Ectomycorrhizal diversity at five different tree species in forests of the Taunus Mountains in Central Germany

Ectomycorrhizal fungi were investigated on five different forest tree species growing in pure stands on the south slope of the Taunus Mountains, which are situated at the northern end of the Rhine rift valley in Central Germany. Mycorrhizal fungi accompanying the genus Xerocomus were identified and their frequencies counted. Using ITS markers, 22 different fungal species were identified down to species level and 6 down to genus level. On European beech (Fagus sylvatica) 16 fungal species and 4 genera were identified and on Sessile oak (Quercus petraea) 16 ectomycorrhizal species and 2 genera were determined. On both deciduous trees we observed exclusively: Cortinarius subsertipes, Genea hispidula, Lactarius quietus, Tylopilus felleus and a Melanogaster genus. On Norway spruce (Picea abies) we identified 13 different mycorrhizal species and 3 different genera, on Silver fir (Abies alba) 12 species and 3 genera, and in association with European larch (Larix decidua) 11 species and 3 genera. On these conifers Cortinarius anomalus, Lactarius necator and a Piloderma genus occurred exclusively. Comparisons with published data of ectomycorrhizal diversity on the same five tree species, growing in different areas of Germany and Europe, led to the conclusion that there is relative site specificity for ectomycorrhizal communities. Upper soil compartments of the stands investigated in the Taunus Mountainssuffer from soil acidification (pH-H20 ~3.7 to ~4.8). However, a clear correlation between upper soil pH-values and fungal diversity was not observed. On the other hand, nitrate concentrations in upper soil compartments (~26 to ~91 kgNO3-/ha) were higher in older stands as compared to younger ones. Higher nitrate concentrations in upper soils correlated with lower numbers of mycorrhizal individuals.

Isolation, Expression and Characterization of rbc L Gene from Ulva prolifera J. Agardh(Ulvophyceae, Chlorophyta)

Ulva prolifera is a typical green alga in subtidal areas and can grow tremendously fast. A highly efficient Rubisco enzyme which is encoded by Up Rbc L gene may contribute to the rapid growth. In this study, the full-length Up Rbc L open reading frame(ORF) was identified, which encoded a protein of 474 amino acids. Phylogenetic analysis of Up Rbc L sequences revealed that Chlorophyta had a closer genetic relationship with higher plants than with Rhodophyta and Phaeophyta. The two distinct residues(aa11 and aa91) were presumed to be unique for Rubisco catalytic activity. The predicted three-dimensional structure showed that one α/β-barrel existed in the C-terminal region, and the sites for Mg^(2+)coordination and CO_2 fixation were also located in this region. Gene expression profile indicated that Up Rbc L was expressed at a higher level under light exposure than in darkness. When the culture temperature reached 35℃, the expression level of Up Rbc L was 2.5-fold lower than at 15℃, and the carboxylase activity exhibited 13.8-fold decrease. Up Rbc L was heterologously expressed in E. coli and was purified by Ni^(2+) affinity chromatography. The physiological and biochemical characterization of recombinant Rubisco will be explored in the future.

The Role of Phosphorylation in Redox Regulation of Photosynthesis Genes psaA and psbA during Photosynthetic Acclimation of Mustard

The long-term response （LTR） to light-quality gradients improves performance and survival of plants in dense stands. It involves redox-controlled transcriptional regulation of the plastome-encoded genes psaAB （encoding the P700 apoproteins of photosystem I） and psbA （encoding the D1 protein of photosystem II） and requires the action of plastidlocalized kinases. To study the potential impact of phosphorylation events on plastid gene expression during the LTR, we analyzed mustard seedlings acclimated to light sources favoring either photosystem I or photosystem II. Primer extension analyses of psaA transcripts indicate that the redox regulation occurs at the principal bacterial promoters, suggesting that the plastid encoded RNA polymerase （PEP） is the target for redox signals. Chloroplast protein fractions containing PEP and other DNA-binding proteins were purified from mustard via heparin-Sepharose chromatography. The biochemical properties of these fractions were analyzed with special emphasis on promoter recognition and specificity, phosphorylation state, and kinase activity. The results demonstrate that the LTR involves the action of small DNA-binding proteins; three of them exhibit specific changes in the phosphorylation state. Auto-phosphorylation assays, in addition, exhibit large differences in the activity of endogenous kinase activities. Chloroplast run-on transcription experiments with the kinase inhibitor H7 and the reductant DTT indicate that phosphorylation events are essential for the mediation of redox signals toward psaA and psbA transcription initiation, but require the synergistic action of a thiol redox signal. The data support the idea that redox signals from the thylakoid membrane are linked to gene expression via phosphorylation events; however, this mediation appears to require a complex network of interacting proteins rather than a simple phosphorelay.

Protein Disulfide Isomerase 2 of Chlamydomonas reinhardtii Is Involved in Circadian Rhythm Regulation

Protein disulfide isomerases （PDIs） are known to play important roles in the folding of nascent proteins and in the formation of disulfide bonds. Recently, we identified a PDI from Chlamydomonas reinhardtii （CrPDI2） by a mass spectrometry approach that is specifically enriched by heparin affinity chromatography in samples taken during the night phase. Here, we show that the recombinant CrPDI2 is a redox-active protein. It is reduced by thioredoxin reductase and catalyzes itself the reduction of insulin chains and the oxidative refolding of scrambled RNase A. By immunoblots, we confirm a high-amplitude change in abundance of the heparin-bound CrPDI2 during subjective night. Interestingly, we find that CrPDI2 is present in protein complexes of different sizes at both day and night. Among three identified interac- tion partners, one （a 2-cys peroxiredoxin） is present only during the night phase. To study a potential function of CrPDI2 within the circadian system, we have overexpressed its gene. Two transgenic lines were used to measure the rhythm of phototaxis~ In the transgenic strains, a change in the acrophase was observed. This indicates that CrPDI2 is involved in the circadian signaling pathway and, together with the night phase-specific interaction of CrPDI2 and a peroxiredoxin, these findings suggest a close coupling of redox processes and the circadian clock in C. reinhardtii.

Identification of Early Nuclear Target Genes of Plastidial Redox Signals that Trigger the Long- Term Response of Arabidopsis to Light Quality Sh∽ts

Natural illumination conditions are highly variable and because of their sessile life style, plants are forced to acclimate to them at the cellular and molecular level. Changes in light intensity or quality induce changes in the reduction/oxidation （redox） state of the photosynthetic electron chain that acts as a trigger for compen- satory acclimation responses comprising functional and structural adjustments of photosynthesis and metabolism. Such responses include redox-controlled changes in plant gene expression in the nucleus and organelles. Here we describe a strategy for the identification of early redox-regulated genes （ERGs） in the nucleus of the model organism Arabidopsis thaliana that respond significantly 30 or 60 min after the generation of a reduction signal in the photosynthetic electron transport chain. By comparing the response of wild-type plants with that of the acclimation mutant stn7, we could specifically identify ERGs. The results reveal a significant impact of chloroplast redox signals on distinct nuclear gene groups including genes for the mitochondrial electron transport chain, tetrapyrrole biosynthesis, carbohydrate metabolism, and signaling lipid synthesis. These expression profiles are clearly different from those observed in response to the reduction of photosynthetic electron transport by high light treatments. Thus, the ERGs identified are unique to redox imbalances in photosynthetic electron transport and were then used for analyzing potential redox-responsive cis-elements, trans-factors, and chromosomal regulatory hot spots. The data identify a novel redox-responsive element and indicate extensive redox control at transcriptional and chromosomal levels that point to an unprecedented impact of redox signals on epigenetic processes.

The Heme-Binding Protein SOUL3 of Chlamydomonas reinhardtii Influences Size and Position of the Eyespot

The flagellated green alga Chlamydomonas reinhardtii has a primitive visual system, the eyespot. It is situated at the cells equator and allows the cell to phototax. In a previous proteomic analysis of the eyespot, the SOUL3 protein was identified among 202 proteins. Here, we investigate the properties and functions of SOUL3. Heterologously expressed SOUL3 is able to bind specifically to hemin. In C. reinhardtii, SOUL3 is expressed at a constant level over the diurnal cycle, but forms protein complexes that differ in size during day and night phases. SOUL3 is primarily localized in the eyespot and it is situated in the pigment globule layer thereof. This is in contrast to the channelrhodopsin photoreceptors, which are localized in the plasma membrane region of the eyespot. Knockdown lines with a significantly reduced SOUL3 level are characterized by mislocalized eyespots, a decreased eyespot size, and alterations in phototactic behavior. Mislocalizations were either anterior or posterior and did not affect association with acetylated microtubules of the daughter four-membered rootlet. Our data suggest that SOUL3 is involved in the organization and placement of the eyespot within the cell.

Telling Duckweed Apart:Genotyping Technologies for the Lemnaceae

The family of Lemnaceae,commonly called duckweed,comprise five genera and 37 species of aquaticmonocotyledonous plants that are endemic to lakes,ponds and brackish water bodies on most continents. They are angiospermsthat have the highest biomass production rate and have been used to remediate wastewater from both municipal andindustrial sources. With these attractive features,duckweed is poised to be a novel agricultural platform that can complementcurrent terrestrial crops. In addition,their remarkable ability to adapt to diverse climates and conditions may provide newunderstanding of stress tolerance and genome adaptability mechanisms. However,one of the first challenges facing theseapplications is to establish reliable and rapid methods for identification of closely related strains that may have very differentbehaviour or properties. This review article summarizes the state of the arts for this endeavour and provides the outlook for thisquest in the near future.