Multiple Fission Research Articles

Morphology engineering for enhanced production of medium-chain-length polyhydroxyalkanoates in Pseudomonas mendocina NK-01

Polyhydroxyalkanoates (PHAs) can be produced by microorganisms from renewable resources and are regarded as promising bioplastics to replace petroleum-based plastics. A medium-chain-length PHAs (mcl-PHA)-producing strain Pseudomonas mendocina NK-01 was isolated previously by our lab and its whole-genome sequence is currently available. Morphology engineering of manipulating cell morphology-related genes has been applied for enhanced accumulation of the intracellular biopolymer short-chain-length PHAs (scl-PHA). However, it has not yet been reported to improve the yield of mcl-PHA by morphology engineering so far. In this work, several well-characterized cell morphology-related genes, including the cell fission ring (Z-ring) location genes minCD, peptidoglycan degradation gene nlpD, actin-like cytoskeleton protein gene mreB, Z-ring formation gene ftsZ, and FtsZ inhibitor gene sulA, were intensively investigated for their impacts on the cell morphology and mcl-PHA accumulation by gene knockout and overexpression in P. mendocina NKU, a upp knockout mutant of P. mendocina NK-01. For a minCD knockout mutant P. mendocina NKU-∆minCD, the average cell length was obviously increased and the mcl-PHA production was improved. However, the nlpD knockout mutant had a shorter cell length and lower mcl-PHA yield compared with P. mendocina NKU. Overexpression of mreB in P. mendocina NKU resulted in spherical cells. When ftsZ was overexpressed in P. mendocina NKU, the cell division was accelerated and the mcl-PHA titer was improved. Furthermore, mreB, ftsZ, or sulA was overexpressed in P. mendocina NKU-∆minCD. Consequently, the mcl-PHA titers were all increased compared with P. mendocina NKU-∆minCD carrying the empty vector. The multiple fission pattern was finally achieved in ftsZ-overexpressing NKU-∆minCD. In this work, improved production of mcl-PHA in P. mendocina NK-01 has been achieved by morphology engineering. This work provides an alternative strategy to enhance mcl-PHA accumulation in mcl-PHA-producing strains.

Effects of cyclin-dependent kinase activity on the coordination of growth and the cell cycle in green algae at different temperatures.

The progression of the cell cycle in green algae dividing by multiple fission is, under otherwise unlimited conditions, affected by the growth rate, set by a combination of light intensity and temperature. In this study, we compared the cell cycle characteristics of Desmodesmus quadricauda at 20 °C or 30 °C and upon shifts between these two temperatures. The duration of the cell cycle in cells grown under continuous illumination at 20 °C was more than double that at 30 °C, suggesting that it was set directly by the growth rate. Similarly, the amounts of DNA, RNA, and bulk protein content per cell at 20 °C were approximately double those of cells grown at the higher temperature. For the shift experiments, cells grown at either 20 °C or 30 °C were transferred to darkness to prevent further growth, and then cultivated at the same or the other temperature. Upon transfer to the lower temperature, fewer nuclei and daughter cells were produced, and not all cells were able to finish the cell cycle by division, remaining multinuclear. Correspondingly, cells placed in the dark at the higher temperature divided faster into more daughter cells than the control cells. These differences correlated with shifts in the preceding cyclin-dependent kinase activity, suggesting that cell cycle progression was not related to growth rate or cell biomass but correlated with cyclin-dependent kinase activity.

Polyploid polynuclear consecutive cell-cycle enables large genome-size in Haematococcus pluvialis

Studies revealed and keep revealing a staggering diversity of life-cycle patterns and genome-sizes in microalgae. Even within a single order like the Volvocales with Volvox carteri, Chlamydomonas reinhardtii, and Dunaliella salina, differences are significant. Additional knowledge gaps in microalgal physiology and genetics hinder our understanding of their evolution and valorization. Herein, we present for the first time, the multiple fission cell-cycle with corresponding DNA quantities of the economically important Volvocales Haematococcus pluvialis. Further, we provide two straightforward protocols for efficient culture synchronization, based on filtration or skimming of zoospores. The cell-cycle and nuclear DNA content where monitored by flow cytometry and fluorescence microscopy after staining with dsDNA dye PICO- or SYBR-green®. The average nuclear DNA content was 3.6–4.1 and 22.6–25.2 pg in zoospores/coenobial daughter-cells and aplanospores, respectively. We found a single DNA replication step for all coenobial daughter cells prior mitosis, followed by one consecutive cytokinesis. Thus, our results suggest a new Haematococcus-type Cn cell-cycle with unusual polyploid and polynuclear phases. This cell-cycle regime might allow for the large genome-size of Haematococcus and is consistent with the mutational-hazard hypothesis for Volvocales microalgae.

A model suite of green algae within the Scenedesmaceae for investigating contrasting desiccation tolerance and morphology.

Microscopic green algae inhabiting desert microbiotic crusts are remarkably diverse phylogenetically, and many desert lineages have independently evolved from aquatic ancestors. Here we worked with five desert and aquatic species within the family Scenedesmaceae to examine mechanisms that underlie desiccation tolerance and release of unicellular versus multicellular progeny. Live cell staining and time-lapse confocal imaging coupled with transmission electron microscopy established that the desert and aquatic species all divide by multiple (rather than binary) fission, although progeny were unicellular in three species and multicellular (joined in a sheet-like coenobium) in two. During division, Golgi complexes were localized near nuclei, and all species exhibited dynamic rotation of the daughter cell mass within the mother cell wall at cytokinesis. Differential desiccation tolerance across the five species, assessed from photosynthetic efficiency during desiccation/rehydration cycles, was accompanied by differential accumulation of intracellular reactive oxygen species (ROS) detected using a dye sensitive to intracellular ROS. Further comparative investigation will aim to understand the genetic, ultrastructural and physiological characteristics supporting unicellular versus multicellular coenobial morphology, and the ability of representatives in the Scenedesmaceae to colonize ecologically diverse, even extreme, habitats.

Evidencing genome downsizing as a result of multiple robertsonian translocations in the subfamily allioideae (Amararyllidaceae)

Large genomes and a history of chromosome rearrangements have made the subfamily Allioideae an interesting model to understand chromosome evolution. The majority of its genera show high karyotype diversity mainly due to polyploidy and Robertsonian translocations (RT) events. The three lineages of Allioideae present different levels of RT, evidenced by metacentric-rich karyotypes observed in Allieae and Tulbaghieae, and a variable proportion between metacentric/acrocentrics in Gilliesieae. Extreme condition of centric fissions occurs in the genera Ipheion, Zoellnerallium and Miersia, whose results in karyotypes formed almost exclusively by acrocentric chromosomes. In this work we investigate how RT drives genome size and chromosome number variation in species of Allioideae. For this, we estimated the genome size by flow cytometry and compiled chromosome number data for 37 species of Allioideae. We employed comparative phylogenetic methods in a Bayesian tree based on ITS1-5.8S-ITS2 sequences. Genome size ranged from 1C = 9.03 picograms (pg) in Ipheon uniflorum to 1C = 54.97 pg in Leucocoryne ixioides. Our analyses revealed that metacentric-rich karyotype is the plesiomorphic state in the subfamily Allioideae and RT as a driving force in genome size variation. This is demonstrated in the relationships Leucocoryne (2n = 14M+4A) /Zoellnerallium (2n = 8M+16A) and Tristagma (2n = 6M+2A)/Ipheion (2n = 14A) with a strong reduction of DNA content (49.5% and 61.47%, respectively) associated to multiple fissions. These changes in Zoellnerallium and Ipheon genomes occurred independently around 12 myr. This relationship was not observed in the Chilean species Speea humilis (2n = 8M+2SM+2A)/ Miersia chilensis (2n = 2SM+18A) with recent differentiation (5 mya), suggesting that multiple centric fusions may require additional evolutionary time to decrease genome size. Our phylogenetic independent contrast analysis of genome size and chromosome number revealed that these traits have a negative correlation (r²=0.18 and p=0.01). This corroborates a tendency of decrease in genome size after events that increase chromosome number such as fissions. Possibly, changes in chromosome linkage may increase meiotic recombination frequency, which might lead to higher probability of unequal crossing over. This mechanism could associate the loss of repetitive sequences to centric fission, explaining downsizing in Allioideae acrocentric-rich genomes.

Evolution of cytokinesis-related protein localization during the emergence of multicellularity in volvocine green algae

BackgroundThe volvocine lineage, containing unicellular Chlamydomonas reinhardtii and differentiated multicellular Volvox carteri, is a powerful model for comparative studies aiming at understanding emergence of multicellularity. Tetrabaena socialis is the simplest multicellular volvocine alga and belongs to the family Tetrabaenaceae that is sister to more complex multicellular volvocine families, Goniaceae and Volvocaceae. Thus, T. socialis is a key species to elucidate the initial steps in the evolution of multicellularity. In the asexual life cycle of C. reinhardtii and multicellular volvocine species, reproductive cells form daughter cells/colonies by multiple fission. In embryogenesis of the multicellular species, daughter protoplasts are connected to one another by cytoplasmic bridges formed by incomplete cytokinesis during multiple fission. These bridges are important for arranging the daughter protoplasts in appropriate positions such that species-specific integrated multicellular individuals are shaped. Detailed comparative studies of cytokinesis between unicellular and simple multicellular volvocine species will help to elucidate the emergence of multicellularity from the unicellular ancestor. However, the cytokinesis-related genes between closely related unicellular and multicellular species have not been subjected to a comparative analysis.ResultsHere we focused on dynamin-related protein 1 (DRP1), which is known for its role in cytokinesis in land plants. Immunofluorescence microscopy using an antibody against T. socialis DRP1 revealed that volvocine DRP1 was localized to division planes during cytokinesis in unicellular C. reinhardtii and two simple multicellular volvocine species T. socialis and Gonium pectorale. DRP1 signals were mainly observed in the newly formed division planes of unicellular C. reinhardtii during multiple fission, whereas in multicellular T. socialis and G. pectorale, DRP1 signals were observed in all division planes during embryogenesis.ConclusionsThese results indicate that the molecular mechanisms of cytokinesis may be different in unicellular and multicellular volvocine algae. The localization of DRP1 during multiple fission might have been modified in the common ancestor of multicellular volvocine algae. This modification may have been essential for the re-orientation of cells and shaping colonies during the emergence of multicellularity in this lineage.

Amblyospora khaliulini (Microsporidia: Amblyosporidae): Investigations on its life cycle and ecology in Aedes communis (Diptera: Culicidae) and Acanthocyclops vernalis (Copepoda: Cyclopidae) with redescription of the species.

A multi-year study was conducted to examine the natural ecology of the microsporidium Amblyospora khaliulini and more fully characterize parasite development and histopathology in all stages of its primary mosquito host, Aedes communis and intermediate copepod host, Acanthocyclops vernalis with redescription of the species. A. khaliulini exhibits polymorphic development, produces three morphologically and functionally distinct spores, and is both horizontally and vertically transmitted. Development in A. vernalis is restricted to females, occurs within the ovaries and results in death of the host. Development is haplophasic with division by binary and multiple fission producing rosette-shaped sporogonial plasmodia and conical uninucleate spores that are orally infectious to Ae. communis larvae. Both sexes are equally susceptible and infections are confined to testes in males and ovaries in females. Initial stages of development include uninucleate schizonts that undergo karyokinesis forming diplokaryotic meronts that divide repeatedly by binary fission. Sporogony occurs in both host sexes, but sporogenesis does not progress normally in adult males and elliptical, thin walled binucleate spores that function in vertical transmission of the microsporidium via infection of the ovaries and eggs are formed in adult females only. Development of vertically acquired infections in larval Ae. communis hosts occurs within fat body tissue, leads to the production of meiospores in male hosts only and results in death during the 4th larval stadium. Initial development is characterized by merogonial multiplication of diplokarya by synchronous binary division producing additional diplokarya. The cessation of merogony and the onset of sporogony are characterized by the simultaneous secretion of a sporophorous vesicle and meiotic division of diplokarya resulting in the formation of octonucleate sporonts that undergo cytokinesis and sporogenesis to form eight uninucleate, broadly ovoid meiospores enclosed within a sporophorous vesicle. The natural prevalence of patent vertically acquired fat body infections in field populations of Ae. communis ranged from 1.6% to 3.6%. Yearly infection rates in A. vernalis copepods ranged from 57.1% to 15.0%. Prevalence rates of horizontally acquired infections in emerging adult Ae. communis ranged from 69.0% to 11.9% in males and 50.0% to 16.4% in females.

Evaluation of neutron flux and fission rate distributions inside the JSI TRIGA Mark II reactor using multiple in-core fission chambers

Within the bilateral project between the CEA Cadarache and the Jožef Stefan Institute (JSI) a wide variety of measurements using multiple fission chambers simultaneously inside the reactor core were performed. The fission rate axial profiles were measured at different positions in the reactor core and at different control rod configurations. A relative comparison of the calculated fission rates using the MCNP code and the measured fission rates was performed. In general the agreement between the measurements and calculations is good, with the deviations within the uncertainties. For better observation and understanding of the neutron flux redistribution due to the control rod movement, the neutron flux and fission rate had been calculated through the entire reactor core for different control rod configurations. The detector position with minimum signal variations due to the regulating and compensating control rod movement during normal operation was determined. The minimum variation is optimal in case we want to reliably determine the reactor power without influence of the regulating and compensating control rod positions.

New Megasphaera-like microfossils reveal their reproductive strategies

New occurrence of Ediacaran Weng’an Biota-type microfossil assemblage has been discovered from the base of the Dengying Formation in Zhenba County, Shaanxi Province, South China. The assemblage is dominated by spherical microfossils ranging from 200 to 1200μm in diameters. Based on the size and morphological features, two genera and three species have been identified, i.e. Megasphaera inornata, M. ornata and Spiralicellula bulbifera. Morphological analyses reveal three types of reproduction, i.e. binary fission, budding, and multiple fission, which are widely distributed among protist and metazoan lineages. Besides, a physiological phenomenon of “ornament growth” has been discovered among M. ornata. These again raise difficulties in determining biological affinities of Megasphaera-like microfossils. The new microfossil assemblage shares the most common elements of embryo-like microfossils with the Weng’an biota. However, those Large Acanthomorphic Acritarchs (LAAs) with great capacity for biostratigraphic correlation are not present. The age of the new assemblage is constrained within the Ediacaran, contemporary with or slightly younger than the Weng’an Biota by regional correlation. The new finding not only extends the geographical distribution of the Ediacaran phosphatized biotas in South China, but also provides valuable fossil materials for studying ontogenesis and evolution of early microscopic organisms and taphonomy of phosphatized soft-bodied fossil Lagerstätte.

A multiplex culture system for the long-term growth of fission yeast cells.

Maintenance of long‐term cultures of yeast cells is central to a broad range of investigations, from metabolic studies to laboratory evolution assays. However, repeated dilutions of batch cultures lead to variations in medium composition, with implications for cell physiology. In Saccharomyces cerevisiae, powerful miniaturized chemostat setups, or ministat arrays, have been shown to allow for constant dilution of multiple independent cultures. Here we set out to adapt these arrays for continuous culture of a morphologically and physiologically distinct yeast, the fission yeast Schizosaccharomyces pombe, with the goal of maintaining constant population density over time. First, we demonstrated that the original ministats are incompatible with growing fission yeast for more than a few generations, prompting us to modify different aspects of the system design. Next, we identified critical parameters for sustaining unbiased vegetative growth in these conditions. This requires deletion of the gsf2 flocculin‐encoding gene, along with addition of galactose to the medium and lowering of the culture temperature. Importantly, we improved the flexibility of the ministats by developing a piezo‐pump module for the independent regulation of the dilution rate of each culture. This made it possible to easily grow strains that have different generation times in the same assay. Our system therefore allows for maintaining multiple fission yeast cultures in exponential growth, adapting the dilution of each culture over time to keep constant population density for hundreds of generations. These multiplex culture systems open the door to a new range of long‐term experiments using this model organism. © 2017 The Authors. Yeast published by John Wiley & Sons, Ltd.

Characterization of the coccoid cyanobacterium Myxosarcina sp. KIOST-1 isolated from mangrove forest in Chuuk State, Federated States of Micronesia

Mangrove forests are known to be inhabited by diverse symbiotic cyanobacterial communities that are capable of N2 fixation. To investigate its biodiversity, root sediments were collected from a mangrove forest in Chuuk State, Federated States of Micronesia (FSM), and an entangled yellow-brown coccoid cyanobacterium was isolated. The isolated cyanobacterium was reproduced by multiple fission and eventually produced baeocytes. Phylogenetic analysis revealed that the isolate was most similar to the genera Myxosarcina and Chroococcidiopsis in the order Pleurocapsales. Compositions of protein, lipid and carbohydrate in the cyanobacterial cells were estimated to be 19.4 ± 0.1%, 18.8 ± 0.4% and 31.5 ± 0.1%, respectively. Interestingly, total fatty acids in the isolate were mainly composed of saturated fatty acids and monounsaturated fatty acids, whereas polyunsaturated fatty acids were not detected. Based on the molecular and biochemical characteristics, the isolate was finally classified in the genus Myxosarcina, and designated as Myxosarcina sp. KIOST-1. These results will contribute to better understanding of cyanobacterial biodiversity in the mangrove forest in FSM as well as the genus Myxosarcina, and also will allow further exploitation of its biotechnological potential on the basis of its cellular characteristics.

Evaluating Marinichlorella kaistiae KAS603 cell size variation, growth and TAG accumulation resulting from rapid adaptation to highly diverse trophic and salinity cultivation regimes

The ability of a microalgal species to adapt to changes in cultivation environment is likely to be beneficial for a successful biofuel/bioproduct production system, because the species could maintain high yields under diverse seasonal or cultivation conditions. Examining factors that enable culturing flexibility in a single species could provide clearer insights than when comparing different species because it will reduce interspecies variability. Marinichlorella kaistiae KAS603 is an excellent model to evaluate mechanisms involved in adaptation to different cultivation regimes. We have studied cell growth, size, triacylglycerol (TAG) accumulation and life cycle stages in this multiple fission dividing strain under a wide variety of conditions, ranging from autotrophic growth in freshwater to mixotrophic growth in fresh and seawater, and to autotrophic growth under saline and hypersaline conditions. Such conditions influence the division mode of the strain, which is linked to biomass and TAG yield. Based on lab and pilot plant experiments, we have discovered that the fastest TAG accumulation takes place under mixotrophy in freshwater, highest yield (culture density plus TAG) condition occurs under mixotrophy in sea water, and the best outdoor culture condition to achieve growth with fewer invasive species is hypersaline natural seawater. In addition to characterizing growth and TAG accumulation characteristics under a wide variety of cultivation conditions, this work sets the stage for investigation into the mechanisms that enable diverse cultivation adaptations, and contribute to the development of this environmentally flexible microalga as a production feedstock.

Cell reproductive patterns in the green alga Pseudokirchneriella subcapitata (=Selenastrum capricornutum) and their variations under exposure to the typical toxicants potassium dichromate and 3,5-DCP.

Pseudokirchneriella subcapitata is a sickle-shaped freshwater green microalga that is normally found in unicellular form. Currently, it is the best known and most frequently used species of ecotoxicological bioindicator because of its high growth rate and sensitivity to toxicants. However, despite this organism’s, our knowledge of its cell biology—for example, the patterns of nuclear and cytoplasmic division in the mitotic stage—is limited. Although it has been reported that P. subcapitata proliferates by popularity forming four daughter cells (autospores) through multiple fission after two nuclear divisions, here, we report two additional reproductive patterns by which two autospores are formed by binary fission (“two-autospore type”) and eight autospores are formed by multiple fission (“eight-autospore type”). Moreover, we found that cell reproductive patterns differed markedly with the culture conditions or with exposure to either of two typical toxicants, potassium dichromate (K2Cr2O7) and 3,5-dichlorophenol (3,5-DCP). The eight-autospore type occurred at the highest frequency in the early phase of culture, but it disappeared under 3,5-DCP at 2.0 mg/L. Under 0.3 mg/L K2CrO7 (Cr(VI)) the eight-autospore type took substantially longer to appear than in control culture. The two-autospore type occurred only in the late phase of culture. To our knowledge, this is the first detailed evaluation of the reproductive patterns of P. subcapitata, which changed dramatically in the presence of toxicants. These findings suggest that observation of the reproductive patterns of P. subcapitata will help to elucidate different cell reactions to toxicants.

Cell size for commitment to cell division and number of successive cell divisions in multicellular volvocine green algae Tetrabaena socialis and Gonium pectorale.

Volvocine algae constitute a green algal lineage comprising unicellular Chlamydomonas, four-celled Tetrabaena, eight to 32-celled Gonium, and others up to Volvox spp., which consist of up to 50,000 cells. These algae proliferate by multiple fissions with cellular growth up to several fold in size and subsequent successive cell divisions. Chlamydomonas reinhardtii cells produce two to 32 daughter cells by one to five divisions, depending on cellular growth in the G1 phase. By contrast, in this study, we found that Tetrabaena socialis and Gonium pectorale cells mostly produced four and eight daughter cells by two and three successive divisions, respectively. In contrast to C. reinhardtii, which is committed to cell division when the cell has grown two-fold, T. socialis and G. pectorale are committed only when the cells have grown four- and eight-fold, respectively. Thus, our results suggest that evolutionary changes in cellular size for commitment largely contributes to the emergence and evolution of multicellularity in volvocine algae.

A compact multi-plate fission chamber for the simultaneous measurement of 233U capture and fission cross-sections

233 U plays the essential role of fissile nucleus in the Th-U fuel cycle. A particularity of 233 U is its small neutron capture cross-section which is about one order of magnitude lower than the fission cross-section on average. Therefore, the accuracy in the measurement of the 233 U capture cross-section essentially relies on efficient capture-fission discrimination thus a combined setup of fission and γ-detectors is needed. At CERN n_TOF the Total Absorption Calorimeter (TAC) coupled with compact fission detectors is used. Previously used MicroMegas (MGAS) detectors showed significant γ-background issues above 100 eV coming from the copper mesh. A new measurement campaign of the 233 U capture cross-section and alpha ratio is planned at the CERN n_TOF facility. For this measurement, a novel cylindrical multi ionization cell chamber was developed in order to provide a compact solution for 14 active targets read out by 8 anodes. Due to the high specific activity of 233 U fast timing properties are required and achieved with the use of customized electronics and the very fast ionizing gas CF4 together with a high electric field strength. This paper describes the new fission chamber and the results of the first tests with neutrons at GELINA proving that it is suitable for the 233 U measurement.

Models of cell division initiation in Chlamydomonas: A challenge to the consensus view

We develop and compare two models for division initiation in cells of the unicellular green alga Chlamydomonas reinhardtii, a topic that has remained controversial in spite of years of empirical work. Achieving a better understanding of C. reinhardtii cell cycle regulation is important because this species is used in studies of fundamental eukaryotic cell features and in studies of the evolution of multicellularity. C. reinhardtii proliferates asexually by multiple fission, interspersing rapid rounds of symmetric division with prolonged periods of growth. Our Model 1 reflects major elements of the current consensus view on C. reinhardtii division initiation, with cells first growing to a specific size, then waiting for a particular time prior to division initiation. In Model 2, our proposed alternative, growing cells divide when they have reached a growth-rate-dependent target size. The two models imply a number of different empirical patterns. We highlight these differences alongside published data, which currently fall short of unequivocally distinguishing these differences in predicted cell behavior. Nevertheless, several lines of evidence are suggestive of more Model 2-like behavior than Model 1-like behavior. Our specification of these models adds rigor to issues that have too often been worked out in relation to loose, verbal models and is directly relevant to future development of informative experiments.

Point-by-Point model calculation of the prompt neutron multiplicity distributionν(A)forU238(n,f)at incident neutron energies ranging from 1 MeV to 80 MeV

Prompt neutron multiplicity distributions \ensuremath{\nu}($A$) are generally required for prompt emission correction of double energy ($2E$) measurements of fission fragments in order to determine pre-neutron fragment properties. The lack of experimental \ensuremath{\nu}($A$) data especially at higher incident neutron energy imposes the use of prompt emission models to predict \ensuremath{\nu}($A$). At incident neutron energies ($\mathit{En}$) where multiple fission chances are involved, the Point-by-Point (PbP) model of prompt emission is able to provide the individual \ensuremath{\nu}($A$) of the compound nuclei of the main and secondary nucleus chains that are undergoing fission at a given $\mathit{En}$. The total \ensuremath{\nu}($A$) are obtained by averaging these individual \ensuremath{\nu}($A$) over the fission chance probabilities (expressed as total and partial fission cross-section ratios). An indirect validation of the total \ensuremath{\nu}($A$) results is proposed. At high $\mathit{En}$ (above 70 MeV) the PbP results of individual \ensuremath{\nu}($A$) of the first few nuclei of the main and secondary nucleus chains exhibit an almost linear increase. This shape is explained by the damping of shell effects entering the superfluid expression of the level-density parameters. They tend to approach the asymptotic values for a great part of the fragments. This fact leads to a smooth and almost linear increase of fragment excitation energy with the fragment mass number that is reflected in a smooth and almost linear behavior of individual \ensuremath{\nu}($A$). The comparison of the present results with those of the GEF code reveals different shapes of \ensuremath{\nu}($A$) as well as different total average neutron multiplicity as a function of the $\mathit{En}$. At high $\mathit{En}$ the PbP calculations definitely reflect the influence of the almost linear shape of individual \ensuremath{\nu}($A$) of the first few nuclei of the U and Pa chains. The differences between the total \ensuremath{\nu}($A$) obtained by averaging the PbP results of individual \ensuremath{\nu}($A$) over fission cross-section ratios of different evaluations are insignificant.

Origin of Amniote Sex Chromosomes: An Ancestral Super-Sex Chromosome, or Common Requirements?

The diversity of sex chromosomes among amniotes is the product of independent evolution of different systems in different lineages, defined by novel sex-determining genes. Convergent evolution is very common, suggesting that some genes are particularly adept at taking on a sex-determining role. Comparative gene mapping, and more recently whole genome sequencing, have now turned up other surprising relationships; different regions of the amniote genome that have become sex determining in some taxa seem to share synteny, or share sequence, in others. Is this, after all, evidence that these regions were once linked in a super-sex chromosome that underwent multiple fission in different ways in different amniote lineages? Or does it signify that special properties of sex chromosomes (paucity of active genes, low recombination, epigenetic regulation to achieve dosage compensation) predispose particular chromosomes to a sex-determining role?

Enhanced production of polyhydroxybutyrate by multiple dividing E. coli.

BackgroundMost bacteria are grown in a binary fission way meaning a bacterial cell is equally divided into two. Polyhydroxyalkanoates (PHA) can be accumulated as inclusion bodies by bacteria. The cell division way and morphology have been shown to play an important role in regulating the bacterial growth and PHA storages.ResultsThe common growth pattern of Escherichia coli was changed to multiple fission patterns by deleting fission related genes minC and minD together, allowing the formation of multiple fission rings (Z-rings) in several positions of an elongated cell, thus a bacterial cell was observed to be divided into more than two daughter cells at same time. To further improve cell growth and PHA production, some genes related with division process including ftsQ, ftsL, ftsW, ftsN and ftsZ, together with the cell shape control gene mreB, were all overexpressed in E. coli JM109 ∆minCD. The changing pattern of E. coli cell growth and morphology resulted in more cell dry weights (CDW) and more than 80 % polyhydroxybutyrate (PHB) accumulation increases compared to its binary fission control grown under the same conditions.ConclusionsThis study clearly demonstrated that combined over-expression genes ftsQ, ftsW, ftsN, ftsL and ftsZ together with shape control gene mreB in multiple division bacterial E. coli JM109 ∆minCD benefited PHA accumulation. Our study provides useful information on increasing the yield of PHA by changing the cell division pattern and cell morphology of E. coli.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0531-6) contains supplementary material, which is available to authorized users.

Engineering the growth pattern and cell morphology for enhanced PHB production by Escherichia coli.

E. coli JM109∆envC∆nlpD deleted with genes envC and nlpD responsible for degrading peptidoglycan (PG) led to long filamentous cell shapes. When cell fission ring location genes minC and minD of Escherichia coli were deleted, E. coli JM109∆minCD changed the cell growth pattern from binary division to multiple fissions. Bacterial morphology can be further engineered by overexpressing sulA gene resulting in inhibition on FtsZ, thus generating very long cellular filaments. By overexpressing sulA in E. coli JM109∆envC∆nlpD and E. coli JM109∆minCD harboring poly(3-hydroxybutyrate) (PHB) synthesis operon phbCAB encoded in plasmid pBHR68, respectively, both engineered cells became long filaments and accumulated more PHB compared with the wild-type. Under same shake flask growth conditions, E. coli JM109∆minCD (pBHR68) overexpressing sulA grown in multiple fission pattern accumulated approximately 70% PHB in 9g/L cell dry mass (CDM), which was significantly higher than E. coli JM109∆envC∆nlpD and the wild type, that produced 7.6 g/L and 8g/L CDM containing 64 % and 51% PHB, respectively. Results demonstrated that a combination of the new division pattern with elongated shape of E. coli improved PHB production. This provided a new vision on the enhanced production of inclusion bodies.