The chlorophyll fluorescence induction curves from mesophyll and guard cell chloroplasts of Saxifiaga cernmua, including both the fast (O to P, the transients involved in the rise in variable fluorescence) and slow (P to steady state fluorescence due to quenching) components, were characterized over a range of excitation intensities using microspectrophotometry (with epi-lumination) equipped with apertures designed to eliminate cross contamination of the fluorescence signal between the two chloroplast types. At low excitation intensities, the fast fluorescence kinetics from guard cell plastids showed an extended I to D phase and a more rapid appearance of P while minimal quenching from P to steady state fluorescence was observed compared to the transients from mesophyll chloroplasts suggesting a lower activity of photochemical (electron movement via carriers between donor and acceptor sites) and nonphotochemical (such as membrane conformational changes) events which regulate the fluorescence induction curve kinetics. As the excitation intensity was increased, the quenching rates of guard cells were faster at initiating conditions for photophosphorylation and the fast and slow fluorescence kinetics from guard cells resembled those of the mesophyll cells.Guard cell chloroplasts of S. ceraua from intact epidermal peels showed a low temperature (77 K) fluorescence emission spectrum having three major peaks (at 685, 695, and 730 nanometers when excited at 440 nanometers) which were qualitatively similar to those in the spectrum obtained from mesophyll tissue.These data suggest that S. cernua guard cell chloroplast photosystems I and II contribute to light-dependent stomatal activity only at high light intensities.

Armillaria mellea, a phytopathogenic fungus, is the only member of the Agaricales (Basidiomycetes) whose fertile vegetative phase in nature is thought to be diploid, rather than dikaryotic. To examine the vegetative ploidy of A. mellea, we used the DNA-binding antibiotic, mithramycin, for fluorometry of in situ nuclear DNA. The measurements of nuclear DNA content indicated that strains derived from single basidiospores of A. mellea were haploid and that strains derived from matings of isolates of single spores were diploid. These data confirm the results of earlier genetic experiments, which show haploidy and diploidy in unmated and mated strains, respectively. Nuclear DNA measurements in known haploid and diploid strains of Aspergillus nidulans confirmed the validity of our protocol.

The antioxidant beta-mercaptoethanol (BME) used in conjunction with the permanent mountant DPX (DPX-BME) retarded fluorescent fading of mithramycin, acridine orange and Hoechst 33258 stained chicken erythrocytes, each to a varying degree. The initial fluorescence of all dyes examined was more intense with DPX-BME than with DPX alone. Specimens mounted in DPX-BME showed strong fluorescence and excellent morphology; if kept in the dark, they could be stored indefinitely without deterioration. Retarding fading of fluorescence with DPX-BME faciliated quantitation of DNA using fluorescence cytophotometry.

The relative nuclear DNA contents of haploid, diploid, and benomyl-induced segregants of diploid strains of the phytopathogenic fungus Armillaria mellea were measured by mithramycin staining and fluorescence photometry. The diploid strains, originally recovered from sexually compatible matings of haploid strains, were heterozygous at mating-type and auxotrophic marker loci. The somatic segregants examined here were derived by treatment of the diploid strains with the fungicide benomyl in previous studies. As expected, the diploid strains had approximately twice as much nuclear DNA as the haploid strains. Most segregants had near-haploid DNA contents and no detectable heterozygosity at the marker loci; these strains were most likely true haploids. Other segregants with near-haploid DNA contents were heterozygous at a marker locus indicating that they were aneuploid. A minority of segregants had near-diploid DNA contents and may have been either aneuploid or diploid.Key words: basidiomycetes, mithramycin, parasexuality.

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