Evolution of density‐dependent movement during experimental range expansions

Range expansions and biological invasions are prime examples of transient processes that are likely impacted by rapid evolutionary changes. As a spatial process, range expansions are driven by dispersal and movement behaviour. Although it is widely accepted that dispersal and movement may be context‐dependent, for instance density‐dependent, and best represented by reaction norms, the evolution of density‐dependent movement during range expansions has received little experimental attention. We therefore tested current theory predicting the evolution of increased movement at low densities at range margins using highly replicated and controlled range expansion experiments across multiple genotypes of the protist model system Tetrahymena thermophila. Although rare, we found evolutionary changes during range expansions even in the absence of initial standing genetic variation. Range expansions led to the evolution of negatively density‐dependent movement at range margins. In addition, we report the evolution of increased intrastrain competitive ability and concurrently decreased population growth rates in range cores. Our findings highlight the importance of understanding movement and dispersal as evolving reaction norms and plastic life‐history traits of central relevance for range expansions, biological invasions and the dynamics of spatially structured systems in general.     

Modeling Tetrahymena thermophila growth and protease production

Oxygen concentrations stimulated growth (maximum number of cells) and protease secretion by Tetrahymena thermophila. Agitation and aeration conditions for growth and protease secretion were optimised by a central composite design. The best optimised combination was a stirrer speed of 338 rpm and an aeration of 1 vvm. Journal of Industrial Microbiology & Biotechnology (2000) 25, 58–61. Received 24 September 1999/ Accepted in revised form 06 March 2000     

Temperature Effects on Maturity Periods in Tetrahymena pyriformis Syngen 1*

Cells of Tetrahymena pyriformis syngen 1 grown at 30 C after conjugation achieve sexual maturity more quickly than do cells grown at 19 C, whether time is measured in numbers of cell divisions or in terms of absolute time. This result is achieved regardless of the temperature at which conjugation and nuclear reorganization occur. These observations differ from those of other workers investigating Paramecium, and suggest that the long term “chronometer” is more tightly coupled to cell division in Paramecium multimicronucleatum and Paramecium caudatum than in Tetrahymena pyriformis.     

Inter-strain Variability of Structural Proteins in Tetrahymena*

SYNOPSIS The high molecular weight proteins found in isolated pellicles of Tetrahymena have been compared in several individual strains within the genus using SDS polyacrylamide gel electrophoresis. Three variants of the B-protein of epiplasm (MW 174,000; 155,000; 145,000) and 2 of the C-protein (MW 140,000; 122,000) were found among the strains examined. No variation was observed in the major kinetodesmal fiber protein (MW 250,000). The variation found between strains in the proteins of a structure which is (as far as we know) the same in all strains indicates a disjunction between evolutionary change at the 2 levels of organization. The taxonomic implications of the observed variation in structural proteins in Tetrahymena are discussed.     

Antiproliferative activity of L-asparaginase of Tetrahymena pyriformis on human breast cancer cell lines

Purified L-asparaginase of Tetrahymena pyriformis is a multi-subunit enzyme exhibiting protein kinase activity as well. The enzyme's L-asparaginase activity is affected by its phosphorylation state. Both native and dephosphorylated L-asparaginase show antiproliferative activity on three breast cancer cell lines (T47D, BT20 and MCF-7) and on Walker 256 cells. These cells do not possess measurable L-asparaginase or L-asparagine synthetase activity. When T47D cells are treated for different times with L-asparaginase and then placed in fresh medium, the growth of cells treated for 1, 3, or 6 hours is initiated and parallels control curve, while the growth of cells treated for 24 or 48 hours with L-asparaginase stays at the same inhibitory level (24 h treatment) or continues to drop (48 h treatment). Addition of D-asparagine, a competitive inhibitor of T. pyriformis L-asparaginase, counteracts the antiproliferative activity of L-asparaginase, indicating that L-asparaginase and not the kinase activity is responsible for that effect.     

Monstrous Tetrahymena with Intraclonal Variation in Structure Produced by Hereditary Modification of Normal Cells

SYNOPSIS Monstrous Tetrahymena pyriformis strain GL may be isolated after exposure of normal cells to numerous heat shocks, to flattening on agar or gelatin plates, or to viscous solutions of methyl cellulose. It is shown that in some cases the abnormalities are inherited and that this results in clones where the cells are different from each other and have various abnormalities with respect to cortical pattern, swimming and feeding behavior, and generation time. Furthermore, it is shown that these cells are produced rather than selected by the experimental treatments. Evidence is presented that growth without division is important for production of the abnormal organisms. The basis of the inheritance of the abnormalities is discussed.     

Effect of phorbol 12-myristate 13-acetate (PMA) on the phosphoinositol (PI) system in Tetrahymena. Study of the 32P incorporation and breakdown of phospholipids

Phorbol 12-myristate 13-acetate (PMA) treatment elicited an increased ³²P incorporation into phospholipids namely phosphatadyl-inositol (PI); phosphatidyl-inositol-4-phosphate (PIP); phosphatidyl-inositol-4,5-bis-phosphate (PIP₂); phosphatidyl-acid (PA); phosphatidyl-choline (PC) and phosphatidyl-ethanolamine (PE) particularly at the 20–30th min after treatment. The ratio of members of the phosphoinositol system, especially PIP and PI, related to the total phospholipid content was increased. PMA (2 × 10^?7 M ) was the most effective of the three concentrations tested. The results call attention to the presence of a working phosphoinositol system in Protozoa.     

Thymidylate Synthetase as a Chemotherapeutic Target in the Treatment of Avian Coccidiosis*

SYNOPSIS. Thymidylate synthetase (E.C.2.1.1.45) has been demonstrated in unsporulated oocysts of Eimeria tenella. The properties of this enzyme have also been investigated in Tetrahymena pyriformis, as a protozoan model, and 7-day-old chick embryo, as a host model. The enzymes from E. tenella and chick embryo were inhibited by all concentrations of MnCl₂ and MgCl₂ tested. Tetrahymena pyriformis thymidylate synthetase was stimulated by low concentrations of both these cations but was inhibited by high concentrations. Subsequent data refer to chick embryo, E. tenella and T. pyriformis respectively: the apparent K_m was 5.89 μM, 5.94 μM, and 0.53 M for the substrate dUMP: and 5.13 μM, 1.10 μM and 4.65 μM, respectively for the cofactor N⁵N¹⁰-methylenetetrahydrofolate. The pH optimum for the enzyme from both chick embryo and T. pyriformis was 8.0, with Tris-HCl buffer; activity of E. tenella thymidylate synthetase was still increasing at pH 8.2. The E. tenella enzyme was found to have a molecular weight of 4.6–4.9 × 10⁵ daltons. The effects of nucleotides, inhibitors, and the omission of assay components on each enzyme are presented. Thymidylate synthetase from E. tenella is not greatly different from that of chick embryo, but does not resemble the enzyme from T. pyriformis. A case for using thymidylate synthetase as a chemotherapeutic target in the treatment of Eimeria infections remains. Indeed Eimeria may be considered as a model for infections caused by other protozoan parasites, such as Toxoplasma and Plasmodium, provided that suitable inhibitors can be found that are not toxic to the host.     

RanGTPase激活蛋白RanGAP在嗜热四膜虫细胞中的定位及功能

RanGTPase激活蛋白(RanGTPase-activatingprotein,RanGAP)和Ran相互作用,提高了RanGTPase水解GTP的效率.RanGAP参与细胞内核质运输、纺锤体组装、核膜重建和异染色质的组装.生物进化过程中,不同生物的RanGAP表现出结构和功能的多样性.本研究从嗜热四膜虫大核基因组中鉴定出1个保守的RanGTPase激活蛋白基因RanGAP(TTHERM_00766430).实时荧光定量PCR表明,RanGAP在四膜虫营养生长、饥饿和有性生殖过程中均有表达,且在有性生殖4~6h表达水平最高.免疫荧光定位表明,在营养生长期、饥饿期及有性生殖的早期,RanGAP定位于细胞质中;在有性生殖后期,RanGAP定位于凋亡的大核中.过表达RanGAP的细胞增殖速率下降,大核分裂和胞质缢缩异常,产生无大核细胞.敲减RanGAP的细胞大核形态异常,细胞增殖速率下降,无丝分裂受到抑制,进而产生无大核细胞.RanGAP的过表达或敲除分别引起四膜虫RAN1,RanBP1和RCC1基因的表达下调或上调.结果表明,RanGAP通过Ran信号通路调控了嗜热四膜虫无性生殖过程中大核的无丝分裂,并可能参与了有性生殖过程中亲本大核的凋亡.     

Secretome analysis of thermophilic mould Myceliophthora thermophila cultivated on rice straw and hydrolysis of lignocellulosic biomass for bioethanol production

Abstract

Myceliophthora thermophila encodes for large number of carbohydrate-active enzymes (CAZymes) involved in lignocellulosic biomass degradation. The mould was grown on rice straw in solid state fermentation at pH 5.0 and 45?°C that produced high levels of cellulolytic and xylanolytic enzymes i.e. 2218.12, 515.23, 478.23, 13.34?U/g DMR for xylanase, CMCase, FPase and β-glucosidase, respectively. The secretome analysis of M. thermophila BJAMDU5 by mass spectroscopy, described 124 different proteins with majority of CAZymes consisting of glycosyl hydrolases (GH), lytic polysaccharide mono-oxygenases (LPMO), carbohydrate esterases (CE) and polysaccharide lyases (PL). Furthermore, the enzyme cocktail of the mould was evaluated for hydrolysis of steam treated rice straw that produced 184.59?mg/g substrate reducing sugars after 24?h, which was used for production of bioethanol by using fast fermenting yeast Saccharomyces cerevisiae resulting in high production of bioethanol.