Photo and hormonal control of meristem identity in the Arabidopsis flower mutants apetala2 and apetala1.

We have analyzed the contributions of phytochrome and gibberellin signal transduction to the control of flower meristem identity in the Arabidopsis mutants apetala1 (ap1) and apetala2 (ap2). ap1 flowers are partially defective for the establishment of flower meristem identity and are characterized by the production of ectopic secondary or axillary flowers and by branching. Axillary flower production is also induced in ap2-1 flowers by short-day photoperiod and is suppressed by hy1, a mutation blocking phytochrome activity. The production of axillary flower by ap2-1 is also suppressed by exogenous gibberellins and by spindly (spy), a mutation that activates basal gibberellin signal transduction in hormone-independent manner. Ectopic axillary flower production and floral branching by ap1 flowers are also suppressed by spy. We conclude that gibberellins promote flower meristem identity and that the inflorescence-like traits of ap2-1 and ap1-1 flowers are due in part to SPY gene activity.     

Asymptotic distribution of stage-grouped population models

Matrix models are often used to predict the dynamics of size-structured or age-structured populations. The asymptotic behaviour of such models is defined by their malthusian growth rate lambda, and by their stationary distribution w that gives the asymptotic proportion of individuals in each stage. As the coefficients of the transition matrix are estimated from a sample of observations, lambda and w can be considered as random variables whose law depends on the distribution of the observations. The goal of this study is to specify the asymptotic law of lambda and w when using the maximum likelihood estimators of the coefficients of the transition matrix. We prove that lambda and w are asymptotically normal, and the expressions of the asymptotic variance of lambda and of the asymptotic covariance matrix of w are given. The convergence speed of lambda and w towards their asymptotic law is studied using simulations. The results are applied to a real case study that consists of a Usher model for a tropical rain forest in French Guiana. They permit to assess the number of trees to measure to get a given precision on the estimated asymptotic diameter distribution, which is an important information on tropical forest management.     

Sequential fractionation of sediment phosphate

By means of sequential extractions with Ca-NTA and EDTA, a separation was performed between Fe(OOH) P and CaC0₃P in a few sediments; the remaining fraction, considered to be organic phosphate, was quantified as well. We found that with the commonly used method of extraction with NaOH and H₂S0₄, less Fe(OOH) P and much more CaC0₃ P was found than with the chelating extractants. The organic phosphate pool in live and dead algal material and in some mud samples was partly hydrolysed and therefore recovered as inorganic phosphates with classical extractions. The difference between chelating extractants and the classical ones is discussed.Abbreviations o-P: ortho phosphate (or its concentration) - org-P: organic phosphate - extr-P: extractable sediment bound phosphate - extr-Fe: extractable sediment bound iron - Fe(OOH) P: iron bound, sediment phosphate - CaCO₃ P: calcium bound, sediment phosphate - org-C: organic sediment bound carbon     

Effect of the apterous56f mutation on N-acetyltransferase and alkaline phosphatase activities in Drosophila melanogaster females

Alkaline phosphatase (AP) and N-acetyltransferase (NAT) activities were studied in 1-day-old Drosophila melanogaster females of the apterous56f (ap56f) strain, having an elevated level of the juvenile hormone (JH) and a decreased level of dopamine as a result of the mutation, and in the Canton S ancestral wild-type strain in the normal conditions and upon an experimental increase in JH titer. The AP and NAT activities in ap56f females were significantly lower than in Canton S females in the norm. JH application increased the AP activity of mutant females to the level characteristic to JH-treated wild-type females.     

Carbohydrate status in roots of two soybean cultivars: A possible parameter to explain different efficiencies concerning phosphate uptake

The carbohydrate levels in the roots of two soybean cultivars (Century, Woodworth) were determined during the early vegetative growth stage. Based on Michaelis-Menten kinetics Century was characterized as being more efficient at phosphate uptake when compared to Woodworth. The experiments were carried out at normal nutrition (+P treatment) as well as under a condition of moderate phosphate stress (–P treatment). The stress situation was created by withdrawal of phosphate from the nutrient solution for five or nine days. Compared to the +P-treatment the deprivation of phosphate led to a reduction of more than 50% in the phosphate content of the plant tissue.The carbohydrates in the roots exhibited differences between both cultivars with Century generally having higher sucrose levels. During the morning the +P treatment of Century exhibited 10 mg g^-1 and in the afternoon 16 mg g^-1 sucrose (plant dry weight basis). This compares to 8 mg g^-1 sucrose (morning) and 13 mg g^-1 sucrose (afternoon) for Woodworth. Phosphate stress increased the levels of hexoses (glucose, fructose) in both varieties significantly. The sucrose levels remained higher for the Century variety even during phosphate stress. We conclude that the shoot of the Century variety supplies its root with an increased amount of energy in form of carbohydrates compared to the Woodworth variety. This may enable the plant to take up phosphate more efficiently. It is also remarkable that the Century variety reduces the pH of the nutrient medium more rapidly. This active proton deposition may create a high membrane potential which is known to be necessary for phosphate uptake.     

Statistical theory of combinatorial libraries of folding proteins: energetic discrimination of a target structure

A self-consistent theory is presented that can be used to estimate the number and composition of sequences satisfying a predetermined set of constraints. The theory is formulated so as to examine the features of sequences having a particular value of Delta=E(f)-(u), where E(f) is the energy of sequences when in a target structure and (u) is an average energy of non-target structures. The theory yields the probabilities w(i)(alpha) that each position i in the sequence is occupied by a particular monomer type alpha. The theory is applied to a simple lattice model of proteins. Excellent agreement is observed between the theory and the results of exact enumerations. The theory provides a quantitative framework for the design and interpretation of combinatorial experiments involving proteins, where a library of amino acid sequences is searched for sequences that fold to a desired structure.     

Estimation of molecular clocks for ITS and 28S rDNA in Erysiphales

Based on a comparative phylogenetic analysis of Goloviomyces and their host tribes of the Asteraceae, we speculate that Golovinomyces first acquired parasitism to the Asteraceae after migration of the family into the Northern Hemisphere and before the divergence of the tribe Carduaeae. The divergence time of the Carduaeae is estimated to be 25.2Myr ago based on the molecular clock of rbcL sequences of the Asteraceae. When 25.2Myr is given at the node of the first split of the phylogenetic tree of Golovinomyces, nucleotide substitution rates of the Erysiphales are calculated to be 2.52 × 10^–9 per site per year (0.01D = 3.97Myr) in the ITS region and 6.5 × 10^–10 per site per year (0.01D = 15.4Myr) in the D1 and D2 regions of the 28S rDNA.     

Dynamics of stage-structure predator-prey systems under density-dependent effect and mortality

We develop a four dimensional predator-prey system in continuous time with stage-structure for both the communities. The reproduction rate of the prey and the transition rate for the predator, in our model, are assumed to be density-dependent. The stability results for the coexisting equilibrium are obtained by making use of Routh–Hurwitz criteria. Because of the density-dependent effects, numerical simulations are applied in complex situations. We observe that increasing values of the coefficients linked with density-dependent term promote the stability of the coexisting steady state. Our main focus is to understand the variation of stocks when mortality rates on different stage classes are increased. We verified that stable stock on mature predator increases with its increasing mortality rate in three different modeling frameworks. However, no such positive effect on the biomass of the immature predator occurs when immature predators are removed, culled or harvested. Therefore, we could conclude that the appearance of hydra effect on many unstructured predator-prey models is due to the mortality of the mature predator only. No hydra effect is also detected when mature prey is removed in several situations we discussed. Overall, the obtained results are new and could be interesting contribution in theoretical ecology.     

Sympatry drives colour and song evolution in wood-warblers (Parulidae)

Closely related species often exhibit similarities in appearance and behaviour, yet when related species exist in sympatry, signals may diverge to enhance species recognition. Prior comparative studies provided mixed support for this hypothesis, but the relationship between sympatry and signal divergence is likely nonlinear. Constraints on signal diversity may limit signal divergence, especially when large numbers of species are sympatric. We tested the effect of sympatric overlap on plumage colour and song divergence in wood-warblers (Parulidae), a speciose group with diverse visual and vocal signals. We also tested how number of sympatric species influences signal divergence. Allopatric species pairs had overall greater plumage and song divergence compared to sympatric species pairs. However, among sympatric species pairs, plumage divergence positively related to the degree of sympatric overlap in males and females, while male song bandwidth and syllable rate divergence negatively related to sympatric overlap. In addition, as the number of species in sympatry increased, average signal divergence among sympatric species decreased, which is likely due to constraints on warbler perceptual space and signal diversity. Our findings reveal that sympatry influences signal evolution in warblers, though not always as predicted, and that number of sympatric species can limit sympatry''s influence on signal evolution.     

Mutational spectrum induced in Saccharomyces cerevisiae by the carcinogen N-2-acetylaminofluorene

The spectrum of mutations induced by the carcinogen N-2-acetylaminofluorene (AAF) was analysed in Saccharomyces cerevisiae using a forward mutation assay, namely the inactivation of the URA3 gene. The URA3 gene, carried on a yeast/bacterial shuttle vector, was randomly modified in vitro using N-acetoxy-N-2-acetylaminofluorene (N-AcO-AAF) as a model reactive metabolite of the carcinogen AAF. The binding spectrum of AAF to the URA3 gene was determined and found to be essentially random, as all guanine residues reacted about equally well with N-AcO-AAF. Independent Ura^– mutants were selected in vivo after transformation of the modified plasmid into a ura3 yeast strain. Plasmid survival decreased as a function of AAF modification, leading to one lethal hit (37% relative survival) for an average of 50 AAF adducts per plasmid molecule. At this level of modification the mutation frequency was equal to 70 × 10^–4, i.e. 50-fold above the background mutation frequency. UV irradiation of the yeast cells did not further stimulate the mutagenic response, indicating the lack of an SOS-like mutagenic response in yeast. Sequence analysis of the URA3 mutants revealed 48% frameshifts, 44% base substitutions and 8 % complex events. While most base substitutions (74%) were found to be targeted at G residues where AAF is known to form covalent C8 adducts, frameshift mutations were observed at GC base pairs in only 24% of cases. Indeed, more than 60% of frameshift events occurred at sequences such as 5-(A/T)_nG-3 where a short (n = 2 or 3) monotonous run of As or Ts is located on the 5' side of a guanine residue. We refer to these mutations as semi-targeted events and present a potential mechanism that explains their occurrence.     

Biological activity of galactoglucomannan-derived oligosaccharides

A mixture of galactoglucomannan-derived oligosaccharides (GGMOs), degree of polymerization 4–8, (1.2 M and 12 M) stimulated the viability of spruce [Picea abies (L.) Karst] embryos predominately on media supplemented with indole-3-acetic acid: zeatin (0.011, 10.01 mg · 1^-1), at pH 5.O. Their effects on the development and morphogenesis of embryos were dependent on the culture conditions used. These GGMOs also improved the viability of spruce protoplasts when applied at the same concentrations in combination with 1-naphthaleneacetic acid, and to a lesser extent with 2,4-dichlorophenoxyacetic acid at pH 3.8. Viability was also maintained in the presence of GGMOs when the growth hormones were absent; however, the efficiency of protoplast division was low.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - GGMOs galactoglucomannan-derived oligosaccharides - IAA indole-3-acetic acid - zeatin 6-[4-hydroxy-3-methyl-but-2-enylamino]purine This research was supported by the Slovak Grant Agency for Science.     

The point of maximum cell water volume excursion in case of presence of an impermeable solute

A relativistic permeability model of cell osmotic response (Cryobiology 40:64-83; 41:366-367) is applied to a two-solute system with one impermeable solute. The use of the normalized water volume (w), and the amount of intracellular permeable solute (x), which is the product of the water volume and intracellular osmolality (y), as the main variables allowed us to obtain a homogeneous differential equation dx(Delta)/dw(Delta)=f(x(Delta)/w(Delta)), where w(Delta)=w-w(f), x(Delta)=x-x(f), and f refers to the final (equilibrium) values. The solution of this equation is an explicit function, w(Delta)=g(x(Delta)), which is given in the text. This approach allows us to obtain an analytical (exact) expression of the water volume at the moment of the maximum excursion (water extremum w(m)). Results are compared with numeration of basic osmotic equations and with approximation given in (Cryobiology 40:64-83). Assumption that, dw/dt approximately 0 gives good approximations of the kinetics of water and permeable CPA after the point of maximum volume excursion (the slow phase of osmotic response). Practical aspects of the relativistic permeability approach are also discussed.     

Osmotic Forces and Gap Junctions in Spreading Depression: A Computational Model

In a computational model of spreading depression (SD), ionic movement through a neuronal syncytium of cells connected by gap junctions is described electrodiffusively. Simulations predict that SD will not occur unless cells are allowed to expand in response to osmotic pressure gradients and K⁺ is allowed to move through gap junctions. SD waves of [K⁺]_out 25 to 60 mM moving at 2 to 18 mm/min are predicted over the range of parametric values reported in gray matter, with extracellular space decreasing up to 50%. Predicted waveform shape is qualitatively similar to laboratory reports. The delayed-rectifier, NMDA, BK, and Na⁺ currents are predicted to facilitate SD, while SK and A-type K⁺ currents and glial activity impede SD. These predictions are consonant with recent findings that gap junction poisons block SD and support the theories that cytosolic diffusion via gap junctions and osmotic forces are important mechanisms underlying SD.     

Characterization of serine/threonine protein phosphatases in RINm5F insulinoma cells

This study investigates the occurrence and regulation of serine/threonine protein phosphatases (PPases) in insulin-secreting RINm5F insulinoma cells. PPases types 1 and 2A were identified in crude RINm5F cell homogenates by both enzymatic assay and Western blot analysis. We then characterized and compared the inhibitory actions of several compounds isolated from cyanobacteria, marine dinoflagellates and marine sponges, (viz. okadaic acid, microcystin-LR, calyculin-A and nodularin) cation-independent PPase activities in RINm5F cell homogenates. It was found that okadaic acid was the least potent inhibitor (IC₅₀10^–9M, IC₁₀₀10^–6M), while the other compounds exhibited IC₅₀ values of 5·10^–10 M and IC₁₀₀ 5·10^–9 M. The findings indicate that the inhibitory substances employed in this study may be used pharmacologically to investigate the role of serine/threonine PPases in RINm5F cell insulin secretion, a process that is likely to be regulated to a major extent by protein phosphorylation.     

CD31 (PECAM-1) exists as a dimer and is heavily N-glycosylated.

CD31 (PECAM-1) is a highly abundant cell surface glycoprotein expressed on hemopoietic and endothelial cells where it functions as a homophilic adhesion and signaling receptor. Since dimerization and appropriate glycosylation are important features in the regulation of cell surface interactions and signal transduction, we studied the pattern of glycosylation as well as the ability of CD31 to undergo dimerization, both in solution and when expressed on cell membranes. CD31 is heavily glycosylated, with an approximate carbohydrate content of 21%. Nineteen neutral and thirteen sialylated glycans were identified. Ultracentrifugation analysis showed that soluble recombinant CD31 exists in equilibrium between a monomer and a dimer with an approximate dissociation constant of 12.5 microM. Chemical cross-linking studies of both soluble and membrane-expressed CD31 confirmed that CD31 exists as a dimer. These studies suggest that, like E-cadherin, PECAM-dimerization is likely to play a role in CD31 adhesion and signaling.     

Activation of rabbit oocytes: the impact of the Ca2+ signal regime on development

Postfertilization manipulation of mammalian embryos results in various developmental alterations. To determine whether the manipulation of the Ca2+ regime causing oocyte activation is a valuable experimental means in helping understand the biological process by which embryos integrate signals from outside and later regulate gene expression, we linked Ca2+ signal parameters i.e. amplitude, number and frequency, with the efficiency and quality of postimplantation development. Freshly ovulated rabbit oocytes were subjected to repetitive and modulated Ca2+ influx. The results provide three major pieces of information. Firstly, the Ca2+ stimulus is the most efficient signal activating mammalian eggs when it is applied in a repetitive manner, the amplitude being the crucial factor. Secondly, the dynamics of early cleavage does not appear to be determined by either the frequency or the amplitude of modulation of the Ca2+ signal that activates the oocyte. Thirdly, amplitude and temporal modulation of the Ca2+ signal in the early minutes influences the developmental performance and the morphology of the rabbit parthenogenetic conceptus at day 11.5 of pregnancy. The results demonstrate the importance of epigenetic events during postfertilization as well as the possible uses of Ca2+ modulation in studying long term developmental effects.