Effects of TNT contaminated soil on vegetation at an explosive range by probing UPLC-qTOF MS profiling method.

Three tree species (Wild olive, Stinkwood and Cape Holy) and a shrub (Dovyalis caffra) were each potted in 20 L pots in order to evaluate the effect of 1,3,5-trinitrotoluene (TNT)-contaminated soil on vegetation. TNT contamination was established by dissolving flake TNT in acetone at 300 and 600 mg per kilogram soil concentrations. One pot for every species was left uncontaminated as control elements. A set of 16 samples, four contaminated, four uncontaminated aerial parts and their corresponding soils, were gathered. These were processed and subjected to a solid phase extraction method to isolate analytes of interest. A laboratory analytical method was applied using ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-qTOF MS). For the UPLC-qTOF MS a gradient for the mobile phase was found which allowed the profiling and separation of metabolites in the aerial parts of the vegetation. This method allowed identification and quantification of major changes caused by TNT contaminated soil on vegetation. The Synapt High Definition Mass Spectrometer SYNAPT HDMS G1 was operated using the electrospray ionisation (ESI) technique in both positive and negative mode. A clear comparison of profiles was achieved and this has been demonstrated by the distinct newly-formed metabolites in the TNT contaminated vegetation understudy. The results have also shown that the chlorophyll region in the contaminated profile was also affected by the uptake of TNT degradation products. This has been observed in the contaminated profiles of Wild olive, Stinkwood and Cape Holly extracts indicating enhanced nutrient availability.

The effect of winter stress on Ilex aquifolium L.previously fumigated with ozone.

European Holly (Ilex aquifolium) received either charcoal-filtered air (CFA) or CFA with 70 nl l(-1) ozone added for 7 h day(-1) over a 28 day period. Plants were then transferred into cooling incubators for hardening (4 degrees C day/2 degrees C night; day length 12 h) for 7 days and then to the frosting stage (2 degrees C day and -5, -10 or -15 degrees C night) for 4 days. The plants were then placed in ambient conditions. Treatment produced significant differences in chlorophyll fluorescence data. Stomatal conductance was significantly higher for the ozone treatments though both showed a general decline over all temperature regimes. Ozone also significantly increased electrolyte leakage and reduced winter survival. These results show that ambient concentrations of ozone can reduce the tolerance of I. aquifolium to freezing stress, which may have serious implications for its establishment and survival.

Micropropagation of Ilex dumosa (Aquifoliaceae) from nodal segments in a tissue culture system.

Micropropagation of Ilex dumosa var. dumosa R. ("yerba señorita") from nodal segments containing one axillary bud was investigated. Shoot regeneration from explants of six-year-old plants was readily achieved in 1/4 strength Murashige and Skoog medium (1/4 MS) plus 30 gr x L(-1) sucrose and supplemented with 4.4 microM BA. Further multiplication and elongation of the regenerated shoots were obtained by subculture in a fresh medium of similar composition with 1.5 gr x L(-1) sucrose. Rooting induction from shoots were achieved in two steps: 1) 7 days in 1/4 MS (30 gr x L(-1) sucrose, 0.25% Phytagel) with 7.3 microM IBA and 2) 21 days in the same medium without IBA and 20 microM of cadaverine added. Regenerated plants were successfully transferred to soil. This micropropagation schedule can be implemented in breeding programs of Ilex dumosa.

Micropropagation of Ilex dumosa (Aquifoliaceae) from nodal segments in a tissue culture system

Micropropagation of Ilex dumosa var. dumosa R. ("yerba señorita") from nodal segments containing one axillary bud was investigated. Shoot regeneration from explants of six-year-old plants was readily achieved in 1/4 strength Murashige and Skoog medium (1/4 MS) plus 30 gr x L(-1) sucrose and supplemented with 4.4 microM BA. Further multiplication and elongation of the regenerated shoots were obtained by subculture in a fresh medium of similar composition with 1.5 gr x L(-1) sucrose. Rooting induction from shoots were achieved in two steps: 1) 7 days in 1/4 MS (30 gr x L(-1) sucrose, 0.25% Phytagel) with 7.3 microM IBA and 2) 21 days in the same medium without IBA and 20 microM of cadaverine added. Regenerated plants were successfully transferred to soil. This micropropagation schedule can be implemented in breeding programs of Ilex dumosa.

Micropropagation of Ilex dumosa (Aquifoliaceae) from nodal segments in a tissue culture system

Micropropagation of Ilex dumosa var. dumosa R. ("yerba señorita") from nodal segments containing one axillary bud was investigated. Shoot regeneration from explants of six-year-old plants was readily achieved in 1/4 strength Murashige and Skoog medium (1/4 MS) plus 30 gr x L(-1) sucrose and supplemented with 4.4 microM BA. Further multiplication and elongation of the regenerated shoots were obtained by subculture in a fresh medium of similar composition with 1.5 gr x L(-1) sucrose. Rooting induction from shoots were achieved in two steps: 1) 7 days in 1/4 MS (30 gr x L(-1) sucrose, 0.25% Phytagel) with 7.3 microM IBA and 2) 21 days in the same medium without IBA and 20 microM of cadaverine added. Regenerated plants were successfully transferred to soil. This micropropagation schedule can be implemented in breeding programs of Ilex dumosa. (AU)