Host genotype and age shape the leaf and root microbiomes of a wild perennial plant.

Bacteria living on and in leaves and roots influence many aspects of plant health, so the extent of a plant's genetic control over its microbiota is of great interest to crop breeders and evolutionary biologists. Laboratory-based studies, because they poorly simulate true environmental heterogeneity, may misestimate or totally miss the influence of certain host genes on the microbiome. Here we report a large-scale field experiment to disentangle the effects of genotype, environment, age and year of harvest on bacterial communities associated with leaves and roots of Boechera stricta (Brassicaceae), a perennial wild mustard. Host genetic control of the microbiome is evident in leaves but not roots, and varies substantially among sites. Microbiome composition also shifts as plants age. Furthermore, a large proportion of leaf bacterial groups are shared with roots, suggesting inoculation from soil. Our results demonstrate how genotype-by-environment interactions contribute to the complexity of microbiome assembly in natural environments.

Collection of aboveground community and species-specific plant biomass from the Jena Experiment (time series since 2002).

This data set comprises time series of aboveground community plant biomass (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) and species-specific biomass from the sown species of several experiments at the field site of a large grassland biodiversity experiment (the Jena Experiment; see further details below). Aboveground community biomass was normally harvested twice a year just prior to mowing (during peak standing biomass twice a year, generally in May and August; in 2002 only once in September) on all experimental plots in the Jena Experiment. This was done by clipping the vegetation at 3 cm above ground in up to four rectangles of 0.2 x 0.5 m per large plot. The location of these rectangles was assigned by random selection of new coordinates every year within the core area of the plots. The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: individual species for the sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. Sown plant community biomass was calculated as the sum of the biomass of the individual sown species. The data for individual samples and the mean over samples for the biomass measures on the community level are given. Overall, analyses of the community biomass data have identified species richness as well as functional group composition as important drivers of a positive biodiversity-productivity relationship. The following series of datasets are contained in this collection: 1. Plant biomass form the Main Experiment: In the Main Experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). 2. Plant biomass from the Dominance Experiment: In the Dominance Experiment, 206 grassland plots of 3.5 x 3.5 m were established from a pool of 9 species that can be dominant in semi-natural grassland communities of the study region. In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 3, 4, 6, and 9 species). 3. Plant biomass from the monoculture plots: In the monoculture plots the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species like the other experiments in May 2002. All plots were maintained by bi-annual weeding and mowing.

Aboveground community and species-specific plant biomass from the Jena Experiment (Dominance Experiment, year 2008)

This data set contains aboveground community plant biomass (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) and species-specific biomass from the sown species of the dominance experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the dominance experiment, 206 grassland plots of 3.5 x 3.5 m were established from a pool of 9 plant species that can be dominant in semi-natural grassland communities of the study region. In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 3, 4, 6, and 9 species). Plots were maintained by bi-annual weeding and mowing. Aboveground community biomass was harvested twice in May and August 2008 on all experimental plots of the dominance experiment. This was done by clipping the vegetation at 3 cm above ground in two rectangles of 0.2 x 0.5 m per experimental plot. The location of these rectangles was assigned by random selection of coordinates within the central area of the plots (excluding an outer edge of 50cm). The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: individual species for the sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material, and remaining plant material that could not be assigned to any category. All biomass was dried to constant weight (70°C, >= 48 h) and weighed. Sown plant community biomass was calculated as the sum of the biomass of the individual sown species. The mean of both samples per plot and the individual measurements are provided in the data file. Overall, analyses of the community biomass data have identified species richness and the presence of particular species as an important driver of a positive biodiversity-productivity relationship.

Aboveground community and species-specific plant biomass from the Jena Experiment (Main Experiment, year 2009)

This data set contains aboveground community biomass in 2009 (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) and species-specific biomass from the sown species of the main experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. Aboveground community biomass was harvested twice in 2009 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the main experiment. This was done by clipping the vegetation at 3 cm above ground in three rectangles of 0.2 x 0.5 m per large plot. The location of these rectangles was assigned prior to each harvest by random selection of coordinates within the core area of the plots (i.e. the central 10 x 15 m). The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: individual species for the sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. Sown plant community biomass was calculated as the sum of the biomass of the individual sown species. The data for individual samples and the mean over samples for all biomass measures are given. Overall, analyses of the community biomass data have identified species richness as well as functional group composition as important drivers of a positive biodiversity-productivity relationship.

Aboveground community and species-specific plant biomass from the Jena Experiment (Main Experiment, year 2010)

This data set contains aboveground community biomass in 2010 (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) and species-specific biomass from the sown species of the main experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. Aboveground community biomass was harvested twice in 2010 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the main experiment. This was done by clipping the vegetation at 3 cm above ground in two rectangles of 0.2 x 0.5 m per large plot. The location of these rectangles was assigned prior to each harvest by random selection of coordinates within the core area of the plots (i.e. the central 10 x 15 m). The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: individual species for the sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. Sown plant community biomass was calculated as the sum of the biomass of the individual sown species. The data for individual samples and the mean over samples for all biomass measures are given. Overall, analyses of the community biomass data have identified species richness as well as functional group composition as important drivers of a positive biodiversity-productivity relationship.

Aboveground plant biomass from the Jena Experiment (Monocultures, year 2003)

This data set contains aboveground plant biomass in 2003 (Sown plant community, Weed plant community, and Dead plant material; all measured in biomass as dry weight) of the monoculture plots of a large grassland biodiversity experiment (the Jena Experiment). In the monoculture plots the biomass of the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species. These 60 species comprising the species pool of the Jena Experiment belong to four functional groups (grasses, legumes, tall and small herbs). Plots were sown in May 2002 and are since maintained by bi-annual weeding and mowing. Aboveground plant biomass was harvested twice in 2003 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the monocultures. This was done by clipping the vegetation at 3 cm above ground in 2 rectangles of 0.2 x 0.5 m per plot. The location of these rectangles was assigned prior to each harvest by random selection of coordinates within the core area of the plots (i.e. excluding an outer edge of 0.5 m). The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: sown plant species, weed plant species (species not sown at the particular plot), and detached dead plant material (i.e., dead plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. The data for individual subsamples (i.e. rectangles) and the mean over samples for all biomass measures are given.

Aboveground plant biomass from the Jena Experiment (Monocultures, year 2010)

This data set contains aboveground plant biomass in 2010 (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) of the monoculture plots of a large grassland biodiversity experiment (the Jena Experiment). In the monoculture plots the biomass of the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species. One of the replicate plots per species was given up after the vegetation period of 2007 for all but the nine species belonging also to the so called dominance experiment in Jena. These nine species are: Alopecurus pratensis, Anthriscus sylvestris, Arrhenatherum elatius, Dactylis glomerata, Geranium pratense, Poa trivialis, Phleum pratense, Trifolium repens and Trifolium pratense.In 2010 plot size was reduced to 1 x 1 m. These 60 species comprising the species pool of the Jena Experiment belong to four functional groups (grasses, legumes, tall and small herbs). Plots were sown in May 2002 and are since maintained by bi-annual weeding and mowing. Aboveground plant biomass was harvested twice in 2010 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the monocultures. This was done by clipping the vegetation at 3 cm above ground in 1 rectangle of 0.2 x 0.5 m per plot. The location of this rectangle was in the center of the plot area. The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed.

Aboveground plant biomass from the Jena Experiment (Monocultures, year 2005)

This data set contains aboveground plant biomass in 2005 (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) of the monoculture plots of a large grassland biodiversity experiment (the Jena Experiment). In the monoculture plots the biomass of the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species. These 60 species comprising the species pool of the Jena Experiment belong to four functional groups (grasses, legumes, tall and small herbs). Plots were sown in May 2002 and are since maintained by bi-annual weeding and mowing. Aboveground plant biomass was harvested twice in 2005 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the monocultures. This was done by clipping the vegetation at 3 cm above ground in 2 rectangles of 0.2 x 0.5 m per plot. The location of these rectangles was assigned prior to each harvest by random selection of coordinates within the core area of the plots (i.e. excluding an outer edge of 0.5 m). The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. The data for individual subsamples (i.e. rectangles) and the mean over samples for all biomass measures are given.

Aboveground plant biomass from the Jena Experiment (Monocultures, year 2006)

This data set contains aboveground plant biomass in 2006 (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) of the monoculture plots of a large grassland biodiversity experiment (the Jena Experiment). In the monoculture plots the biomass of the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species. These 60 species comprising the species pool of the Jena Experiment belong to four functional groups (grasses, legumes, tall and small herbs). Plots were sown in May 2002 and are since maintained by bi-annual weeding and mowing. Aboveground plant biomass was harvested twice in 2006 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the monocultures. This was done by clipping the vegetation at 3 cm above ground in 2 rectangles of 0.2 x 0.5 m per plot. The location of these rectangles was assigned prior to each harvest by random selection of coordinates within the core area of the plots (i.e. excluding an outer edge of 0.5 m). The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. The data for individual subsamples (i.e. rectangles) and the mean over samples for all biomass measures are given.

Aboveground plant biomass from the Jena Experiment (Monocultures, year 2007)

This data set contains aboveground plant biomass in 2007 (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) of the monoculture plots of a large grassland biodiversity experiment (the Jena Experiment). In the monoculture plots the biomass of the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species. These 60 species comprising the species pool of the Jena Experiment belong to four functional groups (grasses, legumes, tall and small herbs). Plots were sown in May 2002 and are since maintained by bi-annual weeding and mowing. Aboveground plant biomass was harvested twice in 2007 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the monocultures. This was done by clipping the vegetation at 3 cm above ground in 2 rectangles of 0.2 x 0.5 m per plot. The location of these rectangles was assigned prior to each harvest by random selection of coordinates within the core area of the plots (i.e. excluding an outer edge of 0.5 m). The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. The data for individual subsamples (i.e. rectangles) and the mean over samples for all biomass measures are given.

Aboveground plant biomass from the Jena Experiment (Monocultures, year 2008)

This data set contains aboveground plant biomass in 2008 (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) of the monoculture plots of a large grassland biodiversity experiment (the Jena Experiment). In the monoculture plots the biomass of the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species. One of the replicate plots per species was given up after the vegetation period of 2007 for all but the nine species belonging also to the so called dominance experiment in Jena. These nine species are: Alopecurus pratensis, Anthriscus sylvestris, Arrhenatherum elatius, Dactylis glomerata, Geranium pratense, Poa trivialis, Phleum pratense, Trifolium repens and Trifolium pratense.In 2008 plot size was reduced to 2.5 x 2.5 m. These 60 species comprising the species pool of the Jena Experiment belong to four functional groups (grasses, legumes, tall and small herbs). Plots were sown in May 2002 and are since maintained by bi-annual weeding and mowing. Aboveground plant biomass was harvested twice in 2008 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the monocultures. This was done by clipping the vegetation at 3 cm above ground in 2 rectangles of 0.2 x 0.5 m per plot. The location of these rectangles was assigned prior to each harvest by random selection of coordinates within the core area of the plots (i.e. excluding an outer edge of 0.5 m). The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. The data for individual subsamples (i.e. rectangles) and the mean over samples for all biomass measures are given.

Aboveground plant biomass from the Jena Experiment (Monocultures, year 2009)

This data set contains aboveground plant biomass in 2009 (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) of the monoculture plots of a large grassland biodiversity experiment (the Jena Experiment). In the monoculture plots the biomass of the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species. One of the replicate plots per species was given up after the vegetation period of 2007 for all but the nine species belonging also to the so called dominance experiment in Jena. These nine species are: Alopecurus pratensis, Anthriscus sylvestris, Arrhenatherum elatius, Dactylis glomerata, Geranium pratense, Poa trivialis, Phleum pratense, Trifolium repens and Trifolium pratense.In 2008 plot size was reduced to 2.5 x 2.5 m. These 60 species comprising the species pool of the Jena Experiment belong to four functional groups (grasses, legumes, tall and small herbs). Plots were sown in May 2002 and are since maintained by bi-annual weeding and mowing. Aboveground plant biomass was harvested twice in 2009 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the monocultures. This was done by clipping the vegetation at 3 cm above ground in 2 rectangles of 0.2 x 0.5 m per plot. The location of these rectangles was in the center of the plot area. The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. The data for individual subsamples (i.e. rectangles) and the mean over samples for all biomass measures are given.

Aboveground plant biomass from the Jena Experiment (Monocultures, year 2002)

This data set contains aboveground plant biomass in 2002 (Sown plant community; measured in biomass as dry weight) of the monoculture plots of a large grassland biodiversity experiment (the Jena Experiment). In the monoculture plots the biomass of the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species. These 60 species comprising the species pool of the Jena Experiment belong to four functional groups (grasses, legumes, tall and small herbs). Plots were sown in May 2002 and are since maintained by bi-annual weeding and mowing. Aboveground plant biomass was harvested twice in 2002 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the monocultures. This was done by clipping the vegetation at 3 cm above ground in 2 rectangles of 0.2 x 0.5 m per plot. The location of these rectangles was assigned prior to each harvest by random selection of coordinates within the core area of the plots (i.e. excluding an outer edge of 0.5 m). The positions of the rectangles within plots were identical for all plots. From the harvested biomass only the separated biomass of the sown plant species was kept. All biomass was dried to constant weight (70°C, >= 48 h) and weighed. The data for individual subsamples (i.e. rectangles) and the mean over samples for all biomass measures are given.

Aboveground plant biomass from the Jena Experiment (Monocultures, year 2004)

This data set contains aboveground plant biomass in 2004 (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) of the monoculture plots of a large grassland biodiversity experiment (the Jena Experiment). In the monoculture plots the biomass of the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species. These 60 species comprising the species pool of the Jena Experiment belong to four functional groups (grasses, legumes, tall and small herbs). Plots were sown in May 2002 and are since maintained by bi-annual weeding and mowing. Aboveground plant biomass was harvested twice in 2004 just prior to mowing (during peak standing biomass in early June and in late August) on all experimental plots of the monocultures. This was done by clipping the vegetation at 3 cm above ground in 2 rectangles of 0.2 x 0.5 m per plot. The location of these rectangles was assigned prior to each harvest by random selection of coordinates within the core area of the plots (i.e. excluding an outer edge of 0.5 m). The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. The data for individual subsamples (i.e. rectangles) and the mean over samples for all biomass measures are given.

Structure and lipid interactions of membrane-associated glycosyltransferases : Cationic patches and anionic lipids regulate biomembrane binding of both GT-A and GT-B enzymes

This thesis concerns work on structure and membrane interactions of enzymes involved in lipid synthesis, biomembrane and cell wall regulation and cell defense processes. These proteins, known as glycosyltransferases (GTs), are involved in the transfer of sugar moieties from nucleotide sugars to lipids or chitin polymers. Glycosyltransferases from three types of organisms have been investigated; one is responsible for vital lipid synthesis in Arabidopsis thaliana (atDGD2) and adjusts the lipid content in biomembranes if the plant experiences stressful growth conditions. This enzyme shares many structural features with another GT found in gram-negative bacteria (WaaG). WaaG is however continuously active and involved in synthesis of the protective lipopolysaccharide layer in the cell walls of Escherichia coli. The third type of enzymes investigated here are chitin synthases (ChS) coupled to filamentous growth in the oomycete Saprolegnia monoica. I have investigated two ChS-derived MIT domains that may be involved in membrane interactions within the endosomal pathway. From analysis of the three-dimensional structure and the amino-acid sequence, some important regions of these very large proteins were selected for in vitro studies. By the use of an array of biophysical methods (e.g. Nuclear Magnetic Resonance, Fluorescence and Circular Dichroism spectroscopy) and directed sequence analyses it was possible to shed light on some important details regarding the structure and membrane-interacting properties of the GTs. The importance of basic amino-acid residues and hydrophobic anchoring segments, both generally and for the abovementioned proteins specifically, is discussed. Also, the topology and amino-acid sequence of GT-B enzymes of the GT4 family are analyzed with emphasis on their biomembrane association modes. The results presented herein regarding the structural and lipid-interacting properties of GTs aid in the general understanding of glycosyltransferase activity. Since GTs are involved in a high number of biochemical processes in vivo it is of outmost importance to understand the underlying processes responsible for their activity, structure and interaction events. The results are likely to be useful for many applications and future experimental design within life sciences and biomedicine.