Growth,fluorescence, photosynthetic O<Subscript>2</Subscript> production and pigment content of salt adapted cultures of <Emphasis Type="Italic">Arthrospira</Emphasis> (<Emphasis Type="Italic">Spirulina</Emphasis>) <Emphasis Type="Italic">platensis</Emphasis>

The effect of salt concentration (NaCl) on growth, fluorescence, photosynthetic activities and pigment content of the cyanobacterium Arthrospira platensis has been investigated over 15 days. It has been observed that high NaCl concentration induces an increase of the growth, photosynthetic efficiency (α), phycobilin/chlorophyll ratio and a slight decrease of dark respiration and compensation points. Moreover, high NaCl concentration enhances photosystem II (PSII) activity compared to photosystem I (PSI). Results show that the phycobilin-PSII energy transfer compared to the chlorophyll-PSII (F_695,600/F_695,440) increases. However, data obtained about the maximal efficiency of PSII photochemistry are controversial. Indeed, the Fv/Fm ratio decreases in salt adapted cultures, while at the same time the trapping flux per PSII reaction center (TR₀/RC) and the probability of electron transport beyond Q_A (₀) remain unchanged at the level of the donor and the acceptor sites of PSII. This effect can be attributed to the interference of phycobilin fluorescence with Chl a when performing polyphasic transient measurements.     

Biotransformation of polychlorinated dibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxin by <Emphasis Type="Italic">Coprinellus</Emphasis> species

A degradation experiment on dibenzo-p-dioxin (DD) and 2,7-dichlorodibenzo-p-dioxin (2,7-DCDD) was carried out using basidiomycetous fungi belonging to the genera Coprinus, Coprinellus, and Coprinopsis. Some species showed a high rate of decrease in DD for the 2-week test period. Among them, Coprinellus disseminatus showed the highest ability to decrease the DD level, close to 100% by the end of 2 weeks. Further examination showed that Coprinellus disseminatus and Coprinellus micaceus, belonging to the genus Coprinellus, were able to metabolize 2,7-DCDD to a monohydroxylated compound, probably mediated by the P450 system. The metabolism of chlorinated DD by fungi capable of living in soil conditions is reported here for the first time.     

Preservation methods of <Emphasis Type="Italic">Tolypothrix tenuis</Emphasis> for use as a cyanobacterial fertilizer

The suitability of the cyanobacterium Tolypothrix tenuis as a potential biofertilizer was evaluated by measuring cell viability in stored samples preserved by various methods, including freezing at −20°C, freeze-drying, desiccation as flakes and immobilization in alginate beads. The viability recovery and the retained viability index (RVI₁₀) were used as indicators of cell survival after 3 and 15 months storage. The highest recovery level (85%) was obtained by freeze-drying using skimmed milk as the resuspension solution for long-term storage. Dried alginate beads showed a better cell survival (RVI₁₀ 0.25) than air-dried flakes (RVI₁₀ −0.63) after 15 months storage. However, desiccated flakes previously treated with Ca²⁺ and Mg²⁺ ions improved cell survival capacity at the longest period assayed (RVI₁₀ 0.08), extending cell viability by 9 months compared to dried-powder material.     

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.     

Predation by <Emphasis Type="Italic">Nesidiocoris tenuis</Emphasis> on <Emphasis Type="Italic">Bemisia tabaci</Emphasis> and injury to tomato

Tomato is the most important vegetable crop in Spain. The mirid bug Nesidiocoris tenuis (Reuter) commonly appears in large numbers in protected and open-air tomato crops where little or no broad-spectrum insecticides are used. Nesidiocoris tenuis is known to be a predator of whiteflies, thrips and several other pest species. However, it is also considered a pest because it can feed on tomato plants, causing necrotic rings on stems and flowers and punctures in fruits. Our objectives were to evaluate predation by N. tenuis on sweetpotato whitefly Bemisia tabaci Gennadius under greenhouse conditions and establish its relationship to N. tenuis feeding on tomato. Two different release rates of N. tenuis were compared with an untreated control (0, 1 and 4 N. tenuis plant⁻¹) in cages of 8 m². Significant reductions of greater than 90% of the whitefly population and correspondingly high numbers of N. tenuis were observed with both release rates. Regression analysis showed that necrotic rings on foliage caused by N. tenuis were best explained by the ratio of B. tabaci nymphs:N. tenuis as predicted by the equation y = 15.086x − 0.6359.

An Oxidized Ergosterol from <Emphasis Type="Italic">Pleurotus cystidiosus</Emphasis> Active Against Anthracnose Causing <Emphasis Type="Italic">Colletotrichum</Emphasis><Emphasis Type="Italic">gloeosporioides</Emphasis>

This study was undertaken to study the antifungal activity of Pleurotus cystidiosus against Colletotrichum gloeosporioides. This was achieved by fractionating the mushroom, P. cystidiosus initially to acetone (A), dichloromethane (D), and hexane (H) and studying the antifungal activity using the standard poisoned food technique. All the test solutions used were in the concentration of 20,000 ppm. The percentage inhibition of extracts A, D, and H was 12, 7, and 0.4%, respectively. Antifungal assay guided fractionation of the most active extract A resulted in four fractions; A1, A2, A3, and A4 having 12, 22, 0, and 17% percentage inhibitions, respectively. Fractions A2 and A4 were selected for further purifications. Normal phase column chromatography of A2 gave A2-1, A2-2, A2-3, and A2-4, with percentage inhibitions 7, 5, 26, and 13%, respectively. The fraction with the highest inhibitory activity (A2-3) was further separated using the Chromatotron and a single compound (A2-3-13) with 41% inhibition was isolated. Structure elucidation of this compound using 1D and 2D NMR spectroscopy proved this compound to be 3beta, 5alpha, 6beta-trihydroxyergosta-7,22-diene.     

<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.     

<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.     

Evidence for Recombination in the Microcystin Synthetase (<Emphasis Type="Italic">mcy</Emphasis>) Genes ofToxic Cyanobacteria <Emphasis Type="Italic">Microcystis</Emphasis><Emphasis Type="Bold">spp</Emphasis>

Recombination has been suggested to be an important factor for the genetic variation of bacterial genes, but few studies have dealt with intragenic recombination between the same or closely related species of cyanobacteria. Here we provide strong evidence for recombination in the microcystin synthetase (mcy) gene cluster of the toxic cyanobacteria Microcystis spp. This gene cluster contains 10 genes (mcyA to J) that encode a mixed polyketide synthase (PKS)/nonribosomal peptide synthetase (NRPS) complex. mcy gene sequences were determined for four selected regions (within mcyA, D, G, and J) within the mcy gene cluster from 1 Canadian and 10 Asian toxic Microcystis and compared with previously published mcy sequences. Split decomposition analysis indicated a reticulate phylogeny of mcyA, and several potential recombination tracts of mcyA were identified by the RDP analysis and a runs test implemented in GENECONV. In contrast, no recombination was detected in the mcyD, G, and J sequences. However, discrepancies among the four mcy gene genealogies were evident from the results of independent split decomposition analyses, which were further supported by incongruence length difference (ILD) tests. Taken together, these findings suggest that both intragenic and intergenic recombination within the mcy gene cluster contributes to the genetic diversity of the mcy genes of Microcystis spp.This article contains online supplementary material.     

The combined use of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> and <Emphasis Type="Italic">Nesidiocoris tenuis</Emphasis> against the tomato borer <Emphasis Type="Italic">Tuta absoluta</Emphasis>

Since Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) was first detected at the end of 2006 in the Mediterranean Basin, several endemic natural enemies have been reported to prey on this exotic pest. The predator Nesidiocoris tenuis Reuter (Hemiptera: Miridae) can regulate T. absoluta populations, because it is able to prey efficiently on T. absoluta eggs. Furthermore, previous studies have demonstrated that first-instar larvae of T. absoluta are highly susceptible to Bacillus thuringiensis (Bt) treatments. In this work, we tested the combination of both approaches under greenhouse conditions. B. thuringiensis formulations were sprayed weekly for two months, three months or throughout the growing cycle, and in all cases, one N. tenuis per plant was also released. Control plants were completely destroyed by the infestation levels reached by T. absoluta. In contrast, all treatments based on B. thuringiensis treatments and releases of N. tenuis reduced leaf damage by more than 97% when compared to the untreated control, with no significant differences among them. Furthermore, yield in the control plants was significantly reduced when compared with all Bt–N. tenuis treatments. Our results demonstrate that when B. thuringiensis treatments are applied immediately after the initial detection of T. absoluta on plants, they do not interfere with N. tenuis establishment in the crop because T. absoluta eggs are available. According to our data, treatments with B. thuringiensis later in the growing season would no longer be necessary because mirids alone would control the pest.     

Antibacterial activity of three South Indian seagrasses, <Emphasis Type="Italic">Cymodocea serrulata</Emphasis>, <Emphasis Type="Italic">Halophila ovalis</Emphasis> and <Emphasis Type="Italic">Zostera capensis</Emphasis>

The antibacterial properties of the three seagrasses namely Cymodocea serrulata, Halophila ovalis and Zostera capensis were tested against the human pathogens Staphylococcus aureus, Bacillus cereus, B. subtilis, Escherichia coli, Salmonella paratyphi, Salmonella typhimurium and Micrococcus luteus, using six different solvents namely, petroleum ether, chloroform, ethyl acetate, acetone, methanol and water. Ethyl acetate and methanol extracts showed maximum activity against most of the pathogens when compared to other solvents. Experiments are underway to isolate active compound(s) implicated in controlling the growth of the pathogens in vitro.     

<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.     

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.     

Growth characteristics and growth modeling of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> and <Emphasis Type="Italic">Planktothrix agardhii</Emphasis> under iron limitation

Although iron is a key nutrient for algal growth just as are nitrogen and phosphorus in aquatic systems, the effects of iron on algal growth are not well understood. The growth characteristics of two species of cyanobacteria, Microcystis aeruginosa and Planktothrix agardhii, in iron-limited continuous cultures were investigated. The relationships between dissolved iron concentration, cell quota of iron, and population growth rate were determined applying two equations, Monod’s and Droop’s equations. Both species produced hydroxamate-type siderophores, but neither species produced catechol-type siderophores. The cell quota of nitrogen for both M. aeruginosa and P. agardhii decreased with decreasing cell quota of iron. The cell quota of phosphorus for M. aeruginosa decreased with decreasing cell quota of iron, whereas those for P. agardhii did not decrease. Iron uptake rate was measured in ironlimited batch cultures under different degrees of iron starvation. The results of the iron uptake experiments suggest that iron uptake rates are independent of the cell quota of iron for M. aeruginosa and highly dependent on the cell quota for P. agardhii. A kinetic model under iron limitation was developed based on the growth characteristics determined in our study, and this model predicted accurately the algal population growth and iron consumption. The model simulation suggested that M. aeruginosa is a superior competitor under iron limitation. The differences in growth characteristics between the species would be important determinants of the dominance of these algal species.     

Role of Arabidopsis <Emphasis Type="Italic">ABF1</Emphasis>/<Emphasis Type="Italic">3</Emphasis>/<Emphasis Type="Italic">4</Emphasis> during <Emphasis Type="Italic">det1</Emphasis> germination in salt and osmotic stress conditions

Key message

Arabidopsis det1 mutants exhibit salt and osmotic stress resistant germination. This phenotype requires HY5, ABF1, ABF3, and ABF4.

Abstract

While DE-ETIOLATED 1 (DET1) is well known as a negative regulator of light development, here we describe how det1 mutants also exhibit altered responses to salt and osmotic stress, specifically salt and mannitol resistant germination. LONG HYPOCOTYL 5 (HY5) positively regulates both light and abscisic acid (ABA) signalling. We found that hy5 suppressed the det1 salt and mannitol resistant germination phenotype, thus, det1 stress resistant germination requires HY5. We then queried publically available microarray datasets to identify genes downstream of HY5 that were differentially expressed in det1 mutants. Our analysis revealed that ABA regulated genes, including ABA RESPONSIVE ELEMENT BINDING FACTOR 3 (ABF3), are downregulated in det1 seedlings. We found that ABF3 is induced by salt in wildtype seeds, while homologues ABF4 and ABF1 are repressed, and all three genes are underexpressed in det1 seeds. We then investigated the role of ABF3, ABF4, and ABF1 in det1 phenotypes. Double mutant analysis showed that abf3, abf4, and abf1 all suppress the det1 salt/osmotic stress resistant germination phenotype. In addition, abf1 suppressed det1 rapid water loss and open stomata phenotypes. Thus interactions between ABF genes contribute to det1 salt/osmotic stress response phenotypes.

     

<Emphasis Type="Italic">Wolinella succinogenes</Emphasis> response to ox-bile stress

The bacterium Wolinella succinogenes is the only known species of its genus. It was first isolated from cow ruminal fluid, and in cattle, it dwells in the reticulum and rumen compartments of the stomach. The global protein response of W. succinogenes to ox-bile was investigated with the aim to understand bile-tolerance mechanisms of the bacterium. Bacteria were grown in liquid media supplemented with different bile concentrations to determine its effects on growth and morphology. Proteomic analyses served to identify 14 proteins whose expression was modulated by the presence of 0.2% bile. Quantitative real-time PCR analyses of the expression of selected genes were employed to obtain independent confirmation of the proteomics data. Proteins differentially expressed revealed metabolic pathways involved in the adaptation of W. succinogenes to bile. The data suggested that bile stress elicited complex physiological responses rather than just specific pathways, and identified proteins previously unknown to be involved in the adaptation of bacteria to bile.     

In Vitro Susceptibility of <Emphasis Type="Italic">C. albicans</Emphasis> and <Emphasis Type="Italic">C. neoformens</Emphasis> to Potential Metabolites from <Emphasis Type="Italic">Streptomycetes</Emphasis>

Forty two Streptomycetes isolates from soils of Kodachadri region in Western ghats were recovered by soil dilution technique. Cross streak method was followed for primary screening of antifungal activity. Positive isolates were subjected to secondary screening by cold extraction of fermentation broth in butanol solvent. Six isolates exhibited broad spectrum antifungal activity against all the tested yeast pathogens like Candida albicans, Candida lipolytica, Cryptococcus neoformens and Saccharomyces cerevisiae. One isolate showed excellent antifungal activity against all test organisms with maximum zone of inhibition 60 mm each incase of C. neoformens and C. albicans. Partial characterization of antifungal metabolite by TLC resulted in a purple spot with an R_f value 0.50. The UV absorption spectra at 218 nm indicated possible chemical nature of the active metabolite as polyene group and purity was assessed by analytical HPLC.     

Salt tolerance of two aquatic macrophytes, <Emphasis Type="Italic">Pistia stratiotes</Emphasis> and <Emphasis Type="Italic">Salvinia molesta</Emphasis>

The physiological responses to NaCl salinity were investigated in two floating aquatic macrophytes, Pistia stratiotes L. and Salvinia molesta L. With the increasing NaCl concentration a decrease in chlorophyll and carotenoid contents was recorded in Salvinia as compared to Pistia. Also a greater increase in H₂O₂ accumulation and lipid peroxidation was observed in the shoot and root tissues of Salvinia as compared to Pistia. The superoxide dismutase, glutathione reductase, catalase and guaiacol peroxidase activities, and ascorbate and glutathione contents increased in Salvinia and Pistia shoot and root tissues in response to NaCl.     

Growth and ion uptake in <Emphasis Type="Italic">Annona muricata</Emphasis> and <Emphasis Type="Italic">A. squamosa</Emphasis> subjected to salt stress

The effects of treatment with NaCl (3, 100 and 300 mM) for 1, 2, 3 and 7 d on plant growth and ion accumulation were analyzed in 2-week and 8-week-old Annona muricata and A. squamosa plants. Fresh mass and root growth inhibition were directly related to the increase in salinity, particularly for A. squamosa. Two-weeks old seedlings were sensitive to 100 and 300 mM NaCl particularly after 7 d, whereas 8-week-old plants were shown to be more resistant to NaCl even at 300 mM NaCl. Na⁺ and Cl^– mostly accumulated in young leaves. Our results suggest that A. squamosa is more sensitive than A. muricata to salt stress and that older seedlings of both species are more tolerant than younger seedlings.