Deborah M.E. Pearce

Characterisation of programmed cell death during aerenchyma formation induced by ethylene or hypoxia in roots of maize ( Zea mays L.)

Abstract. Aerenchyma is a tissue type characterised by prominent intercellular spaces which enhance flooding tolerance in some plant species by facilitating gas diffusion between roots and the aerial environment. Aerenchyma in maize roots forms by collapse and death of some of the cortical cells in a process that can be promoted by imposing oxygen shortage or by ethylene treatment. Maize roots grown hydroponically in 3% oxygen, 1 μl l−1 ethylene or 21% oxygen (control) were analysed by a combination of light and electron microscopy. Use of in-situ terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) suggested internucleosomal cleavage of DNA. However, chromatin condensation detectable by electron microscopy was preceded by cytoplasmic changes including plasma membrane invagination and the formation of vesicles, in contrast to mammalian apoptosis in which chromatin condensation is the first detectable event. Later, cellular condensation, condensation of chromatin and the presence of intact organelles surrounded by membrane resembling apoptotic bodies were observed. All these events were complete before cell wall degradation was apparent. Therefore, aerenchyma formation initiated by hypoxia or ethylene appears to be a form of programmed cell death that shows characteristics in part resembling both apoptosis and cytoplasmic cell death in animal cells.

Methane oxidation in a peatland core

We made an experiment on a 30 cm diameter core of Sphagnum-dominated vegetation and peat to estimate the parameters controlling methane oxidation during movement to the ambient air: 13CH4 was added at the water table, and excess 13CO2 appeared in the gas space above the core. At 20°C in otherwise undisturbed conditions, ∼22% of CH4 was oxidized to CO2 during passage up through the overlying 10-cm thick unsaturated peat and plants. We simulated the experiment, with seven parameters: transfer coefficients in water, in the gas phase, and through the container wall; the rate of CH4 and of CO2 generation; and the two parameters of a hyperbolic relation between CH4 concentration and the rate of CH4 oxidation. We optimized these parameters to fit the experimental results, and then were able to generalize to any temperature (0°−25°C) and any depth (0-55 cm) of water table. Changing temperature has important effects on the proportion of CH4 oxidized.

A simplified method for determining 1-aminocyclopropane-1-carboxylic acid (ACC) in plant tissues using a mass selective detector

A sensitive and specific method is described for the routine assay of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) in 100–200 mg fresh weight samples of green or etiolated tissue. The method involves high performance liquid chromatography (HPLC) and gas chromatography linked to mass spectrometry (GCMS) and uses 14C-labelled ACC as an internal standard, N-benzoyl n-propyl ACC as an easily prepared derivative for HPLC and GCMS, and N-benzoyl isobutyl ACC as an internal standard for GCMS. The procedure is faster and safer than an existing GCMS method and more specific and reliable than indirect assays widely in use. The method has been used to measure ACC in maize roots, young leaves of cucumber, and aerobic or anaerobic seedlings of rice.