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The DNA relative content in nuclei from several Solanum species, which were used as partners for somatic hybridization, were determined using a flow cytometry method. The nuclei were isolated mechanically or via protoplasts from leaves of in vitro grown plants. In the case of S. nigrum as well as S. tuberosum cv. Bzura and dihaploid clone H8105, the nuclei were also obtained from suspension cultured cells by lysis of protoplasts. The source and the method of nuclei isolation affected the pattern of nuclear DNA in the genotypes studied. The mesophyll nuclei showed two distinct peaks on the DNA histograms, whereas the DNA peaks produced by cell suspension nuclei were broad and less distinct. The DNA content in the nuclei, calculated from the DNA histograms of the samples and a DNA standard historgam (Trout Red Blood Cells, having DNA content of 5.05 pg per nucleus), were much lower in mesophyll nuclei than in those obtained from the cell suspension for the same genotypes. The results are discussed in respect of the genetic instability of Solanum genotypes. The usefulness of a flow cytometry approach in somatic hybridization research is also discussed.
High yields of viable protoplasts were obtained from the mesophyll leaf tissue of young seedlings of Lycopersicon glandulosum grown in vitro at 80 µmol s-1m-2 irradiance and 23°-24°C. The isolated protoplasts were cultured first on a modified semiliquid medium of Tan et al. (1987 b), on which they formed microcalli, and afterwards on a greening medium consisting of MS micro/macro nutrients, Nitsch vitamins and 0.5 mg/l BAP + 0.05 mg/l NAA. The green budding calli were in turn transferred to the same medium with BAP and NAA replaced by 2.0 mg/l zeatin + 0.1 mg/l IAA, on which they developed shoots. The regenerated shoots were rooted in 1/3 MS medium without growth regulators, and subsequently planted into pots in the greenhouse. The whole process of regeneration from the moment of protoplast isolation up to the flowering of regenerated plants lasted ~6 months. The regenerated tomatoes were morphologically identical to their parental source plants.
The ultrastructure of chloroplasts was studied in mesophyll cells of the leaves of silver birch (Betula pendula) showing interveinal chlorosis or premature yellowing, in comparison with leaves without symptoms or exhibiting symptoms of natural senescence. The leaves were collected between May 26 to June 7 and additionally in the September 10-12 from the upper part of the crown, from increments of the past four years. No major difference in ultrastructure of chloroplasts was found between spongy and palisade mesophyll cells. The following senescencerelated changes were observed in chloroplasts of prematurely yellowed leaves and showing inteveinal chlorosis: reduced chloroplast size, degeneration of the membrane systems of thylakoids and increased electron density of plastoglobuli. The most electron dark globules (lipid droplets) were found together with starch grains in cells of spongy mesophyll of leaves showing interveinal chlorosis. Abnormal, spherical and rounded chloroplasts with electron-dark inside of thylakoids or the electron-dark stroma between thylakoids were found only in yellowed and chlorotic leaves in spring.
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