Comparing developmental stability in unisexual and bisexual rock lizards of the genus Darevskia.

Parthenogenetic species are usually considered to be short-lived due to the accumulation of adverse mutations, lack of genetic variability, and inability to adapt to changing environment. If so, one may expect that the phenotype of clonal organisms may reflect such genetic and/or environmental stress. To test this hypothesis, we compared the developmental stability of bisexual and parthenogenetic lizards of the genus Darevskia. We assessed asymmetries in three meristic traits: ventral, preanal, and supratemporal scales. Our results suggest that the amount of ventral and preanal asymmetries is significantly higher in clones compared with their maternal, but not paternal, progenitor species. However, it is questionable, whether this is a consequence of clonality, as it may be considered a mild form of outbreeding depression as well. Moreover, most ventral asymmetries were found in the bisexual species Darevskia valentini. We suggest that greater differences in asymmetry levels among bisexuals may be, for instance, a consequence of the population size: the smaller the population, the higher the inbreeding and the developmental instability. On the basis of the traits examined in this study, the parthenogens do not seem to be of significantly poorer quality.

Experimental assessment of social interactions in two species of the genus Teratoscincus (Gekkota).

Social organization of many reptile species that are rare in the wild remains rather unexplored due to difficulties when setting experiments in the field. Behavioral analysis of standard social situations in laboratory conditions is considered an indirect method to reveal social behavior in the field. We studied two rare species of geckos, Teratoscincus scincus and Teratoscincus keyserlingii, inhabiting sand dunes of Uzbekistan and Eastern Iran. A series of experiments was carried out to quantify responses in social interactions among conspecific adults as well as reactions of these towards conspecific/heterospecific subadults and juveniles. We also assessed the effect of species and sex on recorded behavior. Finally, the reaction to threat stimuli simulating predator attack was analyzed. The species effect was recorded only in the response to a simulated predatory attack: T. scincus typically escaped whereas larger T. keyserlingii attacked the stimulus. In accordance with the sexual competition hypothesis, agonistic interactions were nearly exclusively confined to male-male encounters while females were tolerant to each other. Male-female encounters regularly resulted in mating attempts, which suggests that females are not strongly selective in choosing partners. Therefore, male aggression can be linked to mate guarding or territoriality. Adults' lack of interest in immature geckos may indicate generalized tolerance of adults towards young.

Expression of the fission yeast cell cycle regulator cdc25 induces de novo shoot formation in tobacco: evidence of a cytokinin-like effect by this mitotic activator.

During the last decade, the cell cycle and its control by cyclin-dependent kinases (CDKs) has been extensively studied in eukaryotes. The regulation of CDK activity includes, among others, its activation by Cdc25 phosphatase at G2/M. However, within the plant kingdom studies of this regulation have lagged behind and a plant cdc25 homologue has not been identified yet. Here, we report on the effects of transformation of tobacco (Nicotiana tabacum L., cv. Samsun) with fission yeast (Schizosaccharomyces pombe) cdc25 (Spcdc25) on de novo plant organ formation, a process dependent on rate and orientation of cell division. On shoot-inducing medium (low 1-naphthylacetic acid (NAA), high 6-benzylaminopurine (BAP)) the number of shoots formed on internode segments cultured from transgenic plants was substantially higher than in the non-transformed controls. Anatomical observations indicated that the shoot formation process was accelerated but with no changes in the quality and sequence of shoot development. Surprisingly, and in contrast to the controls, when on root-inducing medium (high NAA, low BAP) cultured segments from transgenic plants failed to initiate hardly any roots. Instead, they continued to form shoots at low frequencies. Moreover, in marked contrast to the controls, stem segments from transgenic plants were able to form shoots even without the addition of exogenous growth regulators to the medium. The results indicate that Spcdc25 expression in culture tobacco stem segments mimicked the developmental effects caused by an exogenous hormone balance shifted towards cytokinins. The observed cytokinin-like effects of Spcdc25 transformation are consistent with the concept of an interaction between cell cycle regulators and phytohormones during plant development.

Denaturing RNA electrophoresis in TAE agarose gels.

Current methods of analytical RNA electrophoresis are based on the utilization of either complicated laboratory instrumentation or toxic, carcinogenic, or expensive chemicals. We suggest here the use of classical Tris-acetate-ethylenediamine tetraacetic acid (TAE) agarose gels combined with prior denaturation of RNA samples in hot formamide for the electrophoretic separation of RNA species. We present a brief comparison of the proposed TAE/formamide method with the most common 3-(N-morpholino)propanesulfonic acid/formaldehyde agarose gel protocol and show that both methods produce comparable results for size determination of RNA molecules and subsequent Northern blotting of gels. In addition to purified RNA samples, the robustness of the TAE/formamide protocol is demonstrated by its suitability for the analysis of RNA quality in crude yeast cell lysates containing large amounts of proteins, DNA, and other contaminating molecules. We therefore propose the TAE/formamide agarose electrophoresis as a rapid, simple, and cheaper alternative to current methods of RNA electrophoresis. Additionally, another benefit is the reduced exposure of laboratory personnel to hazardous chemicals.

Tobacco BY-2 cells expressing fission yeast cdc25 bypass a G2/M block on the cell cycle.

The mitotic inducer gene from Schizosaccharomyces pombe, Spcdc25, was used as a tool to investigate regulation of G2/M in higher plants using the BY-2 (Nicotiana tabacum) cell line as a model. Spcdc25-expressing BY-2 cells exhibited a reduced mitotic cell size through a shortening of the G2 phase. The cells often formed isodiametric double files both in BY-2 cells and in cell suspensions derived from 35S::Spcdc25 tobacco plants. In Spcdc25-expressing cells, the tobacco cyclin-dependent kinase, NtCDKB1, showed high activity in early S phase, S/G2 and early M phase, whereas in empty vector cells CDKB1 activity was transiently high in early S phase but thereafter remained lower. Spcdc25-expressing cells also bypassed a block on G2/M imposed by the cytokinin biosynthetic inhibitor lovastatin (LVS). Surprisingly, cytokinins were at remarkably low levels in Spcdc25-expressing cells compared with the empty vector, explaining why these cells retained mitotic competence despite the presence of LVS. In conclusion, synchronised Spcdc25-expressing BY-2 cells divided prematurely at a small cell size, and they exhibited premature, but sustained, CDKB1 activity even though endogenous cytokinins were virtually undetectable.