Comparison of Kinetic Properties Between Plant and Fungal Amine Oxidases

Abstract

Kinetic properties of novel amine oxidases isolated from a mold Aspergillus niger AKU 3302 were compared to those of typical plant amine oxidase from pea seedling (EC 1.4.3.6). Pea amine oxidase showed highest affinity with diamines, such as putrescine and cadaverine, while fungal enzymes oxidized preferably n-hexylamine and tyramine. All enzymes were inhibited by carbonyl reagents, copper chelating agents, some substrate analogs and alkaloids, but there were quite significant differences in the sensitivity and inhibition modes. Aminoguanidine, which strongly inhibited pea amine oxidases showed only little effect on fungal enzymes. Substrate analogs such as 1,5-diamino-3-pentanone and l-amino-3-phenyl-3-propanone, which were potent competitive inhibitors of pea amine oxidases, inhibited fungal enzymes much more weakly and non competitively. Also various alkaloids behaving as competitive inhibitors of pea amine oxidases inhibited the fungal enzymes non competitively. Very surprising was the potent inhibition of fungal enzymes by artificial substrates of pea amine oxidases, E- and Z-1,4-diamino-2-butene. The relationships between the different inhibition modes and possible binding at the active site are discussed.     

Habitat Heterogeneity,Coexistence and Habitat Utilization in Five Backswimmer Species (Notonecta spp.; Hemiptera,Notonectidae)

In this study, we present data on life history, distribution and coexistence among five backswimmer species, Notonecta glauca, N. lutea, N. maculata, N. obliqua and N. reuteri, from 72 waters in a study area on the Swedish west coast. Two temporal life history patterns were found; species reproducing in the autumn (N. reuteri and N. lutea) and those reproducing in the spring (N. glauca N. obliqua and N. maculata). Female backswimmers were generally larger than conspecific males, except for N. lutea, in which the sexes were similar in size. The sex ratio of N. glauca, N. lutea and N. reuteri did not differ from 1/1. The most abundant species was N. glauca (49% of the waters inhabited by backswimmers) and then N. obliqua (19%), N. maculata (14%), N. lutea (10%) and N. reuteri (8%). In all, 972 individuals were recorded. The species were sequenced, with reference to their habitat use (from species occurring in temporal habitats to more stable and complex ones), as follows; N. maculata – N. glauca – N. obliqua – N. reuteri – N. lutea. N. glauca had the widest habitat distribution pattern. Two species combinations in pools were most commonly found and the five species were never found together. Possible competition among adults, according to their habitat distribution, may occur for: N. glauca with all species; N. obliqua mainly with N. glauca; N. lutea mainly with N. reuteri; N. reuteri mainly with N. lutea and N. glauca; N. maculata mainly with N. glauca. If body size is an important factor in the competitive ability between individuals, not merely species but also sex and temporal aspects have to be considered. The most intense competition should then exist between N. lutea and N. reuteri females.     

Floral structure and pollen morphology of two zinc violets (Viola lutea ssp. calaminaria and V. lutea ssp. westfalica) indicate their taxonomic affinity to Viola lutea

Two zinc violets, the yellow form of the Aachen–Liège area and the blue morph of Blankenrode in western Westphalia, have very restricted occurrence on heavy metal waste heaps. Their taxonomic affinities have not been finally resolved. The flower micromorphological analysis presented here indicates that both zinc violets are closely related to the alpine Viola lutea, in line with our earlier published molecular data, but not with the conclusions of other authors. The zinc violets are classed at the rank of subspecies as V. lutea: ssp. calaminaria for the yellow zinc violet and ssp. westfalica for its blue counterpart. Although the violets examined (V. lutea, V. lutea ssp. calaminaria, V. lutea ssp. westfalica) are closely related, there is no evidence that V. lutea ssp. westfalica is a descendent of V. tricolor. Here we provide the most detailed information on generative organ structure in the four violets studied.     

Genetic relatedness among Filobasidiella species

The three accepted species of Filobasidiella, F. neoformans, F. depauperata, and F. lutea, are compared morphologically and by molecular analysis. Sequences of the internally transcribed spacer (ITS) and the small subunit (SSU) gene of the ribosomal RNA (rRNA) gene cluster were obtained, and analysed by Neighbor-joining and Maximum parsimony methods. The three species of Filobsidiella are shown to form a single monophyletic clade, rooted by Tremella mesenterica. F. lutea was recovered as a distinct, but closely related taxon with the Filobasidiella clade. This is the first report of DNA sequences from herbarium specimens of F. lutea.This revised version was published online in October 2005 with corrections to the Cover Date.     

Genotypic diversity and distribution of Ostreobium quekettii within scleractinian corals

The green filamentous endolithic alga Ostreobium quekettii resides inside skeletons of scleractinian corals in close proximity with their tissue and plays a role in the viability of the coral and its associates. This study examined the distribution and diversity of O. quekettii within scleractinian corals from the Red Sea (Eilat, Gulf of Aqaba), using a molecular phylogenetic marker. The massive coral species Porites lutea and Goniastrea perisi were sampled from a depth range of 6–55 m, and ribulose 1,5-bisphosphate carboxylase large subunit gene (rbcL) DNA sequence of the alga was amplified and analyzed for diversity and distribution of ecological patterns. This work reveals that O. quekettii has at least seven different clades distributed along a depth gradient in the examined scleractinian corals. Among the seven identified clades, four were found only in P. lutea, while the other two clades are found in both P. lutea and G. perisi. Goniastrea perisi colonies at depth of 30 m had a distinct O. quekettii clade that was absent in P. lutea. It is obvious from this study that the green endolithic alga O. quekettii is not a single genotype as previously considered but a complex of genotypes and that this differentiation is of ecological significance.     

Construction, Overexpression, and Purification of Arthrobacter globiformis Amine Oxidase–Strep-Tag II Fusion Protein

The copper-containing amine oxidase from Arthrobacter globiformis has been expressed and purified as a fusion protein with a C-terminal Strep-tag II peptide. This tag facilitates the rapid purification of the enzyme on a large scale using the StrepTactin POROS medium. For example, we have demonstrated that 50 mg of protein can be obtained in 2 days from 2 L of Escherichia coli. The purified fusion protein displays turnover and spectroscopic properties that are essentially identical to those of the wild-type enzyme. Given the location of the C-terminus in four amine oxidase crystal structures, this strategy should be quite general for the rapid purification of amine oxidases from multiple sources.     

Litter-associated bacteria and fungi – a comparison of biomass and communities across lakes and plant species

1. We investigated the importance of lake water chemistry and substrate properties in regulating microbial decomposer communities on macrophyte litter. Ten lakes of differing water chemistry, including such variables as nutrient concentration, pH and dissolved organic carbon, were sampled in October 2003. Water chemistry was analysed, and litter from the macrophytes Phragmites australis, Schoenoplectus lacustris and Nuphar lutea was collected from both above and below the water surface. 2. The three plant species differed widely in carbon and nitrogen content. The aerial parts of P. australis had highest C : N ratio (mean value 125), while the lowest values were found in leaves of N. lutea (19.8). 3. Fungal carbon ranged from 0.15 to 6.4 mg g^?1 dry weight (DW), and was higher on aerial than on submerged plant parts. Fungal biomass was highest on S. lacustris and lowest on N. lutea. Denaturing gradient gel electrophoresis revealed no differences in the number of fungal taxa between plant species or plant parts, with the limitation that no molecular analysis was possible for N. lutea. 4. In contrast, bacteria were most abundant on N. lutea, but showed no significant difference between leaf and stem parts. The number of bacterial taxa was highest on the submerged parts of P. australis. 5. The correlations between microbial variables and the properties of lakes and litter were analysed using multivariate statistics. In a principal component analysis, litter properties explained 78% of the variation in microbial variables. In contrast, redundancy analysis revealed that the explanatory power of lake water chemistry was only 20%, indicating that the properties of the growth substratum were more important than those of the lake water for the attached microbial communities.     

Chemical Compositions and Antioxidant Activities of Essential Oils Extracted from the Petals of Three Wild Tree Peony Species and Eleven Cultivars

The aim of this study was to investigate the essential oil (EO) compositions and antioxidant activities from petals of three wild tree peony species (Paeonia delavayi, P. lutea, and P. rockii) and eleven P. suffruticosa cultivars from different cultivar groups. The EOs yields varied from 0.63% to 1.25% (v/v) among samples when using supercritical CO₂ extraction. One hundred and sixty‐three components were detected by GC/MS; and among them, linalool oxide, (Z)‐5‐dodecen‐1‐yl acetate, nonadecane, (Z)‐5‐nonadecene, heneicosane, phytol, and linoleic acid ethyl ester were dominant. According to hierarchical cluster analysis, principal component analysis and correspondence analysis, P. lutea, P. delavayi, and ‘High Noon’ were clustered in a group described as having a refreshing herbal‐like note due to high rates of phytol and linalool oxide. Notably, P. lutea and P. delavayi also had strong DPPH and ABTS radical scavenging activities. These results suggest that P. lutea and P. delavayi are the most promising candidates as useful sources of fragrances and natural antioxidants.     

Functional expression of amine oxidase from <Emphasis Type="Italic">Aspergillus niger</Emphasis> (AO-I) in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

The aim of this work was to prepare recombinant amine oxidase from Aspergillus niger after overexpressing in yeast. The yeast expression vector pDR197 that includes a constitutive PMA1 promoter was used for the expression in Saccharomyces cerevisiae. Recombinant amine oxidase was extracted from the growth medium of the yeast, purified to homogeneity and identified by activity assay and MALDI-TOF peptide mass fingerprinting. Similarity search in the newly published A. niger genome identified six genes coding for copper amine oxidase, two of them corresponding to the previously described enzymes AO-I a methylamine oxidase and three other genes coding for FAD amine oxidases. Thus, A. niger possesses an enormous metabolic gear to grow on amine compounds and thus support its saprophytic lifestyle.     

<Emphasis Type="Italic">Sinningia lutea</Emphasis> (Gesneriaceae), a new species from Southern Brazil

Sinningia lutea (Gesneriaceae) is described and illustrated as a new species from the Pampa Biome, in Southern Brazil. In the literature, this species has hitherto been confused with S. allagophylla from which it can be easily separated using a set of floral and vegetative features (especially corolla color and size and leaf shape). An artificial key to separate S. lutea from S. allagophylla and S. curtiflora (another sympatric species, with some morphological similarities) is presented. Some ecological remarks on the differential distribution of S. lutea and S. allagophylla are also made.     

Inhibitors of Plant Copper Amineoxidases

Abstract

In this review, inhibitors of plant copper amine oxidases from Lens esculenta seedlings, Pisum sativum seedlings, and Euphorbia characias latex are described. Reversible competitive inhibitors and non-competitive inhibitors, irreversible active-site directed inhibitors and mechanism-based inactivators are reviewed in regard to their mechanisms of action.     

Nelumbonaceae:Systematic position and species diversification revealed by the complete chloroplast genome

<正>Nelumbonaceae is a morphologically unique family of angiosperms and was traditionally placed in Nymphaeales;more recently,it was placed in Proteales based on molecular data,or in an order of its own,Nelumbonales. To determine the systematic position of the family and to date the divergence time of the family and the divergence time of its two intercontinentally disjunct species,we sequenced the entire chloroplast genome of Nelumbo lutea and most of the chloroplast genes of,N.nucifera.We carried out phylogenetic and molecular dating analyses of the two species and representatives of 47 other plant families,representing the major lineages of angiosperms, using 83 plastid genes.The N.lutea genome was 163 510 bp long,with a total of 130 coding genes and an overall GC content of 38%.No significant structural differences among the genomes of N.lutea,Nymphaea alba, and Platanus occidentalis were observed.The phylogenetic relationships based on the 83 plastid genes revealed a close relationship between Nelumbonaceae and Platanaceae.The divergence times were estimated to be 109 Ma between the two families and 1.5 Ma between the two Nelumbo species.The estimated time was only slightly longer than the age of known Nelumbo fossils,suggesting morphological stasis within Nelumbonaceae.We conclude that Nelumbonaceae holds a position in or close to Proteales.We further conclude that the two species of Nelumbo diverged recently from a common ancestor and do not represent ancient relicts on different continents.     

Evolution of glutamate dehydrogenase genes: Evidence for two paralogous protein families and unusual branching patterns of the archaebacteria in the universal tree of life

Summary The existence of two families of genes coding for hexameric glutamate dehydrogenases has been deduced from the alignment of 21 primary sequences and the determination of the percentages of similarity between each pair of proteins. Each family could also be characterized by specific motifs. One family (Family 1) was composed of gdh genes from six eubacteria and six lower eukaryotes (the primitive protozoan Giardia lamblia, the green alga Chlorella sorokiniana, and several fungi and yeasts). The other one (Family 11) was composed of gdh genes from two eubacteria, two archaebacteria, and five higher eukaryotes (vertebrates). Reconstruction of phylogenetic trees using several parsimony and distance methods confirmed the existence of these two families. Therefore, these results reinforced our previously proposed hypothesis that two close but already different gdh genes were present in the last common ancestor to the three Ur-kingdoms (eubacteria, archaebacteria, and eukaryotes). The branching order of the different species of Family I was found to be the same whatever the method of tree reconstruction although it varied slightly according the region analyzed. Similarly, the topological positions of eubacteria and eukaryotes of Family II were independent of the method used. However, the branching of the two archaebacteria in Family II appeared to be unexpected: (1) the thermoacidophilic Sulfolobus solfataricus was found clustered with the two eubacteria of this family both in parsimony and distance trees, a situation not predicted by either one of the contradictory trees recently proposed; and (2) the branching of the halophilic Halobacterium salinarium varied according to the method of tree construction: it was closer to the eubacteria in the maximum parsimony tree and to eukaryotesin distance trees. Therefore, whatever the actual position of the halophilic species, archaebacteria did not appear to be monophyletic in these gdh gene trees. This result questions the firmness of the presently accepted interpretation of previous protein trees which were supposed to root unambiguously the universal tree of life and place the archaebacteria in this tree. Offprint requests to: B. Labedan     

Phylogenetic systematics of Sternbergia (Amaryllidaceae) based on plastid and ITS sequence data

The phylogenetics of Sternbergia (Amaryllidaceae) were studied using DNA sequences of the plastid ndhF and matK genes and nuclear internal transcribed spacer (ITS) ribosomal region for 38, 37 and 32 ingroup and outgroup accessions, respectively. All members of Sternbergia were represented by at least one accession, except S. minoica and S. schubertii, with additional taxa from Narcissus and Pancratium serving as principal outgroups. Sternbergia was resolved and supported as sister to Narcissus and composed of two primary subclades: S. colchiciflora sister to S. vernalis, S. candida and S. clusiana, with this clade in turn sister to S. lutea and its allies in both Bayesian and bootstrap analyses. A clear relationship between the two vernal flowering members of the genus was recovered, supporting the hypothesis of a single origin of vernal flowering in Sternbergia. However, in the S. lutea complex, the DNA markers examined did not offer sufficient resolving power to separate taxa, providing some support for the idea that S. sicula and S. greuteriana are conspecific with S. lutea. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 166 , 149–162.     

GENETIC STRUCTURE OF THE FROGS GEOCRINIA LUTEA AND GEOCRINIA ROSEA REFLECTS EXTREME POPULATION DIVERGENCE AND RANGE CHANGES,NOT DISPERSAL BARRIERS

I describe the genetic structure of two frog species, Geocrinia rosea and Geocrinia lutea, using allozyme electrophoresis to understand population structure and thereby possible mechanisms of divergence and speciation. The sampling regimes represented the entire range of both species and provided replicated tests of the impact of ridges, rivers, and dry forest on gene flow. Geocrinia rosea and G. lutea were highly genetically subdivided (F_ST = 0.69, 0.64, respectively). In the extreme, there were fixed allelic differences between populations that were only 4 km (G. rosea) or 1.25 km (G. lutea) apart. In addition to localized divergence, two-dimensional scaling of genetic distance allowed the recognition of broad-scale genetic groups, each consisting of several sample sites. Patterns of divergence were unrelated to the presence of ridges, rivers, or dry forest. I argue that range contraction and expansion, combined with extreme genetic divergence in single, isolated populations, best accounts for the genetic structure of these species.     

Unusual modification patterns in the transfer ribonucleic acids of archaebacteria

The modification patterns of the transfer RNAs of ten archaebacteria (Halobacterium volcanii, Halococcus morrhuae, Methanobacterium bryantii, Methanobrevibacter smithii, Methanococcus vannielii, Methanococcus voltae, Methanomicrobium mobile, Methanosarcina barkeri, Thermoplasma acidophilum, andSulfolobus acidocaldarius) were analyzed by two-dimensional thin-layer chromatography of the³²P-labeled nucleotides. All species lack ribothymidine and 7-methylguanosine, and dihydrouridine is absent from all butM. barkeri. Pseudouridine, 2′-O-methylcytidine, 1-methylguanosine, andN ²,N ²-dimethylguanosine are present in all of them; except forM. barkeri andT. acidophilum, all haveN ²-methylguanosine. All, exceptH. volcanii andH. morrhuae, contain 1-methyladenosine, and these two organisms andS. acidocalderius only contain 5-methylcytidine. The transfer RNA modification patterns of the archaebacteria are distinct from those of typical eubacteria (Escherichia coli) and typical eukaryotes (Saccharomyces cerevisiae), although they are somewhat more similar to the latter than the former.     

Comparative study of leaf architecture and cuticles of Nelumbo changchangensis from the Eocene of Hainan Island,China, and the two extant species of Nelumbo (Nelumbonaceae)

Fossil leaves of Nelumbo changchangensis, collected from the Eocene of Hainan Island, China, were studied and compared with those of the extant species of Nelumbo, N. nucifera Gaertn. and N. lutea Willd. The fossil leaves have all the specialized features of extant Nelumbo in leaf architecture, except that the organization of the areolae looks much more irregular than that of extant Nelumbo. Comparisons of the cuticle and epicuticular ultrastructure indicate that: (1) N. changchangensis resembles N. nucifera in that anticlinal cell walls of the lower epidermis are straight along the major veins and near leaf bases and are shallowly undulate with U‐ to V‐shaped undulations inside the areolae; (2) N. changchangensis differs from N. lutea in that anticlinal cell walls of the lower epidermis of the latter are deeply undulate with U‐, V‐ to reversed Ω‐shaped undulations inside the areolae; and (3) epicuticular wax crystals are more densely distributed on the leaves of N. changchangensis and N. nucifera than they are in N. lutea. These findings shed significant light on the cuticle differentiation of fossil and extant Nelumbo species. The morphometric comparisons indicate that almost all the synapomorphies of extant Nelumbo were already present by the Eocene, © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2016, 180 , 123–137.     

Effects of growth temperature and winter duration on leaf phenology of a spring ephemeral (Gagea lutea) and a summergreen forb (Maianthemum dilatatum)

Effects of growth temperature and winter duration on leaf longevity were compared between a spring ephemeral, Gagea lutea, and a forest summergreen forb, Maianthemum dilatatum. The plants were grown at day/night temperatures of 25/20°C and 15/10°C after a chilling treatment for variable periods at 2°C. The temperature regime of 25/20°C was much higher than the mean air temperatures for both species in their native habitats. Warm temperature of 25/20°C and/or long chilling treatment shortened leaf longevity in G. lutea, but not in M. dilatatum. The response of G. lutea was consistent with that reported for other spring ephemerals. Air temperature increases as the vegetative season progresses. The decrease in leaf longevity in G. lutea under warm temperature condition ensures leaf senescence in summer, an unfavorable season for its growth. This also implies that early leaf senescence could occur in years with early summers. Warm spring temperatures have been shown to accelerate the leafing-out of forest trees. The decrease in leaf longevity due to warm temperature helps synchronize the period of leaf senescence roughly with the time of the forest canopy leaf-out. Prolonged winter due to late snowmelt has been shown to shorten the vegetative period for spring ephemerals. The decrease in leaf longevity due to long chilling treatment would correspond with this shortened vegetative period.     

Population genetic structure of the South American species Hypochaeris lutea (Asteraceae)

The genus Hypochaeris has a recent evolutionary history caused by long‐distance dispersal in conjunction with adaptive radiation in the South American continent. Hypochaeris lutea is a perennial herb that grows mostly at altitudes of around 1000 m in cold swamps of the southern regions of Brazil. We investigated the amplified fragment length polymorphism (AFLP) in 270 individuals representing 11 Brazilian populations of H. lutea to elucidate the population genetic structure of this species. The frequencies of polymorphic loci and gene diversity ranged from 83.42% to 91.66% and from 0.26 to 0.34, respectively. Analysis of molecular variance revealed that most of the genetic variability was found within (76.67%) rather than among (23.3%) populations, agreeing with the pattern of genetic distribution within and among populations observed in other allogamous species of Hypochaeris. A Mantel test showed no correlation between genetic and geographic distances when all populations were considered. Simulations performed using a Bayesian approach consistently identified two clusters with different admixture proportions of individuals, as also revealed by a UPGMA dendrogram of populations. The pattern of genetic structure observed in H. lutea is consistent with a process of successive colonization events by long‐distance dispersal resembling the rapid and recent radiation that has been proposed to explain the origin of the South American species of Hypochaeris.     

Pollination ofNuphar (Nymphaeaceae) in Europe: Flies and bees rather thanDonacia beetles

Nuphar (Nymphaeaceae) comprises a small holarctic group of aquatic perennials whose flowers are pollinated by flies, bees, and beetles. We studied pollination in different populations of the two European speciesN. lutea andN. pumila in Norway and in Germany. Flowers are self-compatible and protogynous, preventing automatic selfing, and insect pollination is required for seed production. Sympatric populations were studied in Vest-Agder county in Norway to determine whetherN. lutea andN. pumila have the same or different pollinators. Allopatric populations ofN. lutea in Germany and Norway were then compared to determine whether their pollinator spectra differ as would be expected in an open flower with seemingly easily accessible pollen and nectar. Results of the present as well as previous studies of the pollination ofN. lutea andN. pumila show that both species are mainly pollinated by flies, including apparentNuphar specialists, such as the scatophagidHydromyza livens and the ephydridsHydrellia andNotiphila, the last also a long-known pollinator ofN. advena in Florida. Pollinator overlap between sympatric heterospecific populations was small, while allopatric conspecific populations had similar visitor and pollinator spectra. We found no evidence of pollination byDonacia beetles as reported from some North American populations ofNuphar. Dedicated to Univ.-Prof. DrF. Ehrendorfer on the occasion of his 70th birthday