Development of a pulsed uniform supersonic gas expansion system based on an aerodynamic chopper for gas phase reaction kinetic studies at ultra-low temperatures.

A detailed description of a new pulsed supersonic uniform gas expansion system is presented together with the experimental validation of the setup by applying the CRESU (French acronym for Cinétique de Réaction en Ecoulement Supersonique Uniforme or Reaction Kinetics in a Uniform Supersonic Flow) technique to the gas-phase reaction of OH radicals with 1-butene at ca. 23 K and 0.63 millibars of helium (carrier gas). The carrier gas flow, containing negligible mixing ratios of OH-precursor and 1-butene, is expanded from a high pressure reservoir (337 millibars) to a low pressure region (0.63 millibars) through a convergent-divergent nozzle (Laval type). The novelty of this experimental setup is that the uniform supersonic flow is pulsed by means of a Teflon-coated aerodynamic chopper provided with two symmetrical apertures. Under these operational conditions, the designed Laval nozzle achieves a temperature of (22.4 ± 1.4) K in the gas jet. The spatial characterization of the temperature and the total gas density within the pulsed uniform supersonic flow has also been performed by both aerodynamical and spectroscopic methods. The gas consumption with this technique is considerably reduced with respect to a continuous CRESU system. The kinetics of the OH+1-butene reaction was investigated by the pulsed laser photolysis/laser induced fluorescence technique. The rotation speed of the disk is temporally synchronized with the exit of the photolysis and the probe lasers. The rate coefficient (k(OH)) for the reaction under investigation was then obtained and compared with the only available data at this temperature.

Evolutionary diversification of clades of squamate reptiles.

We analysed the diversification of squamate reptiles (7488 species) based on a new molecular phylogeny, and compared the results to similar estimates for passerine birds (5712 species). The number of species in each of 36 squamate lineages showed no evidence of phylogenetic conservatism. Compared with a random speciation-extinction process with parameters estimated from the size distribution of clades, the alethinophidian snakes (2600 species) were larger than expected and 13 clades, each having fewer than 20 species, were smaller than expected, indicating rate heterogeneity. From a lineage-through-time plot, we estimated that a provisional rate of lineage extinction (0.66 per Myr) was 94% of the rate of lineage splitting (0.70 per Myr). Diversification in squamate lineages was independent of their stem age, but strongly related to the area of the region within which they occur. Tropical vs. temperate latitude exerted a marginally significant influence on species richness. In comparison with passerine birds, squamates share several clade features, including: (1) independence of species richness and age; (2) lack of phylogenetic signal with respect to clade size; (3) general absence of exceptionally large clades; (4) over-representation of small clades; (5) influence of region size on clade size; and (6) similar rates of speciation and extinction. The evidence for both groups suggests that clade size has achieved long-term equilibrium, suggesting negative feedback of species richness on the rate of diversification.

Genetic analysis of Agrobacterium tumefaciens susceptibility in Brassica oleracea.

The genetic control and heritability of Agrobacterium tumefaciens susceptibility was investigated using a doubled haploid (DH) mapping population of Brassica oleracea and the associated RFLP map. Preliminary studies were carried out by analysis of an 8 x 8 diallel, for which the parental lines were selected to include a range of susceptibilities to A. tumefaciens. The variation observed within the diallel was attributed to both additive and dominant gene effects, with additive gene effects being more important. A broad sense heritability value of 0.95 suggested that 95% of the observed variation was due to genetic effects, with just 5% attributed to non-genetic or environmental effects. A high narrow-sense heritability value of 0.79 suggested that 79% of this trait was controlled by additive gene effects and, therefore, the potential to introduce this trait into breeding material is high. Fifty-nine DH lines from the mapping population were screened for susceptibility towards A. tumefaciens. Variation in susceptibility was observed across the population. The results of the DH screen were entered into the mapping programme MAPQTL and a highly significant quantitative trait loci (QTL) associated with susceptibility to A. tumefaciens was identified on linkage group 09. The use of substitution lines covering this region confirmed the location of this QTL. This work shows that susceptibility to A. tumefaciens is a heritable trait, and the transfer of susceptibility into resistant lines is demonstrated. These findings may help to overcome genotype restrictions to genetic transformation.

Genetic analysis of in vitro shoot regeneration from cotyledonary petioles of Brassica oleracea.

Diallel analysis was used to investigate the genetic control of in vitro shoot regeneration in Brassica oleracea. Twelve doubled haploid (DH) lines, selected to include a range of genotypes with differing shoot regeneration potentials, were crossed reciprocally to produce 132 F(1) and 12 selfed, DH families. Cotyledonary petioles from 4-day-old seedlings, from all families, were excised and maintained on MS medium supplemented with 2 mg/l BAP. Explants were scored after 44 days for both the presence or absence of shoots and the number of regenerating shoots per explant. Diallel analysis showed both shoot regeneration and the production of multiple shoots to be controlled by additive and dominant gene effects, with additive effects being more important. Additive gene effects accounted for 71% and 77% of the genetic variation observed within the diallel for shoot regeneration and multiple shoot regeneration, respectively. By investigating the shoot regeneration potential of subsequent backcross and F(2) populations, the ability to introduce and increase shoot regeneration potential into otherwise recalcitrant lines was demonstrated.