Genome size and polyploidy variation in the tropical hardwood genusTerminalia (Combretaceae)

Chromosome complements and 2C DNA amounts of six species ofTerminalia have been studied.Terminalia oliveri, T. myriocarpa andT. arjuna are diploid (2n = 24),T. chebula andT. bellirica are tetraploid (2n = 48),T. muelleri shows a triploid number (2n = 36). Two well demarcated groups of species are recognizable on the basis of chromosome length and 2C DNA values which range from 3.60 pg (T. oliveri) to 12.80 pg (T. bellirica) showing a 3.5-fold difference. Differences of DNA per basic genome or per chromosome are greatest (1.97-fold) betweenT. oliveri andT. arjuna. Two species groups (1)T. oliveri andT. chebula, and (2)T. myriocarpa, T. arjuna, T. muelleri, T. bellirica, therefore are well differentiated by DNA per basic genome, irrespective of polyploidy. The mean values of the two groups are 1.81 pg and 3.34 pg, respectively, showing a 1.84-fold difference. Within diploids and tetraploids there is 1.97-fold and 1.76-fold variation, respectively.     

Genome size in gymnosperms

The DNA 2C and per chromosome values of 57 species belonging to 22 genera of gymnosperms have been analysed. The overall range is 12-fold with a modal value of about 30.0 pg.Cycadales exhibit a 2-fold difference. AmongConiferales with a 4-fold variation, thePinaceae have higher mean DNA contents as well as a greater range and diversity than other families. Remarkable interspecific differences are found inCycas, Picea, Larix, Pinus, Callitris, Cupressus, andChamaecyparis. Despite this, there is a constancy of basikaryotypes within these genera.Gnetum shows a distinctly low DNA value.     

Nuclear DNA amounts in 112 species of tropical hardwoods -- new estimates

The 4C DNA values of 112 species, belonging to 37 families have a range from 0.83 pg (Bixa orellana) to 15.54 pg (Thryallis angustifolia), showing a 18.72-fold variation. The genome size varies from 0.21 pg (Bixa orellana) to 3.32 (Thespesia populnea), with a 15.8-fold difference. The Bombacaceae has the minimum range (1.08-fold) of variation, while the maximum (5.0-fold) is shown by the Fabaceae. The Boraginaceae, Lauraceae, Malpighiaceae, and Malvaceae generally have higher 4C DNA values of > 10 pg, while the Bixaceae, Caricaceae, Oxalidaceae, and Santalaceae have lower values of < 2.0 pg. These data add further to our knowledge on variation in DNA amount in tropical hardwoods.     

Evolution of genome size inAllium (Alliaceae)

The 4C DNA amounts of 86 species fromAllium subgg.Allium, Rhizirideum, Bromatorrhiza, Melanocrommyum, Caloscordum andAmerallium show a 8.35-fold difference ranging from 35.60 pg (A. ledebourianum, 2n = 16) to 297.13 pg (A. validum 2n = 56). At diploid level the difference is 3.57-fold betweenA. ledebourianum (35.60 pg) andA. ursinum (127.14 pg). This shows that a significant loss and/or gain of DNA has occurred during evolution. On average subgg.Rhizirideum andAllium have less DNA amount than subgg.Melanocrommyum andAmerallium. The distribution of nuclear DNA amounts does not show discontinuous pattern and regular groups. The evolution of genome size has been discussed in relation to polyploidy and genomes, heterochromatin, adaptive changes in morphological characteristics, phenology and ecological factors, and infrageneric classification.     

DNA CONTENT AND EVOLUTION IN THE MICROSERIDINAE

Relative 2C nuclear DNA contents were microphotometrically determined from nuclei isolated from eight species of Microseris, four species of Agoseris, and Phalacroseris Bolanderi. The thirteen species are diploid (2n = 18) western North American members of the subtribe Microseridinae, tribe Cichorieae, of the family Compositae. A 7.7-fold variation in DNA content was detected. Phalacroseris has the highest DNA content and Agoseris heterophylla has the lowest. Within the genera Microseris and Agoseris, a 2.8- and 3.1-fold range in DNA content was detected. The higher values were from perennial species, and the lower values were from annual inbreeding species. Both evolutionary increases and decreases in nuclear DNA content have apparently occurred during the differentiation of the subtribe.     

Chemosystematics of theRutaceae: Comments on the interpretation ofda Silva & al.

The revision of theRutaceae into 17 provisional tribes, based primarily on the distribution of secondary metabolites (da Silva & al. 1988) is critically reviewed. In three areas where sufficient phytochemical data is available, i.e. the proto-Rutaceae, (provisional tribesZanthoxylum, Phellodendron, Toddalia, andEuodia pro parte), the AfricanToddalioideae sensuEngler, (provisional tribesEuodia pro parte,Acronychia), andClauseneae sensuSwingle, (provisional tribesClausena, Glycosmis, Micromelum, Merrillia), it is shown that the proposals made byda Silva & al. are seriously flawed. It is suggested that for other areas of the family insufficient phytochemical information is available to justify these proposals. In a wider context it is suggested that this approach, based on only one set of characters and on a wholly insufficient data base, is unhelpful to the task of producing a new classification of the family.     

Correlations between 2C DNA values and habit inCassia (Leguminosae:Caesalpinioideae)

Ten species of the genusCassia show a range of 2C DNA amounts from 1.30 to 2.54 pg at the same ploidy level. Remarkably, a distinct 2-fold increase is depicted by an arboreal speciesC. excelsa while the rest comprising of herbs, trees and shrubs have a range from 1.30 to 1.47 pg. These form a natural grouping with respect to mean DNA amounts which differ by 0.05 pg in the herbs, trees and shrubs respectively.     

Comparative microcalorimetric and dilatometric analysis of the interactions of quinacrine,chloroquine, and ethidium bromide with DNA

The enthalpies of binding of chloroquine and quinacrine to DNA at different molar ratios of drug to DNA and at different ionic strengths have been measured. The limiting values obtained with quinacrine fall in the range found for typical intercalating agents (e.g., ethidium, proflavin, adriamycin), whereas the value obtained with chloroquine is always zero, independent of the ratio of drug to DNA and ionic strength. The dilatometric measurements performed on the same systems and on the ethidium–DNA system show that when ethidium and quinacrine bind to DNA at low drug/DNA ratios, a volume decrease of about 16 mL/mol of bound drug occurs. No change in volume is observed when the two drugs bind to DNA through external, electrostatic forces. The volume change can be attributed to the loss of structured water around hydrophobic moieties of the drug molecules, following intercalation. In contrast, chloroquine binding to DNA at low drug/DNA ratios is characterized by a volume change distinctly smaller than that shown by quinacrine. The low ΔV_B and ΔH_B values shown by chloroquine are discussed in terms of the mechanism of interaction with DNA.     

DNA amount, latitude, and crop plant distribution

Avdulov and Stebbins noted a tendency for species with large chromosomes in several angiosperm groups or families (including the Gramineae, Commelinaceae, Liliales, Polemoniales and the Leguminosae) to be localized in distribution to temperate latitudes. As chromosome size and DNA content are closely correlated, the distribution of species with large DNA amounts per chromosome, or per diploid genome, might expected to be similarly localized. This hypothesis was tested using samples of crop species with large ranges of DNA amounts from the Gramineae and the Leguminosae. For instance, the mean DNA amount per chromosome for the sample of cereal grain species showed about a 36-fold range from 0·033 picograms (pg) in Eragrostis tef to 1·186 pg in Secale cereale, while for the sample of pulse crops the range was about 70-fold from 0·032 pg in Lablab niger to 2·225 pg in Vicia faba. The results for cereal grain crops, cultivated pasture grasses and pulse crops show that cultivation of species with high DNA amounts per diploid genome tends to be localized in temperate latitudes, or to seasons and regions at lower latitudes where the conditions approximate to those normally found in temperate latitudes. Moreover, man has shown a strong tendency to choose species for cultivation with increasingly lower DNA amounts at successively lower latitudes. Thus, there is a cline for DNA amount and latitude. This cline is exhibited independently by both C₃ and C₄ crop species, and by both annuals and perennials and hence is independent of life cycle type and the taxonomic distribution of C₃ and C₄ photosynthesis. The cline is apparently a natural phenomenon which man has modified and exaggerated in agriculture. It is suggested, therefore, that interspecific variation in DNA amount between angiosperm species may have adaptive significance affecting the distribution of both crop and non-crop species. The cline might be caused either by variation in DNA amount per se, or by variation in some factor(s) correlated with DNA amount. The factor(s) causally responsible for the cline, and their mode of action should be investigated since they may have implications for agriculture and plant breeding.     

Ecophysiology and distribution of the endemic leafless spurgeEuphorbia aphylla and the introducedE. tirucalli (Euphorbiaceae,Euphorbia sect.Tirucalli) in the Canary Islands

The leafless spurgeEuphorbia aphylla (Euphorbiaceae), an endemic species restricted to several of the Canary Islands where it inhabits coastal and arid localities, expresses Crassulacean Acid Metabolism (CAM) when it is subjected to summer drought. A flexible CAM is consistent with the general ecology of the species. It is the only member of sect.Tirucalli native to the Canary Islands and is at the north-western edge of the section's biogeographical range. The other members of the section have a paleotropic distribution and are found throughout Africa. Many of them are regarded as obligate CAM plants, includingE. tirucalli which was used as a comparison in ecophysiological experiments examining the response ofE. aphylla to drought and temperature.     

Conifer genome sizes of 172 species,covering 64 of 67 genera,range from 8 to 72 picogram

Nuclear genome size of conifers as measured by flow cytometry with propidium iodide was investigated, striving to collect at least a single species from each genus. 64 out of 67 genera and 172 species were measured. Of the 67 genera, 21 are reported here for the first time and the same is true for 76 species. This nearly doubles the number of measured genera and adds 50% to the number of analyzed species. Conifers have chromosome numbers in the range of n = (7)10–12(19). However, the nuclear DNA content (2C‐value) is shown here to range from 8.3 to 71.6 picogram. The largest genome contains roughly 6 × 10¹⁰ more base pairs than the smallest genome. Genome sizes are evaluated and compared with available taxonomic treatments. For the mainly (sub)tropical Podocarpaceae small genome sizes were found with a 2C‐value of only 8–28 pg, with 13.5 pg on average. For the Taxaceae 2C‐values from 23–60 pg were determined. Not surprisingly, the genus Pinus with 97 species (39 species measured here) has a broad range with 2C = 38–72 pg. A factor of 2 difference is also found in the Cupressaceae (136 species) with nuclear DNA contents in the range 18–35 pg. Apart from the allohexaploid Sequoia, ploidy plays a role only in Juniperus and some new polyploids are found. The data on genome size support conclusions on phylogenetic relationships obtained by DNA sequencing. Flow cytometry is applicable even to young plants or seeds for the monitoring of trade in endangered species.     

Regulation of proliferation of UM-SCV-1A and UM-SCV-6 vulvar carcinoma cells by cytokines

 The biology and pathogenesis of vulvar carcinoma are poorly understood at present. In order to understand this disease better, we have used recently developed squamous cell carcinoma lines of the vulva as models. Two cell lines originating from two individuals (UM-SCV-1A and UM-SCV-6) were cultured in vitro in 10% fetal calf serum. The effects of interleukins 10 and 13, interferons α and γ, granulocyte/macrophage-growth-stimulating factor (GM-CSF), tumor necrosis factor α (TNFα), and transforming growth factor β (TGFβ) on the proliferation of the cells was investigated by using radioactively labelled uridine as tracer. In addition, an investigation on the molecular structure of extracted cellular DNA was carried out to investigate whether programmed cell death (apoptosis) would be inducible by any of the factors. In UM-SCV-1A cells, interleukin-10 (IL-10) and interleukin-13 (IL-13) caused an approximately 12-fold decrease in DNA synthesis in cells cultured for 72 h (P<0.001), while GM-CSF had no significant effect. TGFβ showed a significant inhibitory effect on deoxyuridine incorporation (P<0.001), which was 2.0- and 4.2-fold at 48 h and 72 h, respectively. TFGα showed a 1.2-fold inhibitory effect on DNA synthesis at 48 h (P<0.01) and a 1.5-fold inhibition at 72 h (P<0.05). Interferon γ (IFNγ) showed an inhibitory effect on DNA synthesis (1.3-fold; P<0.01). In UM-SCV-6 cells, both IL-10 and IL-13 showed inhibitory effects on deoxyuridine incorporation (1.3- and 1.4-fold at 48 h, respectively; P<0.001) that were even more pronounced at 72 h (2.4- and 2.5-fold respectively; P<0.001). IFNγ caused a 3.6-fold inhibition of DNA synthesis by UM-SCV-6 cells at 72 h (P<0.001). Both TFGβ and TNFα inhibited uridine incorporation (3.0- and 1.6-fold at 48 h, respectively; 2.7-fold at 72 h for both factors). GM-CSF inihibited DNA synthesis by UM-SCV-6 cells 1.3- 2.0-fold at 48 h and 72 h, respectively. In dose/response analyses, the effect of INFα on DNA synthesis was inhibitory in both cell lines at 48 h, while stimulatory effects were observed at 72 h. Electrophoretic analyses of DNA isolated from cells cultured in the presence or absence of different factors did not reveal DNA fragmentation. All cytokines, with the exception of IFNα, showed inhibitory effects on DNA synthesis by vulvar carcinoma cells. Of the factors studied, the recently described interleukins 10 and 13 showed potent inhibition of cell growth, encouraging further investigation on the molecular mechanisms of the observed inhibition. Apoptosis does not seem to be induced in the two vulvar carcinoma cell lines by any of the cytokines studied. Received: 26 March 1996 / Accepted: 5 December 1996     

Biotransformation of hydroquinone to arbutin in plant <Emphasis Type="Italic">in vitro</Emphasis> cultures — preliminary results

Cells from Echinacea purpurea (L.) Moench. (Asteraceae), Exacum affine Balf. f. (Gentianaceae), Melittis melissophyllum L. (Lamiaceae), Ruta graveolens L. and Ruta graveolens ssp. divaricata (Tenore) Gams. (Rutaceae) agitating cultures perform a biotransformation reaction on exogenously supplied hydroquinone into its β-D-glucoside — arbutin, product with valuable medicinal and cosmetic properties. The maximum content of arbutin (determined by HPLC) in the biomass from investigated cultures is 4.01; 3.44; 1.79; 2.48 and 5.07 g/100 g d.w., respectively. Nothing but Ammi majus L. (Apiaceae) cultures contain trace amounts of the product. Arbutin is accumulated in cells; it is occasionally found in media only in vestigial amounts. In most of the investigated cultures the efficiency of the biotransformation process is about 60 %.     

Labaditin,a cyclic peptide with rich biotechnological potential: preliminary toxicological studies and structural changes in water and lipid membrane environment

Cyclic peptides isolated from the plants of the Euphorbiaceae family have been largely studied due to their rigid conformation, which is considered significant for biologic activity. The peptide Labaditin (L₀) and its open chain analogs (L₁) were synthesized by the solid-phase peptide synthesis technique (Fmoc/tBu), and purified to elucidate its interaction with membrane models. A shift in λ_max emission and Stern–Volmer constants values indicate that both tryptophans migrate to a more apolar environment, with L₁ decreasing less than L₀. A circular dichroism (CD) study revealed that L₀ was kept unstructured in aqueous media as much as in the presence of dipalmitoilphosphatidylcholine liposomes. The thermodynamic studies by differential calorimetry (DSC) show a ΔH increase (50 and 18 kcal/mol, for L₀ and L₁, respectively) with peptide concentrations, which is indicative of lipids associating with peptides, resulting in the inability of the lipids to participate in the main transition. Therefore, all CD, DSC, and fluorescence data suggest a greater L₀ membrane insertion. A probable mechanism for Labaditin interaction is based initially on the hydrophobic interaction of the peptide with the lipid membrane, conformational change, peptide adsorption on the lipid surface, and internalization process. Peptide’s antibacterial effect was also evaluated and revealed that only L₀ showed reduction in viability in Gram-positive bacteria while no effects to the Gram-negative.     

Transient electric birefringence of the bacteriophages T3 and T7

Electric birefringence measurements of suspensions of T3 and T7 bacteriophages in 10^?2 M phosphate buffer, pH 6.9, show that there is a difference in their rotational diffusion coefficient. The values corrected to 25°C and water viscosity are D_25,w = 4630 ± 130 sec^?1 and D_25,w = 5290 ± 260 sec^?1 for T3 and T7, respectively. The value obtained from shell model calculations (according to Filson and Bloomfield) is D_25,w = 4500 ± 600 sec^?1. The apparent permanent dipole moments are 4.5 × 10^?26 C·m and 1.7 × 10^?26 C·m for T3 and T7, respectively. For both phage particles the intrinsic optical anisotropy is +7.2 × 10^?3. It is shown that this anisotropy is mainly due to the DNA molecule inside the head of the phage. Its positive value means that there exists an excess orientation of the DNA helix perpendicular to the symmetry axis of the particle. For T7 an unexpectedly large increase of Δn_s and K_sp occurs at a glycerol concentration of about 30% (v/v). This increase is interpreted as being caused by a change of the shape of the particle and/or a change in the secondary structure of the DNA inside the head of the bacteriophage.     

Ionic strength effects on macroion diffusion and excess light-scattering intensities of short DNA rods

Quasielastic and static light-scattering measurements were made on DNA isolated from chicken erythrocyte mononucleosomes as a function of ionic strength between 6 × 10^?4 and 1.0M. A transition from single-exponential autocorrelation functions to markedly non-single-exponential decays was observed around 10^?2M ionic strength and was accompanied by a large decrease in the excess light-scattering intensity. Autocorrelation functions recorded below 10^?2M salt were well fit by the sum of two exponential relaxation which differed by as much as 100-fold in time constants. Apparent diffusion coefficients for the fast and slow processes plateaued around 10^?3M with numerical values approximately 10-fold and 1/10, respectively, of the translational diffusion coefficient for mononucleosome DNA at high ionic strength. This behavior is similar to that observed with poly(L -lysine), for which the slow decay has been associated with a transition to an extraordinary phase. The strong and complex salt dependence observed here illustrates potential difficulties in deriving structural information from scattering by polyions at low ionic strength.     

Aspects of the fossil record and phylogeny of the familyRutaceae (Zanthoxyleae,Toddalioideae)

A comparative analysis of the seed morphology and anatomy of fossil and extantRutaceae (mainlyZanthoxyleae andToddalioideae) is presented. This allows to place the most important fossil taxa in a time-table and on paleogeographical maps. A phylogenetic scheme demonstrates the postulated historical relationships ofEvodia, Zanthoxylum, Fagara, Rutaspermum, Acronychia, Toddalia, Fagaropsis, andPhellodendron.     

Introduction of axial chirality in a planar aromatic ligand results in chiral recognition with DNA

Dimerization of a hydroxycarbazole produces an axially chiral biaryl, BICOL ( 2 ). One enantiomer (R)‐ 2 , is capable of enantioselective binding to different polymorphs of DNA. The biaryl (R)‐ 2 was shown by fluorescence and circular dichroism to induce a shift of Z‐DNA to B‐DNA. The opposite enantiomer (S)‐ 2 shows no specific binding. The significant difference in behaviour between the two enantiomers (S)‐ 2 and (R)‐ 2 is in line with molecular modelling studies which show two very different binding geometries between the enantiomers with each polymorph of DNA. Copyright © 2010 John Wiley & Sons, Ltd.     

Theory of Helix-Coil Transition on DNA-Ligand Complexes: The Effect to Two Types of Interaction of Ligand on the Parameters of Transition

Abstract

The effect of ligand interacting with native DNA by two types on the parameters of helix-coil transition in homopolymers is considered using the most probable distribution method (Yu.S. Lazurkin et al., Biopolymers 1970). It is shown that at a small relative concentration of ligand the melting enthalpy (ΔH) of DNA may be obtained from the universal formula which contains only values directly known from the experiments. It is shown that the formula for the change of melting temperature and width of melting range depending on the total ligand concentration in solution is converted into the corresponding formulae which are defined for the case when only one type of interaction of ligand and DNA is considered.     

High-affinity microtubule protein-higher organism DNA complexes. Many-fold enrichment in repetitive mouse DNA sequences comprised of satellite DNAs

We have examined aspects of the interaction of cycled microtubule protein preparations with ³⁵S-labeled mouse DNA tracer in a competition system with unlabelled competitor E. coli or mouse DNA. The nitrocellulose filter binding assay was used to measure interaction by scintillation counting. DNA molecular weight affected the levels of filter retained ³⁵S-labelled mouse tracer DNA. Filter retention levels increased if ³⁵S-labelled mouse DNA tracer size was increased, and the filter binding level decreased if competitor DNA size was increased. There was a sizeable, reproducible difference in the ³⁵S-labelled mouse DNA tracer binding level of about 1% when E. coli or mouse DNA competitors were compared. Mouse DNA more effectively competed with ³⁵S-labelled mouse DNA for microtubule protein binding than did E. coli DNA, suggesting that a small class of higher-organism DNA sequences interacts very strongly with microtubule protein. From other studies we know this to be the MAP fraction (Marx, K.A. and Denial, T. (1984) in The Molecular Basis of Cancer (Rein, R., ed.), Alan R. Liss, New York, in the press; and Villasante, E., Corces, V.G., Manso-Martinez, R. and Avila, J. (1981) Nucleic Acids Res. 9, 895–908). We find that this difference in competitor DNA strength is qualitatively similar under high-stringency conditions (0.5 M NaCl, high competitor [DNA]) we developed for examining high-affinity complexes. Under high-stringency conditions we isolated 1.2% and 0.6% of ³⁵S-labelled mouse DNA at 4200 and 350 bp respective sizes as nitrocellulose filter bound DNA-protein complexes. At both molecular weights these high-affinity DNA sequences, isolated from the filters, were shown to be significantly enriched in repetitive DNA sequences by S₁ nuclease solution reassociation kinetics. The kinetics are consistent with about a 4-fold mouse satellite DNA enrichment as well as enrichment in other repetitious DNA sequence classes. The high molecular weight filter-bound DNA samples were sedimented to equilibrium in CsCl buoyant density gradients and found to contain primarily mouse satellite DNA density sequences (1.691 g/cm³) with some minor fractions at other density positions (1.670, 1.682, 1.705, 1.740, 1.760 g/cm³) similar to those observed by our laboratory in previous investigations of micrococcal nuclease-resistant chromatin (Marx, K.A. (1977) Biochem. Biophys. Res. Commun. 78, 777–784). That the high-affinity microtubule-bound DNA was some 3–5-fold enriched in mouse satellite sequences was demonstrated by its characteristic BstNI restriction enzyme cleavage pattern