Euplotes Cytoskeleton: Tubulin and Microtubular Systems in Interphase1,2

The Euplotes cytoskeleton obtained by treatment with Triton X-100 in a microtubule-stabilizing buffer has been studied by electron microscopy and SDS-polyacrylamide electrophoresis. An antiserum has been prepared using the tubulin band from preparative gels of Euplotes cytoskeletons as an antigen. This antiserum reacts with the tubulin from different ciliated protozoa but fails to recognize the vertebrate tubulin by immunoblotting. Immunoblotting studies have demonstrated a slower electrophoretic mobility for the α-tubulin of Euplotes and Oxytricha than that of Paramecium and Tetrahymena. A cytoplasmic microtubular network in Euplotes has been revealed by indirect immunoftuorescence using both an anti-α-tubulin monoclonal antibody directed against chick brain tubulin and an antiserum raised against the tubulin of Euplotes.     

Phylogenomic species delimitation and host‐symbiont coevolution in the fungus‐farming ant genus Sericomyrmex Mayr (Hymenoptera: Formicidae): ultraconserved elements (UCEs) resolve a recent radiation

Ants in the Neotropical genus Sericomyrmex Mayr cultivate fungi for food. Both ants and fungi are obligate, coevolved symbionts. The taxonomy of Sericomyrmex is problematic because the morphology of the worker caste is generally homogeneous across all of the species within the genus, species limits are vague, and the relationships between them are unknown. We used ultraconserved elements (UCEs) as genome‐scale markers to reconstruct evolutionary history and to infer species boundaries in Sericomyrmex. We recovered an average of ～990 UCE loci for 88 Sericomyrmex samples from across the geographical range of the genus as well as for five outgroup taxa. Using maximum likelihood and species‐tree approaches, we recovered nearly identical topologies across datasets with 50–95% matrix completeness. We identify nine species‐level lineages in Sericomyrmex, including two new species. This is less than the previously described 19 species, even accounting for two species for which we had no UCE samples, which brings the total number of Sericomyrmex species to 11. Divergence‐dating analyses recovered 4.3 Ma as the crown‐group age estimates for Sericomyrmex, indicating a recent, rapid radiation. We also sequenced mitochondrial cytochrome oxidase subunit I (COI) for 125 specimens. Resolution and support for clades in our COI phylogeny are weak, indicating that COI is not an appropriate species‐delimitation tool. However, taxa within species consistently cluster together, suggesting that COI is useful as a species identification (‘DNA barcoding’) tool. We also sequenced internal transcribed spacer (ITS) and large subunit (LSU) for 32 Sericomyrmex fungal cultivars. The fungal phylogeny confirms that Sericomyrmex fungi are generalized higher‐attine cultivars, interspersed with Trachymyrmex‐associated fungal species, indicating cultivar sharing and horizontal transfer between these two genera. Our results indicate that UCEs offer immense potential for delimiting and resolving relationships of problematic, recently diverged species.     

Isolation of polymorphic microsatellite loci from the flea beetle Phyllotreta nemorum L. (Coleoptera: Chrysomelidae)

Ten microsatellite markers for the flea beetle Phyllotreta nemorum were developed using di‐ and trinucleotide repeat‐enriched libraries. Each of these primer pairs were characterized on 96 individuals. Expected heterozygosities ranged between 0.11 and 0.84 and the number of alleles ranged between two and 14 per locus. These microsatellite markers are the first published for any Phyllotreta species.     

Comparative study of behavioural tests in the SOD1G93A mouse model of amyotrophic lateral sclerosis

In preclinical trials, a sensitive functional test is required to detect changes in the motor behaviour of the SOD1G93A mouse model of amyotrophic lateral sclerosis (ALS). We evaluated changes in body weight and motor impairment in behavioural tests, such as the rotarod, the hanging-wire test and the treadmill, of transgenic and wild type mice. We found differences in detection of the onset of symptoms and progression of the disease between the different tests assessed. Moreover, the data showed significant gender differences in the motor behaviour of this mouse model. The rotarod and the hanging-wire test were more sensitive to detect early motor impairment. Moreover, the results suggested that the rotarod and hanging-wire became the most accurate tests rather than treadmill to characterise the ALS disease phenotype.     

Role of aquaporins in root water transport of ectomycorrhizal jack pine (Pinus banksiana) seedlings exposed to NaCl and fluoride

Effects of ectomycorrhizal (ECM) fungus Suillus tomentosus on water transport properties were studied in jack pine (Pinus banksiana) seedlings. The hydraulic conductivity of root cortical cells (L_pc) and of the whole root system (L_pr) in ECM plants was higher by twofold to fourfold compared with the non‐ECM seedlings. HgCl₂ had a greater inhibitory effect on L_pc in ECM compared with non‐ECM seedlings, suggesting that the mercury‐sensitive, aquaporin (AQP)‐mediated water transport was largely responsible for the differences in L_pc between the two groups of plants. L_pc was rapidly and drastically reduced by the 50 mm NaCl treatment. However, in ECM plants, the initial decline in L_pc was followed by a quick recovery to the pre‐treatment level, while the reduction of L_pc in non‐ECM seedlings progressed over time. Treatments with fluoride reduced L_pc by about twofold in non‐ECM seedlings and caused smaller reductions of L_pc in ECM plants. When either 2 mm KF or 2 mm NaF were added to the 50 mm NaCl treatment solution, the inhibitory effect of NaCl on L_pc was rapidly reversed in both groups of plants. The results suggest that AQP‐mediated water transport may be linked to the enhancement of salt stress resistance reported for ECM plants.     

Encystment in the Hypotrichous Ciliate Paraurostyla weissei: Ultrastructure and Cytochemistry1

The cyst wall of Paraurostyla weissei consists of four morphologically distinct layers. It shows an ultrastructure and composition similar to that of the previously described kinetosome-resorbing cysts, and its cytoplasm displays characteristics of “urostylid-type” cysts. Therefore it is possible to consider it a “transition ciliate” between Stichotrichina and Sporadotrichina.     

Variation in sexual reproduction in orchids and its evolutionary consequences: a spasmodic journey to diversification

The great taxonomic diversity of the Orchidaceae is often attributed to adaptive radiation for specific pollinators driven by selection for outcrossing. However, when one looks beyond the product to the process, the evidence for selection is less than overwhelming. We explore this problem by discussing relevant aspects of orchid biology and asking which aspects of reproduction explain the intricate pollination mechanisms and diversification of this family. We reaffirm that orchids are primarily pollination limited, the severity of which is affected by resource constraints. Fruit set is higher in temperate than in tropical species, and in species which offer pollinator rewards than those that do not. Reproductive success is skewed towards few individuals in a population and effective population sizes are often small. Population structure, reproductive success and gene flow among populations suggest that in many situations genetic drift may be as important as selection in fostering genetic and morphological variation in this family. Although there is some evidence for a gradualist model of evolutionary change, we believe that the great diversity in this family is largely a consequence of sequential and rapid interplay between drift and natural selection.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 84 , 1–54.