Classification of electronically generated phantom targets by an Atlantic bottlenose dolphin (Tursiops truncatus).

Animal behavior experiments require not only stimulus control of the animal's behavior, but also precise control of the stimulus itself. In discrimination experiments with real target presentation, the complex interdependence between the physical dimensions and the backscattering process of an object make it difficult to extract and control relevant echo parameters separately. In other phantom-echo experiments, the echoes were relatively simple and could only simulate certain properties of targets. The echo-simulation method utilized in this paper can be used to transform any animal echolocation sound into phantom echoes of high fidelity and complexity. The developed phantom-echo system is implemented on a digital signal-processing board and gives an experimenter fully programmable control over the echo-generating process and the echo structure itself. In this experiment, the capability of a dolphin to discriminate between acoustically simulated phantom replicas of targets and their real equivalents was tested. Phantom replicas were presented in a probe technique during a materials discrimination experiment. The animal accepted the phantom echoes and classified them in the same manner as it classified real targets.

Transgene copy number can be positively or negatively associated with transgene expression.

Two different types of T-DNA insert were found in tobacco plants transformed with Agrobacterium tumefaciens. High-expressing (H) types had one copy of the T-DNA at a locus and produced high expression of the transgene uidA, as measured by uidA RNA levels and beta-glucuronidase activity; low-expressing (L) types had inverted repeats of the T-DNA at a locus and produced low uidA expression. H-types from different transformants acted additively, and cross-fertilization between two different homozygous transformants with H-type inserts produced F1 plants with GUS activity that equalled the parents and individual F2 plants with 50%, 100%, 150% and 200% of parental values. However, the L-type inserts worked in trans to suppress uidA expression from H-type inserts when both were present in the same genome. Hence when a transformant homozygous for the L-type insert was crossed to one homozygous for the H-type, all plants in the F1 and F2 generations with both types of insert had low GUS activity while F2 segregants that only had the H-type inserts had high GUS activity again. Suppression of the H-type gene was associated with increased methylation of the insert. Particle acceleration was used to introduce further copies of uidA into tissues of the transformants. Regardless of the promoter used, those plants with endogenous L-type inserts showed none of the distinct loci of GUS activity readily visible in material with no inserts, showing that L-type inserts could suppress not only the uidA expression of genomic homologues, but also of copies added in vitro.

The effect of T-DNA copy number, position and methylation on reporter gene expression in tobacco transformants.

Inter-transformant variability in the expression of introduced genes was studied in the R1 and R2 generations of 10 tobacco transformants, produced by Agrobacterium-mediated transformation. In replicated and physiologically equivalent material, tranformants showed considerable variability in the expression of the reporter gene uidA as shown by transcript levels and beta-glucuronidase (GUS) activity. However, homozygous R2 material could be investigated for seven of the transformants and among these, and in one line in which two inserts could segregate independently, this inter-transformant variability was reduced to simple bimodal expression. The two levels of expression for GUS activity in leaves were high or low (approximately 2.5 or 0.3 nmol cm-2 min-1 respectively), with no continuous variation. Transformants in the high group had single T-DNA insertions, while those in the low group had multiple T-DNA insertions, at the same or different loci. Within each group, although T-DNA was apparently integrated at different sites in the plant genome, there was no evidence of position effects. GUS activity levels of the transformants were very similar in the field and in environmentally controlled conditions under high or low light. Plants with multiple insertions and low expression also tended to have increased methylation of the integrated T-DNA.

Genetic variability in the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase and its small subunit mRNA in pea.

The amount of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) was studied during greening in 12Pisum sativum L. genotypes. The proportion of mRNA coding for the small subunit of Rubisco (SSU mRNA) was also monitored by hybridization with a cDNA (complementary DNA) probe from one of the nuclear genes coding for SSU (rbcS). Both the SSU mRNA and Rubisco (in g·FW)(-1)) contents rapidly increased in all genotypes on illumination of dark-grown seedlings. Natural genetic variability was found in the amounts of SSU mRNA, Rubisco· (g FW)(-1), total RNA·(g FW)(-1) and mRNA·(µg total RNA)(-1). Differences among genotypes in SSU mRNA were apparently the result of differences in the rate of light-induced accumulation of therbcS gene transcripts. Genotype means for SSU mRNA and Rubisco· (g FW)(-1) amounts during greening were significantly correlated (r=0.788;P<0.01). This indicates a relationship between genetic differences in the rate of light-induced accumulation of therbcS gene transcripts and the Rubisco amount, and establishes a link between natural genetic variability at the molecular and the physiological levels. Genotypic variability in SSU mRNA during greening was also positively correlated to the Rubisco content per unit leaf area in fully greened leaves. Although Southern-blot analysis indicated that there was also natural genetic variability in the copy number of therbcS genes, this difference in copy number could not account for the differences in SSU mRNA production.

Genotype- and promoter-induced variability in transient ß-glucuronidase expression in pea protoplasts.

Leaf mesophyll protoplasts isolated from pea (Pisum sativum L.) genotypes Century and PI244253 showed transient expression of ß-glucuronidase (GUS) when electroporated with plasmid DNA containing various promoter-leader sequence constructs driving the GUS gene. The optimum conditions for transient expression were: using protoplasts isolated from leaf material that had been kept in the dark for 90 h; electroporating at 250 V and 960 µF; and using 125 µg of calf thymus carrier DNA and 75 µ of plasmid DNA. PI244253 had 5 to 20 times the GUS activity levels of Century. Similar levels of transient expression were obtained using either the nopaline synthase or cauliflower mosaic virus 35S (35S) promoters. These levels were lower than that obtained using a duplicated 35S promoter derivative. The presence of an untranslated coat protein mRNA leader sequence from alfalfa mosaic virus between each promoter and the GUS gene resulted in increased GUS activity. Leaf mesophyll protoplasts and root protoplasts of PI244253 did not differ in levels of transient expression.