and the elevation of a subspecies to species (P. curtiflorus), bringing the total number of species to eight, which are keyed and described, complete with nomenclature and type citations.     

New combinations and typifications in <Emphasis Type="Italic">Bistorta</Emphasis>, <Emphasis Type="Italic">Persicaria</Emphasis>, <Emphasis Type="Italic">Polygonum,</Emphasis> and <Emphasis Type="Italic">Rumex</Emphasis> (Polygonaceae)

New combinations are proposed in anticipation of the Polygonaceae treatment in the forthcoming volume of Intermountain Flora: Polygonum kelloggii var. esotericum, P. kelloggii var. watsonii , Rumex densiflorus var. pycnanthus , R. salicifolius var. utahensis, and R. occidentalis var. tomentellus. Typifications are proposed to facilitate ongoing studies in Polygonaceae and to maintain current usage.     

Physiological characterisation of <Emphasis Type="Italic">acuB</Emphasis> deletion in <Emphasis Type="Italic">Aspergillus niger</Emphasis>

The acuB gene of Aspergillus niger is an ortholog of facB in Aspergillus nidulans. Under carbon-repression conditions, facB is repressed, thereby preventing acetate metabolism when the repressing carbon source is present. Even though facB is reported to be repressed directly by CreA, it is believed that a basal level of FacB activity exists under glucose-repressive conditions. In the present study, the effect of deletion of acuB on the physiology of A. niger was assessed. Differences in organic acid and acetate production, enzyme activities and extracellular amino and non-amino organic acid production were determined under glucose-repressing and -derepressing conditions. Furthermore, consumption of alternative carbon sources (e.g. xylose, citrate, lactate and succinate) was investigated. It was shown that AcuB has pleiotropic effects on the physiology of A. niger. The results indicate that metabolic pathways that are not directly involved in acetate metabolism are influenced by acuB deletion. Clear differences in organic acid consumption and production were detected between the ∆acuB and reference strain. However, the hypothesis that AcuB is responsible for basal AcuA activity necessary for activation of acetate metabolic pathways, even during growth on glucose, could not be confirmed. The experiments demonstrated that also when acuB was deleted, no acetate was formed. Therefore, AcuB cannot be the only activator of AcuA, and another control mechanism has to be available for activating AcuA.     

Acute stress hyporesponsive period in nestling Thin-billed prions <Emphasis Type="Italic">Pachyptila belcheri</Emphasis>

When confronted with acute stressors, vertebrates show a highly conserved evolved sequence of physiological, hormonal and behavioural responses, including the activation of the hypothalamic–pituitary–adrenal axis. Many young vertebrates show a stress hyporesponsive period, where they exhibit a reduced glucocorticoid response. Here, we analyzed the stress response of nestling Thin-billed prions Pachyptila belcheri and compared chicks with different previous experience with capture and handling. We found that chicks had a stress response, but baseline and peak levels were below those measured in adults. The stress response of the chicks was rapid and followed by fast recovery, such that the total amount of corticosterone released in response to handling was very much lower in chicks than adults. These results indicate that nestling Thin-billed prions exhibit a stress hyporesponsive period. This was not due to habituation, as CORT measurements at baseline and elevated levels were similar in chicks handled daily and naïve chicks. The comparison with other published studies showed that the stress response of chicks usually peaks earlier and lower than in adults, and researchers should take care to measure stress-induced levels at an appropriate sampling time.     

<Emphasis Type="Italic">In vitro</Emphasis> minimum growth for conservation of <Emphasis Type="Italic">Drosophyllum lusitanicum</Emphasis>

The present paper reports a protocol for minimum growth conservation of Drosophyllum lusitanicum (L.) Link. in vitro. Double-node cuttings were maintained for 4, 8 and 12 months at 5 or 25 °C in the dark. The effects of sucrose either alone at 5, 20, 30, 40 and 60 g dm⁻³ or at 20, 40 and 60 g dm⁻³ in combination with 20 g dm⁻³ mannitol, on survival and post-storage shoot multiplication efficiency were investigated. The cultures could effectively be conserved under minimum growth at 5 °C for 8 months on Murashige and Skoog’s medium supplemented with 60 g dm⁻³ sucrose, 20 g dm⁻³ mannitol and 0.91 μM zeatin. Following extended conservation, the cultures could be successfully regenerated into new shoots, and they were morphologically similar to those of non-stored controls.     

Peanut RNA Helicase <Emphasis Type="Italic">AhRH47</Emphasis> Sustains Protein Synthesis Under Stress and Improves Stress Adaptation in <Emphasis Type="Italic">Arabidopsis</Emphasis>

Stress responsive RNA helicases are involved in translation initiation sustain protein synthesis. In this study, a stress responsive DEAD box RNA helicase, AhRH47 from peanut cDNA library was identified and characterised during stress. In silico analysis of AhRH47 showed the nine conserved motifs characteristic of an RNA helicase. The phylogenetic and amino acid sequence alignment analyses revealed that AhRH47 is highly homologous to an important DEAD box RNA helicase (eIF4A), which is involved in translation initiation. AhRH47 is stress responsive, being highly expressed under salinity and moisture stress, which is induced to a lesser extent under PEG and ABA treatments. Constitutive overexpression of AhRH47 in Arabidopsis conferred enhanced tolerance to salinity and mannitol-induced stresses. In addition, the transgenic plants showed improved tolerance under moisture stress and exhibited improved recovery growth on stress alleviation. Overexpressing plants showed increased ¹⁴C-labelled amino acids incorporation in to protein especially under stress condition. The results suggest AhRH47 transgenic lines maintained higher protein synthesis under stress and thus improved adaptation to osmotic and desiccation stresses.