Phenology of cryptomonads and the CRY1 lineage in a coastal brackish lagoon (Vistula Lagoon,Baltic Sea)

Cryptomonadales have acquired their plastids by secondary endosymbiosis. A novel clade—CRY1—has been discovered at the base of the Cryptomonadales tree, but it remains unknown whether it contains plastids. Cryptomonadales are also an important component of phytoplankton assemblages. However, they cannot be readily identified in fixed samples, and knowledge on dynamics and distribution of specific taxa is scarce. We investigated the phenology of the CRY1 lineage, three cryptomonadales clades and a species Proteomonas sulcata in a brackish lagoon of the Baltic Sea (salinity 0.3–3.9) using fluorescence in situ hybridization. A newly design probe revealed that specimens of the CRY1 lineage were aplastidic. This adds evidence against the chromalveolate hypothesis, and suggests that the evolution of cryptomonadales’ plastids might have been shorter than is currently assumed. The CRY1 lineage was the most abundant cryptomonad clade in the lagoon. All of the studied cryptomonads peaked in spring at the most freshwater station, except for P. sulcata that peaked in summer and autumn. Salinity and concentration of dissolved inorganic nitrogen most significantly affected their distribution and dynamics. Our findings contribute to the ecology and evolution of cryptomonads, and may advance understanding of evolutionary relationships within the eukaryotic tree of life.     

Targeted gene exchange in plant cells mediated by a zinc finger nuclease double cut

Genome modification by homology‐directed repair (HDR) is an attractive tool for the controlled genetic manipulation of plants. Here, we report the HDR‐mediated gene exchange of expression cassettes in tobacco BY‐2 cells using a designed zinc finger nuclease (ZFN). The target contained a 7‐kb fragment flanked by two ZFN cutting sites. That fragment was replaced with a 4‐kb donor cassette, which integrates gene markers for selection (kanamycin resistance) and for scoring targeting (red fluorescent protein, RFP). Candidates resulting from cassette exchange were identified by molecular analysis of calli generated by transformation via direct DNA delivery. The precision of HDR‐mediated donor integration was evaluated by Southern blot analysis, sequencing of the integration locus and analysis of RFP fluorescence by flow cytometry. Screening of 1326 kanamycin‐resistant calli yielded 18 HDR events, 16 of which had a perfect cassette exchange at the insert junction and 13 of which produced functional RFP. Our results demonstrate that ZFN‐based HDR can be used for high frequency, precise, targeted exchange of fragments of sizes that are commercially relevant in plants.     

A new mode of SAM domain mediated oligomerization observed in the CASKIN2 neuronal scaffolding protein

Background

CASKIN2 is a homolog of CASKIN1, a scaffolding protein that participates in a signaling network with CASK (calcium/calmodulin-dependent serine kinase). Despite a high level of homology between CASKIN2 and CASKIN1, CASKIN2 cannot bind CASK due to the absence of a CASK Interaction Domain and consequently, may have evolved undiscovered structural and functional distinctions.

Results

We demonstrate that the crystal structure of the Sterile Alpha Motif (SAM) domain tandem (SAM1-SAM2) oligomer from CASKIN2 is different than CASKIN1, with the minimal repeating unit being a dimer, rather than a monomer. Analytical ultracentrifugation sedimentation velocity methods revealed differences in monomer/dimer equilibria across a range of concentrations and ionic strengths for the wild type CASKIN2 SAM tandem and a structure-directed double mutant that could not oligomerize. Further distinguishing CASKIN2 from CASKIN1, EGFP-tagged SAM tandem proteins expressed in Neuro2a cells produced punctae that were distinct both in shape and size.

Conclusions

This study illustrates a new way in which neuronal SAM domains can assemble into large macromolecular assemblies that might concentrate and amplify synaptic responses.

     

Effects of Heart Rate Variability Biofeedback on EEG Alpha Asymmetry and Anxiety Symptoms in Male Athletes: A Pilot Study

Heart rate variability biofeedback (HRV-BFB) has been shown as useful tool to manage stress in various populations. The present study was designed to investigate whether the biofeedback-based stress management tool consisting of rhythmic breathing, actively self-generated positive emotions and a portable biofeedback device induce changes in athletes’ HRV, EEG patterns, and self-reported anxiety and self-esteem. The study involved 41 healthy male athletes, aged 16–21 (mean 18.34 ± 1.36) years. Participants were randomly divided into two groups: biofeedback and control. Athletes in the biofeedback group received HRV biofeedback training, athletes in the control group didn’t receive any intervention. During the randomized controlled trial (days 0–21), the mean anxiety score declined significantly for the intervention group (change-4 p < 0.001) but not for the control group (p = 0.817). In addition, as compared to the control, athletes in biofeedback group showed substantial and statistically significant improvement in heart rate variability indices and changes in power spectra of both theta and alpha brain waves, and alpha asymmetry. These changes suggest better self-control in the central nervous system and better flexibility of the autonomic nervous system in the group that received biofeedback training. A HRV biofeedback-based stress management tool may be beneficial for stress reduction for young male athletes.     

Plant genome editing in the European Union—to be or not to be—a GMO

New plant-breeding techniques have been boosting plant breeding, since only a few years but already first promising products are pushing to the market. In contrast to this, in many countries, the current Directives regulating genetically modified organisms have been established more than 25 years ago, especially in the European Union being based on clear differentiation between transgenic plants and conventional breeding. Therefore, the question arises if these Directives are suitable to face the new challenge of genetic engineering or if there is a need for updated regulations.     

Some growth responses of apple seedlings to abscisic acid and growth stimulators

Antonovka seedlings were obtained from the embryos soaked in abscisic acid (ABA) alone, gibberellin A₄₊₇, benzyladenine or the mixtures of these regulators. The inhibitory effect of ABA on the growth of the seedlings was of temporary duration. The height of the 12 week-old seedlings was dependent only on the degree of seed after-ripening and not on the ABA treatment. The growth stimulators studied were unable to overcome the inhibitory effect of ABA observed in the early stages of seedling growth. The growth stimulators did not substitute, for cold treatment of apple seeds which is the only treatment so far known to overcome the dwarf conditions of the seedlings obtained from non-after-ripened embryos. ABA greatly suppressed the early stages of apple seedling growth (up to 6 weeks) during which they resembled physiological dwarfs. The highest concentrations of ABA (2·0 μg ml^?1) greatly modified the root system of the seedlings, and produced a larger percentage of seedlings with unbranched roots.     

Suppression of the release of prostaglandin-like substances by hydrocortisone in vivo

Muscular exercise of the dog's hind leg evokes the release of prostaglandin-like substances / PG-like substances/ into femoral venous blood. The release of PG-like substances detected by the bioassay method was significantly greater in adrenalectomized as compared to normal dogs. To test the possibility that this difference may be related to the deficiency of adrenocortical secretion in adrenalectomized dogs, the effect of hydrocortisone / HC / and aldosterone / AS / upon the release of PG-like substances induced by muscular work of the dog's hind leg was investigated. The doses of HC and AS infused intravenously or intraarterially were close to the range of physiological secretion rate of these hormones. HC suppressed the release of PG-like material by 30 to 60%, whereas AS had no effect upon the rate and duration of the release. The rate of removal of exogenous PGE₂ in the hind limb circulation was not influenced by HC, suggesting that the diminution of PG release by HC results from the suppression of PG generation rather than from the enhancement of degradation. It is suggested that inhibitory effect of HC upon the rate of the release of PG-like substances may be related to the membrane-stabilizing properties of this hormone. The difference in the intensity of the release of PG-like substances between normal and adrenalectomized dogs suggests that, at least in some conditions, the release of endogenous PGs from tissues may be influenced by the state of adrenocortical activity.