Effects of flower dimorphism and light environment on arbuscular mycorrhizal colonisation in a cleistogamous herb

Although it is known that floral dimorphism contributes to the maintenance of mixed breeding systems, the consequences of producing progeny of a contrasting genetic background and seeds with differential resource allocation has been practically ignored regarding establishment of belowground organisms–plant interactions. This article evaluates the combined effect of floral dimorphism with cross type and light environment on interactions between Ruellia nudiflora and arbuscular mycorrhizal fungi (AMF). R. nudiflora produces cleistogamous (CL) flowers that exhibit obligate self‐pollination and chasmogamous (CH) flowers with facultative self‐ (CHs) or cross‐ (CHc) pollination. We evaluated the establishment of the plant–AMF interaction in progeny derived from each floral type, under two light conditions (shaded versus open). We established different scenarios depending on the existence of inbreeding depression (ID) and whether the differential resource allocation (DRA) to CH and CL flowers affected the R. nudiflora–AMF interaction. We predicted that under shaded light conditions there might be an intensification of ID, having a negative effect on AMF colonisation. The percentages of hyphae and vesicles in the harvested roots was significantly higher in the shaded plants (F ≥ 4.11, P < 0.05), while progeny of CHc and CHs presented a higher percentage of hyphae and vesicle colonisation compared to CL progeny (F = 15.26, P < 0.01). The results show that DRA to CH flowers and light availability both determines the establishment of R. nudiflora–AMF interaction. The results also suggest that even under stressful light conditions, endogamy does not affect this interaction, which may explain the success of R. nudiflora as an invasive species.     

A folate independent role for cytosolic HPPK/DHPS upon stress in Arabidopsis thaliana

Cytosolic HPPK/DHPS (cytHPPK/DHPS) in Arabidopsis is a functional enzyme with activity similar to its mitochondrial isoform. Genomic complementation of the cytHPPK/DHPS knockout mutant with the wild type gene led to a complete rescue of the stress sensitive mutant phenotype in seed germination tests under abiotic stress conditions. Moreover, over-expression of the gene resulted in higher germination rate under stress as compared to the wild-type, confirming its role in stress resistance. Analysis of folates in seedlings, inflorescence and dry seeds showed unchanged levels in the wild-type, mutant and over-expressor line, upon stress and normal conditions, suggesting a role for cytHPPK/DHPS distinct from folate biosynthesis and a folate-independent stress resistance mechanism. This apparently folate-independent mechanism of stress resistance points towards a possible role of pterins, since the product of HPPK/DHPS is dihydropteroate.     

Protein isolates from jumbo squid (Dosidicus gigas) by pH-shift processing

There is a growing interest in adding value to the jumbo squid Dosidicus gigas fishery. This work describes two extraction procedures for processing muscle to obtain protein isolates with suitable functional properties. The effect on muscle protein solubility and protein recovery of combining freezing and grinding raw materials during storage was evaluated. Processes are based on extraction of protein at acid or alkaline pH and subsequent iso-electric precipitation. About 85% of the initial muscle protein was solubilized at pH 3 and 11. Regardless of the pH used for extraction, about 90% of the protein was obtained after precipitation at pH 5.5. The total yield from both procedures was 75%. Treatments during storage did not significantly affect solubility and yield of protein. Wastewater contained negligible amounts of protein and may be reused. Processing by acid and alkaline extraction are feasible alternatives for obtaining protein isolates either from fresh or frozen squid muscle, which is an important consideration when choosing the most appropriate and inexpensive method to scale up this technology.     

Endocannabinoids potentiate synaptic transmission through stimulation of astrocytes

Endocannabinoids and their receptor CB1 play key roles in brain function. Astrocytes express CB1Rs that are activated by endocannabinoids released by neurons. However, the consequences of the endocannabinoid-mediated neuron-astrocyte signaling on synaptic transmission are unknown. We show that endocannabinoids released by hippocampal pyramidal neurons increase the probability of transmitter release at CA3-CA1 synapses. This synaptic potentiation is due to CB1R-induced Ca(2+) elevations in astrocytes, which stimulate the release of glutamate that activates presynaptic metabotropic glutamate receptors. While endocannabinoids induce synaptic depression in the stimulated neuron by direct activation of presynaptic CB1Rs, they indirectly lead to synaptic potentiation in relatively more distant neurons by activation of CB1Rs in astrocytes. Hence, astrocyte calcium signal evoked by endogenous stimuli (neuron-released endocannabinoids) modulates synaptic transmission. Therefore, astrocytes respond to endocannabinoids that then potentiate synaptic transmission, indicating that astrocytes are actively involved in brain physiology.     

Evaluation of properties of several cheese-ripening fungi for potential biotechnological applications

Strains of Penicillium camemberti and Penicillium roqueforti were tested for properties that could be important for future biotechnological applications of these fungi. Penicillium camemberti CECT 2267 and P. roqueforti NRRL 849 showed the most promising performances in terms of growth, protoplast production, and protoplast regeneration abilities. Transformation of these strains with a plasmid containing gene encoding phleomycin resistance showed that they also have a high transformation frequency. In addition, both strains showed low extracellular proteolytic activity. Thus, these strains have all the characteristics to make them suitable for future genetic improvement, recombinant protein production, and other potential biotechnological applications.     

Hairpin RNA derived from viral NIa gene confers immunity to wheat streak mosaic virus infection in transgenic wheat plants

Wheat streak mosaic virus (WSMV), vectored by Wheat curl mite, has been of great economic importance in the Great Plains of the United States and Canada. Recently, the virus has been identified in Australia, where it has spread quickly to all major wheat growing areas. The difficulties in finding adequate natural resistance in wheat prompted us to develop transgenic resistance based on RNA interference (RNAi). An RNAi construct was designed to target the nuclear inclusion protein ‘a’ (NIa) gene of WSMV. Wheat was stably cotransformed with two plasmids: pStargate‐NIa expressing hairpin RNA (hpRNA) including WSMV sequence and pCMneoSTLS2 with the nptII selectable marker. When T₁ progeny were assayed against WSMV, ten of sixteen families showed complete resistance in transgenic segregants. The resistance was classified as immunity by four criteria: no disease symptoms were produced; ELISA readings were as in uninoculated plants; viral sequences could not be detected by RT‐PCR from leaf extracts; and leaf extracts failed to give infections in susceptible plants when used in test‐inoculation experiments. Southern blot hybridization analysis indicated hpRNA transgene integrated into the wheat genome. Moreover, accumulation of small RNAs derived from the hpRNA transgene sequence positively correlated with immunity. We also showed that the selectable marker gene nptII segregated independently of the hpRNA transgene in some transgenics, and therefore demonstrated that it is possible using these techniques, to produce marker‐free WSMV immune transgenic plants. This is the first report of immunity in wheat to WSMV using a spliceable intron hpRNA strategy.     

A Polarization-Independent SPR Fiber Sensor

The resonance of surface plasma waves in metallic layers is a strongly polarization-dependent phenomenon by the very nature of the physical effect responsible of that resonance. This implies the necessity of polarization-controlling elements to be added to any operative surface-plasmon-resonance-based sensor. A fully symmetrical, circular-section double deposition of a metallic and a dielectric layer on a uniform-waist tapered optical fiber (SymDL-UWT) permits us to completely eliminate the dependence on polarization of the plasmon excitation, with the corresponding operative advantages and basic theoretical consequences. We depict the fabrication process of these transducers, which is based on the use of a simple and efficient rotating element developed by us, and show the characteristics of the produced devices. No such device has been depicted up to date. As our experimental results show, this kind of devices can be considered a very good option for the development of simple, compact, and efficient chemical and biological sensors.     

2-DE based proteomic analysis of Saccharomyces cerevisiae wild and K+ transport-affected mutant (trk1,2) strains at the growth exponential and stationary phases

By using a 2-DE based workflow, the proteome of wild and potassium transport mutant trk1,2 under optimal growth potassium concentration (50 mM) has been analyzed. At the exponential and stationary phases, both strains showed similar growth, morphology potassium content, and Vmax of rubidium transport, the only difference found being the Km values for this potassium analogue transport, higher for the mutant (20 mM) than for the wild (3–6 mM) cells.Proteins were buffer-extracted, precipitated, solubilized, quantified, and subjected to 2-DE analysis in the 5–8 pH range. More differences in protein content (37–64 mg g^? 1 cell dry weight) and number of resolved spots (178–307) were found between growth phases than between strains. In all, 164 spots showed no differences between samples and a total of 105 were considered to be differential after ANOVA test. 171 proteins, corresponding to 71 unique gene products have been identified, this set being dominated by cytosolic species and glycolitic enzymes. The ranking of the more abundant spots revealed no differences between samples and indicated fermentative metabolism, and active cell wall biosynthesis, redox homeostasis, biosynthesis of amino acids, coenzymes, nucleotides, and RNA, and protein turnover, apart from cell division and growth. PCA analysis allowed the separation of growth phases (PC1 and 2) and strains at the stationary phase (PC3 and 4), but not at the exponential one. These results are also supported by clustering analysis. As a general tendency, a number of spots newly appeared at the stationary phase in wild type, and to a lesser extent, in the mutant. These up-accumulated spots corresponded to glycolitic enzymes, indicating a more active glucose catabolism, accompanied by an accumulation of methylglyoxal detoxification, and redox-homeostasis enzymes. Also, more extensive proteolysis was observed at the stationary phase with this resulting in an accumulation of low Mr protein species.     

Combined phospholipid biomarker-16S rRNA gene denaturing gradient gel electrophoresis analysis of bacterial diversity and physiological status in an intertidal microbial mat

A combined lipid biomarker-16S rRNA gene denaturing gradient gel electrophoresis analysis was used to monitor changes in the physiological status, biomass, and microbial composition of a microbial mat. In the morning hours, an increase in the biomass of layers containing a high density of phototrophs and a decrease in the growth rate in the deep layers were observed. The combined approach also revealed differences in major groups of microorganisms, including green nonsulfur, gram-positive, and heterotrophic bacteria.     

Effect of caffeine on in vivo processing of alkylated bases in proliferating plant cells

DNA damage was induced by either 2 mM ethylmethanesulfonate or 1 Gy of gamma-irradiation in Allium cepa L. root meristems. The percentage of DNA that migrated towards the anode during microelectrophoresis after alkali denaturation (pH approximately 13.5) of the isolated nuclei (comet assay) reflects the amount of single strand breaks present in them. There was some DNA migration (12.8+/-2.4%) in untreated roots. This percentage doubled at the end of 1.5 h treatment with the mono-functional alkylating agent 2 mM ethylmethanesulfonate, and trebled after a single exposure to 1 Gy of gamma-rays. A proportion of the DNA migration caused by these two treatments was reversed (repaired) by a 2 h long period of in vivo recovery. However, when 5 mM caffeine was applied after removal of the alkylating agent, the amount of DNA migrating to the comet tail over the same 2 h period was almost double that at the onset of recovery. In both control and irradiated nuclei, caffeine also increased the initial level of DNA migration in the comet assay, but to a lesser extent. These results indicate that caffeine increases the DNA damage that accumulates during the processing of alkylated bases and, to a lesser extent, of the DNA bases damaged by gamma-irradiation. Thus, the potentiation effect of caffeine on induced chromosomal damage may not just be due to caffeine-induced cancellation of the G2 checkpoint, but also to a direct effect this methylxantine has on the processing of DNA damage.