The phytochelatin synthase from Nitella mucronata (Charophyta) plays a role in the homeostatic control of iron(II)/(III).

Although some charophytes (sister group to land plants) have been shown to synthesize phytochelatins (PCs) in response to cadmium (Cd), the functional characterization of their phytochelatin synthase (PCS) is still completely lacking. To investigate the metal response and the presence of PCS in charophytes, we focused on the species Nitella mucronata. A 40 kDa immunoreactive PCS band was revealed in mono-dimensional western blot by using a polyclonal antibody against Arabidopsis thaliana PCS1. In two-dimensional western blot, the putative PCS showed various spots with acidic isoelectric points, presumably originated by post-translational modifications. Given the PCS constitutive expression in N. mucronata, we tested its possible involvement in the homeostasis of metallic micronutrients, using physiological concentrations of iron (Fe) and zinc (Zn), and verified its role in the detoxification of a non-essential metal, such as Cd. Neither in vivo nor in vitro exposure to Zn resulted in PCS activation and PC significant biosynthesis, while Fe(II)/(III) and Cd were able to activate the PCS in vitro, as well as to induce PC accumulation in vivo. While Cd toxicity was evident from electron microscopy observations, the normal morphology of cells and organelles following Fe treatments was preserved. The overall results support a function of PCS and PCs in managing Fe homeostasis in the carophyte N. mucronata.

Growth and Oxidative Stress of Brittlewort (Nitella pseudoflabellata) in Response to Cesium Exposure.

The present study evaluated the impact of cesium ((133)Cs) at four concentrations (0, 0.001, 0.01, and 0.1 mg L(-1)) on growth, concentrations of chlorophyll and carotenoid pigments, and oxidative stress responses in the charophyte, Nitella pseudoflabellata, over 30 days. Oxidative stress was quantified by measuring anti-oxidant enzyme activities and H2O2 content. When compared with the control, significantly elevated activity levels of the anti-oxidative enzymes ascorbic peroxidase, catalase and guaiacol peroxidase were observed at 0.1 mg L(-1) (all p < 0.05), even though the H2O2 level was not significantly elevated. Carotenoid and chlorophyll a and b pigment levels were significantly reduced (all p < 0.05) at Cs exposures of 0.01 and 0.1 mg L(-1). Photosynthetic efficiency (i.e., Fv/Fm) was significantly reduced (p < 0.05) at Cs concentrations ≥0.001 mg L(-1). Significant reduction (p < 0.05) of plant growth (i.e., shoot length) was also observed after 1 week of exposure at Cs concentrations ≥0.001 mg L(-1). Our results suggested that Cs exposure reduced plant growth and affected plant functioning via activating the defense mechanism against oxidative stress in Nitella.

A survey of PPR proteins identifies DYW domains like those of land plant RNA editing factors in diverse eukaryotes.

The pentatricopeptide repeat modules of PPR proteins are key to their sequence-specific binding to RNAs. Gene families encoding PPR proteins are greatly expanded in land plants where hundreds of them participate in RNA maturation, mainly in mitochondria and chloroplasts. Many plant PPR proteins contain additional carboxyterminal domains and have been identified as essential factors for specific events of C-to-U RNA editing, which is abundant in the two endosymbiotic plant organelles. Among those carboxyterminal domain additions to plant PPR proteins, the so-called DYW domain is particularly interesting given its similarity to cytidine deaminases. The frequency of organelle C-to-U RNA editing and the diversity of DYW-type PPR proteins correlate well in plants and both were recently identified outside of land plants, in the protist Naegleria gruberi. Here we present a systematic survey of PPR protein genes and report on the identification of additional DYW-type PPR proteins in the protists Acanthamoeba castellanii, Malawimonas jakobiformis, and Physarum polycephalum. Moreover, DYW domains were also found in basal branches of multi-cellular lineages outside of land plants, including the alga Nitella flexilis and the rotifers Adineta ricciae and Philodina roseola. Intriguingly, the well-characterized and curious patterns of mitochondrial RNA editing in the slime mold Physarum also include examples of C-to-U changes. Finally, we identify candidate sites for mitochondrial RNA editing in Malawimonas, further supporting a link between DYW-type PPR proteins and C-to-U editing, which may have remained hitherto unnoticed in additional eukaryote lineages.

Microtubule-dependent motility and orientation of the cortical endoplasmic reticulum in elongating characean internodal cells.

Motility of the endoplasmic reticulum (ER) is predominantly microtubule- dependent in animal cells but thought to be entirely actomyosin-dependent in plant cells. Using live cell imaging and transmission electron microscopy to examine ER motility and structural organization in giant internodal cells of characean algae, we discovered that at the onset of cell elongation, the cortical ER situated near the plasma membrane formed a tight meshwork of predominantly transverse ER tubules that frequently coaligned with microtubules. Microtubule depolymerization increased mesh size and decreased the dynamics of the cortical ER. In contrast, perturbing the cortical actin array with cytochalasins did not affect the transverse orientation but decreased mesh size and increased ER dynamics. Our data suggest that myosin-dependent ER motility is confined to the ER strands in the streaming endoplasm, while the more sedate cortical ER uses microtubule-based mechanisms for organization and motility during early stages of cell elongation. We show further that the ER has an inherent, NEM-sensitive dynamics which can be altered via interaction with the cytoskeleton and that tubule formation and fusion events are cytoskeleton-independent.

Phycoremediation of Chromium (VI) by Nitella and impact of calcium encrustation.

This article discusses the applicability of the Charophyte, Nitella pseudoflabellata in the remediation of Cr (VI) contaminated waters at different calcifying potentials. Its growth was found to be positively correlated with Ca in water (CaW), but marginally significant in the presence of Cr (VI) in water (CrW). High CaW resulted in calcite encrustation on the plant cell wall. CaW was found to be aiding Cr (VI) fixation in the long run, as this correlated positively with both CaW and CrW. However, Ca interfered with passive Cr (VI) accumulation in live plant matter at low CrW concentrations (1mg/g Cr dry weight of plant. Cr (VI) concentrations greater than 0.4 mg/L were too toxic, showing maximum quantum efficiency of PSII photochemistry (F(v)/F(m)) values<0.63. The opposite was noticed (F(v)/F(m)>0.76) when Cr (VI) was less than 0.2mg/L. Elongation curve patterns based on shoot lengths showed similar scenarios. In all cases high CaW units with calcite encrustation found to be least affected by Cr (VI) toxicity. Optimum remediation was obtained using a combination of high Ca and Cr (VI) in the case of passive (short-term) operation and low Ca and Cr (VI) for active (long-term) operation. Under the passive scenario, plants accumulated above 1.2mg/g Cr dry weight whereas in the active case, accumulation was 0.8 mg/g Cr dry weight. We conclude that Nitella-mediated Cr (VI) remediation is a promising technique within the range and conditions investigated.

[Estimation of efficiency of complex-method intestinal therapy using Nitella flexilis test-object for acute intestinal obstruction of various etiology].

The aim of our investigation was to define the presence of correlation between toxicity of intestinal contents and clinical and laboratory indices in the cases of ileus and peritonitis with the use of complex method of enteral therapy. Experiments have been carried out on internodal cells of Nitella Flexilis water plant. Efficiency of the given method (investigation of toxicity of the intestinal content with the use of test-object Nitella Flexilis) has been studied in 25 patients, who underwent correction of morpho-functional status of small intestine. Results of investigation have shown that changes in toxicity of intestinal content reliably reflect the nature and dynamics of the pathobiologic processes proceeding in GIS in treated patients suffering from acute intestinal impassability. Using the method of correction of morpho-functional status of small intestine decreases the role of "Intestinal factor" in formation of endogenous intoxication, which in turn results in early normalization of clinical and laboratory indices, favored process of early post-operational period, decreased rate of lethality and shortened rehabilitation period.

Membrane potential genesis in Nitella cells, mitochondria, and thylakoids.

The resting membrane potential of Nitella cells shifts in parallel with the change in H+ equilibrium potential, but is not equal to the H+ equilibrium potential. The deviation of the membrane potential from the H+ equilibrium potential depends on the extrusion rate of H+ by the electrogenic H+-pump. The activity of the electrogenic H+-pump was formulated in terms of the change in the free energy of ATP hydrolysis. The deviation of membrane potential from the H+ equilibrium potential induces a passive H+ flow. The passive inward H+ current may be coupled with Cl- uptake. The coupling rate of H+,Cl- co-transport was discussed. The membrane potential of mitochondria was electrochemically formulated in terms of oxidation-reduction H2/H+ half-cells spontaneously formed at the inner and outer boundaries of each trans-membrane electron-conducting pathway. The membrane potential formed by a pair of H2/H+ redox cells is pH-sensitive in its nature, but deviates from the H+ equilibrium potential to an extent that depends on the logarithm of the ratio of H2 concentrations at the inner and outer boundaries. The membrane potential of thylakoids is considered to be primarily due to the electromotive force of photocells embedded in the thylakoid membrane, as far as the anode and cathode of each photocell are in contact with the inner and outer solutions, respectively. The light-induced electronic current yields oxygen at the inner boundary and causes an increase in the H2 pool at the outer boundary of the electron-conducting pathway, which has no shunting plastoquinone chain between these two boundaries.