Testing for endocytosis in plants

Summary. For many years endocytosis has been regarded with great scepsis by plant physiologists. Although now generally accepted, care must still be taken with experiments designed to demonstrate endocytic uptake at the plasma membrane. We have taken a critical look at the various agents which are in use as markers for plant endocytosis, pointing out pitfalls and precautions which should be taken. We also take this opportunity to introduce the tyrphostins – tyrosine kinase inhibitors –, which also seem to prevent endocytosis in plants. Correspondence and reprints: Heidelberg Institute for Plant Sciences, University of Heidelberg, Im Neuenheimer Feld 230, 69120 Heidelberg, Federal Republic of Germany.     

The presence of a haloarchaeal type tyrosyl-tRNA synthetase marks the opisthokonts as monophyletic

Lateral gene transfer plays an important role in the evolution of life. Events of ancient gene transfer can transmit genetic novelties to descendent lineages and subsequently shape their genetic systems. We here present the analyses of the gene encoding tyrosyl-tRNA synthetase (tyrRS), which reveal two eukaryotic tyrRS lineages, one including the opisthokonts and the other the remaining eukaryotes. The different origins of tyrRS lineages between the opisthokonts and the remaining eukaryotes indicate a likely case of ancient lateral gene transfer of tyrRS from an archaeon to the opisthokonts, which lends further support for the monophyly of the latter group. Ancient paralogy followed by differential gene loss is an alternative, albeit less parsimonious explanation for the distribution of the two eukaryotic tyrRS types. In either case, the presence of a haloarchaeal tyrRS type in the opisthokonts marks this group as monophyletic. This finding also points to the potential utility of ancient gene transfer events as molecular markers for major organismal lineages.     

Enzymatic Biobleaching of Two Recalcitrant Paper Dyes with Horseradish and Soybean Peroxidase

A stilbene dye (Direct Yellow 11) and a methine dye, Basazol 46L, recalcitrant to common chemical bleaches, were treated with horseradish and soybean peroxidases. Both enzymes were effective at chromophore removal. When compared to laccase in combination with a mediator (ABTS), soybean peroxidase was more effective at oxidative dye removal, especially for the methine dye.     

Bone loss induced by ovariectomy in rats is prevented by gene transfer of parathyroid hormone or an Arg-Gly-Asp-containing peptide

Osteoporosis is a major and growing healthcare concern as the population ages. The genes of both a Arg-Gly-Asp (RGD)-containing peptide and parathyroid hormone (PTH) were used to reduce bone loss induced by ovariectomy (OVX) in rats. Plasmids with either RGD or PTH gene were delivered into the quadriceps of OVX rats. The expression of the genes was detected by RT-PCR and radioimmunoassay. Analysis of bone mineral density, bone mechanical testing and bone mineral content indicated an improvement in bone properties in both RGD-transferred and PTH-transferred rats compared to OVX rats. Gene transfer of either RGD or PTH is therefore a possible approach to prevent bone loss in OVX rats thus providing a potential method to prevent osteoporosis in clinical situations.These authors contributed equally to this work     

Distribution of Bacillus megaterium QM B1551 plasmids among other B. megaterium strains and Bacillus species

Bacillus megaterium QM B1551 contains seven plasmids. Two are small rolling circle plasmids and five are theta-replicating plasmids with cross-hybridizing replicons that define a new family of very homologous yet compatible theta replicons. Previous sequencing of several of the plasmids has shown genes with high similarity to those on the genomes and plasmids of other Gram-positive bacteria. To test the possible distribution of these plasmids, nine other B. megaterium strains and 20 other Bacillus or related species were tested for the presence of similar replicons, and specific flanking DNA by both hybridization and PCR. The theta replicons were widespread among the B. megaterium strains, and two had one or more of the rolling circle plasmids, but none of the plasmid replicon regions were observed in the other Bacillus or related species. It appears from the data that even though some plasmids carry genes suggesting horizontal transfer, their replicons seem to be unique to B. megaterium, or rarely present in related species.     

Environmental signals from leaves--a physiognomic analysis of European vegetation

Leaf physiognomic traits vary predictably along climatic and environmental gradients. The relationships between leaf physiognomy and climate have been investigated on different continents, but so far an investigation based on European vegetation has been missing. A grid data set (0.5 degrees x 0.5 degrees latitude/longitude) has been compiled in order to determine spatial patterns of leaf physiognomy across Europe. Based on distribution maps of native European hardwoods, synthetic chorologic flora lists were compiled for all grid cells. Every synthetic chorologic flora was characterised by 25 leaf physiognomic traits and correlated with 16 climatic parameters. Clear spatial patterns of leaf physiognomy have been observed, which are statistically significant related to certain, temperature-related climate parameters. Transfer functions for several climatic parameters have been established, based on the observed relationships. The study provides evidence that synthetically generated floras represent a powerful tool for analysing spatial patterns of leaf physiognomy and their relationships to climate. The transfer functions from the European data set indicate slightly different relationships of leaf physiognomy and environment compared with results obtained from other continents.     

Structure of cyanobacterial Photosystem I

Photosystem I is one of the most fascinating membrane protein complexes for which a structure has been determined. It functions as a bio-solar energy converter, catalyzing one of the first steps of oxygenic photosynthesis. It captures the light of the sun by means of a large antenna system, consisting of chlorophylls and carotenoids, and transfers the energy to the center of the complex, driving the transmembrane electron transfer from plastoquinone to ferredoxin. Cyanobacterial Photosystem I is a trimer consisting of 36 proteins to which 381 cofactors are non-covalently attached. This review discusses the complex function of Photosystem I based on the structure of the complex at 2.5 Å resolution as well as spectroscopic and biochemical data.     

Structural and functional organization of the peripheral light-harvesting system in Photosystem I

This review centers on the structural and functional organization of the light-harvesting system in the peripheral antenna of Photosystem I (LHC I) and its energy coupling to the Photosystem I (PS I) core antenna network in view of recently available structural models of the eukaryotic Photosystem I–LHC I complex, eukaryotic LHC II complexes and the cyanobacterial Photosystem I core. A structural model based on the 3D homology of Lhca4 with LHC II is used for analysis of the principles of pigment arrangement in the LHC I peripheral antenna, for prediction of the protein ligands for the pigments that are unique for LHC I and for estimates of the excitonic coupling in strongly interacting pigment dimers. The presence of chlorophyll clusters with strong pigment–pigment interactions is a structural feature of PS I, resulting in the characteristic red-shifted fluorescence. Analysis of the interactions between the PS I core antenna and the peripheral antenna leads to the suggestion that the specific function of the red pigments is likely to be determined by their localization with respect to the reaction center. In the PS I core antenna, the Chl clusters with a different magnitude of low energy shift contribute to better spectral overlap of Chls in the reaction center and the Chls of the antenna network, concentrate the excitation around the reaction center and participate in downhill enhancement of energy transfer from LHC II to the PS I core. Chlorophyll clusters forming terminal emitters in LHC I are likely to be involved in photoprotection against excess energy.     

Structure–function studies on the complex iron–sulfur flavoprotein glutamate synthase: the key enzyme of ammonia assimilation

Glutamate synthases are complex iron–sulfur flavoproteins that participate in the essential ammonia assimilation pathway in microorganisms and plants. The recent determination of the 3-dimensional structures of the α subunit of the NADPH-dependent glutamate synthase form and of the ferredoxin-dependent enzyme of Synechocystis sp. PCC 6803 provides a framework for the interpretation of the functional properties of these enzymes, and highlights protein segments most likely involved in control and coordination of the partial catalytic activities of glutamate synthases, which take place at sites distant from each other in space. In this review, we focus on the current knowledge on structure–function relationships in glutamate synthases, and we discuss open questions on the mechanisms of control of the enzyme reaction and of electron transfer among the enzyme flavin cofactors and iron–sulfur clusters.