Seed and vegetative propagation methods for the rare Florida native species Chionanthus pygmaeus (pygmy fringetree)

Chionanthus pygmaeus Small (pygmy fringetree) (Oleaceae) is an endemic and rare Florida species, which has an attractive, small habit giving it great potential for use in managed landscapes. Members of the genus Chionanthus are difficult to propagate via cuttings and possess complex seed dormancies that are not well understood. Conservation of pygmy fringetree and its potential for commercial propagation for use in managed landscapes is contingent on a better understanding of its complex seed dormancy and enhancement of its propagation. I conducted two experiments to assess sexual and asexual propagation methods for pygmy fringetree. The first experiment was conducted to determine what factors are involved in overcoming seed dormancy. Various scarification treatments, which mimicked conditions seeds are exposed to in the wild, were investigated to determine their effects on germination of 20-year-old seeds originally collected from the species’ native range. Treatments included endocarp removal, sulfuric acid, boiling-water, and smoke-water treatments. Prior to treatment initiation, seed viability was estimated to be 12%. Treated seeds went through two cold- and two warm-stratification periods of 4°C and 25°C, respectively, in a dark growth chamber. After 180 days, none of the treatments induced early germination. Seeds were then tested for viability, which was 11%. Seed dormancy of the species is apparently complex, allowing some of the seeds to retain some degree of viability, but without dormancy requirements satisfied. The second experiment was conducted to assess if pygmy fringetree could be successfully propagated via hardwood or root cuttings if the appropriate combination of environmental conditions and hormones were applied. Hardwood and root cuttings were treated with either 1000 ppm IBA talc, 8000 ppm IBA talc, or inert talc. All cuttings were placed on a mist bench in a greenhouse for 9 weeks. Hardwood cuttings were supplemented with bottom heat at 24 °C. No treatments were successful in inducing adventitious root formation. I conclude that pygmy fringetree seeds possess complex dormancy that was not able to be overcome by the treatments utilized. However, this result is confounded by the age of the seeds used in the experiment. I also conclude that vegetative propagation of pygmy fringetree is highly dependent on the time of year cuttings are harvested. Further research of both seed and asexual propagation methods need to be explored before pygmy fringetree can be propagated on a commercial scale.

Improving the stability of an oxime based electrochemical microsensor for organo-phosphate vapor detection

A micro gas sensor has been developed by our group for the detection of organo-phosphate vapors using an aqueous oxime solution. The analyte diffuses from the high flow rate gas stream through a porous membrane to the low flow rate aqueous phase. It reacts with the oxime PBO (1-Phenyl-1,2,3,-butanetrione 2-oxime) to produce cyanide ions, which are then detected electrochemically from the change in solution potential. Previous work on this oxime based electrochemistry indicated that the optimal buffer pH for the aqueous solution was approximately 10. A basic environment is needed for the oxime anion to form and the detection reaction to take place. At this specific pH, the potential response of the sensor to an analyte (such as acetic anhydride) is maximized. However, sensor response slowly decreases as the aqueous oxime solution ages, by as much as 80% in first 24 hours. The decrease in sensor response is due to cyanide which is produced during the oxime degradation process, as evidenced by the cyanide selective electrode. Solid phase micro-extraction carried out on the oxime solution found several other possible degradation products, including acetic acid, N-hydroxy benzamide, benzoic acid, benzoyl cyanide, 1-Phenyl 1,3-butadione, 2-isonitrosoacetophenone and an imine derived from the oxime. It was concluded that degradation occurred through nucleophilic attack by a hydroxide or oxime anion to produce cyanide, as well as a nitrogen atom rearrangement similar to Beckmann rearrangement. The stability of the oxime in organic solvents is most likely due to the lack of water, and specifically hydroxide ions. The reaction between oxime and organo-phosphate to produce cyanide ions requires hydroxide ions, and therefore pure organic solvents are not compatible with the current micro-sensor electrochemistry. By combining a concentrated organic oxime solution with the basic aqueous buffer just prior to being used in the detection process, oxime degradation can be avoided while preserving the original electrochemical detection scheme. Based on beaker cell experiments with selective cyanide sensitive electrodes, ethanol was chosen as the best organic solvent due to its stabilizing effect on the oxime, minimal interference with the aqueous electrochemistry, and compatibility with the current microsensor material (PMMA). Further studies showed that ethanol had a small effect on micro-sensor performance by reducing the rate of cyanide production and decreasing the overall response time. To avoid incomplete mixing of the aqueous and organic solutions, they were pre-mixed externally at a 10:1 ratio, respectively. To adapt the microsensor design to allow for mixing to take place within the device, a small serpentine channel component was fabricated with the same dimensions and material as the original sensor. This allowed for seamless integration of the microsensor with the serpentine mixing channel. Mixing in the serpentine microchannel takes place via diffusion. Both detector potential response and diffusional mixing improve with increased liquid residence time, and thus decreased liquid flowrate. Micromixer performance was studies at a 10:1 aqueous buffer to organic solution flow rate ratio, for a total rate of 5.5 μL/min. It was found that the sensor response utilizing the integrated micromixer was nearly identical to the response when the solutions were premixed and fed at the same rate.