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The diversification of plant cytosolic small heat shock proteins preceded the divergence of mosses.

A cDNA library was constructed with mRNA isolated from heat-stressed cell cultures of Funaria hygrometrica (Bryophyta, Musci, Funariaceae). cDNA clones encoding six cytosolic small heat shock proteins (sHSPs) were identified using differential screening. Phylogenetic analysis of these sHSP sequences with other known sHSPs identified them as members of the previously described higher plant cytosolic class I and II families. Four of the F. hygrometrica sHSPs are members of the cytosolic class I family, and the other two are members of the cytosolic class II family. The presence of members of the cytosolic I and II sHSP families in a bryophyte indicates that these gene families are ancient, and evolved at least 450 MYA. This result also indicates that the plant sHSP gene families duplicated much earlier than did the well-studied phytochrome gene family. Members of the cytosolic I and II sHSP families are developmentally regulated in seeds and flowers in higher plants. Our findings show that the two cytosolic sHSP families evolved before the appearance of these specialized structures. Previous analysis of angiosperm sHSPs had identified class- or family-specific amino acid consensus regions and determined that rate heterogeneity exists among the different sHSP families. The analysis of the F. hygrometrica sHSP sequences reveals patterns and rates of evolution distinct from those seen among angiosperm sHSPs. Some, but not all, of the amino acid consensus regions identified in seed plants are conserved in the F. hygrometrica sHSPs. Taken together, the results of this study illuminate the evolution of the sHSP gene families and illustrate the importance of including representatives of basal land plant lineages in plant molecular evolutionary studies.

Amino Acid Sequence↗

Downstream effects of a hydroelectric reservoir on aquatic plant assemblages.

Macrophytes were studied downstream of the Rophémel hydroelectric dam on the River Rance (Côtes d'Armor Department, western France) to assess the effects of hydroelectric functioning on river macrophyte communities. We studied ten representative sections of the hydro-peaking channel on five occasions in 1995 and 1996, on a 15-km stretch of river. Floristic surveys were carried out on sections 50 m in length, and genera of macroalgae, species of bryophyta, hydrophytes, and emergent rhizophytes were identified. For the aquatic bryophytes and spermatophytes section of our study, we compared our results with 19th century floristic surveys, before the dam was built. During the vegetative growth period, the hydro-peaking frequency was low. The plant richness was highest near the dam. The macrophyte communities were highly modified according to the distance to the dam. The frequency and magnitude of hydro-peaking was related to the aquatic macrophyte richness in an Intermediate Disturbance Hypothesis position. However, the results of the eco-historical comparison with 19th century floristic surveys point to the original nature of the flora found at the site. Some floral patterns, seen during both periods and at an interval of 133 years, were indicative of the ubiquity of the aquatic flora and of the plants" adaptability. This demonstrates the importance of taking river basin history into account in such biological surveys.

Biodiversity↗

Survey of the taxonomic and tissue distribution of microsomal binding sites for the non-host selective fungal phytotoxin, fusicoccin.

The recent identification of the fusicoccin-binding protein (FCBP) in plasma membranes from monocotyledonous and dicotyledonous angiosperms has opened the basis for an elucidation of the toxin's mechanism(s) of action and indicated a widespread occurrence of the FCBP in plants. Results of a detailed taxonomic survey of fusicoccin-binding sites are reported. Binding sites were not found in prokaryotes, animal tissues, fungi and algae including the most direct extant ancestors of the land plants (Coleochaete). From the Psilotales (Psilophytatae) to the monocotyledonous angiosperms, all taxa analyzed possessed high-affinity microsomal fusicoccin-binding sites. A heterogeneous picture emerged for the Bryophyta. Anthoceros crispulus (Anthocerotae), the only hornwort available to study, lacked fusicoccin binding. Within the Hepaticae as well as the Musci, species lacking and species exhibiting toxin binding were found. The binding site thus seems to have emerged very early in the evolution of the land plants. The tissue distribution of fusicoccin-binding sites was studied in Vicia faba L. shoots. All tissues analyzed showed fusicoccin binding, although not to the same extent. On a per-cell basis, guard cells were found to contain, compared to mesophyll cells, a nine-fold higher number of binding sites. Based on cell surface area, the site density is by a factor of 32 higher in guard cells than in mesophyll cells. Tissue specific expression of the binding sites is suggested by these findings.

Animals↗

The leaves of green plants as well as a cyanobacterium, a red alga, and fungi contain insulin-like antigens.

We report the detection of insulin-like antigens in a large range of species utilizing a modified ELISA plate assay and Western blotting. We tested the leaves or aerial parts of species of Rhodophyta (red alga), Bryophyta (mosses), Psilophyta (whisk ferns), Lycopodophyta (club mosses), Sphenopsida (horsetails), gymnosperms, and angiosperms, including monocots and dicots. We also studied species of fungi and a cyanobacterium, Spirulina maxima. The wide distribution of insulin-like antigens, which in some cases present the same electrophoretic mobility as bovine insulin, together with results recently published by us on the amino acid sequence of an insulin isolated from the seed coat of jack bean (Canavalia ensiformis) and from the developing fruits of cowpea (Vigna unguiculata), suggests that pathways depending on this hormone have been conserved through evolution.

Animals↗

Seasonal variations of diacylglyceryl-N,N,N-trimethylhomoserine content in Polypodiophyta.

Diacylglyceryl-N,N,N-trimethylhomoserine (DGTS) belongs to a family of three known betaine lipids. It is synthesized by a number of bacteria, fungi, brown, red and green algae, in addition to lichens, and is also found in higher plants Bryophyta, Equisetophyta and Polydiophyta. It has been determined that DGTS plants can be DGTS-positive or DGTS-negative. The purpose of the present study was to examine the leaves of the ferns Drytopteris filix-mas (L.) Tod. and Pteridium aquilinum (L.) Kuhn, collected at different time periods during a single vegetation season for their DGTS contents. The results obtained show that amounts of DGTS in all of the species examined varies relative to the time periods in which they were collected.

Cycadopsida↗

Antifungal constituents from the Chinese moss Homalia trichomanoides.

Bioautographic assay on TLC plates was adopted to guide the fractionation of the Et2O extract of Homalia trichomanoides (Hedw.) B. S. G., which led to the isolation of the novel p-terphenyl derivative trichomanin (= 4,4''-dihydroxy-1,1':4',1''-terphenyl-2',3',5',6'-tetrayl tetrakis(phenylacetate); 1), together with five known compounds: 3alpha-methoxyserrat-14-en-21beta-ol (2), 3beta-methoxyserrat-14-en-21beta-ol (3), 3beta-methoxyserrat-14-en-21-one (4), atranorin (5), and methyl 2,4-dihydroxy-3,6-dimethylbenzoate (6). Their structures were determined on the basis of spectral data (1D- and 2D-NMR, MS), X-ray crystallographic analysis, and chemical transformation. Compounds 3, 5, and 6 exhibited antifungal activity against Candida albicans, with minimum inhibitory doses (MID) of 2.0, 2.0, and 0.6 microg, respectively.

Antifungal Agents↗

"In-gel patch electrophoresis:" a new method for environmental DNA purification.

Most of the microorganism species are largely untapped and could represent an interesting reservoir of genes useful for biotechnological applications. Unfortunately, a major difficulty associated with the methods used to isolate environmental DNA is related to the contamination of the extracted material with humic substances. These polyphenolic compounds inhibit the DNA processing reactions and severely impede cloning procedures. In this work, we describe a rapid, simple, and efficient method for the purification of genomic DNA from environmental samples: we added a chromatography step directly embedded into an agarose gel electrophoresis. This strategy enabled the DNA extraction from various environmental samples and it appeared that the purity grade was compatible with digestion by restriction enzymes and polymerase chain reaction (PCR) amplifications.

Bryophyta↗

A liquid chromatography-mass spectrometric method for the determination of oak moss allergens atranol and chloroatranol in perfumes.

This paper describes a validated liquid chromatographic-tandem mass spectrometric method for quantitative analysis of the potential oak moss allergens atranol and chloroatranol in perfumes and similar products. The method employs LC-MS-MS with electrospray ionization (ESI) in negative mode. The compounds are analysed by selective reaction monitoring (SRM) of 2 or 3 ions for each compound in order to obtain high selectivity and sensitivity. The method has been validated for the following parameters: linearity; repeatability; recovery; limit of detection; and limit of quantification. The limits of detection, 5.0 ng/mL and 2.4 ng/mL, respectively, for atranol and chloroatranol, achieved by this method allowed identification of these compounds at concentrations below those causing allergic skin reactions in oak-moss-sensitive patients. The recovery of chloratranol from spiked perfumes was 96+/-4%. Low recoveries (49+/-5%) were observed for atranol in spiked perfumes, indicating ion suppression caused by matrix components. The method has been applied to the analysis of 10 randomly selected perfumes and similar products.

Allergens↗

Extraction of caesium from mosses and incorporation into an oxalate crystalline lattice.

The ratios of stable and radioactive isotopes in the environment are not constant and their effects on living organisms are not the same. Whilst radionuclides can be monitored by measuring their radioactivity levels, it is also important to determine the stable isotopes. The method described in this paper represents a simple and cheap process by which to extract caesium from moss, without significant destruction of the plant, and to incorporate the isolated caesium into a water-soluble crystal. Ammonium oxalate and phosphoric acid are water-soluble substances at 5% concentration but, when the solutions are combined, crystallisation occurs. Crystals were estimated to contain 68.30% O, 20.98% C, 4.68% H, 6.04% N and less than 0.01% P, corresponding to NH4HC2O4 x H2C2O4 x 2H2O (C4H11NO10). The crystal mass did not vary with the species of moss, although the effectiveness of caesium extraction did. Typically, 73.0 +/- 6.1% of the solubilised caesium was entrapped within the crystals.

Bryophyta↗

Using delta15N values to characterise the nitrogen nutrient pathways from intensive animal units.

Previous studies on foliar delta15N values, in certain bryophytes, have indicated signature similarities to source pollutants. The object of this study was to investigate the effect further, by examining the mechanisms whereby isotopic fractionation occurs in systems such as atmospheric ammonia (NH3), throughfall, vegetation and soil. Measurements taken in and around point emission sources will then be used to characterise the various fractionation effects associated with these N transformations, as well as to demonstrate some of the issues associated with using delta15N values as pollution indicators. The atmospheric dispersion model UK-ADMS has also been used to model atmospheric delta15NH3 emissions, with signatures exhibiting marked negative shifts immediately downwind of an agricultural NH3 source. Similar dispersion patterns were mapped for NH3 concentration data illustrating the link between these two forms of measurement.

Air Pollutants↗

Moss-associated methylobacteria as phytosymbionts: an experimental study.

Methylotrophic bacteria inhabit the surface of plant organs, but the interaction between these microbes and their host cells is largely unknown. Protonemata (gametophytes) of the moss Funaria hygrometrica were cultivated in vitro under axenic conditions and the growth of the protonemal filaments recorded. In the presence of methylobacteria (different strains of Methylobacterium), average cell length and the number of cells per filament were both enhanced. We tested the hypothesis that auxin (indole-3-acetic acid, IAA), secreted by the epiphytic bacteria and taken up by the plant cells, may in part be responsible for this promotion of protonema development. The antiauxin parachlorophenoxyisobutyric acid (PCIB) was used as a tool to analyze the role of IAA and methylobacteria in the regulation of cell growth. In the presence of PCIB, cell elongation and protonema differentiation were both inhibited. This effect was compensated for by the addition of different Methylobacterium strains to the culture medium. Biosynthesis and secretion of IAA by methylobacteria maintained in liquid culture was documented via a colorimetric assay and thin layer chromatography. Our results support the hypothesis that the development of Funaria protonemata is promoted by beneficial phytohormone-producing methylobacteria, which can be classified as phytosymbionts.

Bryophyta↗

Microhabitat selection in the simple oribatid community dwelling in epilithic moss cover (Acari: Oribatida).

The moss cover of a roof was studied as the model of a simple habitat divided into microhabitats by the members of a community of saprophagous mites. This community consisted of two species of oribatid mites: Scutovertex minutus and Trichoribates trimaculatus. They were extracted from moss onto moist paper, and subsequently, their mobility, responses to moisture fluctuation, and food selection were tested in laboratory experiments. For the nutritional biology, the microanatomy of their alimentary tract was examined according to the system of histological characteristics formulated in the laboratory of the author (type of food, digestive activity of gut walls, etc.). The paraplast sections of the mites were stained by Masson triple stain for these purposes. Moreover, the enzymological tests (chitinase and cellulase activities) were performed to detail the digestive processes. Such an approach was applied to field-sampled specimens as well as to those in the laboratory experiments. These above-mentioned data were discussed with respect to microhabitat selection, vertical and horizontal distribution, and dispersal ability of these two oribatid mites sharing this habitat. These two species differ in several characteristics from each other and these differences resulted in their different microhabitat choices and role in the habitat as a whole.

Acari↗

Accuracy and underlying mechanisms of shifting movements in cellists.

Accuracy of shifting movements between two notes was examined in nine cellists (intermediate-professional skill levels). Three pairs of notes separated by different distances were tested under the same movement rate. Finger position on the string was measured by a circuit. Angular velocities of the left upper arm and forearm were measured by two angular velocity sensors; thus elbow angular velocity during shifts was estimated. Results showed that with increased elbow velocity and shifting distance endpoint variability stayed constant. The force of gravity assisted elbow extension during shifts toward higher pitched notes compared to flexion towards lower pitched notes, but faster movement velocity did not result in increased landing variability. Performance for note E on the A string was found to be less variable than other notes, suggesting that physical cue from the cello body geometry was used as a landmark for finger position. Cutaneous feedback from the thumb when hitting the body-neck junction enabled faster elbow extension velocity compared to shifts towards other notes. Cellists who showed higher performance accuracy also showed higher perceptual ability and performance proficiency. These results suggest that long-term over-training of fast and accurate movements enables musicians to maintain accuracy and variability across different movement distances and velocities. Higher perceptual ability and performance proficiency are correlated with increased accuracy but not lower variability, indicating although perceptual ability and performance proficiency are important for pitch accuracy, movement variability is still constrained by the capacity of the motor system, which is highly fine-tuned and different than non-musicians.

Analysis of Variance↗

Origin, evolution, and metabolic role of a novel glycolytic GAPDH enzyme recruited by land plant plastids.

NAD-specific glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a cytosolic marker enzyme of eukaryotes (GapC; EC 1.2.1.12). Land plants possess an additional NADP+-dependent enzyme (EC 1.2.1.13) within their chloroplasts which is composed of two subunits, GapA and GapB. Another plastid GAPDH enzyme (GapCp) was recently discovered in gymnosperms and ferns. This novel GapCp is closely related to cytosolic GapC and displays glycolytic NAD+ cosubstrate specificity. Here we show that this new gene GapCp is also present and actively expressed in angiosperms, mosses, and liverworts. Phylogenetic analyses of the available GapC and GapCp sequences suggest that the gene duplication giving rise to GapCp occurred in ancestral charophyte algae. The data are also consistent with a monophyletic origin of charophytes and land plants and further support the view that land plants arose from a Coleochaete-like green alga. Northern hybridizations were employed to study the expression of the genes GapCp, GapC, GapA, and GapB in green and nongreen tissues from pepper (Capsicum annuum). The results demonstrate that GapCp mRNAs are mainly expressed in red pepper fruit and roots, in which the transcript levels of photosynthetic GapA and GapB are downregulated. This suggests that in flowering plants GapCp plays a specific role in glycolytic energy production of nongreen plastids such as chromoplasts and leukoplasts and that angiosperms may be the only land plants where glycolysis is absent in green chloroplasts.

Amino Acid Sequence↗

Diversification of ftsZ during early land plant evolution.

The plastid division proteins FtsZ are encoded by a small nuclear gene family in land plants. Although it has been shown for some of the gene products that they are imported into plastids and function in plastid division, the evolution and function of this gene family and their products remain to be unraveled. Here we present two new ftsZ genes from the moss Physcomitrella patens and compare the genomic structure of members of the two plant ftsZ gene families. Comparison of sequence features and phylogenetic analyses confirm the presence of two clusters of paralogues in land plants and demonstrate that these genes were duplicated before the divergence of mosses, ferns and seed plants.

Amino Acid Sequence↗

Organization and expression of the GSK3/shaggy kinase gene family in the moss Physcomitrella patens suggest early gene multiplication in land plants and an ancestral response to osmotic stress.

GSK3/Shaggy kinases are involved in a wide range of fundamental processes in animal development and metabolism. In angiosperm plants, these kinases are encoded by moderate-sized gene families, which appear to have a complex set of functions. Here, we present the characterization of five members of the GSK3/Shaggy gene family in the bryophyte Physcomitrella patens. The P. patens GSK3/Shaggy kinases (PpSK) are organized in a group of closely related paralogues with respect to their gene sequence and structure. Indeed, a phylogenetic analysis of the GSK3/Shaggy kinase sequences from plants and animals showed that the five PpSK proteins are monophyletic, and closer to subgroups I and IV described in angiosperms. Expression analyses performed by quantitative real-time RT-PCR on a wide range of growing conditions showed that PpSK genes responded only to either desiccation, PEG or sorbitol. As demonstrated by both inductions of marker genes and protonemal cell plasmolyses, these treatments resulted in a hyperosmotic stress. Altogether, these data suggest that (1) GSK3/Shaggy kinase gene multiplication occurred early in plant evolution, before the separation between bryophytes and vascular plants, and (2) both gene loss and duplication occurred in the ancestor of P. patens along with functional gene diversification in angiosperms. However, conservation of the transcriptional responses between Physcomitrella and Arabidopsis suggests the identification of an ancestral response of the GSK3/Shaggy kinases genes to osmotic stress.

Amino Acid Sequence↗